CoChat
Sign Up
Cited by 340RNA Research and SplicingGenetics and Neurodevelopmental DisordersRNA regulation and disease

The neurogenetics of alternative splicing

Open access
Vuong, Celine K., Black, Douglas L., Zheng, Sika·2016·Nature Reviews Neuroscience
DOI 10.1038/nrn.2016.27PMC PMC4861142
Open this paper in CoChat — free account

Abstract

Alternative precursor-mRNA splicing is a key mechanism for regulating gene expression in mammals and is controlled by specialized RNA-binding proteins. The misregulation of splicing is implicated in multiple neurological disorders. We describe recent mouse genetic studies of alternative splicing that reveal its critical role in both neuronal development and the function of mature neurons. We discuss the challenges in understanding the extensive genetic programmes controlled by proteins that regulate splicing, both during development and in the adult brain.

AI snapshot

Written from the paper’s full text.

TL;DR

This review synthesizes mouse genetic studies showing that alternative splicing regulators control neuronal development, migration, synaptogenesis, synaptic function, and survival, and that their misregulation contributes to neurological and neurodegenerative disease. It highlights both the power of conditional knockouts and isoform-rescue experiments and the challenges of pleiotropy, paralog redundancy, and linking specific splicing events to phenotypes.

Key findings

  • Alternative splicing regulators such as PTBP1/PTBP2, SRRM4, NOVA1/2, RBFOX1/2, KHDRBS, MBNL2, nELAVL, SCNM1, TDP43/FUS, and the U2 snRNA gene Rnu2-8 have critical roles across neurogenesis, neuronal migration, synaptogenesis, synaptic function, and neurodegeneration.
  • Mouse mutants show severe phenotypes: pan-neuronal Ptbp1 loss causes progressive ependymal cell loss, hydrocephaly, and death by 10 weeks; only 15% of Srrm4Δ7–8 mice survive beyond birth; Rbfox2−/− mice show increased mortality with frequent hydrocephaly at 1 month; Nova1/2 double-knockout mice are born paralysed and die from respiratory failure; NMF291 Rnu2-8 mutants lose cerebellar granule neurons beginning at postnatal week 4, develop tremors at 8 weeks, and progress to truncal ataxia by 12 weeks.
  • Isoform-specific rescue experiments link particular splicing events to phenotypes: a Dab1Δ7bc transgene rescues the migration defect for a subset of layer II–IV Nova2−/− neurons, and restoring Z+ Agrn expression in Nova1/2 double-knockout motor neurons rescues acetylcholine receptor clustering, nerve terminal apposition, neuromuscular junction morphology, and muscle responses to stimuli, but not paralysis or early mortality.
  • Mis-splicing is implicated in disease: TDP43 is a major component of cytoplasmic inclusions found in 95% of sporadic ALS and FTLD cases; MBNL2 loss produces myotonic dystrophy-like CNS phenotypes; RBFOX1 mutations are identified in epilepsy and autism spectrum disorder; and Scnm1 modifies the severity of the Scn8a medJ splicing mutation.

Methods

  • Review article surveying recent mouse genetic studies of individual splicing regulators in the mammalian nervous system, with a focus on phenotypic analyses rather than extensively reviewed genome-wide methods.
  • Evidence base includes mouse germline, conditional, and pan-neuronal knockouts, transgenic rescues, forward genetic mutants, and cited transcriptome/CLIP studies; no new experimental data are reported.
  • Main analysis is a narrative synthesis of regulator-by-regulator phenotypes across neurogenesis, migration, synaptogenesis, synaptic function, and neurodegeneration.

Limitations

  • Stated in the paper: the neurogenetics of splicing is still in its infancy; only a few of many hundreds of potential splicing regulatory proteins are beginning to be analysed; pleiotropic or lethal phenotypes obscure later functions; paralogous regulators often show partial redundancy that mutes single-gene effects; how altered splicing changes encoded protein function is usually unknown; the large number of targets makes it difficult to link a phenotype to a particular splicing event; and RNA-binding proteins also affect polyadenylation, translation, and stability, with indirect or secondary effects possible.
  • Evident from the text: as a review, it reports no primary data or systematic meta-analysis, so conclusions depend on cited mouse models and translational relevance to human disease is often inferred rather than directly tested.
  • Evident from the text: many cited phenotypes come from different mutant alleles, genetic backgrounds, and developmental windows, limiting direct comparability across studies.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1038/nrn.2016.27

Cites

Statistics

Nothing to recompute.

Claims the paper makes

0 of 244 cited sources checked · 0 supported · 0 flagged

  • Premisecitation

    The mammalian nervous system makes extensive use of splicing regulation to generate specialized protein isoforms that affect all aspects of neuronal development and function.

    “the mammalian nervous system makes extensive use of splicing regulation to generate specialized protein isoforms that affect all aspects of neuronal development and function1–4.”
    Quote in the paperSubstantial claim0 of 4 sources checked
    • cites [1] Li, Q., Lee, J.-A. & Black, D. L. Neuronal regulation of alternative pre-mRNA splicing. Nat. Rev. Neurosci. 8, 819–831 (2007). source

      Not checked in the free run.

    • cites [2] Zheng, S. & Black, D. L. Alternative pre-mRNA splicing in neurons: growing up and extending its reach. Trends Genet. 29, 442–448 (2013). source

      Not checked in the free run.

    • cites [3] Raj, B. & Blencowe, B. J. Alternative splicing in the mammalian nervous system: recent insights into mechanisms and functional roles. Neuron 87, 14–27 (2015). source

      Not checked in the free run.

    • cites [4] Darnell, R. B. RNA protein interaction in neurons. Annu. Rev. Neurosci. 36, 243–270 (2013). source

      Not checked in the free run.

  • Premisenone given

    Splicing regulators can exert different effects on a target transcript depending on their binding position, modification by signalling pathways and interactions with cofactors.

    “These proteins are structurally diverse and can exert different effects on a target transcript depending on their binding position, their modification by signalling pathways and their interactions with cofactors.”
    Quote in the paperNot a substantial claim
  • Premisecitation

    Reduced PTBP1 expression enhances miR-124 repression of the REST complex.

    “Reduced PTBP1 expression also enhances miR-124 repression of the REST (repressor element 1-silencing transcription factor) complex16”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [16] Xue, Y. et al. Direct conversion of fibroblasts to neurons by reprogramming PTB-regulated microRNA circuits. Cell 152, 82–96 (2013). This study demonstrates that PTBP1 acts as a master regulator of cell identity by driving a posttranscriptional regulatory programme that inhibits neuronal differentiation. source

      Not checked in the free run.

  • Premisecitation

    SRRM4 is unique for its brain-specific expression.

    “but is unique for its brain-specific expression29.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [29] Calarco, J. A. et al. Regulation of vertebrate nervous system alternative splicing and development by an SR-related protein. Cell 138, 898–910 (2009). source

      Not checked in the free run.

  • Premisecitation

    Alternative splicing patterns are regulated by specialized pre-mRNA binding proteins that alter spliceosome assembly at specific splice sites.

    “Alternative splicing patterns (BOX 1) are regulated by specialized pre-mRNA binding proteins that alter spliceosome assembly at specific splice sites5–8”
    Quote in the paperSubstantial claim0 of 4 sources checked
    • cites [5] Will, C. L. & Lührmann, R. Spliceosome structure and function. Cold Spring Harb. Perspect. Biol. 3, a003707 (2011). source

      Not checked in the free run.

    • cites [6] Matera, A. G. & Wang, Z. A day in the life of the spliceosome. Nat. Rev. Mol. Cell. Biol. 15, 108–121 (2014). source

      Not checked in the free run.

    • cites [7] Fu, X.-D. & Ares, M. Context-dependent control of alternative splicing by RNA-binding proteins. Nat. Rev. Genet. 15, 689–701 (2014). source

      Not checked in the free run.

    • cites [8] Lee, Y. & Rio, D. C. Mechanisms and regulation of alternative pre-mRNA splicing. Annu. Rev. Biochem. 84, 291–323 (2015). source

      Not checked in the free run.

  • May combine two claims. 81
  • Classified as probably not a substantial claim. 20

  • May combine two claims.

    Premisecitation

    PTBP1 is abundant in neural stem cells and progenitors, and on mitotic exit its expression is sharply reduced by the induction of miR-124.

    “PTBP1 is abundant in neural stem cells and progenitors, but on mitotic exit its expression is sharply reduced by the induction of the neuronal microRNA miR-124 (REF. 15).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [15] Makeyev, E. V., Zhang, J., Carrasco, M. A. & Maniatis, T. The microRNA miR-124 promotes neuronal differentiation by triggering brain-specific alternative pre-mRNA splicing. Mol. Cell 27, 435–448 (2007). This study demonstrates that an important splicing regulator, PTBP1, is controlled during neuronal development by the microRNA miR-124. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S4 (based).

    Premisenone given

    Alternative splicing patterns change dramatically as cells progress along the neuronal lineage.

    “Alternative splicing patterns change dramatically as cells progress along the neuronal lineage.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Each PTBP contains four RNA recognition motif domains that together bind extended CU-rich sequences.

    “Each PTBP contains four RNA recognition motif domains that together bind to extended CU-rich sequences13,14.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [13] Keppetipola, N., Sharma, S., Li, Q. & Black, D. L. Neuronal regulation of pre-mRNA splicing by polypyrimidine tract binding proteins, PTBP1 and PTBP2. Crit. Rev. Biochem. Mol. Biol. 47, 360–378 (2012). source

      Not checked in the free run.

    • cites [14] Spellman, R. et al. Regulation of alternative splicing by PTB and associated factors. Biochem. Soc. Trans. 33, 457–460 (2005). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationcitation

    Loss of ependymal cells in pan-neuronal Ptbp1 mutants is restricted to the dorsal telencephalon, indicating variable roles for PTBP1 across brain regions.

    “The loss of ependymal cells is restricted to the dorsal telencephalon, indicating variable roles for PTBP1 across brain regions.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [24] Shibasaki, T. et al. PTB deficiency causes the loss of adherens junctions in the dorsal telencephalon and leads to lethal hydrocephalus. Cereb. Cortex 23, 1824–1835 (2013). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    Mutation of an individual splicing regulator often leads to a phenotype with high pleiotropy and/or lethality owing to its extensive target set.

    “the mutation of an individual regulator often leads to a phenotype that either exhibits a high degree of pleiotrophy and/or is lethal owing to its extensive target set.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    Splicing regulators often have paralogues whose partially redundant function reduces the phenotypic impact of single gene mutations.

    “splicing regulators often have paralogues, the partially redundant function of which reduces the phenotypic impact of single gene mutations.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisenone given

    PTBP2 is found in neurons, myoblasts and spermatocytes.

    “PTBP2 (nPTB or brPTB) is found in neurons, myoblasts and spermatocytes.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationcitation

    PTBP1 loss may induce precocious differentiation of radial glial cells into neurons, depleting the radial glial cell pool that later gives rise to ependymal cells.

    “PTBP1 loss may induce the precocious differentiation of radial glial cells into neurons, thereby depleting the radial glial cell pool that later gives rise to ependymal cells25”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [25] Spassky, N. et al. Adult ependymal cells are postmitotic and are derived from radial glial cells during embryogenesis. J. Neurosci. 25, 10–18 (2005). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Germline deletion of Srrm4 results in fewer Pax6+ cells in the ventricular zone and fewer postmitotic NeuN+ neurons.

    “the germline deletion of Srrm4 results in fewer Pax6+ cells in the ventricular zone and fewer postmitotic NeuN+ neurons35”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [35] Quesnel-Vallières, M., Irimia, M., Cordes, S. P. & Blencowe, B. J. Essential roles for the splicing regulator nSR100/SRRM4 during nervous system development. Genes Dev. 29, 746–759 (2015). References 29, 30, 31 and 35 lay out a compelling description of a splicing regulatory network controlled by a neuronal splicing factor. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    Nearly all mammalian multi-exon genes produce multiple mRNA isoforms through alterations in the choice of splice sites.

    “Nearly all mammalian multi-exon genes produce multiple mRNA isoforms through alterations in the choice of splice sites to produce proteins of different structures and functions, or to alter mRNA localization, translation or decay.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Repression of Ptbp2 exon 10 by PTBP1 leads to nonsense-mediated mRNA decay of the Ptbp2 transcript and prevents its expression in PTBP1+ cells.

    “The repression of exon 10 leads to nonsense-mediated mRNA decay (NMD) of the Ptbp2 transcript and prevents its expression in PTBP1+ cells15,20,21.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [15] Makeyev, E. V., Zhang, J., Carrasco, M. A. & Maniatis, T. The microRNA miR-124 promotes neuronal differentiation by triggering brain-specific alternative pre-mRNA splicing. Mol. Cell 27, 435–448 (2007). This study demonstrates that an important splicing regulator, PTBP1, is controlled during neuronal development by the microRNA miR-124. source

      Not checked in the free run.

    • cites [20] Boutz, P. L. et al. A post-transcriptional regulatory switch in polypyrimidine tract-binding proteins reprograms alternative splicing in developing neurons. Genes Dev. 21, 1636–1652 (2007). source

      Not checked in the free run.

    • cites [21] Spellman, R., Llorian, M. & Smith, C. W. J. Crossregulation and functional redundancy between the splicing regulator PTB and its paralogs nPTB and ROD1. Mol. Cell 27, 420–434 (2007). References 15, 20 and 21 provide an early example of cross-regulation by splicing regulators. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    SRRM4 promotes splicing of the REST4 isoform, which has reduced transcriptional repression activity.

    “SRRM4 promotes splicing of the REST4 isoform, which lacks four of the nine zinc fingers found in the full-length protein31 and has reduced transcriptional repression activity32–34.”
    Quote in the paperSubstantial claim0 of 4 sources checked
    • cites [31] Raj, B. et al. Cross-regulation between an alternative splicing activator and a transcription repressor controls neurogenesis. Mol. Cell 43, 843–850 (2011). source

      Not checked in the free run.

    • cites [32] Palm, K., Metsis, M. & Timmusk, T. Neuron-specific splicing of zinc finger transcription factor REST/NRSF/XBR is frequent in neuroblastomas and conserved in human, mouse and rat. Brain Res. Mol. Brain Res. 72, 30–39 (1999). source

      Not checked in the free run.

    • cites [33] Shimojo, M., Lee, J. H. & Hersh, L. B. Role of zinc finger domains of the transcription factor neuron-restrictive silencer factor/repressor element-1 silencing transcription factor in DNA binding and nuclear localization. J. Biol. Chem. 276, 13121–13126 (2001). source

      Not checked in the free run.

    • cites [34] Tabuchi, A. et al. REST4-mediated modulation of REST/NRSF-silencing function during BDNF gene promoter activation. Biochem. Biophys. Res. Commun. 290, 415–420 (2002). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S4 (based).

    Premisenone given

    Splicing defects are increasingly implicated in neurological and neurodegenerative diseases.

    “Splicing defects are being increasingly implicated in neurological and neurodegenerative diseases”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisenone given

    Different neuronal cell types often express different combinations of splicing regulators and show distinct susceptibility to their mutation.

    “different neuronal cell types often express different combinations of regulators and show a distinct susceptibility to their mutation.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    SRRM4 can antagonize PTBP activity, and the regulatory programmes for these two proteins significantly overlap.

    “suggesting that SRRM4 can antagonize PTBP activity, and the regulatory programmes for these two proteins significantly overlap.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Knockdown of Srrm4 in the developing mouse cortex inhibits neuronal differentiation and leads to accumulation of Pax6+ progenitor cells in the ventricular zone and depletion of differentiated cells from the cortical plate.

    “Knockdown of Srrm4 in the developing mouse cortex inhibits neuronal differentiation and leads to the accumulation of Pax6+ progenitor cells in the ventricular zone and the depletion of differentiated cells from the cortical plate31.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [31] Raj, B. et al. Cross-regulation between an alternative splicing activator and a transcription repressor controls neurogenesis. Mol. Cell 43, 843–850 (2011). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Mice with pan-neuronal loss of Ptbp1 have grossly normal brain morphology at an early age but show progressive loss of ependymal cells from the lateral ventricles with hydrocephaly and die by 10 weeks after birth.

    “Mice with pan-neuronal loss of Ptbp1 have grossly normal brain morphology at an early age, but show a progressive loss of ependymal cells from the lateral ventricles with hydrocephaly and die by 10 weeks after birth24.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [24] Shibasaki, T. et al. PTB deficiency causes the loss of adherens junctions in the dorsal telencephalon and leads to lethal hydrocephalus. Cereb. Cortex 23, 1824–1835 (2013). source

      Not checked in the free run.

  • May combine two claims.

    Premisenone given

    Each regulatory protein can affect many RNA targets, and each transcript is usually targeted by multiple regulators.

    “Although each regulatory protein can affect many different RNA targets, each transcript is usually targeted by multiple regulators (FIG. 1).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Cultured Ptbp2−/− embryonic cortical neurons initially appear normal in plating efficiency and neurite outgrowth, but exhibit progressive cell death beginning in the second week.

    “cultured Ptbp2−/− embryonic cortical neurons initially appear normal in plating efficiency and neurite outgrowth, but exhibit progressive cell death beginning in the second week.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [64] Li, Q. et al. The splicing regulator PTBP2 controls a program of embryonic splicing required for neuronal maturation. eLife 3, e01201 (2014). References 63 and 64 demonstrate that PTBP2 controls a transition from fetal to adult splicing programmes that is required for proper neuronal maturation and survival. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Bv/bv mutants show deafness, head tossing and circling.

    “Bv/bv mutants show deafness, head tossing and circling69.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [69] Deol, M. S. & Gluecksohn-Waelsch, S. The role of inner hair cells in hearing. Nature 278, 250–252 (1979). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    In wild-type neurons, NOVA2 represses both exon 7b and 7c of the Dab1 transcript, and the resulting DAB1 protein isoform DAB1Δ7bc is subject to ubiquitylation upon Reelin activation.

    “In wild-type neurons, NOVA2 represses both exon 7b and 7c of the Dab1 transcript, and the resulting DAB1 protein isoform DAB1Δ7bc is subject to ubiquitylation upon Reelin activation43–45.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [43] Rice, D. S. et al. Disabled-1 acts downstream of Reelin in a signaling pathway that controls laminar organization in the mammalian brain. Development 125, 3719–3729 (1998). source

      Not checked in the free run.

    • cites [44] Arnaud, L., Ballif, B. A. & Cooper, J. A. Regulation of protein tyrosine kinase signaling by substrate degradation during brain development. Mol. Cell. Biol. 23, 9293–9302 (2003). source

      Not checked in the free run.

    • cites [45] Bock, H. H., Jossin, Y., May, P., Bergner, O. & Herz, J. Apolipoprotein E receptors are required for reelin-induced proteasomal degradation of the neuronal adaptor protein Disabled-1. J. Biol. Chem. 279, 33471–33479 (2004). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    PTBP1 restricts PSD95 protein expression to neurons, whereas PTBP2 controls the temporal induction of PSD95 late in neuronal maturation.

    “PTBP1 restricts PSD95 protein expression to neurons, whereas PTBP2 controls the temporal induction of PSD95 late in neuronal maturation.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    RBFOX binding upstream of, or within, an alternative exon typically inhibits exon inclusion, whereas downstream binding usually promotes splicing.

    “RBFOX binding upstream of, or within, an alternative exon typically inhibits exon inclusion, whereas downstream binding usually promotes splicing48.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [48] Kuroyanagi, H. Fox-1 family of RNA-binding proteins. Cell. Mol. Life Sci. 66, 3895–3907 (2009). source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    RBFOX proteins all bind the RNA sequence element (U)GCAUG via a single RNA recognition motif domain.

    “The RBFOX proteins all bind the RNA sequence element (U)GCAUG via a single RNA recognition motif domain.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Emx1-Ptbp2−/− brains begin to show cortical atrophy as early as P5, and by P15 the cortex shows massive cell death and is almost completely degenerated.

    “Emx1–Ptbp2−/− brains begin to show cortical atrophy as early as P5, and by P15 the cortex shows massive cell death and is almost completely degenerated.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [64] Li, Q. et al. The splicing regulator PTBP2 controls a program of embryonic splicing required for neuronal maturation. eLife 3, e01201 (2014). References 63 and 64 demonstrate that PTBP2 controls a transition from fetal to adult splicing programmes that is required for proper neuronal maturation and survival. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    All three Rbfox genes are broadly expressed in the brain, with individual neuronal cell types expressing different combinations at different developmental times.

    “All three Rbfox genes are broadly expressed in the brain, with individual neuronal cell types expressing different combinations at different developmental times51–55.”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [51] McKee, A. E. et al. A genome-wide in situ hybridization map of RNA-binding proteins reveals anatomically restricted expression in the developing mouse brain. BMC Dev. Biol. 5, 14 (2005). source

      Not checked in the free run.

    • cites [52] Underwood, J. G., Boutz, P. L., Dougherty, J. D., Stoilov, P. & Black, D. L. Homologues of the Caenorhabditis elegans Fox-1 protein are neuronal splicing regulators in mammals. Mol. Cell. Biol. 25, 10005–10016 (2005). source

      Not checked in the free run.

    • cites [53] Gehman, L. T. et al. The splicing regulator Rbfox1 (A2BP1) controls neuronal excitation in the mammalian brain. Nat. Genet. 43, 706–711 (2011). This study demonstrates the role of the RBFOX1 splicing regulator in controlling neuronal excitation, and the involvement of alternative splicing in epileptogenesis. Along with reference 55, this study exemplifies how families of paralogous splicing regulators can have either redundant or separate functions depending on the brain region. source

      Not checked in the free run.

    • cites [54] Hammock, E. A. D. & Levitt, P. Developmental expression mapping of a gene implicated in multiple neurodevelopmental disorders, A2bp1 (Fox1). Dev. Neurosci. 33, 64–74 (2011). source

      Not checked in the free run.

    • cites [55] Gehman, L. T. et al. The splicing regulator Rbfox2 is required for both cerebellar development and mature motor function. Genes Dev. 26, 445–460 (2012). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Pan-neuronal Rbfox2 knockout mice show increased mortality with frequent hydrocephaly at 1 month of age.

    “Pan-neuronal Rbfox2 knockout mice show increased mortality with frequent hydrocephaly at 1 month of age55.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [55] Gehman, L. T. et al. The splicing regulator Rbfox2 is required for both cerebellar development and mature motor function. Genes Dev. 26, 445–460 (2012). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Srrm4Δ7-8 mice show no gross morphological phenotype during embryonic development, but most die from respiratory failure within a few hours of birth.

    “Srrm4Δ7–8 mice show no gross morphological phenotype during embryonic development, but most die from respiratory failure within a few hours of birth.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [35] Quesnel-Vallières, M., Irimia, M., Cordes, S. P. & Blencowe, B. J. Essential roles for the splicing regulator nSR100/SRRM4 during nervous system development. Genes Dev. 29, 746–759 (2015). References 29, 30, 31 and 35 lay out a compelling description of a splicing regulatory network controlled by a neuronal splicing factor. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    The in vivo programme of neuronal induction contains multiple fail-safe mechanisms, with PTBP1 and SRRM4 serving opposing roles in reinforcing the robustness of the regulatory network.

    “suggest that the in vivo programme of neuronal induction contains multiple fail-safe mechanisms, with PTBP1 and SRRM4 serving opposing roles in reinforcing the robustness of the regulatory network.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Nova1−/− mice appear normal at birth but die in the second postnatal week with motor neuron apoptosis, profound motor failure and action-induced tremors.

    “Nova1−/− mice appear normal at birth, but die in the second postnatal week with motor neuron apoptosis, profound motor failure and action-induced tremors37,38.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [37] Yang, Y. Y., Yin, G. L. & Darnell, R. B. The neuronal RNA-binding protein Nova-2 is implicated as the autoantigen targeted in POMA patients with dementia. Proc. Natl Acad. Sci. USA 95, 13254–13259 (1998). source

      Not checked in the free run.

    • cites [38] Jensen, K. B. et al. Nova-1 regulates neuron-specific alternative splicing and is essential for neuronal viability. Neuron 25, 359–371 (2000). source

      Not checked in the free run.

  • May combine two claims.

    Premiseown data

    Rbfox2−/− mice have a severely affected cerebellum with substantially reduced size and a loss of foliation.

    “The cerebellum is severely affected, with a substantially reduced size and a loss of foliation.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premiseown data

    Rbfox2−/− Purkinje cells show a substantial delay in migration and increased cell death, resulting in a disorganized Purkinje cell layer.

    “The Rbfox2−/− Purkinje cells show a substantial delay in migration and increased cell death, resulting in a disorganized Purkinje cell layer (FIG. 3).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Downregulation of PTBP1 in neural stem/progenitor cells as they exit mitosis induces the expression of PTBP2, which is required for neuronal development and survival.

    “The downregulation of PTBP1 in neural stem/progenitor cells as they exit mitosis induces the expression of PTBP2, which is required for neuronal development and survival.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisenone given

    NOVA1 and NOVA2 each contain three K homology (KH)-type RNA-binding domains and bind clusters of YCAY elements.

    “The two paralogues NOVA1 and NOVA2 each contain three K homology (KH)-type RNA-binding domains and bind clusters of YCAY elements.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationcitation

    Progenitor cell proliferation and radial glia morphology are largely unaffected in Nova2-null mice, suggesting a defect in neuronal migration rather than subtype specification.

    “Progenitor cell proliferation and radial glia morphology are largely unaffected, suggesting a defect in neuronal migration rather than subtype specification.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [39] Yano, M., Hayakawa-Yano, Y., Mele, A. & Darnell, R. B. Nova2 regulates neuronal migration through an RNA switch in disabled-1 signaling. Neuron 66, 848–858 (2010). This paper beautifully demonstrates how NOVA2-dependent alternative splicing regulates the function of a receptor protein important for neuronal migration in the cortex and highlights the spatial specificity of splicing regulation through the use of knockout and transgenic mice. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Overexpression of either PTBP1 or PTBP2 in mature neurons inhibits exon 18 inclusion, PSD95 protein expression and excitatory synapse formation.

    “Overexpression of either PTBP1 or PTBP2 in mature neurons inhibits exon 18 inclusion, PSD95 protein expression and excitatory synapse formation67.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [67] Zheng, S. et al. PSD-95 is post-transcriptionally repressed during early neural development by PTBP1 and PTBP2. Nat. Neurosci. 15, 381–S1 (2012). This study describes how an alternative splicing event can control the overall expression of a neuronal-specific gene, and how PTBP1 and PTBP2 control splicing programmes required for synapse formation. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Ptbp2 mutant mice are paralysed and unresponsive to touch at birth and show perinatal lethality with respiratory failure.

    “These mice are paralysed and unresponsive to touch at birth and show perinatal lethality with respiratory failure, possibly owing to a loss of innervation to the diaphragm.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [63] Licatalosi, D. D. et al. Ptbp2 represses adult-specific splicing to regulate the generation of neuronal precursors in the embryonic brain. Genes Dev. 26, 1626–1642 (2012). source

      Not checked in the free run.

    • cites [64] Li, Q. et al. The splicing regulator PTBP2 controls a program of embryonic splicing required for neuronal maturation. eLife 3, e01201 (2014). References 63 and 64 demonstrate that PTBP2 controls a transition from fetal to adult splicing programmes that is required for proper neuronal maturation and survival. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Srrm4−/− mice have fewer late-born, upper-layer neurons and more early-born, lower-layer neurons.

    “Srrm4−/− mice have fewer late-born, upper-layer neurons and more early-born, lower-layer neurons”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [35] Quesnel-Vallières, M., Irimia, M., Cordes, S. P. & Blencowe, B. J. Essential roles for the splicing regulator nSR100/SRRM4 during nervous system development. Genes Dev. 29, 746–759 (2015). References 29, 30, 31 and 35 lay out a compelling description of a splicing regulatory network controlled by a neuronal splicing factor. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    NOVA1 is mainly expressed in the hindbrain and ventral spinal cord, whereas NOVA2 is predominant in the forebrain and dorsal spinal cord.

    “NOVA1 is mainly expressed in the hindbrain and ventral spinal cord, whereas NOVA2 is predominant in the forebrain and dorsal spinal cord, with some overlapping expression in portions of the midbrain and hindbrain37.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [37] Yang, Y. Y., Yin, G. L. & Darnell, R. B. The neuronal RNA-binding protein Nova-2 is implicated as the autoantigen targeted in POMA patients with dementia. Proc. Natl Acad. Sci. USA 95, 13254–13259 (1998). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationcitation

    In Nova2−/− neurons, abnormal inclusion of exons 7b and 7c produces a more stable isoform that may antagonize the activity of DAB1Δ7bc.

    “In Nova2−/− neurons, the abnormal inclusion of exons 7b and 7c produces a more stable isoform that may antagonize the activity of DAB1Δ7bc46,47.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [46] Feng, L., Allen, N. S., Simo, S. & Cooper, J. A. Cullin 5 regulates Dab1 protein levels and neuron positioning during cortical development. Genes Dev. 21, 2717–2730 (2007). source

      Not checked in the free run.

    • cites [47] Simo, S., Jossin, Y. & Cooper, J. A. Cullin 5 regulates cortical layering by modulating the speed and duration of Dab1-dependent neuronal migration. J. Neurosci. 30, 5668–5676 (2010). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Nova1/2 dKO mice, but not single-knockout mice, show fewer acetylcholine receptor clusters and a loss of apposition between AChR clusters and phrenic nerve terminals.

    “the dKO mice, but not the single-knockout mice, show fewer acetylcholine receptor (AChR) clusters and a loss of apposition between AChR clusters and phrenic nerve terminals”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [38] Jensen, K. B. et al. Nova-1 regulates neuron-specific alternative splicing and is essential for neuronal viability. Neuron 25, 359–371 (2000). source

      Not checked in the free run.

    • cites [73] Ruggiu, M. et al. Rescuing Z+ agrin splicing in Nova null mice restores synapse formation and unmasks a physiologic defect in motor neuron firing. Proc. Natl Acad. Sci. 106, 3513–3518 (2009). This reference reports a good example of using transgene rescue to understand the function of alternatively spliced isoforms and to dissect a portion of a highly pleiotropic phenotype. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    SAM68 is found in both the nucleus and the cytosol of many cell types and affects a variety of cellular processes, including splicing in the nucleus.

    “SAM68 is found in both the nucleus and the cytosol of many cell types81 and affects a variety of cellular processes, including splicing in the nucleus82,83.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [81] Lukong, K. E. & Richard, S. Sam68, the KH domain-containing superSTAR. Biochim. Biophys. Acta 1653, 73–86 (2003).

      Not checked in the free run.

    • cites [82] Matter, N., Herrlich, P. & König, H. Signal-dependent regulation of splicing via phosphorylation of Sam68. Nature 420, 691–695 (2002). source

      Not checked in the free run.

    • cites [83] Grange, J. et al. Somatodendritic localization and mRNA association of the splicing regulatory protein Sam68 in the hippocampus and cortex. J. Neurosci. Res. 75, 654–666 (2004). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Nova1/2 dKO mice are born paralysed and die from respiratory failure indicative of NMJ defects.

    “Nova1/2 dKO mice are born paralysed and die from respiratory failure indicative of NMJ defects73.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [73] Ruggiu, M. et al. Rescuing Z+ agrin splicing in Nova null mice restores synapse formation and unmasks a physiologic defect in motor neuron firing. Proc. Natl Acad. Sci. 106, 3513–3518 (2009). This reference reports a good example of using transgene rescue to understand the function of alternatively spliced isoforms and to dissect a portion of a highly pleiotropic phenotype. source

      Not checked in the free run.

  • May combine two claims.

    Premisenone given

    Hundreds of exons are mis-spliced in the Mbnl2−/− brain and overlap significantly with those known to be mis-spliced in myotonic dystrophy.

    “Hundreds of exons are mis-spliced in the Mbnl2−/− brain and overlap significantly with those known to be mis-spliced in myotonic dystrophy.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationcitation

    The paralysis phenotype and early mortality are unchanged by the Z+ Agrn transgene, indicating contributions of additional NOVA targets.

    “The paralysis phenotype and early mortality are unchanged by the transgene, indicating the contributions of additional NOVA targets, possibly in other brain regions.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [73] Ruggiu, M. et al. Rescuing Z+ agrin splicing in Nova null mice restores synapse formation and unmasks a physiologic defect in motor neuron firing. Proc. Natl Acad. Sci. 106, 3513–3518 (2009). This reference reports a good example of using transgene rescue to understand the function of alternatively spliced isoforms and to dissect a portion of a highly pleiotropic phenotype. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    SAM68 and SLM1 are both expressed in Purkinje cells and probably have redundant roles in their development.

    “SAM68 and SLM1 are both expressed in Purkinje cells and probably have redundant roles in their development.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The KHDRBS family has three members: SAM68, SLM1 and SLM2.

    “The KHDRBS family has three members: SRC-associated in mitosis 68 kDa protein (SAM68; also known as KHDRBS1), SLM1 (also known as KHDRBS2) and SLM2 (also known as KHDRBS3)80.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [80] Di Fruscio, M., Chen, T. & Richard, S. Characterization of Sam68-like mammalian proteins SLM-1 and SLM-2: SLM-1 is a Src substrate during mitosis. Proc. Natl Acad. Sci. USA 96, 2710–2715 (1999). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    In cerebellar granule neurons, KCl depolarization and kainic acid treatment increase SAM68 phosphorylation at Ser20 via CamKIV activation, leading to increased repression of Nrxn1 exon 20.

    “In cerebellar granule neurons, both KCl depolarization and kainic acid treatment increase SAM68 phosphorylation at Ser20 via CamKIV activation, leading to increased repression of Nrxn1 exon 20 and changes in trans-synaptic interactions94”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [94] Iijima, T. et al. SAM68 regulates neuronal activity-dependent alternative splicing of neurexin-1. Cell 147, 1601–1614 (2011). This paper and the preceding references describe how a key determinant of synaptogenesis is regulated at the level of splicing. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    SLM1 and SLM2 are more restricted to the nervous system and have distinct expression patterns.

    “SLM1 and SLM2 are more restricted to the nervous system and have distinct expression patterns.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisenone given

    Germline deletion of Mbnl2 results in abnormal sleep patterns, memory loss and learning deficits.

    “Germline deletion of Mbnl2 results in abnormal sleep patterns, memory loss and learning deficits.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisereasoning

    NMDAR-mediated responses and pattern-induced long-term potentiation are impaired in Mbnl2-null mice.

    “NMDAR-mediated responses and pattern-induced long-term potentiation are impaired in Mbnl2-null mice.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Slm1 and Slm2 single-null mice are viable with no apparent behavioural or anatomical defects.

    “Slm1 and Slm2 single-null mice are viable with no apparent behavioural or anatomical defects84,87,98.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [84] Iijima, T., Iijima, Y., Witte, H. & Scheiffele, P. Neuronal cell type-specific alternative splicing is regulated by the KH domain protein SLM1. J. Cell Biol. 204, 331–342 (2014). source

      Not checked in the free run.

    • cites [87] Traunmüller, L., Bornmann, C. & Scheiffele, P. Alternative splicing coupled nonsense-mediated decay generates neuronal cell type-specific expression of SLM proteins. J. Neurosci. 34, 16755–16761 (2014). source

      Not checked in the free run.

    • cites [98] Ehrmann, I. et al. The tissue-specific RNA binding protein T-STAR controls regional splicing patterns of neurexin pre-mRNAs in the brain. PLoS Genet. 9, e1003474 (2013). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Long-term potentiation of excitatory post-synaptic currents is unchanged in Nova2-null hippocampus, as are basal excitatory and inhibitory synaptic transmission.

    “By contrast, long-term potentiation of excitatory post-synaptic currents is unchanged, as are basal excitatory and inhibitory synaptic transmission.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [100] Huang, C. S. et al. Common molecular pathways mediate long-term potentiation of synaptic excitation and slow synaptic inhibition. Cell 123, 105–118 (2005). source

      Not checked in the free run.

  • May combine two claims.

    Premisenone given

    Although all three MBNLs are expressed in the brain, only Mbnl2-null mice exhibit obvious central nervous system phenotypes.

    “Although all three MBNLs are expressed in the brain, only Mbnl2-null mice exhibit obvious central nervous system phenotypes.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Restoring Z+ Agrn expression in dKO motor neurons via a transgene rescues AChR clustering, nerve terminal apposition, NMJ morphology and muscle responses to stimuli.

    “restoring Z+ Agrn expression in dKO motor neurons via a transgene rescues AchR clustering, nerve terminal apposition, NMJ morphology and muscle responses to stimuli73.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [73] Ruggiu, M. et al. Rescuing Z+ agrin splicing in Nova null mice restores synapse formation and unmasks a physiologic defect in motor neuron firing. Proc. Natl Acad. Sci. 106, 3513–3518 (2009). This reference reports a good example of using transgene rescue to understand the function of alternatively spliced isoforms and to dissect a portion of a highly pleiotropic phenotype. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    NRXNβ AS4- isoforms preferentially bind neuroligin 1 at glutamatergic synapses, whereas NRXNβ AS4+ isoforms preferentially bind NLGN2 at GABAergic and glycinergic synapses.

    “NRXNβ AS4– isoforms preferentially bind neurolignin 1 (B) concentrated at glutamatergic synapses, whereas NRXNβ AS4+ isoforms preferentially bind NLGN2(A) at GABAergic and glycin-ergic synapses93”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [93] Chih, B., Gollan, L. & Scheiffele, P. Alternative splicing controls selective trans-synaptic interactions of the neuroligin-neurexin complex. Neuron 51, 171–178 (2006). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Germline Sam68-null mice have deficits of bone metabolism, sexual organ development, motor coordination and motor learning.

    “Germline Sam68-null mice have deficits of bone metabolism, sexual organ development, motor coordination and motor learning95,96”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [95] Richard, S. et al. Ablation of the Sam68 RNA binding protein protects mice from age-related bone loss. PLoS Genet. 1, e74 (2005). source

      Not checked in the free run.

    • cites [96] Lukong, K. E. & Richard, S. Motor coordination defects in mice deficient for the Sam68 RNA-binding protein. Behav. Brain Res. 189, 357–363 (2008). source

      Not checked in the free run.

  • May combine two claims.

    Premisereasoning

    Sam68/Slm1 dKO mice are similar to Sam68 single-null mice, except for additional defects in cerebellar foliation and scattered ectopic Purkinje cells in the molecular layer.

    “Sam68/Slm1 dKO mice were similar to Sam68 single-null mice, except for additional defects in cerebellar foliation and scattered ectopic Purkinje cells in the molecular layer.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    nELAVLs are target antigens in patients with paraneoplastic neurological disorders.

    “Like NOVA, nELAVLs are target antigens in patients with paraneo-plastic neurological disorders121,122.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [121] Szabo, A. et al. HuD, a paraneoplastic encephalomyelitis antigen, contains RNA-binding domains and is homologous to Elav and Sex-lethal. Cell 67, 325–333 (1991). source

      Not checked in the free run.

    • cites [122] Sakai, K. et al. A hippocampal protein associated with paraneoplastic neurologic syndrome and small cell lung carcinoma. Biochem. Biophys. Res. Commun. 199, 1200–1208 (1994). source

      Not checked in the free run.

  • May combine two claims.

    Premisereasoning

    Elavl3-null mice show spontaneous cortical hypersynchrony and non-convulsive electrographic seizures.

    “These mice also show spontaneous cortical hypersynchrony and non-convulsive electrographic seizures.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Pan-neuronal deletion of Rbfox1 leads to increased susceptibility to spontaneous seizures and kainic acid-induced seizures.

    “Pan-neuronal deletion of Rbfox1 (Rbfox1loxp/loxp; Nestin-Cre) leads to increased susceptibility to spontaneous seizures and seizures induced by kainic acid”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [53] Gehman, L. T. et al. The splicing regulator Rbfox1 (A2BP1) controls neuronal excitation in the mammalian brain. Nat. Genet. 43, 706–711 (2011). This study demonstrates the role of the RBFOX1 splicing regulator in controlling neuronal excitation, and the involvement of alternative splicing in epileptogenesis. Along with reference 55, this study exemplifies how families of paralogous splicing regulators can have either redundant or separate functions depending on the brain region. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The ELAVL family consists of four highly homologous members.

    “The ELAVL (also known as Hu) family of proteins consists of four highly homologous members109–111”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [109] Good, P. J. A conserved family of elav-like genes in vertebrates. Proc. Natl Acad. Sci. USA 92, 4557–4561 (1995). source

      Not checked in the free run.

    • cites [110] Brennan, C. M. & Steitz, J. A. HuR and mRNA stability. Cell. Mol. Life Sci. 58, 266–277 (2001). source

      Not checked in the free run.

    • cites [111] Hinman, M. N. & Lou, H. Diverse molecular functions of Hu proteins. Cell. Mol. Life Sci. 65, 3168–3181 (2008). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    nELAVLs bind introns and Elavl3/4 dKO brains show hundreds of splicing changes.

    “recent genome-wide profiling analyses have revealed intronic binding of nELAVLs and hundreds of splicing changes in Elavl3/4 dKO brains.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    ELAVL1 is widely expressed in non-neuronal tissues.

    “ELAVL1 (also known as HuR or HuA) is widely expressed in non-neuronal tissues”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [51] McKee, A. E. et al. A genome-wide in situ hybridization map of RNA-binding proteins reveals anatomically restricted expression in the developing mouse brain. BMC Dev. Biol. 5, 14 (2005). source

      Not checked in the free run.

    • cites [119] Okano, H. J. & Darnell, R. B. A hierarchy of Hu RNA binding proteins in developing and adult neurons. J. Neurosci. 17, 3024–3037 (1997). source

      Not checked in the free run.

    • cites [120] Pascale, A., Amadio, M. & Quattrone, A. Defining a neuron: neuronal ELAV proteins. Cell. Mol. Life Sci. 65, 128–140 (2008). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Rbfox1+/−; Rbfox2−/− brains show dramatic splicing changes at Scn8a exons 5 and 18, including a twofold decrease in exon18A that decreases functional Nav1.6.

    “the Rbfox1+/−; Rbfox2−/− brains show dramatic splicing changes at both exons 5 and 18, including a twofold decrease in exon18A that decreases the amount of functional Nav1.6 (REF. 55).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites REF. 55

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    In a C57Bl/6J background, the same medJ mutation produces only 5% correctly spliced transcript and mice show progressive paralysis and lethality by 1 month of age.

    “The same mutation in a C57Bl/6J background produces only 5% correctly spliced transcript and the mice show progressive paralysis and lethality by 1 month of age151.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [151] Kearney, J. A. et al. Molecular and pathological effects of a modifier gene on deficiency of the sodium channel Scn8a (Nav1.6). Hum. Mol. Genet. 11, 2765–2775 (2002). source

      Not checked in the free run.

  • May combine two claims.

    Premisenone given

    Rbfox1−/− brains show decreased 5b and increased 5a inclusion.

    “The Rbfox1−/− brains show decreased 5b and increased 5a inclusion.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    The medJ mutation has a four base pair deletion in the 5′ splice site of intron 3 and causes skipping of both exons 2 and 3 in a majority of Scn8a transcripts.

    “This mutation, medJ, does not alter the coding sequence of Scn8a but instead has a four base pair deletion in the 5′ splice site of intron 3, which causes skipping of both exons 2 and 3 in a majority of Scn8a transcripts”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [148] Kohrman, D. C., Harris, J. B. & Meisler, M. H. Mutation detection in the med and medJ alleles of the sodium channel Scn8a. Unusual splicing due to a minor class AT-AC intron. J. Biol. Chem. 271, 17576–17581 (1996). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    Rbfox proteins have roles in both cerebellar development and mature function.

    “The early and late phenotypes of Rbfox mutation demonstrate roles for these proteins in both cerebellar development and mature function.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    medJ mice show hindlimb paralysis, muscle atrophy and degeneration of Purkinje cells.

    “The medJ mice show hindlimb paralysis, muscle atrophy and the degeneration of Purkinje cells149.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [149] Sidman, R. L., Cowen, J. S. & Eicher, E. M. Inherited muscle and nerve diseases in mice: a tabulation with commentary. Ann. NY Acad. Sci. 317, 497–505 (1979). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Targeted deletion of Scnm1 in the C3H strain confirmed that SCNM1 affects both the splicing of the Scn8amedJ transcript and the mouse phenotype.

    “targeted deletion of Scnm1 in the C3H strain confirmed that SCNM1 affects both the splicing of the Scn8amedJ transcript and the mouse phenotype152,153.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [152] Sprunger, L. K., Escayg, A., Tallaksen-Greene, S., Albin, R. L. & Meisler, M. H. Dystonia associated with mutation of the neuronal sodium channel Scn8a and identification of the modifier locus Scnm1 on mouse chromosome 3. Hum. Mol. Genet. 8, 471–479 (1999). source

      Not checked in the free run.

    • cites [153] Howell, V. M. et al. A targeted deleterious allele of the splicing factor SCNM1 in the mouse. Genetics 180, 1419–1427 (2008). References 151–153 demonstrate how a small change in the amount of a particular transcript isoform can dramatically affect neuronal function, and highlight the action of splicing regulators as phenotypic modifiers. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Inclusion of Scn8a exon 18N introduces a premature termination codon and leads to nonsense-mediated decay.

    “Inclusion of 18N introduces a premature termination codon and leads to nonsense-mediated decay”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [146] Plummer, N. W., McBurney, M. W. & Meisler, M. H. Alternative splicing of the sodium channel SCN8A predicts a truncated two-domain protein in fetal brain and non-neuronal cells. J. Biol. Chem. 272, 24008–24015 (1997). source

      Not checked in the free run.

    • cites [147] O'Brien, J. E. et al. Rbfox proteins regulate alternative splicing of neuronal sodium channel SCN8A. Mol. Cell. Neurosci. 49, 120–126 (2012). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    The severity of the medJ phenotype is affected by genetic background and correlates with the amount of correctly spliced transcript.

    “The severity of the medJ phenotype was found to be affected by genetic background and to correlate with the amount of correctly spliced transcript152.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [152] Sprunger, L. K., Escayg, A., Tallaksen-Greene, S., Albin, R. L. & Meisler, M. H. Dystonia associated with mutation of the neuronal sodium channel Scn8a and identification of the modifier locus Scnm1 on mouse chromosome 3. Hum. Mol. Genet. 8, 471–479 (1999). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    In the C3H background, medJ mice live a normal lifespan with dystonia and ataxia, with 10% of Scn8a transcripts correctly spliced.

    “In the C3H background, medJ mice live a normal lifespan with dystonia and ataxia, with 10% of Scn8a transcripts correctly spliced.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [151] Kearney, J. A. et al. Molecular and pathological effects of a modifier gene on deficiency of the sodium channel Scn8a (Nav1.6). Hum. Mol. Genet. 11, 2765–2775 (2002). source

      Not checked in the free run.

    • cites [152] Sprunger, L. K., Escayg, A., Tallaksen-Greene, S., Albin, R. L. & Meisler, M. H. Dystonia associated with mutation of the neuronal sodium channel Scn8a and identification of the modifier locus Scnm1 on mouse chromosome 3. Hum. Mol. Genet. 8, 471–479 (1999). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Rbfox2−/− Purkinje cells form a proper layer but show decreased dendritic arborization and irregular, less frequent spontaneous action potentials.

    “Rbfox2−/− Purkinje cells eventually form a proper layer, but show decreased dendritic arborization in the molecular layer, as well as irregular and less frequent spontaneous action potentials.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [55] Gehman, L. T. et al. The splicing regulator Rbfox2 is required for both cerebellar development and mature motor function. Genes Dev. 26, 445–460 (2012). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    In medJ/C3H mice, replacement of fetal Nav1.2 is delayed and Nav1.6 at the nodes of Ranvier reaches only 10-20% of wild-type levels.

    “In medJ/C3H mice, the replacement of fetal Nav1.2 is delayed and the amount of Nav1.6 at the nodes of Ranvier reaches only 10–20% of that seen in wild-type mice.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [151] Kearney, J. A. et al. Molecular and pathological effects of a modifier gene on deficiency of the sodium channel Scn8a (Nav1.6). Hum. Mol. Genet. 11, 2765–2775 (2002). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Not all U2 genes are the same; individual genes within a cluster can show temporal and cell-type-specific patterns of expression.

    “These data demonstrate that not all U2 genes are the same, but that individual genes within a cluster can show temporal and cell-type-specific patterns of expression.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premiseown data

    Both wild-type and mutant Rnu2-8 RNA are selectively expressed in the cerebellum and increase in expression after granule neuron maturation.

    “both wild-type and mutant Rnu2-8 RNA are selectively expressed in the cerebellum and increase in expression after granule neuron maturation.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premiseown data

    When highly expressed, the mutant U2 snRNA decreases overall splicing efficiency and affects alternative splicing patterns.

    “When highly expressed, the mutant U2 snRNA decreases the overall splicing efficiency and affects alternative splicing patterns.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Fus-null mice die soon after birth.

    “Fus-null mice die soon after birth179”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [179] Hicks, G. G. et al. Fus deficiency in mice results in defective B-lymphocyte development and activation, high levels of chromosomal instability and perinatal death. Nat. Genet. 24, 175–179 (2000). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Both ALS-associated mutations and changes in wild-type TDP43 expression alter a large programme of alternative exons in mutant mice.

    “Both the ALS-associated mutations and the changes in wild-type TDP43 expression alter a large programme of alternative exons in mutant mice170,174,175.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [170] Polymenidou, M. et al. Long pre-mRNA depletion and RNA missplicing contribute to neuronal vulnerability from loss of TDP-43. Nat. Neurosci. 14, 459–468 (2011). source

      Not checked in the free run.

    • cites [174] Tollervey, J. R. et al. Characterizing the RNA targets and position-dependent splicing regulation by TDP-43. Nat. Neurosci. 14, 452–458 (2011). source

      Not checked in the free run.

    • cites [175] Arnold, E. S. et al. ALS-linked TDP-43 mutations produce aberrant RNA splicing and adult-onset motor neuron disease without aggregation or loss of nuclear TDP-43. Proc. Natl Acad. Sci. 110, E736–E745 (2013).

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    TDP43 and FUS regulate distinct groups of alternative exons and target different sets of mRNAs in the cytoplasm.

    “TDP43 and FUS regulate distinct groups of alternative exons and target different sets of mRNAs in the cytoplasm178,181”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [178] Rogelj, B. et al. Widespread binding of FUS along nascent RNA regulates alternative splicing in the brain. Sci. Rep. 2, 603 (2012). source

      Not checked in the free run.

    • cites [181] Fujioka, Y. et al. FUS-regulated region- and cell-type-specific transcriptome is associated with cell selectivity in ALS/FTLD. Sci. Rep. 3, 2388 (2013). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    U2 snRNA binds to the branch point during spliceosome assembly and then forms base pairs with U6 snRNA to become a key portion of the spliceosome catalytic centre for all major class introns.

    “U2 snRNA binds to the branch point during spliceosome assembly and then forms base pairs with the U6 snRNA to become a key portion of the spliceosome catalytic centre for all major class introns6,8,183,184”
    Quote in the paperSubstantial claim0 of 4 sources checked
    • cites [6] Matera, A. G. & Wang, Z. A day in the life of the spliceosome. Nat. Rev. Mol. Cell. Biol. 15, 108–121 (2014). source

      Not checked in the free run.

    • cites [8] Lee, Y. & Rio, D. C. Mechanisms and regulation of alternative pre-mRNA splicing. Annu. Rev. Biochem. 84, 291–323 (2015). source

      Not checked in the free run.

    • cites [183] Wahl, M. C., Will, C. L. & Lührmann, R. The spliceosome: design principles of a dynamic RNP machine. Cell 136, 701–718 (2009). source

      Not checked in the free run.

    • cites [184] Black, D. L. et al. Mechanisms of alternative pre-messenger RNA splicing. Annu. Rev. Biochem. 72, 291–336 (2003). source

      Not checked in the free run.

  • May combine two claims.

    Premiseown data

    NMF291 mice develop tremors at 8 weeks, progressing to truncal ataxia by 12 weeks.

    “and develop tremors at 8 weeks, progressing to truncal ataxia by 12 weeks.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [182] Jia, Y., Mu, J. C. & Ackerman, S. L. Mutation of a U2 snRNA gene causes global disruption of alternative splicing and neurodegeneration. Cell 148, 296–308 (2012). This paper demonstrates that mutation of a small RNA component of the spliceosome core can lead to a highly specific neurodegenerative phenotype. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    FUS is a widely expressed, predominantly nuclear RNA-binding protein.

    “Another ALS and FTLD gene, FUS156, is a widely expressed, predominantly nuclear RNA-binding protein176,177”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [176] Iko, Y. et al. Domain architectures and characterization of an RNA-binding protein, TLS. J. Biol. Chem. 279, 44834–44840 (2004). source

      Not checked in the free run.

    • cites [177] Zinszner, H., Sok, J., Immanuel, D., Yin, Y. & Ron, D. TLS (FUS) binds RNA in vivo and engages in nucleo-cytoplasmic shuttling. J. Cell. Sci. 110, 1741–1750 (1997). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    In myelodysplastic syndromes, mutations in several components of the core splicing apparatus cause very specific splicing defects in particular transcripts and lead to a highly tissue-specific phenotype.

    “mutations in several components of the core splicing apparatus, such as U2 small nuclear ribonucleoprotein auxiliary factor 2 (U2AF65), splicing factor 3B, subunit 1 (SF3B1) and other proteins, were found to cause very specific splicing defects in particular transcripts and to lead to a highly tissue-specific phenotype185–187.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [185] Yoshida, K. et al. Frequent pathway mutations of splicing machinery in myelodysplasia. Nature 478, 64–69 (2011). source

      Not checked in the free run.

    • cites [186] Lindsley, R. C. & Ebert, B. L. Molecular pathophysiology of myelodysplastic syndromes. Annu. Rev. Pathol. 8, 21–47 (2013). source

      Not checked in the free run.

    • cites [187] Yoshida, K. & Ogawa, S. Splicing factor mutations and cancer. Wiley Interdiscip. Rev. RNA 5, 445–459 (2014). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    ALS-like phenotypes are seen in mice with partial depletion of TDP43 by RNA interference or with targeted deletion of Tdp43 in motor neurons.

    “ALS-like phenotypes are seen in mice with partial depletion of TDP43 by RNA interference, or with targeted deletion of Tdp43 in motor neurons”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [163] Kraemer, B. C. et al. Loss of murine TDP-43 disrupts motor function and plays an essential role in embryogenesis. Acta Neuropathol. 119, 409–419 (2010). source

      Not checked in the free run.

    • cites [164] Sephton, C. F. et al. TDP-43 is a developmentally regulated protein essential for early embryonic development. J. Biol. Chem. 285, 6826–6834 (2010). source

      Not checked in the free run.

    • cites [165] Wu, L.-S. et al. TDP-43, a neuro-pathosignature factor, is essential for early mouse embryogenesis. Genesis 48, 56–62 (2010). source

      Not checked in the free run.

  • May combine two claims.

    Premisenone given

    In the RNA-binding protein knockouts analysed so far, heterozygous mice develop largely normally, but some splicing targets are still altered by the reduction in regulatory protein dose.

    “In the RNA-binding protein knockouts so far analysed, the heterozygous mice develop largely normally, but some splicing targets are still altered by the reduction in regulatory protein dose.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based). May combine two claims.

    Premisenone given

    RNA-binding proteins can antagonize each other or synergize in RNA binding.

    “RNA-binding proteins can antagonize each other or synergize in RNA binding.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    RNA-binding proteins have been connected to mis-splicing in neuropsychiatric disorders, including epilepsy, autism spectrum disorder, inherited ataxias and schizophrenia.

    “Other work has connected RNA-binding proteins to mis-splicing in neuropsychiatric disorders, including epilepsy, autism spectrum disorder, inherited ataxias and schizophrenia.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Many RNA-binding proteins controlling splicing choices also affect the choice of poly-A site and can be found in the cytoplasm, where they control translation or stability of target transcripts through binding in 3′ UTRs.

    “many RNA-binding proteins controlling splicing choices also affect the choice of poly-A site and can also be found in the cytoplasm, where they control the translation or stability of target transcripts through binding in 3′ UTRs197”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [197] Lee, J.-A. et al. Cytoplasmic Rbfox1 regulates the expression of synaptic and autism-related genes. Neuron 89, 113–128 (2016). source

      Not checked in the free run.

  • May combine two claims.

    Premisenone given

    Splicing regulators extensively modulate each other's activity and control the activity of transcriptional regulators.

    “Splicing regulators extensively modulate each other’s activity, as well as controlling the activity of transcriptional regulators (FIG. 1).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites FIG. 1

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S4 (based). May combine two claims.

    Premisenone given

    Many alternative splicing events are conserved across mammalian or vertebrate species, but the effect of these splicing changes on protein activity is usually unknown.

    “Although many alternative splicing events are conserved across mammalian or vertebrate species, the effect of these splicing changes on protein activity is usually unknown.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    Splicing regulators are rarely entirely redundant and usually show differences in their range of expression.

    “However, regulators are rarely entirely redundant and usually show differences in their range of expression, as seen with the NOVA, nELAVL and RBFOX proteins.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    In cells where paralogous splicing regulators are co-expressed, double mutation often leads to new splicing changes not seen in either single mutant and reveals new phenotypes.

    “In cells where they are co-expressed, double mutation will often lead to new splicing changes that are not seen in either single mutant and reveal new phenotypes.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S4 (based).

    Interpretationnone given

    Splicing programmes provide a highly interconnected layer of regulation that can alter protein activity without easily discernible changes in overall expression.

    “Splicing programmes provide a highly interconnected layer of regulation that can alter protein activity without easily discernible changes in the overall expression.”
    Quote in the paperNot a substantial claim
96 of 96

  • The paper relies on this without support.

    Mouse models of splicing regulator mutations are informative about the mammalian nervous system and human neurogenetics.

    “The biological roles of splicing regulators and the cellular programmes they control are currently being elucidated through mouse genetics.”
    “we survey recent genetic studies of individual splicing regulators and the diverse roles they have in the mammalian nervous system.”
1 of 1

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Mouse experiments indicate RBFOX1 has distinct nuclear and cytoplasmic isoforms with differing functions.

    “Experiments in mouse indicate that RBFOX1 has distinct nuclear and cytoplasmic isoforms with differing functions, the nuclear isoform primarily regulating pre-mRNA alternative splicing, and the cytoplasmic isoform binding to the 3’ UTR to stabilize target transcripts involved in regulation of neuronal excitability (28, 32, 58, 60).”

    From Transcriptome-wide isoform-level dysregulation in ASD, schizophrenia, and bipolar disorder (2018) · cites this paper as [28]

The citation lists haven’t been fetched yet

References 208 · Cited by 340

Authors
Vuong, C. K., Black, D. L., & Zheng, S.
Affiliations
  • University of California, Los Angeles (US)
  • University of California, Riverside (US)
Corresponding author
  • Black, Douglas L.
  • Zheng, Sika
ORCID
Venue
Nature Reviews Neuroscience
Publisher
Springer Science and Business Media LLC
Published
2016-04-20
Volume
17
Issue
5
Pages
265-281
ISSN
  • 1471-003X (ISSN-L)
  • 1471-0048
DOI
10.1038/nrn.2016.27
PMC
PMC4861142
OpenAlex
W2337654115
Semantic Scholar
60dfcfb96501fc0fa9733e5b33070473e3fece9c
Cited by
340
References
208
Citations by year
2026: 18 · 2025: 39 · 2024: 40 · 2023: 44 · 2022: 33 · 2021: 37 · 2020: 39 · 2019: 23 · 2018: 33 · 2017: 26 · 2016: 8
FWCI
2.71
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature reviews. Neuroscience (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Institutes of Health
  • National Institute of Mental Health (K99MH096807, R00MH096807)
  • National Institute of General Medical Sciences (R01 GM 49662, T32GM007185, R01GM049662)
  • National Institute of Neurological Disorders and Stroke (F31NS093923)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetics and Neurodevelopmental Disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
Keywords
alternative splicing, splicing regulators, RNA-binding proteins, neuronal development, neurological disorders, genetic mouse models, gene expression regulation
MeSH terms
Animals, Brain, Humans, Nerve Net, Nerve Tissue Proteins, Neurons, Alternative Splicing, Neurogenesis

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-10.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-10.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review