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Cited by 70Amino Acid Enzymes and MetabolismMetabolomics and Mass Spectrometry StudiesNeuroscience and Neuropharmacology Research

Large neutral amino acid levels tune perinatal neuronal excitability and survival

Open access
Knaus, Lisa S., Basilico, Bernadette, Malzl, Daniel, Gerykova Bujalkova, Maria, Smogavec, Mateja, Schwarz, Lena A., Gorkiewicz, Sarah, Amberg, Nicole +8 more·2023·Cell
DOI 10.1016/j.cell.2023.02.037

Published version of a preprint

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Abstract

Little is known about the critical metabolic changes that neural cells have to undergo during development and how temporary shifts in this program can influence brain circuitries and behavior. Inspired by the discovery that mutations in SLC7A5, a transporter of metabolically essential large neutral amino acids (LNAAs), lead to autism, we employed metabolomic profiling to study the metabolic states of the cerebral cortex across different developmental stages. We found that the forebrain undergoes significant metabolic remodeling throughout development, with certain groups of metabolites showing stage-specific changes, but what are the consequences of perturbing this metabolic program? By manipulating Slc7a5 expression in neural cells, we found that the metabolism of LNAAs and lipids are interconnected in the cortex. Deletion of Slc7a5 in neurons affects the postnatal metabolic state, leading to a shift in lipid metabolism. Additionally, it causes stage- and cell-type-specific alterations in neuronal activity patterns, resulting in a long-term circuit dysfunction.

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Written from the paper’s full text.

TL;DR

The paper profiles the developing mouse cerebral cortex metabolome and shows that large neutral amino acid (LNAA) uptake via Slc7a5 is required perinatally for neuronal lipid metabolism, excitability, survival, and long-term circuit/behavioral function. Deleting Slc7a5 in forebrain neural cells causes stage-specific metabolic reprogramming, postnatal microcephaly, reduced neuronal firing, and persistent behavioral defects, with parallels to human SLC7A5-mutant microcephaly.

Key findings

  • Metabolomic profiling of wild-type mouse cortex at E14.5, P2, and P40 quantified 346 metabolites; 273/346 changed over time. At P2 versus E14.5, 137 metabolites decreased and 60 increased; at P40, 202 metabolites differed from P2. LNAAs/BCAAs showed declining perinatal trajectories.
  • Neural-cell deletion of Slc7a5 (`Slc7a5 fl/fl; Emx1-Cre+`) caused stage-specific LNAA accumulation in P2 cortical tissue but intracellular depletion in isolated cells, plus altered lipid metabolism: reduced glycerophospholipids and increased triacylglycerols. P5 cortical proteomics identified 1,202 deregulated proteins (954 upregulated, 248 downregulated), with lipid-metabolism proteins enriched among upregulated proteins.
  • Mutants were born with normal brain size but developed postnatal microcephaly by P40; cleaved caspase-3 increased at P2 and P5, and MADM mosaic analysis showed fewer `Slc7a5−/−` upper-layer excitatory neurons by P5 and P40. Layer II/III pyramidal neurons showed reduced action potential firing at P6–P7 but not P25–P26, and this excitability defect was cell-autonomous in mosaic-MADM recordings.
  • Adult mutants showed persistent behavioral defects, including hyperactivity, mild gait defects, hind-limb clasping, and sociability/social-memory impairments. In two siblings with compound heterozygous SLC7A5 variants, microcephaly progressed from −3 SD at birth to −5 SD at 7 months (patient 130-1) and from −2.5 SD at birth to −4.5 SD at 6–7 months (patient 130-2).

Methods

  • Mouse cortex was profiled by untargeted metabolomics, targeted intracellular metabolomics, untargeted lipidomics, proteomics, and bulk RNA-sequencing across developmental stages (E14.5, P2, P40 and other perinatal time points). Conditional `Slc7a5 fl/fl` mice were crossed to `Emx1-Cre` for neural-cell deletion and to `Tie2-Cre` for BBB/endothelial deletion; MADM-8 mosaics were used for sparse single-cell knockout and cell-autonomy tests.
  • Analyses included PCA, Gaussian mixture model clustering of metabolite trajectories, KEGG/GO enrichment, Pearson correlation of wild-type versus mutant trajectories (`r < 0.975`), and integration of proteomic, metabolomic, and lipidomic data. Electrophysiology used whole-cell current-clamp recordings from layer II/III pyramidal neurons at P6–P7 and P25–P26; behavior was assessed in adult mice.
  • Human data came from trio whole-exome sequencing of family 130 (two affected siblings and parents), identifying compound heterozygous SLC7A5 variants: c.1124C>T, p.(Pro375Leu) and a novel intragenic deletion of exons 5–10; head circumference was measured longitudinally.

Limitations

  • Stated by the paper: available tools cannot assess metabolic pathways with needed time and spatial resolution, so the study describes tissue-wide metabolic states rather than cell-type-specific or short-lasting changes; causality is challenging because metabolic arms are interconnected; mouse-specific metabolic changes may exist; human brain metabolism is difficult to study because most tools involve in vitro systems.
  • Evident from the text: the mouse deletion is restricted to the Emx1 lineage (excitatory neurons and pallial glia, not BBB endothelium), so effects in other neural cell types are not directly tested; the human evidence is from two siblings, limiting generalizability; and the exact mechanism linking lipid changes to reduced excitability remains unresolved, as the paper notes that transcriptomics did not uncover ion-channel expression changes.

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Crossref · 10.1016/j.cell.2023.02.037

Cites

115 of 115 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

8 of 20 cited sources checked · 5 supported · 0 flagged · accuracy not yet measured

  • Premisecitation

    Deletion of Slc7a5 from the blood-brain barrier in mice does not lead to a reduction in brain size.

    “while patients with SLC7A5 mutations present with microcephaly, the deletion of Slc7a5 from the BBB in mice (Slc7a5 fl/fl ;Tie2-Cre positive [+] mice) does not lead to a reduction in brain size.6”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [6] Tărlungeanu, 2016, Impaired amino acid transport at the blood brain barrier is a cause of autism spectrum disorder, Cell source

      The Results describe the Tie2Cre;Slc7a5fl/fl BBB-deletion mice and report that their overall brain morphology was indistinguishable from controls (Figure S4A). The paper also says the mice appeared normal in size, but that refers to the animals; our copy does not directly report a brain-size measurement or explicitly rule out a reduction in brain size. (LLM, conf=0.78)

      Experimental — accuracy not yet measured
  • Premisecitation

    Mutations in SLC7A5, a transporter of metabolically essential large neutral amino acids, are a rare cause of autism spectrum disorders.

    “We recently identified mutations in the gene SLC7A5, encoding a large neutral amino acid (LNAA) transporter (LAT1), as a rare cause of autism spectrum disorders.6”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [6] Tărlungeanu, 2016, Impaired amino acid transport at the blood brain barrier is a cause of autism spectrum disorder, Cell source
  • Findingcitation

    Slc7a5 mutants show an increased level of Bckdk.

    “Finally, Slc7a5 mutants also show an increased level of Bckdk, a regulator of the BCAA cata-bolic pathway also implicated in the phosphorylation of cytoplasmic Acl, a key enzyme in citrate-based lipogenesis.40”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [40] White, 2018, The BCKDH kinase and phosphatase integrate BCAA and lipid metabolism via regulation of ATP-citrate lyase, Cell Metab. source
  • Premisecitation

    The pathological variants identified in patients are mostly missense mutations that impact the transport capacity, not the expression, of the transporter.

    “Notably, the pathological variants identified in patients are mostly missense mutations that impact the transport capacity, not the expression, of the transporter.6”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [6] Tărlungeanu, 2016, Impaired amino acid transport at the blood brain barrier is a cause of autism spectrum disorder, Cell source
  • Findingown data

    Trio-whole exome sequencing identified compound heterozygous pathogenic variants in SLC7A5 in both siblings: the previously described missense variant c.1124C>T, p.(Pro375Leu) and a novel intragenic deletion of exons 5 to 10.

    “Trio-whole exome sequencing (WES) analysis of both siblings and their parents identified compound heterozygous pathogenic variants in the SLC7A5 gene: the previously described and functionally assessed missense variant c.1124C>T, p.(Pro375Leu) 6 and a novel intragenic deletion of exons 5 to 10.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [6] Tărlungeanu, 2016, Impaired amino acid transport at the blood brain barrier is a cause of autism spectrum disorder, Cell source
  • May combine two claims. 52
  • Classified as probably not a substantial claim. 41
  • The quote only partly states this claim. 2

  • May combine two claims.

    Premisecitation

    The constant features associated with biallelic pathogenic SLC7A5 variants include microcephaly, developmental and motor delay, seizures, and autistic features.

    “The phenotypic comparison of published patients 6 revealed that the constant features associated with biallelic pathogenic SLC7A5 variants include microcephaly, developmental and motor delay, seizures, and autistic features (Table S1).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [6] Tărlungeanu, 2016, Impaired amino acid transport at the blood brain barrier is a cause of autism spectrum disorder, Cell source

      The Results section, “SLC7A5 mutations in patients with ASD and motor delay,” reports affected children with homozygous SLC7A5 variants, autistic features, microcephaly, and motor problems. It does not establish developmental delay or seizures as constant features of those patients; the Discussion mentions seizures in connection with a different disorder. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

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

    Premisecitation

    A shift in the ratio of specific GPL subclasses can affect membrane properties such as its fluidity and curvature, which can further modulate neuronal excitability.

    “In addition, a shift in the ratio of specific GPL subclasses can affect membrane properties such as its fluidity and curvature, 25 which can further modulate neuronal excitability.45,47,71”
    Quote in the paperNot a substantial claim1 of 4 sources checked
    • cites [25] Holthuis, 2014, Lipid landscapes and pipelines in membrane homeostasis, Nature source
    • cites [45] Zheng, 2011, Lipid-dependent gating of a voltage-gated potassium channel, Nat. Commun. source

      The Results section ('Cholesterol effect on KvAP as predicted by lipid gating'; page unavailable) reports that changing membrane lipid composition altered gating of the KvAP potassium channel. The Discussion (page unavailable) likewise proposes that lipids help determine whether voltage-gated channels open or close. The paper does not show that shifts among specific GPL subclasses change membrane fluidity or curvature, or that those changes modulate neuronal excitability; its experiments used a prokaryotic channel in artificial membranes. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [47] Tillman, 2003, Effects of membrane lipids on ion channel structure and function, Cell Biochem. Biophys. source
    • cites [71] Schmidt, 2006, Phospholipids and the origin of cationic gating charges in voltage sensors, Nature source

  • May combine two claims.

    Findingown data

    Trio-WES analysis of both siblings and their parents identified compound heterozygous pathogenic variants in the SLC7A5 gene: the previously described and functionally assessed missense variant c.1124C>T, p.(Pro375Leu) and a novel intragenic deletion of exons 5 to 10.

    “Trio-WES analysis of both siblings and their parents identified compound heterozygous pathogenic variants in the SLC7A5 gene: the previously described and functionally assessed missense variant c.1124C>T, p.(Pro375Leu) 6 and a novel intragenic deletion of exons 5 to 10.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [6] Tărlungeanu, 2016, Impaired amino acid transport at the blood brain barrier is a cause of autism spectrum disorder, Cell source

  • May combine two claims.

    Findingown data

    At P40, 202 metabolites show a different level than at P2, with an approximately equal number of metabolites showing increased or decreased levels.

    “At P40, 202 metabolites show a different level than at P2, with an approximately equal number of metabolites showing increased or decreased levels (Figure 1C 0 ).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The forebrain undergoes significant metabolic remodeling throughout development, with certain groups of metabolites showing stage-specific changes.

    “We found that the forebrain undergoes significant metabolic remodeling throughout development, with certain groups of metabolites showing stage-specific changes”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Among metabolites increasing at P2 compared with E14.5, purine- and pentose phosphate-related metabolites are overrepresented, and among those decreasing, AA-related metabolites are enriched.

    “Enrichment analysis revealed an overrepresentation of purine-and pentose phosphate-related metabolites among those increasing at P2 (compared with E14.5) and enrichment of AA-related metabolites among those decreasing (Figure 1D).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Deletion of Slc7a5 alters the levels of these AAs in cortical tissue only at P2, indicating a function of Slc7a5 in neonatal mice.

    “However, the deletion of Slc7a5 alters the levels of these AAs in cortical tissue only at P2 (Figure 2H), indicating a function of Slc7a5 in neonatal mice.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Comparative untargeted lipidomic analysis of P2 Slc7a5 mutant and control cortical tissue and dissociated cells revealed a specific reduction of GPLs in mutant cortical cells, along with an increase of triacylglycerols (TGs) in Slc7a5 deficient cortical tissue.

    “These analyses revealed a specific reduction of GPLs in mutant cortical cells, along with an increase of triacylglycerols (TGs) in Slc7a5 deficient cortical tissue (Figures 3A and 3B).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The variability across animals of the same age was small, indicating that the observed changes are tightly regulated.

    “The variability across animals of the same age was small (Figure S1A), indicating that the observed changes are tightly regulated.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Deletion of Slc7a5 in neurons affects the postnatal metabolic state, leading to a shift in lipid metabolism.

    “Deletion of Slc7a5 in neurons affects the postnatal metabolic state, leading to a shift in lipid metabolism.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Several of the lipids differentially abundant in Slc7a5 mutants are neuron enriched, while astrocyte-enriched lipids are not affected in mutants.

    “Interestingly, several of the lipids differentially abundant in Slc7a5 mutants are neuron enriched, while astrocyte-enriched lipids are not affected in mutants (Figure 3C).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Those GO terms were reproducible at the mRNA expression level and linked to the reduction of GPLs.

    “Those GO terms were reproducible at the mRNA expression level (Figure S3J; Data S5) and linked to the reduction of GPLs 32 (Figures 3A and 3B).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Compared with E14.5, at P2, the level of 137 metabolites is significantly decreased and that of 60 increased.

    “Compared with E14.5, at P2, the level of 137 metabolites is significantly decreased and that of 60 increased (Figure 1C).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Cluster 1, representing metabolites with ascending trajectories over time, is enriched for neuroactive and cAMP signaling pathways, with the majority steeply increasing between P2 and P40.

    “This analysis revealed a predictable enrichment for neuroactive and cAMP signaling pathways in the cluster representing metabolites with ascending trajectories over time (cluster 1, Figures 1F and 1G), with the majority of the metabolites steeply increasing between P2 and P40.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    PCA of lipid-related metabolites shows that while at E14.5 and P40 mutant and control samples cluster together, P2 samples separate by genotype, suggesting a time-point-specific alteration in lipid composition due to loss of Slc7a5.

    “Specifically, a PCA of all the detected lipid-related metabolites disclosed that while at E14.5 and P40 mutant and control samples cluster together, P2 samples separate by genotype (Figure S3H), thereby suggesting a time-point-specific alteration in lipid composition due to the loss of Slc7a5”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The level of these AAs is higher, not lower, in mutants compared with controls, suggesting that in the absence of Slc7a5, the LNAAs accumulate in the extracellular space and are not consumed by neural cells.

    “Furthermore, the level of these AAs is higher, not lower, in mutants compared with controls (Figure 2H), suggesting that in the absence of Slc7a5, the LNAAs accumulate in the extracellular space and are not consumed by neural cells (Figure 2I).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    GO enrichment analysis returned proteins involved in lipid metabolism as the most significant and numerous among upregulated proteins, while downregulated GO terms are enriched for neuron projection and membrane-associated proteins.

    “Gene ontology (GO) enrichment analysis returned proteins involved in lipid metabolism as being the most significant and numerous among the upregulated proteins (Figure 3E right), while among the downregulated GO terms, we found an enrichment for neuron projection and membrane-associated proteins (Figure 3E left; Data S4).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    At P40, there is a substantial increase in glutamine and glutamate-related metabolites and a decrease in a different set of AAs, disclosing a stage-specific regulation of AA metabolism.

    “At P40, we detected a substantial increase in glutamine and glutamate-related metabolites and a decrease in a different set of AAs (Figure 1E), disclosing a stage-specific regulation of AA metabolism.”
    Quote in the paperSubstantial claim

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

    Findingown data

    Patient 130-2 displayed severe microcephaly, growth retardation, global developmental delay, and autistic features.

    “At genetic counseling, patient 130-2 was 4 years and 4 months old and displayed severe microcephaly (-6 SD), growth retardation (-5 up to -6 SD), global developmental delay (independent sitting possible, no walking, absent speech) and autistic features.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Histological analysis revealed a reduction in the thickness of the cerebral cortex of P40 Slc7a5 mutant animals, with layers II and III being the drivers of this difference.

    “Histological analysis revealed a reduction in the thickness of the cerebral cortex of P40 Slc7a5 fl/fl ;Emx1-Cre+ animals (Figures 4B and 4B 0 ), with layers II and III being the drivers of this difference (Figures S5E-S5G 0 ).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Co-immunoprecipitation of Slc7a5 from neural cells confirmed its interaction with Slc3a2 and excluded interactions with components of the B1/B3 integrin pathway.

    “Indeed, a co-immunoprecipitation experiment of Slc7a5 from neural cells, while confirming its interaction with Slc3a2, 34 excluded interactions with components of the B1/ B3 integrin pathway (Data S6).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [34] Yan, 2019, Structure of the human LAT1–4F2hc heteromeric amino acid transporter complex, Nature source

  • May combine two claims.

    Findingown data

    Slc7a5 fl/fl;Emx1-Cre+ mutants are born with normal brain size, indicating that Slc7a5 is dispensable in the NPCs of the forebrain.

    “In agreement with Slc7a5 expression profile (Figure S1B) and the AA profile, which changes only postnatally, Slc7a5 fl/fl ;Emx1-Cre+ mutants are born with normal brain size (Figures S5B and S5C), indicating that Slc7a5 is dispensable in the NPCs of the forebrain.”
    Quote in the paperSubstantial claim

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

    Findingown data

    The first seizures in patient 130-2 occurred at age 6 years and are controlled with anticonvulsive treatment.

    “The first seizures occurred at the age of 6 years and are controlled with anticonvulsive treatment.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Mosaic-MADM animals present fewer Slc7a5-/- (green) neurons in upper cortical layers (LII-LIV) compared with control (red) cells, while neurons in lower layers (LV and LVI) are not affected by Slc7a5 deletion.

    “Further, mosaic-MADM animals present fewer Slc7a5 À/À (green) neurons in upper cortical layers (LII-LIV) compared with control (red) cells, while neurons in the lower layers (LV and LVI) are not affected by Slc7a5 deletion.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Brain MRI of patient 130-1 showed pontocerebellar and corpus callosum hypoplasia in addition to microcephaly.

    “Brain MRI showed, in addition to microcephaly, pontocerebellar and corpus callosum hypoplasia.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Slc7a5 fl/fl;Emx1-Cre+ mice are born at Mendelian ratios, are viable, and at birth do not display obvious growth defects compared with wild-type littermates.

    “Slc7a5 fl/fl ;Emx1-Cre+ mice are born at Mendelian ratios, are viable and at birth do not display obvious growth defects compared with their wild-type littermates (i.e., Slc7a5 fl/fl ;Emx1-CreÀ or Slc7a5 fl/+ ;Emx1-Cre+ mice) (Figure S5A).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Patients 130-1 and 130-2 are siblings from a non-consanguineous family presenting with the clinical feature of SLC7A5 deficiency.

    “Patients 130-1 and 130-2 are siblings from a non-consanguineous family presenting with the clinical feature of SLC7A5 deficiency (Figure S5P).”
    Quote in the paperSubstantial claim

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

    Findingown data

    Before WES analysis, aCGH provided inconspicuous results for both patients.

    “Before WES analysis an aCGH (Microarry-based Comparative Genomic Hybridization) was carried out, providing inconspicuous results for both patients.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Mutant neurons at P6-P7 display larger amplitude, faster rise time, and more hyperpolarized AP threshold, matching AP features observed in control neurons at P25-P26.

    “Indeed, compared with controls, mutant neurons at P6-P7 display larger amplitude (Figure 6F), faster rise time (Figure 6G), and more hyperpolarized AP threshold (Figure 6H), matching the AP features observed in control neurons at P25-P26”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Patient 130-2 had mild microcephaly at birth (-2.5 SD), progressively deteriorating to -4.5 SD at age 6-7 months.

    “A mild microcephaly was diagnosed at birth (À2.5 SD), progressively deteriorating to À4.5 SD at the age of 6-7 months (Figure 4H).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Patient 130-1 showed microcephaly at birth (HC 31 cm, -3 SD) that deteriorated progressively to -5 SD at 7 months.

    “Patient 130-1 was born at term and showed microcephaly at birth (head circumference (HC) of 31 cm, -3 SD).Birth weight and height were normal.The microcephaly deteriorated progressively to -5 SD (HC of 36.6 cm) at the age of 7 months.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Non-neuronal cells are unaffected in Slc7a5 mutants, indicating that Slc7a5 is important specifically for neuronal functions.

    “In contrast, non-neuronal cells are unaffected (Figures S5L-S5O), indicating that Slc7a5 is important specifically for neuronal functions.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Adult Slc7a5 fl/fl;Emx1-Cre+ mutant animals present with increased horizontal and vertical explorative behavior but no anxiety-like behaviors.

    “In an open field, mutant animals present with increased horizontal (Figures 7A-7B 0 ) and vertical explorative behavior (Figure 7C) but no anxiety-like behaviors (Figure 7D).”
    Quote in the paperSubstantial claim

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

    Findingown data

    Patient 130-1 displayed pronounced muscular hypotonia and motor delay, and developed seizures at age 3 years.

    “The patient displayed pronounced muscular hypotonia and motor delay.At the age of 3 years, the patient developed seizures, which could be successfully treated with valproate.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    At P0, there are no significant changes in the ratio of green to red cells, supporting that Slc7a5 deficiency does not affect the proliferative phase of cortical development.

    “While at P0 we did not observe significant changes in the ratio of green to red cells (Figures 5B and 5B 0 ), supporting that Slc7a5 deficiency does not affect the proliferative phase of cortical development”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The difference in brain size between control and Slc7a5 mutant animals appears during the first postnatal week and remains stable from P10 onward, coinciding with increased Slc7a5 expression levels in neural cells perinatally.

    “By monitoring the brain weight over time, we found that the difference in brain size between control and Slc7a5 mutant animals appears during the first postnatal week (Figure 4C) and remains stable from P10 onward, coinciding with the increased Slc7a5 expression levels in neural cells perinatally (Figures 2A and 2B).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Decay time, resting membrane potential, ISI ratio, and half width are not affected perinatally in LII/LIII pyramidal neurons of Slc7a5 mutant and control animals.

    “Decay time (J), resting membrane potential (K), ISI ratio (L), and half width (M) are not affected perinatally (Slc7a5 fl/fl ;Emx1-CreÀ: n = 22 cells/3 mice; Slc7a5 fl/fl ;Emx1-Cre+: n = 30 cells/3 mice (P6-P7); Slc7a5 fl/fl ;Emx1-CreÀ: n = 5 cells/3 mice; Slc7a5 fl/fl ;Emx1-Cre+: n = 15 cells/3 mice (P25-P26).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    There are significant differences in overall dendritic spine density and/or the ratio of different spine classes of mutant and wild-type LII/LIII pyramidal neurons in the SSCtx at P6-P7 (**p < 0.01, *p < 0.05, ns p > 0.05).

    “Analysis of the overall dendritic spine density (F) and the ratio of the different spine classes (G-I) of mutant and wild-type LII/LIII pyramidal neurons in the SSCtx at P6-P7 (n = 3 mice per genotype, n > 28 dendrites per mouse; **p < 0.01, *p < 0.05, ns p > 0.05; unpaired two-tailed t test).”
    Quote in the paperSubstantial claim

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

    Findingown data

    The number of microglia (Iba1+) in cortical columns of adult mutant and wild-type mice is not significantly different (ns p > 0.05).

    “Immunostaining (N) and quantification (O) of microglia (Iba1+) in cortical columns of adult mutant and wild-type mice (means ± SD; n = 3 animals per genotype; ns p > 0.05; unpaired two-tailed t test; scale bars, 100 mm).”
    Quote in the paperNot a substantial claim

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

    Findingown data

    The number of astrocytes (Gfap+) in cortical columns of adult mutant and wild-type mice is not significantly different (ns p > 0.05).

    “Immunostaining (L) and quantification (M) of astrocytes (Gfap+) in cortical columns of adult mutant and wild-type mice (means ± SD; n = 4 animals per genotype; ns p > 0.05; unpaired two-tailed t test; scale bars, 100 mm).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    In P5 Slc7a5 fl/fl;Emx1-Cre+ cortex, the levels of Atf4, Ire-alpha, and the phosphoEif2a/Eif2a ratio normalized to Gapdh are not significantly different from control (ns p > 0.05).

    “Quantification of (H) Atf4, (I) Ire-alpha expression levels, and (J) phosphoEif2a/Eif2a ratio normalized to Gapdh (n = 4 mice per genotype; ns p > 0.05).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Mutant mice exhibit moderate motor deficits, such as decreased stride and stance length, as well as hind limb clasping behavior.

    “Indeed, mutant mice exhibit moderate motor deficits (Figure7E), such as decreased stride and stance length (Figures 7F-7F 00 ), as well as hind limb clasping behavior (Figures7G and 7G 0).”
    Quote in the paperSubstantial claim

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

    Findingown data

    The brain-to-body weight ratio of newborn (P0-P1) Slc7a5 fl/fl;Emx1-Cre+ and wild-type mice is not significantly different (ns p > 0.05).

    “Images (B) and quantification (B 0 ) of brain-to-body weight ratio of newborn (P0-P1) Slc7a5 fl/fl ;Emx1-Cre+ and wild-type mice (means ± SD; n = 9 animals per genotype; ns p > 0.05; unpaired two-tailed t test; scale bars, 1 mm).”
    Quote in the paperNot a substantial claim

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

    Findingown data

    The two analyses (hydrophilic interaction liquid chromatography and reversed phase chromatography) returned overlapping metabolites.

    “The two analyses returned overlapping metabolites.”
    Quote in the paperNot a substantial claim

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

    Findingown data

    The ratio of oxidized and reduced glutathione is unaffected in cortical cells of mutant mice.

    “The ratio of oxidized and reduced glutathione is unaffected in cortical cells of mutant mice”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Loss of Slc7a5 causes extracellular accumulation and intracellular depletion of LNAAs.

    “Loss of Slc7a5 causes extracellular accumulation and intracellular depletion of LNAAs (bottom).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Slc7a5 fl/fl ;Emx1-Cre+ mice have indications of sociability and social-memory impairments.

    “We found that Slc7a5 fl/fl ;Emx1-Cre+ mice have indications of sociability and social-memory impairments (Figures7H-7K 0).”
    Quote in the paperSubstantial claim

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

    Findingown data

    Brain and body weight of newborn Slc7a5 fl/fl;Emx1-Cre+ and wild-type littermates are not significantly different (ns p > 0.05).

    “Brain and body weight of newborn Slc7a5 fl/fl ;Emx1-Cre+ and wild-type littermates (means ± SD; n = 8 animals per genotype; ns p > 0.05, unpaired two-tailed t test).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The mTOR, UPR, and AMPK pathways are not affected in Slc7a5 deficient mice.

    “The mTOR, UPR, and AMPK pathways are not affected in Slc7a5 deficient mice”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Deletion of Slc7a5 in neurons causes stage- and cell-type-specific alterations in neuronal activity patterns, resulting in long-term circuit dysfunction.

    “Additionally, it causes stage-and cell-type-specific alterations in neuronal activity patterns, resulting in a long-term circuit dysfunction.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    There is a potential defect at the axon initial segment (AIS) and a faster AP backpropagation in the somatodendritic compartment of mutant neurons.

    “highlighting a potential defect at the axon initial segment (AIS) and a faster AP backpropagation in the somatodendritic compartment of mutant neurons (Figures 6I and 6J)”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    All transgenic mouse lines were backcrossed into C57BL/6J background >2 times.

    “All transgenic mouse lines were backcrossed into C57BL/6J background >2 times.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    The reduction of intracellular BCAA levels leads to a compensatory upregulation of several shared enzymes (e.g., Acaa2, Acadm, and Echs1), thereby increasing FA beta-oxidation and causing a drop in FAs.

    “The reduction of the intracellular BCAA levels leads to a compensatory upregulation of several of these shared enzymes (e.g., Acaa2, Acadm, and Echs1), thereby increasing FA b-oxidation and causing a drop in FAs.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    These morphological alterations persist until adulthood and are not associated with changes in excitability.

    “However, these morphological alterations persist until adulthood (Figures S7R and S7S) and, therefore, are not associated with changes in excitability.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    All animal protocols were approved by the Institutional Animal Care and Use Committee at ISTA and by the Bundesministerium für Bildung, Wissenschaft und Forschung, Austria (approval number: BMBWF-66.018/0015-V/3b/2019).

    “All animal protocols were approved by the Institutional Animal Care and Use Committee at ISTA and by the Bundesministerium fu ¨r Bildung, Wissenschaft und Forschung, Austria (approval number: BMBWF-66.018/0015-V/3b/2019).”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Experiments were carried out using littermate pairs of the same sex.

    “Experiments were carried out using littermate pairs of the same sex.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    SLC7A5 mutations likely result in decreased BCAA uptake and lead to microcephaly during the period of cortical refinement and programmed cell death in mice and humans.

    “Therefore, SLC7A5 mutations likely result in decreased BCAA uptake and lead to microcephaly during the period of cortical refinement and programmed cell death in mice and humans.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    We worked to ensure sex balance in the selection of non-human subjects.

    “We worked to ensure sex balance in the selection of non-human subjects.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Written informed consent and collection of data and samples were obtained according to a protocol approved by the Ethics Committee of the Medical University of Vienna (protocol number 1443/2020).

    “For the human subjects (patients and their parents), written informed consent and collection of data and samples were obtained according to a protocol approved by the Ethics Committee of the Medical University of Vienna (protocol number 1443/2020).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). The quote only partly states this claim.

    Study design and conductnone given

    A quality control (QC) sample was prepared by pooling 10 mL of each sample for untargeted metabolomics.

    “10 mL of each sample were pooled and used as a quality control (QC) sample.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    After LC-MS/MS analysis, retention times were verified by standard addition of pure compounds to arbitrarily selected samples, validating experimental retention times with the respective pure substances.

    “After LC-MS/MS analysis, retention times were verified by standard addition of pure compounds to arbitrarily selected samples, validating experimental retention times with the respective pure substances.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    P-values were corrected using the Benjamini-Hochberg procedure implemented in statsmodels 0.13.1.88.

    “P-values were computed using an unpaired two-tailed t-test and corrected using the Benjamini-Hochberg procedure implemented in statsmodels 0.13.1.88”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites statsmodels 0.13.1.88

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

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

    Study design and conductnone given

    EquiSPLASH LIPIDOMIX Quantitative Mass Spec Internal Standards and Metabolomics Amino Acid Mix Standards were added as internal standards to the solvent.

    “EquiSPLASHÔ LIPIDOMIXâ Quantitative Mass Spec Internal Standards (Avanti Polar Lipids) and Metabolomics Amino Acid Mix Standards (Cambridge Isotope Laboratories) were added as internal standards to the solvent.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Both males and females were used for experiments.

    “Both, males and females were used for experiments.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    For all LC-MS/MS transitions, the optimal collision energy was defined by analyzing pure metabolite standards.

    “For all transitions, the optimal collision energy was defined by analyzing pure metabolite standards.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    P5 Slc7a5 fl/fl ;Tie2-Cre+ animals were used to ensure that the precipitated Slc7a5 is derived from neural cells-only.

    “Brief, P5 Slc7a5 fl/fl ;Tie2-Cre+ animals were used to ensure that the precipitated Slc7a5 is derived from neural cells-only.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Compound annotation was performed with a mass accuracy of 3 ppm for precursor masses and 10 ppm for fragment ion masses, searched in public spectral databases and an in-house spectral library.

    “Compound annotation was performed with a mass accuracy of 3 ppm for precursor masses and 10 ppm for fragment ion masses searched in public spectral databases as well as our in-house spectral library.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    A QC sample was analysed after every 5th experimental sample to monitor instrument performance throughout the analytical sequence.

    “A QC sample was analysed after every 5th experimental sample to monitor instrument performance throughout the analytical sequence.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    Emx1-driven Cre recombinase expression in neural cells of the neocortex was verified using the Gt26Sor mtmG reporter mouse line.

    “Emx1-driven Cre recombinase expression in neural cells of the neocortex was verified by utilizing the Gt26Sor mtmG reporter mouse line.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Pooled quality control (QC) samples were prepared by mixing equal aliquots from each processed sample.

    “Pooled quality control (QC) samples were prepared by mixing equal aliquots from each processed sample.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Slc7a5 is expressed in excitatory neurons of different cortical layers during post-natal neurodevelopment (P1, P7, and P21).

    “Slc7a5 expression in excitatory neurons of different cortical layers during post-natal neurodevelopment (P1, P7, and P21); based on single-cell RNA sequencing data (Yuan et al. 73 ; L2/L3, layer II/III excitatory neurons; L4, layer IV excitatory neurons; L5, LV excitatory neurons; L6, layer VI excitatory neurons).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Yuan et al. 73

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

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

    Study design and conductown data

    In the cortical lysates dataset, two out of five mutant samples behaved as outliers and were excluded from further analysis.

    “During the analysis phase, two out of five mutant samples from the cortical lysates dataset behaved as outliers and were excluded from further analysis.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Crude lysates were pre-cleared to eliminate proteins, which unspecifically bind to the beads.

    “Crude lysates were pre-cleared to eliminate proteins, which unspecifically bind to the beads.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    For determination of background signals and subsequent background subtraction, an additional processed blank sample was recorded.

    “For determination of background signals and subsequent background subtraction, an additional processed blank sample was recorded.”
    Quote in the paperNot a substantial claim

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

    Study design and conductcitation

    Raw peak areas were normalized using the probabilistic quotient normalization method as described in Dieterle et al. 95, implemented using pandas 1.3.5 86 and numpy 1.22.4.87.

    “Raw peak areas were normalized using the probabilistic quotient normalization method as described in Dieterle et al. 95 implemented using pandas 1.3.5 86 and numpy 1.22.4.87”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Dieterle et al. 95

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites pandas 1.3.5 86

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites numpy 1.22.4.87

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

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

    Study design and conductnone given

    Where applicable, multiple testing corrections have been performed using the Benjamini-Hochberg procedure.

    “Where applicable, multiple testing corrections have been performed using the Benjamini-Hochberg procedure.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    All experimental samples were measured in a randomized manner.

    “All experimental samples were measured in a randomized manner.”
    Quote in the paperNot a substantial claim

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

    Study design and conductcitation

    Lysates enriched for cell membranes were obtained based on the protocol modified from Lee et al. 97.

    “Lysates enriched for cell membranes were obtained based on the protocol modified from Lee et al. 97”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Lee et al. 97

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

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

    Study design and conductnone given

    QC samples were additionally measured in confirmation mode to obtain additional MS/MS spectra for metabolite identification.

    “QC samples were additionally measured in confirmation mode to obtain additional MS/MS spectra for identification.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Multiple QCs were injected at the beginning of the analysis in order to equilibrate the LC-MS system.

    “Multiple QCs were injected at the beginning of the analysis in order to equilibrate the LC-MS system.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims. The quote only partly states this claim.

    Study design and conductnone given

    Samples were randomly assigned into the autosampler and analyzed in a randomized fashion for untargeted metabolomics.

    “Samples were randomly assigned into the autosampler and injected on the respective phase system.”
    • Also stated in: § Western Blot · plain
      “Samples were analyzed in a randomized fashion and QC samples were additionally measured in confirmation mode to obtain additional MS/MS spectra for identification.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Experimentally obtained retention times were used for the validation of metabolite identifications.

    “Experimentally obtained retention times were used for the validation of metabolite identifications.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    In the co-immunoprecipitation, normal rabbit IgG was used as a control.

    “During the 2hrs pre-clearing step, pre-washed beads were incubated in lysis buffer with either normal rabbit igG control (Cell Signaling Technologies) or the anti-LAT1 (Slc7a5) antibody (25mg of antibody per sample) rotating at 4 C.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    Feature identification was based on accurate mass, isotope pattern, MS/MS fragment scoring and retention time matching to an in-house library (metabolomics) and the MS-DIAL LipidBlast library (lipidomics).

    “Feature identification was based on accurate mass, isotope pattern, MS/MS fragment scoring and retention time matching to an in-house library (metabolomics) and the MS-DIAL LipidBlast library (lipidomics).”
    Quote in the paperNot a substantial claim
84 of 84

  • The paper relies on this without support.

    Little is known about the critical metabolic changes that neural cells have to undergo during development and how temporary shifts in this program can influence brain circuitries and behavior.

    “Little is known about the critical metabolic changes that neural cells have to undergo during development and how temporary shifts in this program can influence brain circuitries and behavior.”

  • The paper relies on this without support.

    It remains largely unknown whether and how the level of LNAAs changes over time in the brain and how fluctuations in their amount may influence neurodevelopment.

    “However, it remains largely unknown whether and how the level of these amino acids (AAs) changes over time in the brain and how fluctuations in their amount may influence the course of neurodevelopment.”

  • The paper relies on this without support.

    Little is known about the metabolic program unfolding during brain development and the specific nutrient dependencies that this entails.

    “In fact, little is known about the metabolic program unfolding during brain development and the specific nutrient dependencies that this entails.”

  • The paper relies on this without support.

    There are little data on how brain metabolism changes immediately after birth.

    “For example, there are little data on how brain metabolism changes immediately after birth.”

  • The paper relies on this without support.

    It is unknown how maturing neurons meet their energy demand at intermediate developmental stages when they lack full glial support.

    “However, it is unknown how maturing neurons meet their energy demand at intermediate developmental stages when they lack full glial support (Figure 1A middle).”

  • The paper relies on this without support.

    Most LNAAs are essential and their presence in the human body depends on dietary intake.

    “Most of the LNAAs are essential; thus, their presence in the human body depends on dietary intake.”

  • The paper relies on this without support.

    BCAAs are substrates of the SLC7A5 transporter.

    “This cluster sparked our interest since BCAAs are substrates of the SLC7A5 transporter.”

  • The paper relies on this without support.

    There is no detailed description of how the levels of various metabolites change in the forebrain over time.

    “Although metabolism is a crucial element of cellular fitness, there is no detailed description of how the levels of various metabolites change in the forebrain over time.”

  • The paper relies on this without support.

    The potential upstream signaling and the pattern of neuronal activity determining this phenomenon remain unclear.

    “However, the potential upstream signaling and the pattern of neuronal activity determining this phenomenon remain unclear.”

  • The paper relies on this without support.

    Disrupting the refinement process can permanently affect brain circuits.

    “Identifying extrinsic and intrinsic factors that can modulate neuronal properties at this developmental stage is critical since disrupting the refinement process can permanently affect brain circuits.”

  • The authors state this as an assumption.

    Quantifications in reversed phase chromatography would align with quantifications in hydrophilic interaction liquid chromatography.

    “Assuming quantifications in RP would align with quantifications in HILIC, we combined HILIC and RP by adding to the HILIC data those metabolites that were detected by RP but not by HILIC.”

  • The paper relies on this without support.

    The metabolic landscape of maturing neurons is largely unknown.

    “The metabolic landscape of maturing neurons is largely unknown.”
12 of 12

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • P2 data integration includes proteomics performed at P5

    Major

    The Figure 3 legend describes the integrated proteomic, metabolomic, and lipidomic data as coming from P2 tissue and cells, but the methods state that the whole-cortex proteomics samples were collected at P5. This matters to the claimed perinatal timing of the integrated metabolic analysis.

  • Figure 5 sample counts name P2 instead of P5

    Minor

    The legend identifies panel C as an analysis at P5, but its sample-count statement assigns the corresponding group to P2. No P2 panel is described in this figure.

  • P25-P26 electrophysiology group counts are reversed

    Minor

    Figure 6B reports 5 control cells and 15 mutant cells at P25-P26. The Figure S7 legend assigns 5 mutant cells and 15 control cells to its P25-P26 analysis.

  • Two different primer sequences share the same identifier

    Minor

    The key resources table assigns two different sequences to the primer named Q_SLC7A5_In7-8F, making the specified forward primer ambiguous.

  • Proteomics sample count conflicts with stated outlier exclusions

    Minor

    Figure S3I reports four animals per genotype for cortical protein expression. The proteomics methods say that two of five mutant cortical-lysate samples were excluded, leaving three mutant samples for analysis.

5 of 5

Cites

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Authors
Knaus, L. S., Basilico, B., Malzl, D., Gerykova Bujalkova, M., Smogavec, M., Schwarz, L. A., Gorkiewicz, S., Amberg, N., Pauler, F. M., Knittl-Frank, C., Tassinari, M., Maulide, N., Rülicke, T., Menche, J., Hippenmeyer, S., & Novarino, G.
Affiliations
  • Institute of Science and Technology Austria (AT)
  • Max Perutz Labs (AT)
  • Medical University of Vienna (AT)
  • University of Vienna (AT)
  • University of Veterinary Medicine Vienna (AT)
Corresponding author
Novarino, Gaia
ORCID
Venue
Cell
Publisher
Elsevier BV
Published
2023-04
Volume
186
Issue
9
Pages
1950-1967.e25
ISSN
0092-8674 (ISSN-L)
DOI
10.1016/j.cell.2023.02.037
OpenAlex
W4361286239
Cited by
70
References
114
Citations by year
2026: 15 · 2025: 26 · 2024: 18 · 2023: 11
FWCI
5.26
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, other-oa)
  • PubMed (publishedVersion)
  • IRIS Research product catalog (Sapienza University of Rome) (submittedVersion, cc-by-nc-nd)
  • Institute of Science and Technology Austria (submittedVersion, cc-by)
Funders
  • European Union
  • European Research Council (715508, 725780)
  • Austrian Science Fund (DK W1232-B24)
  • Horizon 2020 Framework Programme
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Amino Acid Enzymes and Metabolism
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
Keywords
large neutral amino acids, SLC7A5, neuronal excitability, neuronal survival, cortical development, lipid metabolism, neural activity patterns, autism spectrum disorder, metabolite profiling, postnatal development, circuit dysfunction
MeSH terms
Animals, Brain, Female, Humans, Mutation, Neurons, Pregnancy, Amino Acids, Neutral, Large Neutral Amino Acid-Transporter 1, Mice

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 2026-10-23.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-23.

Cited sources are checked by gpt-6-sol.

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.

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