The DOI matches its Crossref record; 80 of the 80 references it cites were found in a registry or an index; 2 of the 65 sources its claims cite were checked for support.
Molecular interrogation of hypothalamic organization reveals distinct dopamine neuronal subtypes
Open accessAbstract
The hypothalamus contains the highest diversity of neurons in the brain. Many of these neurons can co-release neurotransmitters and neuropeptides in a use-dependent manner. Investigators have hitherto relied on candidate protein-based tools to correlate behavioral, endocrine and gender traits with hypothalamic neuron identity. Here we map neuronal identities in the hypothalamus by single-cell RNA sequencing. We distinguished 62 neuronal subtypes producing glutamatergic, dopaminergic or GABAergic markers for synaptic neurotransmission and harboring the ability to engage in task-dependent neurotransmitter switching. We identified dopamine neurons that uniquely coexpress the Onecut3 and Nmur2 genes, and placed these in the periventricular nucleus with many synaptic afferents arising from neuromedin S+ neurons of the suprachiasmatic nucleus. These neuroendocrine dopamine cells may contribute to the dopaminergic inhibition of prolactin secretion diurnally, as their neuromedin S+ inputs originate from neurons expressing Per2 and Per3 and their tyrosine hydroxylase phosphorylation is regulated in a circadian fashion. Overall, our catalog of neuronal subclasses provides new understanding of hypothalamic organization and function.
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67 Own result · 46 Not checked yet · 21 Own interpretation · 14 Own method · 1 Partly supported · accuracy not yet measured
In the paper
“Subsequent differential mRNA expression analysis subdivided Oxt and Avp neurosecretory cells, which project to the posterior lobe of the pituitary4, into 3 and 4 subtypes, respectively”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
What the sources say
- Not checked yetcites [4] Swanson, L.W. & Kuypers, H.G. The paraventricular nucleus of the hypothalamus: cytoarchitectonic subdivisions and organization of projections to the pituitary, dorsal vagal complex, and spinal cord as demonstrated by retrograde fluorescence double-labeling methods. J. Comp. Neurol. 194, 555–570 (1980). source
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“we set out to validate our data by choosing the predicted 4th subtype of dopamine neurons (cluster #11; “dopamine-4 cells”), which despite its small sample size (6 neurons) segregated from other dopamine clusters in our predictive model”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Expression-based prediction of a novel dopamine neuron subtype
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- Rests on an assumption: Small cluster size and lack of positional information can make single-cell RNA-seq data liable to false positives.
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“It is noteworthy that neuromedin S-containing neurons in the SCN co-express Vip (42%)”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Dopamine-4 neurons are entrained by neuromedin S
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In the paper
“Meanwhile, other Vip+ neurons that, however, were Grp+/- and Nms- clustered separately”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Dopamine-4 neurons are entrained by neuromedin S
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In the paper
“we employed multiple immunofluorescence labeling to show the co-existence of VGLUT2 and GAD67 proteins in nerve endings at the median eminence (ME)”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
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“we also identified combinations of genes whose co-expression patterns predicted individual subclusters with a high degree of accuracy”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Molecular validation through novel neuropeptide marks
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“Subsequent histochemistry validated these data by spatially confining neuromedin S production to the SCN (Fig. 7b) 32.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Dopamine-4 neurons are entrained by neuromedin S
What the sources say
- Not checked yetcites [32] Mori, K. et al. Identification of neuromedin S and its possible role in the mammalian circadian oscillator system. EMBO J. 24, 325–335 (2005). source
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In the paper
“We produced a hierarchical dendrogram (Fig. 2a), which segregated neurotransmitter and neuropeptide-containing cells into subclasses”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
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In the paper
“For clusters #22,#23 we note the differential expression of the hypocretin receptor 1 and 2 genes (Hcrtr1, Hcrtr2), establishing their molecular divergence”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
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In the paper
“Our single-cell RNA-seq data allowed the identification of many novel markers with a high degree of subclass selectivity”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Molecular validation through novel neuropeptide marks
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- It may state two separate claims as one. 46
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In the paper
“These newly distinguished neurons are exclusively situated at the medio-posterior subdivision of the periventricular area and receive Nms + synaptic afferents from SCN neurons regulating the circadian clock 32, 48, in addition to the expression of clock genes in the periventricular and Arc dopamine cells themselves 49.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
What the sources say
- Partly supportedcites [32] Mori, K. et al. Identification of neuromedin S and its possible role in the mammalian circadian oscillator system. EMBO J. 24, 325–335 (2005). sourceExperimental — accuracy not yet measured
The Results report NMS expression in the SCN core, and the Discussion suggests that NMS helps regulate the circadian pacemaker within the SCN. Neither passage establishes NMS-positive synaptic input from the SCN to the specified periventricular neurons. The paper also does not establish those neurons’ exclusive location or clock-gene expression in periventricular and arcuate dopamine cells. (LLM, conf=0.83)
- Not checked yetcites [48] DeMaria, J.E., Lerant, A.A. & Freeman, M.E. Prolactin activates all three populations of hypothalamic neuroendocrine dopaminergic neurons in ovariectomized rats. Brain Res. 837, 236–241 (1999). source
- Partly supportedcites [49] Lee, I.T. et al. Neuromedin s-producing neurons act as essential pacemakers in the suprachiasmatic nucleus to couple clock neurons and dictate circadian rhythms. Neuron 85, 1086–1102 (2015). sourceExperimental — accuracy not yet measured
The Discussion section, “Synaptic Transmission from Nms Neurons is Required for SCN Timekeeping” (page unavailable), identifies Nms neurons as SCN pacemakers that regulate circadian rhythms through synaptic transmission. It does not show that the newly distinguished periventricular neurons receive synaptic afferents from them, or establish those neurons’ exclusive location. The clock-gene statement is attached to citation 49, not this source. (LLM, conf=0.84)
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In the paper
“Coincident detection of onecut-3 and enzymatically-active TH phosphorylated at residue Ser40 26 on a Th- GFP background revealed that these dopamine neurons populate the periventricular nucleus of the hypothalamus (Fig. 5d), with their dendritic trees being primarily oriented vertically”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Expression-based prediction of a novel dopamine neuron subtype
What the sources say
- Not checked yetcites [26] Tekin, I., Roskoski, R. Jr., Carkaci-Salli, N. & Vrana, K.E. Complex molecular regulation of tyrosine hydroxylase. J. Neural Transm. 121, 1451–1481 (2014). source
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In the paper
“we revealed that even minute mRNA copy numbers can translate into meaningful biological modalities by showing the co-existence of vasopressin and VGLUT2 (Fig. 3e) and GAD67 (Fig. 3e).”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
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In the paper
“glutamatergic neurons showed an apparent variability in their gene expression for neuropeptides since clusters #33-35 contained Trh, Bdnf and Adcyap1 mRNAs. In contrast, clusters #57-62 exhibited only marginal (if any) expression of genes for polypeptides (particularly Cck)”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
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In the paper
“we broadened our specificity testing to all 62 predicted neuronal clusters and show that, as opposed to redundant Th and Sst expression, both Slc6a3 (DAT) and Onecut3 were predominantly found in this particular cell population”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Expression-based prediction of a novel dopamine neuron subtype
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In the paper
“Nevertheless, several neuronal clusters were dominated by neurons from male (e.g., #55) or female (#6) subjects, suggesting, but not being causal to, gender-specific coding”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Cell-state changes and neuronal identity
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In the paper
“This was particularly notable for clusters #8-11 cells (with low Gad1 and high Gad2) which also harbored Th, Slc18a2 (vesicular monoamine transporter 2, VMAT2) and in some cases, Slc6a3 (encoding dopamine transporter 1, DAT), qualifying them as dopamine neurons and suggesting the existence of dual dopamine/GABA neurotransmitter phenotypes”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
What the sources say
- Not checked yetcites [21] Everitt, B.J., Hökfelt, T., Wu, J.Y. & Goldstein, M. Coexistence of tyrosine hydroxylase-like and gamma-aminobutyric acid-like immunoreactivities in neurons of the arcuate nucleus. Neuroendocrinology 39, 189–191 (1984). source
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In the paper
“The significance of Th and tyrosine hydroxylase protein in these neurons remains unclear since they appear to lack the gene for aromatic amino-acid decarboxylase (AADC”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
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In the paper
“Accordingly, Qrfp- and Npvf-expressing hypothalamic neurons are both glutamatergic and segregate into clusters #35 and #37, respectively”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Molecular validation through novel neuropeptide marks
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“#62-enriched genes included Zfp458, Ppp1r12b, whereas neurons in #61 were enriched in Pou2f2”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
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“Whereas Adcyap1r1 and Oprl1 are promiscuously expressed in most neuronal clusters (data not shown), Nmur2 and Nmbr are highly specific for dopamine-4 neurons”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Dopamine-4 neurons are entrained by neuromedin S
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“we resolved that Qrfp+ and Npvf hypothalamic neurons co-express the genes for orexin/hypocretin (Hcrt; Supplementary Fig. 6b) and galanin (Gal; Supplementary Fig. 6c), respectively.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Molecular validation through novel neuropeptide marks
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“Neurotransmitter identities included glutamatergic (clusters #1-7 and #30-62), dopaminergic (#8-11) and GABAergic (#12-29) cells.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
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“As such, proopiomelanocortin (Pomc gene) depicted a clustering example”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Neuronal heterogeneity in the hypothalamus
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In the paper
“We show that dopamine-4 neurons (A14, cluster #11) express the gene for DAT (Slc6a3; Fig. 6a), contrasting ventrolateral Arc/A12 neurons, but being in Slc6a3 expression similar to the dorsomedial A12 group 28.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Morphological stratification of dopamine neurons in intact brains
What the sources say
- Not checked yetcites [28] Meister, B. & Elde, R. Dopamine transporter mRNA in neurons of the rat hypothalamus. Neuroendocrinology 58, 388–395 (1993). source
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In the paper
“histochemical detection of TH using COLM technology demonstrated that most TH+ axons leaving the A14 area project laterally using the supraoptic decussation and upper lateral pathway and terminate outside the hypothalamus, likely in basolateral and central amygdaloid nuclei”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Morphological stratification of dopamine neurons in intact brains
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“Lightsheet microscopy revealed 474 cells in an adult mouse brain (Fig. 5f, Supplementary Movies 1,2), while conventional neuroanatomy in serial sections projected 500-1,000 cells/brain.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Expression-based prediction of a novel dopamine neuron subtype
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“In comparison, non-magnocellular cells present a more heterogeneous trait, and can be separated into “GABAergic” (Gad1/Gad2/Slc32a1) and “glutamatergic” (Slc17a6) subtypes20.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
What the sources say
- Not checked yetcites [20] Krashes, M.J. et al. An excitatory paraventricular nucleus to AgRP neuron circuit that drives hunger. Nature 507, 238–242 (2014). source
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In the paper
“When quantifying GFP intensity in these TH+ cells, we find that their GFP signal is significantly lower than that of other TH+ cell groups (Supplementary Fig. 7c), while their phospho-Ser40 levels are distinctly elevated”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Expression-based prediction of a novel dopamine neuron subtype
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In the paper
“In turn, these cells lacked neuropeptides that were abundant in other dopamine subclasses like Ghrh1 and Tac1 (Fig. 5b), and partly Npy2r, the gene encoding neuropeptide Y receptor Y2.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Expression-based prediction of a novel dopamine neuron subtype
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In the paper
“our data reveal that pancreatic polypeptide receptor (Npy6r) mRNA is confined to Vip+/Nms+ circadian neurons (cluster #41) leaving Vip-only neurons of the neighboring circadian cluster (#40) unchecked.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
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In the paper
“our analysis of leptin receptor (Lepr) expression whose activity upon binding white fat-derived leptin 41 is associated with satiety, shows that Lepr is a low-expressed gene (1 mRNA copy/per cell) and it is only detected in ~3% of hypothalamic neurons.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
What the sources say
- Not checked yetcites [41] Friedman, J.M. & Halaas, J.L. Leptin and the regulation of body weight in mammals. Nature 395, 763–770 (1998). source
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In the paper
“Significantly, Per2 was also expressed in cluster #42 (Fig. 8c, Supplementary Fig. 8f), where 23% of Per2+ neurons express Nms”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Neuromedin S+ neurons of the circadian pacemaker might modulate dopamine-4 cells
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In the paper
“Moreover, we characterized Lepr mRNA expression in some glutamatergic neurons, corroborating published reports42.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
What the sources say
- Not checked yetcites [42] Scott, M.M. et al. Leptin targets in the mouse brain. J. Comp. Neurol. 514, 518–532 (2009). source
plaincorrelational- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
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In the paper
“we find that neuromedin S co-localizes with vesicle-associated membrane protein 2 (VAMP2), marking presynapses, in close apposition to predominantly proximal dendrites of TH+/onecut-3+ periventricular neurons”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Neuromedin S+ neurons of the circadian pacemaker might modulate dopamine-4 cells
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In the paper
“This mark showed a distinct difference in the rising slope of cumulative distribution of function (CDF; p = 0.0167), suggesting different levels of active TH enzyme at day and night periods”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Neuromedin S+ neurons of the circadian pacemaker might modulate dopamine-4 cells
What the sources say
- Not checked yetcites [26] Tekin, I., Roskoski, R. Jr., Carkaci-Salli, N. & Vrana, K.E. Complex molecular regulation of tyrosine hydroxylase. J. Neural Transm. 121, 1451–1481 (2014). source
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In the paper
“To remove noise prior to clustering, we first used a primary (coarse) filter and selected for genes, which were distinguished by t-test to be enriched in neurons using a false discovery rate (FDR) of 5% (results in 9,013 neuronal genes in total).”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Level 2 analysis of neurons
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Data were from 898 cells, which qualified as neurons based on level 1 clustering.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Level 2 analysis of neurons
plain- Asserted: passes — The authors put it forward as true.
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In the paper
“Drop-seq data processing of 220 neurons as output was performed using the Drop-seq Tools v1.12 software22.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Drop-seq analysis
What the sources say
- Not checked yetcites [22] Macosko, E.Z. et al. Highly parallel genome-wide expression profiling of individual cells using nanoliter droplets. Cell 161, 1202–1214 (2015). source
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“These refinements led to 62 neuronal sub-clusters being discerned.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Level 2 analysis of neurons
plaindirect- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Cells with more than 1,500 molecules/cell (excluding rRNA, mitochondrial RNA and repeats) were analyzed, and resulted in a total of 3,131 cells.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Level 1 analysis
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“This step distinguished 91 neuronal clusters (Fig. 1c).”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Level 2 analysis of neurons
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In the paper
“Neuromedin S is the endogenous agonist at neuromedin U receptor 2, with its production being concentrated in the SCN32.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Dopamine-4 neurons are entrained by neuromedin S
What the sources say
- Not checked yetcites [32] Mori, K. et al. Identification of neuromedin S and its possible role in the mammalian circadian oscillator system. EMBO J. 24, 325–335 (2005). source
plaindirect- Asserted: passes — The authors put it forward as true.
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In the paper
“these data reinforce our conclusion that hypothalamic neurons might either co-release neurotransmitters, segregate them to subsets of synapses or switch between modes of neurotransmission in response to specific inputs”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
What the sources say
- Not checked yetcites [16] Pinto, S. et al. Rapid rewiring of arcuate nucleus feeding circuits by leptin. Science 304, 110–115 (2004). source
- Not checked yetcites [17] Cristino, L. et al. Obesity-driven synaptic remodeling affects endocannabinoid control of orexinergic neurons. Proc. Natl. Acad. Sci. USA 110, E2229–E2238 (2013). source
hedgedconditional- Asserted: passes — The authors put it forward as true.
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In the paper
“Thus, we suggest that A14 dopamine neurons are bona fide neurosecretory cells (see also31), which are integrated into (extra-) hypothalamic neuronal networks.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Morphological stratification of dopamine neurons in intact brains
What the sources say
- Not checked yetcites [31] Goudreau, J.L., Lindley, S.E., Lookingland, K.J. & Moore, K.E. Evidence that hypothalamic periventricular dopamine neurons innervate the intermediate lobe of the rat pituitary. Neuroendocrinology 56, 100–105 (1992). source
plaincorrelational- Asserted: passes — The authors put it forward as true.
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In the paper
“we show that onecut-3+ dopamine neurons form a cell continuum spanning the SCN-Arc domain with an increasing cell gradient caudally, thus qualifying as “A14” dopamine neurons6, 27.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Expression-based prediction of a novel dopamine neuron subtype
What the sources say
- Not checked yetcites [6] Björklund, A., Moore, R.Y., Nobin, A. & Stenevi, U. The organization of tubero-hypophyseal and reticulo-infundibular catecholamine neuron systems in the rat brain. Brain Res. 51, 171–191 (1973). source
- Not checked yetcites [27] Hökfelt, T., Martensson, R., Bjorklund, A., Kleinau, S. & Goldstein, M. Distribution maps of tyrosine-hydroxylase-immunoreactive neurons in the rat brain. in Handbook of Chemical Neuroanatomy (eds. Bjorklund, A. & Hökfelt, T.) 277–379 (Elsevier, 1984).
boostedcorrelational- Asserted: passes — The authors put it forward as true.
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In the paper
“These data also recapitulate the known dichotomy of hypothalamic dopamine neurons releasing their contents either at the ME into the hypophyseal portal system 29 or using DAT to remove superfluous dopamine at central synapses.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Morphological stratification of dopamine neurons in intact brains
What the sources say
- Not checked yetcites [29] Ben-Jonathan, N. & Hnasko, R. Dopamine as a prolactin (PRL) inhibitor. Endocr. Rev. 22, 724–763 (2001). source
boosteddirect- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“we assign these neurons as bona-fide excitatory neuropeptidergic subtypes.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Molecular validation through novel neuropeptide marks
plain- Asserted: passes — The authors put it forward as true.
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In the paper
“The latter neuronal pool also contains GAD21 and is a source of dopamine inhibiting prolactin release 29.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Morphological stratification of dopamine neurons in intact brains
What the sources say
- Not checked yetcites [29] Ben-Jonathan, N. & Hnasko, R. Dopamine as a prolactin (PRL) inhibitor. Endocr. Rev. 22, 724–763 (2001). source
plaindirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“neuromedin S release into the CSF would reconcile this difference and provide an alternative means of long-distance signaling.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Neuromedin S release sites
hedgeddirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Our molecular profiling focused on the central column of the hypothalamus, leaving its most lateral and caudal segments uncharted.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
plain- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“showing that episodic expression of any gene during the lifetime of a neuron might be considered a common feature.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
hedged- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Particular effort was directed towards minimizing the number of animals used and their suffering during experiments.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Animals, tissue preparation and histochemistry
plaindirect- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
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In the paper
“By consensus definition, each neuron is expected to contain at least one fast neurotransmitter, which can be co-released with another neurotransmitter (e.g. dopamine), one or more peptide, gaseous or lipid neuromodulator20, 43–45.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
What the sources say
- Not checked yetcites [20] Krashes, M.J. et al. An excitatory paraventricular nucleus to AgRP neuron circuit that drives hunger. Nature 507, 238–242 (2014). source
- Not checked yetcites [43] Boulland, J.L. et al. Vesicular glutamate and GABA transporters sort to distinct sets of vesicles in a population of presynaptic terminals. Cereb. Cortex 19, 241–248 (2009). source
- Not checked yetcites [44] Dawson, T.M. & Snyder, S.H. Gases as biological messengers: nitric oxide and carbon monoxide in the brain. J. Neurosci. 14, 5147–5159 (1994). source
- Not checked yetcites [45] Jonas, P., Bischofberger, J. & Sandkühler, J. Corelease of two fast neurotransmitters at a central synapse. Science 281, 419–424 (1998). source
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
In the paper
“When comparing single-cell RNA-seq data from cortical/hippocampal vs. hypothalamic neurons, the following differences apply: i) in the cerebral cortex, higher mRNA transcript levels are seen in interneurons, which also contain both Gad1 and Gad2 transcripts, and ii) cortical neurons rarely, if at all, produce mixed excitatory/inhibitory phenotypes 9.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
What the sources say
- Not checked yetcites [9] Zeisel, A. et al. Brain structure. Cell types in the mouse cortex and hippocampus revealed by single-cell RNA-seq. Science 347, 1138–1142 (2015). source
plaincorrelational- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
None of the sources cited for this claim have been checked yet.
In the paper
“For dopamine, parvocellular dorsomedial neurosecretory cells (Th+) in the Arc (A12) give rise to large nerve endings in the ME, amassing 1/3 of all boutons in the lateral external layer 46, and release dopamine for the tonic inhibition of pituitary prolactin release29.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
What the sources say
- Not checked yetcites [46] Ajika, K. & Hökfelt, T. Ultrastructural identification of catecholamine neurones in the hypothalamic periventricular-arcuate nucleus-median eminence complex with special reference to quantitative aspects. Brain Res. 57, 97–117 (1973). source
- Not checked yetcites [29] Ben-Jonathan, N. & Hnasko, R. Dopamine as a prolactin (PRL) inhibitor. Endocr. Rev. 22, 724–763 (2001). source
plaindirect- Asserted: passes — The authors put it forward as true.
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- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
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It may state two separate claims as one.
None of the sources cited for this claim have been checked yet.
In the paper
“This synaptic circuit is significant, since it integrates the circadian pacemaker (characterized by Per genes) 33, 34and hormonal output, particularly diurnally fluctuating dopamine levels in the general circulation 7.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
What the sources say
- Not checked yetcites [33] Zhang, L. et al. A PERIOD3 variant causes a circadian phenotype and is associated with a seasonal mood trait. Proc. Natl. Acad. Sci. USA 113, E1536–E1544 (2016). source
- Not checked yetcites [34] Vitaterna, M.H. et al. Differential regulation of mammalian period genes and circadian rhythmicity by cryptochromes 1 and 2. Proc. Natl. Acad. Sci. USA 96, 12114–12119 (1999). source
- Not checked yetcites [7] Horvath, T.L. Suprachiasmatic efferents avoid phenestrated capillaries but innervate neuroendocrine cells, including those producing dopamine. Endocrinology 138, 1312–1320 (1997). source
plaincorrelational- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
None of the sources cited for this claim have been checked yet.
In the paper
“The reporter lines GAD67+/gfp, GAD67+/gfp::CCKBAC/DsRedT3 52, 53, B6.B6D2-Tg(Th-EGFP)21-31Koba (developed by K. Kobayashi and available from Riken Bioresource Center), Crh-IRES-Cre::B6.Cg-Gt(ROSA)26Sortm6(CAG-ZsGreen 1)Hze/J (Ai6; both GFP reporters were from the Jackson laboratories, #012704 and #007906) and dopamine transporter (Dat1)-Cre::B6.Cg-Gt(ROSA)26Sortm14(CAG-dTomato)Hze/J mice were”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Animals, tissue preparation and histochemistry
What the sources say
- Not checked yetcites [52] Máté, Z. et al. Spatiotemporal expression pattern of DsRedT3/CCK gene construct during postnatal development of myenteric plexus in transgenic mice. Cell Tissue Res. 352, 199–206 (2013). source
- Not checked yetcites [53] Pierreux, C.E., Vanhorenbeeck, V., Jacquemin, P., Lemaigre, F.P. & Rousseau, G.G. The transcription factor hepatocyte nuclear factor-6/Onecut-1 controls the expression of its paralog Onecut-3 in developing mouse endoderm. J. Biol. Chem. 279, 51298–51304 (2004). source
plainn = 3- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim.
In the paper
“When comparing single-cell RNA-seq data from cortical/hippocampal vs. hypothalamic neurons, the following differences apply: i) in the cerebral cortex, higher mRNA transcript levels are seen in interneurons, which also contain both Gad1 and Gad2 transcripts, and ii) cortical neurons rarely, if at all, produce mixed excitatory/inhibitory phenotypes 9.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
What the sources say
- Not checked yetcites [9] Zeisel, A. et al. Brain structure. Cell types in the mouse cortex and hippocampus revealed by single-cell RNA-seq. Science 347, 1138–1142 (2015). source
hedged- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“An additional strength of our dataset is that it relies on both male and female animals kept under ad lib feeding conditions, suggesting that we resolved ground-state Lepr mRNA expression, relevant for behavioral output in both sexes (see also Supplementary Fig. 2a).”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. It may state two separate claims as one.
In the paper
“This result lends significant support to previous data 40 and advances knowledge in receptor-dependent function determination for pancreatic polypeptide.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
What the sources say
- Not checked yetcites [40] Yulyaningsih, E. et al. Pancreatic polypeptide controls energy homeostasis via Npy6r signaling in the suprachiasmatic nucleus in mice. Cell Metab. 19, 58–72 (2014). source
boostedcorrelational- Asserted: passes — The authors put it forward as true.
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- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
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It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“The processing of cells from male and female animals was random to avoid any methodological bias.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Cell capture, imaging, lysis and RNA-seq
plain- Asserted: passes — The authors put it forward as true.
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- Load-bearing: fails — The paper’s conclusions may not depend on it.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
None of the sources cited for this claim have been checked yet.
In the paper
“These might be bioenergetically more favourable than rewiring strategies 16, 17.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
What the sources say
- Not checked yetcites [16] Pinto, S. et al. Rapid rewiring of arcuate nucleus feeding circuits by leptin. Science 304, 110–115 (2004). source
- Not checked yetcites [17] Cristino, L. et al. Obesity-driven synaptic remodeling affects endocannabinoid control of orexinergic neurons. Proc. Natl. Acad. Sci. USA 110, E2229–E2238 (2013). source
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
In the paper
“No statistical methods were used to pre-determine sample sizes but our sample sizes are similar to those reported for the cerebral cortex and hippocampus 9.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Level 1 analysis
What the sources say
- Not checked yetcites [9] Zeisel, A. et al. Brain structure. Cell types in the mouse cortex and hippocampus revealed by single-cell RNA-seq. Science 347, 1138–1142 (2015). source
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Experimental protocols were in accordance with the European Communities Council Directive (86/609/EEC), approved by regional ethical committees and regulated by applicable local laws (Stockholms Norra Djurförsöksetiska Nämnd; N512/12 (Sweden) and Tierversuchsgesetz 2012, BGBI, Nr. 114/2012 (Austria)).”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Animals, tissue preparation and histochemistry
plain- Asserted: passes — The authors put it forward as true.
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- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“This arrangement could contribute to the flexibility of hypothalamic neuronal networks to adapt to ever-changing environmental stimuli.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: fails — It may be too vague to check against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
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Probably not a substantial claim. It may be too vague to check against evidence. The paper’s conclusions may not depend on it.
In the paper
“Our data outline a potential cellular substrate that may participate in dopamine release into the hypophyseal portal system and/or the intermediate lobe for the chronospecific inhibition of pituitary prolactin secretion 29 (Fig. 6g,h).”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
What the sources say
- Not checked yetcites [29] Ben-Jonathan, N. & Hnasko, R. Dopamine as a prolactin (PRL) inhibitor. Endocr. Rev. 22, 724–763 (2001). source
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Capture quality for exclusion of debris or doublets was performed after each capture experiment.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Cell capture, imaging, lysis and RNA-seq
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. The quote only partly states this claim.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Cumulatively, we recognize onecut-3+ A14 dopamine neurons as distinctly different – at the level of molecular make-up and connectivity – from the dorsomedial A12 dopamine group; thus showcasing that single-cell RNA-seq in combination with neuroanatomy can produce a molecular protomap of hypothalamic neuronal organization and aid the future discovery of functionally-segregated neuronal subclasses.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Discussion
plainconditional- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“To this end, we used two independent criteria: the difference in the average and the difference in the fraction of positive (>0) cells (Table S3).”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Dendrogram construction and split point listing
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“and merged clusters that showed a high percentage of similarity, i.e. none of the key markers differed between the two cell populations.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Level 2 analysis of neurons
plain- Asserted: passes — The authors put it forward as true.
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- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Data from all cells that passed visual quality control were imported as molecule counts (including all metadata annotations).”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Level 1 analysis
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Moreover, we omitted clusters that appeared thalamus specific because of the presence of known thalamus-specific homeobox genes and transcription factors.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Level 2 analysis of neurons
hedged- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Only neurons situated ventral to the PVN and <120 μm from the lateral wall of the third ventricle were counted at both hemispheres (Fig. 5d, Supplementary Fig. 7d).”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Quantitative morphometry of periventricular onecut-3+/TH+ neurons
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Next, we manually removed putative artifactual clusters that had formed due to low/variable quality of RNA”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Level 2 analysis of neurons
hedgeddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
None of the sources cited for this claim have been checked yet.
In the paper
“The algorithm has by now been extensively validated using both computational (including comparisons to PCA, and independent-component analysis, tSNE, and GP-LVM) biological validation tools 9, 63–66.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > BackSPIN algorithm
What the sources say
- Not checked yetcites [9] Zeisel, A. et al. Brain structure. Cell types in the mouse cortex and hippocampus revealed by single-cell RNA-seq. Science 347, 1138–1142 (2015). source
- Not checked yetcites [63] Schwanhäusser, B. et al. Global quantification of mammalian gene expression control. Nature 473, 337–342 (2011). source
- Not checked yetcites [64] Islam, S. et al. Quantitative single-cell RNA-seq with unique molecular identifiers. Nat. Methods 11, 163–166 (2014). source
- Not checked yetcites [65] Tsafrir, D. et al. Sorting points into neighborhoods (SPIN): data analysis and visualization by ordering distance matrices. Bioinformatics 21, 2301–2308 (2005). source
- Not checked yetcites [66] Marques, S. et al. Oligodendrocyte heterogeneity in the mouse juvenile and adult central nervous system. Science 352, 1326–1329 (2016). source
boosted- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
In the paper
“The q value was calculated as the p value corrected for multiple testing using horizontal correction across the 62 clusters with the Benjamini-Hochberg procedure 75 to control the false discovery rate (FDR).”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Calculating significance using the Wilcoxon rank-sum test
What the sources say
- Not checked yetcites [75] Ose, A. et al. Functional characterization of mouse organic anion transporting peptide 1a4 in the uptake and efflux of drugs across the blood-brain barrier. Drug Metab. Dispos. 38, 168–176 (2010). source
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“None of the experiments required data collection in a blinded fashion.”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Online Methods > Statistical analysis of histochemical and imaging data
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
The paper relies on this without support.
The hypothalamus lacks distinct layering or stringent anatomical organizational principles.
“In contrast to the cerebellum and cerebral cortex, the hypothalamus lacks distinct layering or other stringent anatomical, repetitive organizational principles.”
The paper relies on this without support.
The degree of molecular diversity among hypothalamic dopamine neurons is unknown.
“However, the degree of molecular diversity amongst dopamine neurons remains unknown.”
The paper relies on this without support.
There is no consensus on or direct correlation between neurotransmitter and neuropeptide expression patterns.
“However, there is no consensus on, or direct correlation between, neurotransmitter-neuropeptide expression patterns.”
The paper relies on this without support.
Small cluster size and lack of positional information can make single-cell RNA-seq data liable to false positives.
“However, the often small cluster size (< 10 cells) and the lack of positional information per se could render single-cell RNA-seq data liable to false positive outcomes.”
The paper relies on this without support.
The hypothalamus has many neuronal subtypes that are functionally different yet intermingled in small volumes, which makes it hard to study.
“In contrast, the study of brain regions that contain many functionally distinct, but positionally interspersed, neuronal subtypes in small volumes, such as the hypothalamus, remains a significant challenge.”
The paper relies on this without support.
Any new method requires extensive validation.
“Any new method requires extensive validation.”
The paper relies on this without support.
Perfusion through the internal carotid and vertebral arteries preserves tissue integrity better than other fixation methods.
“We have applied direct perfusion via the internal carotid and vertebral arteries, which facilitated the preservation of tissue integrity relative to alternative fixation methods.”
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?
In the citing paper
“Transcriptomically derived cell type taxonomies in the adult mammalian brain, with majority of the studies conducted in mouse, have consistently revealed a hierarchical organization of cell types (Fig. 2) (Brain Initiative Cell Census Network, 2021; Macosko et al., 2015; Romanov et al., 2017; Russ et al., 2021; Saunders et al., 2018; Shekhar et al., 2016; Tasic et al., 2016; Tasic et al., 2018;”
From What is a cell type and how to define it? (2022) · cites this paper as Romanov et al., 2017
In the citing paper
“Moreover, single-cell sequencing has the potential to identify cell-type-specific molecular targets for the development of new pharmaceuticals that would affect specific neural circuits rather than the entire brain [69].”
From Translational opportunities for circuit-based social neuroscience: advancing 21st century psychiatry (2021) · cites this paper as [69]
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.
Rated minor: it would not touch a result — a value no result rests on, a p-value that stays on the same side of the significance level, or a conflict that is clerical, resolved elsewhere in the paper, or outside its own findings.
In the paper
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
“Our hierarchical clustering approach also separated 33 clusters of mostly “glutamatergic” modalities (#28-62).”
§ A novel organizing principle of the hypothalamus reveals molecularly segregated periventricular dopamine neurons > Results > Hierarchical clustering reveals neurotransmitter-neuropeptide relationships
“Neurotransmitter identities included glutamatergic (clusters #1-7 and #30-62), dopaminergic (#8-11) and GABAergic (#12-29) cells.”
Citations
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Distinct evolutionary trajectories of neuronal and hair cell nicotinic acetylcholine receptorsPreprint
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One Cell At a Time: A Unified Framework to Integrate and Analyze Single-cell RNA-seq DataPreprint
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Metadata
- Authors
- Romanov, R. A., Zeisel, A., Bakker, J., Girach, F., Hellysaz, A., Tomer, R., Alpár, A., Mulder, J., Clotman, F., Keimpema, E., Hsueh, B., Crow, A. K., Martens, H., Schwindling, C., Calvigioni, D., Bains, J. S., Máté, Z., Szabó, G., Yanagawa, Y., Zhang, M.-D., Rendeiro, A., Farlik, M., Uhlén, M., Wulff, P., Bock, C., Broberger, C., Deisseroth, K., Hökfelt, T., Linnarsson, S., Horvath, T. L., & Harkany, T.
- Affiliations
- Medical University of Vienna (AT)
- Karolinska Institutet (SE)
- Stanford University (US)
- Semmelweis University (HU)
- Science for Life Laboratory (SE)
- UCLouvain (BE)
- University of Calgary (CA)
- HUN-REN Institute of Experimental Medicine (HU)
- Gunma University (JP)
- Austrian Academy of Sciences (AT)
- Christian-Albrechts-Universität zu Kiel (DE)
- ORCID
- Romanov, Roman A
- Zeisel, Amit
- Bakker, Joanne
- Hellysaz, Arash
- Tomer, Raju
- Alpár, Alán
- Mulder, Jan
- Clotman, Frédéric
- Keimpema, Erik
- Hsueh, Brian
- Martens, Henrik
- Calvigioni, Daniela
- Bains, Jaideep S
- Máté, Zoltán
- Szabó, Gábor
- Yanagawa, Yuchio
- Rendeiro, Andre
- Farlik, Matthias
- Uhlén, Mathias
- Wulff, Peer
- Bock, Christoph
- Broberger, Christian
- Deisseroth, Karl
- Hökfelt, Tomas
- Linnarsson, Sten
- Harkany, Tibor
- Venue
- Nature Neuroscience
- Published
- 2016-12-19
- Volume
- 20
- Issue
- 2
- Pages
- 176-188
- ISSN
- 1097-6256 (ISSN-L)
- 1546-1726
- PMC
- PMC7615022
- OpenAlex
- W2562003322
- Cited by
- 502
- References
- 80
- Citations by year
- 2026: 25 · 2025: 54 · 2024: 42 · 2023: 75 · 2022: 58 · 2021: 58 · 2020: 56 · 2019: 37 · 2018: 66 · 2017: 29 · 2016: 1
- FWCI
- 22.91
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Neuroscience (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Digital Access to Libraries (submittedVersion, other-oa)
- Funders
- Semmelweis Egyetem
- Novo Nordisk Fonden (NNF17OC0027294, NNF15OC0015964, NNF14OC0011061, NNF16OC0021482, NNF14OC0010695, NNF15OC0016258)
- National Institute on Aging (P01AG051459)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
- Life Sciences › Neuroscience › Endocrine and Autonomic Systems › Circadian rhythm and melatonin
- Health Sciences › Medicine › Reproductive Medicine › Hypothalamic control of reproductive hormones
- Keywords
- hypothalamus, dopaminergic neurons, neuronal cell types, single-cell RNA sequencing, neuropeptides, neurotransmitters, circadian clock, neuronal organization
- MeSH terms
- Animals, Dopamine, Hypothalamus, Immunohistochemistry, Mice, Inbred C57BL, Mice, Transgenic, Synaptic Transmission, Neuropeptides, Suprachiasmatic Nucleus, Tyrosine 3-Monooxygenase, Neurotransmitter Agents, Dopaminergic Neurons
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.
A language model drew the claims from the paper on 2026-10-07, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).
A second model labelled each claim on 2026-10-07 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).
Cited sources are checked with a language model’s help (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.
What counts as a claim ›How we measure confidence ›How we check this ›
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