Cell type-specific transcriptional programs in mouse prefrontal cortex during adolescence and addiction

May combine two claims.

The neurons express Snap25 and can be divided into excitatory (Slc17a7+) and inhibitory (Gad2+) neurons.

“The neurons express Snap25 and can be divided into excitatory (Slc17a7+) and inhibitory (Gad2+) neurons (Fig. 1c, d).”

May combine two claims.

Excitatory and inhibitory neurons of the PFC exhibit great transcriptional differences, forming distinct cell subtypes.

“Indeed, we found that excitatory and inhibitory neurons of the PFC exhibit great transcriptional differences, forming distinct cell subtypes.”

May combine two claims.

Using the two models (adolescence and drug addiction), the study uncovered transcriptional dynamics in each neuron subtype associated with adaptive and challenge-induced neuroplasticity, highlighting distinct and conserved mechanisms.

“Using the two models (adolescence and drug addiction), we uncovered transcriptional dynamics in each neuron subtype associated with adaptive and challenge-induced neuroplasticity, highlighting distinct and conserved mechanisms.”

May combine two claims.

The excitatory neurons form the largest (52.3%) cell class in the PFC, while the inhibitory neurons comprise a smaller portion (4.3%) of the total populations.

“The excitatory neurons form the largest (52.3%) cell class in the PFC, while the inhibitory neurons comprise a smaller portion (4.3%) of the total populations, consistent with the general excitatory/inhibitory ratio reported in most cortical areas (Supplementary Data 2).”

May combine two claims.

For example, corticotrophin releasing hormone signaling (Exc-1,2,6,7,8), dopamine receptor signaling (Exc-7,8), glutamate receptor signaling (Exc-4,5,7,8,9), CXCR4 signaling (Exc-1 to 9, 13) or IGF-1 signaling (Exc-2,3,6,8,13) selectively impacted only the relevant clusters.

“For example, corticotrophin releasing hormone signaling (known to alter pyramidal neuron excitability29, Exc-1,2,6,7,8) dopamine receptor signaling (Exc-7,8), glutamate receptor signaling (Exc-4,5,7,8,9), CXCR4 signaling (Exc-1 to 9, 13) or IGF-1 signaling (Exc-2,3,6,8,13) selectively impacted only the relevant clusters (Fig. 4e).”

May combine two claims.

Even non-neuronal cells undergo robust changes in transcription, with the highest impact on endothelial cells with 879 genes affected.

“Interestingly, even the non-neuronal cells undergo robust changes in transcription, with the highest impact on endothelial cells with 879 genes affected (Fig. 4c).”

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

Following the same filtering criteria of P60 PFC, the study obtained 10,646 high quality mouse P21 PFC cells.

“Following the same filtering criteria of P60 PFC, we obtained 10,646 high quality (median of 2808 genes and 7384 UMI per-cell) mouse P21 PFC cells.”

May combine two claims.

Unlike excitatory neurons, inhibitory neurons do not show distinct laminar organization, although some types may be differentially enriched within particular cortical layers.

“Unlike excitatory neurons, inhibitory neurons do not show distinct laminar organization, although some types may be differentially enriched within particular cortical layers.”

May combine two claims.

Multi-channel RNA-smFISH revealed that the distinct non-overlapping Pou3f1+ (Exc-10) and Tshz2+ (Exc-12) neurons both express Etv-1, revealing their L5 identity.

“Multi-channel RNA-smFISH revealed that the distinct non-overlapping Pou3f1+ (Exc-10) and Tshz2+ (Exc-12) neurons both express Etv-1, revealing their L5 identity (Fig. 2e).”

May combine two claims.

Across cell types, the highest percentage of affected genes comprised enzymes (~50%), followed by transcription factors (~12-13%) and transporters.

“Interestingly, across cell types, the highest percentage of affected genes comprised enzymes (~50%), followed by transcription factors (~12–13%) and transporters (Supplementary Fig. 6b).”

May combine two claims.

Within excitatory neurons, between 4 (in Exc-10) to 50 (in Exc-8) epigenetic regulators were significantly altered in the various subtypes, with most subtypes having 20-30 epigenetic regulators changed.

“For example, within the excitatory neurons, between 4 (in Exc-10) to 50 (in Exc-8) epigenetic regulators were significantly altered in the various subtypes, with most subtypes having 20–30 epigenetic regulators changed (Supplementary Data 5).”

May combine two claims.

Prmt1, a histone arginine methylase associated with gene activation, is upregulated in most excitatory neuron subtypes; while Yeats4 and Ube2d3 involved in histone acetylation and ubiquitylation, respectively, are downregulated across most excitatory neuron subtypes.

“For example, Prmt1, a histone arginine methylase associated with gene activation31, is upregulated in most excitatory neuron subtypes; while Yeats4 and Ube2d3 involved in histone acetylation and ubiquitylation, respectively, are downregulated across most excitatory neuron subtypes (Fig. 4f).”

• cites [31] Wang, H. et al. Methylation of histone H4 at arginine 3 facilitating transcriptional activation by nuclear hormone receptor. Science 293, 853–857 (2001). sourceNot checked yet

  Not checked in the free run.

• AssumptionRests on an assumption: Altered histone modifiers can regulate gene expression either positively or negatively.

May combine two claims.

As many as 646 genes in mature oligodendrocytes, 288 genes in microglia, 268 genes in astrocytes, 209 genes in newly formed oligodendrocytes, and 53 genes in oligodendrocyte precursor were affected.

“In addition, as many as 646 genes in mature oligodendrocytes, 288 genes in microglia, 268 genes in astrocytes, 209 genes in newly formed oligodendrocytes, and 53 genes in oligodendrocyte precursor were affected (Fig. 4c).”

May combine two claims.

Higher resolution clustering of PFC excitatory neurons identified 26 distinct subtypes, a number comparable to the excitatory neuron subtypes of VISp and ALM reported by Tasic et al.

“To enable a better comparison, we first performed a “higher resolution” clustering of the PFC excitatory neurons, which identified 26 distinct subtypes (Fig. 3a), a number comparable to the excitatory neuron subtypes of VISp and ALM reported by Tasic et al.14.”

• cites Tasic et al.14 Tasic, B. et al. Shared and distinct transcriptomic cell types across neocortical areas. Nature 563, 72–78 (2018). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

The study detects distinct inhibitory neuron populations in PFC and defines characteristic regional markers for their identification.

“While divergent inhibitory neuron populations have been reported in other cortical regions based on different marker combinations14, we detect the distinct populations in PFC and define characteristic regional markers for their identification.”

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

Inhibitory neurons are present in much lower numbers in the PFC, accounting for 4.3% of total PFC cells (at a ratio of 1:12 with excitatory neurons).

“Like most other cortical areas, the inhibitory neurons are present in much lower numbers in the PFC, only accounting for 4.3% of the total PFC cells (at a ratio of 1:12 with excitatory neurons).”

May combine two claims.

PFC-9 (G13) or PFC6,15,16,17 (G11) showed no similarity to any VISp subpopulations, although they grouped with L5-IT projection cells in the ALM.

“For example, PFC-9 (G13) or PFC6,15,16,17 (G11) showed no similarity to any VISp subpopulations, although they grouped with L5-IT projection cells in the ALM (Fig. 3c: PFC labeled in blue, VISp red and ALM purple).”

May combine two claims.

Unique markers could be detected for each of the 26 subtypes, showed specific enrichment on t-SNE plots, and could be validated histologically from RNA-FISH.

“Unique markers could be detected for each of the 26 subtypes (Fig. 3b) that showed specific enrichment on the t-SNE plots (Supplementary Fig. 4a) and could be validated histologically from RNA-FISH (Supplementary Fig. 4b).”

May combine two claims.

Rspo2, a common marker for the G11 members, showed distinctly enriched expression in PFC, but no appreciable signal in VISp.

“Rspo2, a common marker for the G11 members, showed distinctly enriched expression in PFC, but no appreciable signal in VISp (Fig S4c).”

May combine two claims.

Some non-inhibitory neuron populations also appear to express Meis2, making Pbx3 a more selective marker.

“Some non-inhibitory neuron populations also appear to express Meis2 (Supplementary Fig. 3c), making Pbx3 a more selective marker.”

May combine two claims.

PFC clusters 7 and 14 (G7) showed no similarity to the ALM subpopulations but resembled the L6IT-5.

“Similarly, PFC clusters 7 and 14 (G7) showed no similarity to the ALM subpopulations but resembled the L6IT-5 (Fig. 3c, Supplementary Data 3).”

May combine two claims.

The major known inhibitory neuron populations like Sst+, Pavlb+ and Vip+ were detected and could be confirmed from RNA-FISH.

“As expected, the major known inhibitory neuron populations like Sst+, Pavlb+ and Vip+ were detected (Fig. 2g, Supplementary Fig. 3b) and could also be confirmed from RNA-FISH (Supplementary Fig. 3c).”

May combine two claims.

While most excitatory neuron subtypes of PFC have corresponding cell clusters in VISp or ALM group together, clusters that are unique to PFC do exist.

“While most excitatory neuron subtypes of PFC have corresponding cell clusters in VISp or ALM group together, clusters that are unique to PFC do exist.”

May combine two claims.

Using scRNA-seq, the study revealed the comprehensive neuronal subtypes comprising the anatomically defined PFC region and their corresponding cortical layer identity.

“In summary, using scRNA-seq, we revealed the comprehensive neuronal subtypes comprising the anatomically defined PFC region and their corresponding cortical layer identity.”

May combine two claims.

Co-staining for unique markers detected Exc-10 (Pou3f1+) and Exc-12 (Tshz2+) neurons adjacently within the same layer, while Exc-13 (Foxp2+) neuron cluster was detected mainly in a deeper layer.

“Co-staining for unique markers detected the Exc-10 (Pou3f1+) and Exc-12 (Tshz2+) neurons adjacently within the same layer, while Exc-13 (Foxp2+) neuron cluster was detected mainly in a deeper layer (Fig. 2c).”

May combine two claims.

Classification revealed 12 clusters of inhibitory neurons (Inhib-1 to Inhib-12), highlighting functional diversity and specialization.

“Classification revealed 12 clusters (referred as Inhib-1, 2,..12), highlighting functional diversity and specialization (Fig. 2f, Supplementary Fig. 3a).”

May combine two claims.

Nnat, a selective marker for G7, is seen enriched in PFC but depleted in ALM.

“Accordingly, Nnat, a selective marker for G7 is seen enriched in PFC but depleted in ALM (Supplementary Fig. 4d).”

May combine two claims.

Exc-4 (Calb1+/Cux2+) and Exc-6 (Calb1−/Cux2+) belong to the superficial layer L2/3, while cluster Exc-8 and Exc-13 belong to L5 and L6, respectively.

“Notably, Exc-4 (Calb1+/Cux2+) and Exc-6 (Calb1−/Cux2+) belong to the superficial layer L2/3. On the other hand, cluster Exc-8 and Exc-13 belong to L5 and L6, respectively (Supplementary Fig. 2d-table).”

May combine two claims.

Effect was smaller at 48 h, but became more pronounced at 15 day with all excitatory subtypes impacted to some degrees.

“Effect was smaller at 48 h, but became more pronounced at 15 day with all excitatory subtypes impacted to some degrees.”

May combine two claims.

Additional pathways that can support plasticity are also affected in subtypes Exc-7 and 8 (example: ephrin, integrin, Rac, Gastrin/Cck and ultimately, even synaptic LTP), suggesting their greater involvement.

“Additional pathways that can support plasticity are also affected in subtypes Exc-7 and 8 (example: ephrin, integrin, Rac, Gastrin/Cck and ultimately, even synaptic LTP), suggesting their greater involvement”

May combine two claims.

Key neuronal functions like axon guidance, gap junction signaling and vesicular trafficking (clathrin mediated endocytosis) are all affected in these five excitatory neuron clusters that can impact structural as well as electrical properties (through gap junction) of synapses.

“Key neuronal functions like axon guidance, gap junction signaling and vesicular trafficking (clathrin mediated endocytosis) are all affected in these five excitatory neuron clusters that can impact structural as well as electrical properties (through gap junction) of synapses.”

May combine two claims.

At 15 day drug withdrawal, the number of genes affected increased in every single subtype with Exc-2, 6, 7, and 8 showing the greatest effects.

“At 15 day drug withdrawal, the number of genes affected increased in every single subtype with Exc-2, 6, 7, and 8 showing the greatest effects.”

May combine two claims.

Contrary to neurons, non-neurons were impacted most strongly at 48 h time point and subdued at 15-day in general.

“contrary to neurons, non-neurons were impacted most strongly at 48 h time point and subdued at 15-day in general”

May combine two claims.

Genes like Adamts16, Myo18b, Astn2, and Erbb2 exhibit cell cluster-specific or enriched expression and are only associated with bipolar, while some other genes, like Zfp804a and Zfpm2, are associated with both.

“genes like Adamts16, Myo18b, Astn2, and Erbb2 exhibit cell cluster-specific or enriched expression (Fig. 5c) and are only associated with bipolar, while some other genes, like Zfp804a and Zfpm2, are associated with both.”

May combine two claims.

In schizophrenia, excitatory neurons are highly affected, with Exc-9 and 11 as the most relevant cells as these cell clusters express the highest number of schizophrenia relevant genes.

“in schizophrenia, we found that excitatory neurons are highly affected, with Exc-9 and 11 as the most relevant cells as these cell clusters express the highest number of schizophrenia relevant genes”

May combine two claims.

A limited impact on non-neuronal populations was also observed, with the endothelial cells most affected, followed by oligodendrocytes.

“A limited impact on non-neuronal populations was also observed, with the endothelial cells most affected, followed by oligodendrocytes.”

May combine two claims.

Exc-6 and Exc-2 are of L2/3 while Exc-8, 7, and 10 are L5.

“Exc-6 and Exc-2 are of L2/3 while Exc-8, 7, and 10 are L5”

May combine two claims.

Exc-1, 8 and 13 were the most strongly induced, while Exc-6 maintained induction.

“Exc-1, 8 and 13 were the most strongly induced, while Exc-6 maintained induction”

May combine two claims.

About 10 or less genes were altered per excitatory neuron subtype (with no change in Exc-8, 10, 11) except Exc-6 where more than 40 genes changed.

“About 10 or less genes were altered per excitatory neuron subtype (with no change in Exc-8, 10, 11) except Exc-6 where more than 40 genes changed”

May combine two claims.

For schizophrenia, Exc-13 accounts for the maximum number of differential GWAS candidate genes among the neurons, and endothelial cells account for the maximum of the non-neuronal cells.

“for schizophrenia, Exc-13 accounts for the maximum number of differential GWAS candidate genes among the neurons, and the endothelial cells account for the maximum of the non-neuronal cells”

May combine two claims.

A broad impact of cocaine on transcription was observed across PFC, but with selective impacts on different cell populations at different stages.

“A broad impact of cocaine on transcription was observed across PFC, but with selective impacts on different cell populations at different stages”

May combine two claims.

For bipolar disorder, Exc-8 accounts for the most candidates (followed by Exc-13), while endothelial cells for the non-neuronal cells.

“for bipolar disorder Exc-8 accounts for the most candidates (followed by Exc-13), while endothelial cells for the non-neuronal cells”

May combine two claims.

Compared to the excitatory neurons of VISp and ALM, the PFC has 5 and 2 unique neuron subtypes respectively, which can be identified using distinctive markers (like Nnat or Rspo2).

“Compared to the excitatory neurons of VISp and ALM, the PFC has 5 (PFC9, 15, 16, 17) and 2 (PFC7 and 14) unique neuron subtypes respectively (Fig. 3c), which can be identified using distinctive markers (like Nnat or Rspo2)”

May combine two claims.

The impact is negligible during maintenance and dramatically increases during prolonged drug withdrawal.

“the impact is negligible during maintenance and dramatically increases during prolonged drug withdrawal.”

May combine two claims.

The excitatory neurons could be broadly classified into 13 clusters, which could be further separated into 26 clusters with higher resolution.

“The excitatory neurons could be broadly classified into 13 clusters, which could be further separated into 26 clusters with higher resolution.”

May combine two claims.

Transcriptional changes from cocaine-induced plasticity is still significant and impacts multiple neuron subtypes.

“transcriptional changes from cocaine-induced plasticity is still significant and impacts multiple neuron subtypes.”

May combine two claims.

scRNA-seq revealed the identity of many discrete cell types in mouse PFC and defined their unique transcriptional features.

“using scRNA-seq, we revealed the identity of the many discrete cell types in mouse PFC and defined their unique transcriptional features.”

May combine two claims.

Each subtype regulates a combination of these (few common and many unique), with both activators and repressors.

“Each subtype regulates a combination of these (few common and many unique), with both activators and repressors.”

May combine two claims.

Widespread transcriptional adaptations occur in each cell type in early postnatal life during spontaneous experience-dependent plasticity, as well as in later life under compulsive cocaine addiction conditions.

“We demonstrated how widespread transcriptional adaptations occur in each cell type in early postnatal life during spontaneous experience-dependent plasticity, as well as in later life under a compulsive cocaine addiction conditions.”

May combine two claims.

Both Exc-7 and 8 show Drd1 expression and undergo strong transcriptional modulation in subsequent cocaine IVSA experiment.

“both subtypes show Drd1 expression and undergo strong transcriptional modulation in subsequent cocaine IVSA experiment”

May combine two claims.

Drd1 is also expressed by Exc-12 and 13 which are moderately impacted (enrich oxidative phosphorylation or mitochondrial function pathways).

“Drd1is also expressed by Exc-12 and 13 (Supplementary Fig. 9e) which are moderately impacted (enrich oxidative phosphorylation or mitochondrial function pathways).”

May combine two claims.

Some of them (Exc-7, 8, and 13) express dopamine receptor Drd1 and may be directly responsive to dopamine released by VTA neurons projecting to PFC.

“some of them (Exc-7, 8, and 13) express dopamine receptor Drd1 (Supplementary Fig. 9e) and may be directly responsive to dopamine released by VTA neurons projecting to PFC.”

May combine two claims.

Exc-13 records highest number of altered GWAS candidate genes in schizophrenia, and many genes like Hs3tst5 (<−4 fold), Kif5c (<−2 fold) or Wwox (>1.5 fold) are regulated in this subtype during adolescence, suggesting a vulnerable neuron subtype in schizophrenia pathogenesis.

“Exc-13 records highest number of altered GWAS candidate genes in schizophrenia (Fig. 5e), and many genes like Hs3tst5 (<−4 fold), Kif5c (<−2 fold) or Wwox (>1.5 fold) are regulated in this subtype during adolescence, suggesting a vulnerable neuron subtype in schizophrenia pathogenesis.”

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

For some conditions, such as schizophrenia, bipolar disorder or autism, strong influence on excitatory or inhibitory neurons were observed; while for some other disorders (eg. bipolar or Alzheimer's), non-neuronal cells have equal or even greater contribution.

“For some conditions, such as schizophrenia, bipolar disorder or autism, strong influence on excitatory or inhibitory neurons were observed; while for some other disorders (eg. bipolar or Alzheimer’s), non-neuronal cells have equal or even greater contribution”

May combine two claims.

Marcksl1 expression is dynamically regulated during adolescence and is highly expressed in P21 when key functions involving this gene, such as cytoskeletal regulation, protein kinase C signaling or calmodulin signaling, are on the rise.

“Marcksl1, whose expression is dynamically regulated during adolescence (Supplementary Fig. 5a–c), is highly expressed in P21 when key functions involving this gene, such as cytoskeletal regulation, protein kinase C signaling or calmodulin signaling55, are on the rise.”

• cites [55] El Amri, M., Fitzgerald, U. & Schlosser, G. MARCKS and MARCKS-like proteins in development and regeneration. J. Biomed. Sci. 25, 43 (2018). sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S2 (checkable).

Our study presents a comprehensive account of the widespread transcriptional dynamics in the postnatal PFC.

“our study presents a comprehensive account of the widespread transcriptional dynamics in the postnatal PFC.”

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

In total, an expression matrix including 20,718 genes and 27,702 cells was used.

“In total, an expression matrix including 20,718 genes and 27,702 cells was used.”

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

After clustering and manual assessment of double droplets, a total of 24,822 cells were retained (11,886 saline and 12,936 cocaine).

“After clustering and manual assessment of double droplets a total of 24,822 cells were retained (11,886 saline and 12,936 cocaine).”

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

Ptgds is upregulated at P60, is believed to have neurotrophic function and high frontal lobe expression.

“Ptgds upregulated at P60 (Supplementary Fig. 5d), is believed to have neurotrophic function56 and high frontal lobe expression57”

• cites [56] Tada, K., Murakami, M., Kambe, T. & Kudo, I. Induction of cyclooxygenase-2 by secretory phospholipases A2 in nerve growth factor-stimulated rat serosal mast cells is facilitated by interaction with fibroblasts and mediated by a mechanism independent of their enzymatic functions. J. Immunol. 161, 5008–5015 (1998). sourceNot checked yet

  Not checked in the free run.

• cites [57] Unger, T. et al. Transcriptome differences between the frontal cortex and hippocampus of wild-type and humanized presenilin-1 transgenic mice. Am. J. Geriatr. Psychiatry 13, 1041–1051 (2005). sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim.

After removing low quality cells (<800 detected genes and cells having >10% mitochondria transcriptome) and double-droplet cells, a total of 10,646 high quality cells were obtained from P21 PFC.

“After removing low quality cells (<800 detected genes and cells having >10% mitochondria transcriptome) and double-droplet cells, a total of 10,646 high quality cells were obtained from P21 PFC.”

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

Adolescence (P21) to adulthood (P60) in mouse is a critical postnatal period believed to be associated with greatly increased neuroplasticity and manifestation of most neuropsychiatric disorders.

“We then analyzed transcriptional dynamics in each of the neuron subtypes between adolescence8,9 (P21) and adulthood (P60) in mouse, a critical postnatal period believed to be associated with greatly increased neuroplasticity, and manifestation of most neuropsychiatric disorders7,10.”

• cites [7] Nurnberger, J. I., Berrettini, W. & Niculescu, A. B. in The Medical Basis of Psychiatry (eds S. Hossein Fatemi & Paula J. Clayton) 553–600 (Springer New York, 2016). sourceNot checked yet

  Not checked in the free run.

• cites [10] Paus, T., Keshavan, M. & Giedd, J. N. Why do many psychiatric disorders emerge during adolescence? Nat. Rev. Neurosci. 9, 947–957 (2008). sourceNot checked yet

  Not checked in the free run.

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

The PFC encodes and regulates the highest cognitive functions through dynamic integration of sensory-stimulus-driven inputs from multiple brain regions.

“The PFC encodes and regulates the highest cognitive functions (such as learning, memory, judgment, decision-making, emotion, risk assessment or social behavior) through dynamic integration of sensory-stimulus-driven inputs from multiple brain regions2.”

• cites [2] Funahashi, S. & Andreau, J. M. Prefrontal cortex and neural mechanisms of executive function. J. Physiol. Paris 107, 471–482 (2013). sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim.

Major emotional or psychological episodes like chronic stress, trauma, and drug addiction can alter brain function and behavior even much later in life.

“On the other hand, major emotional or psychological episodes like chronic stress, trauma, drug addiction can alter brain function and behavior even much later in life5,6.”

• cites [5] McEwen, B. S. & Morrison, J. H. The brain on stress: vulnerability and plasticity of the prefrontal cortex over the life course. Neuron 79, 16–29 (2013). sourceNot checked yet

  Not checked in the free run.

• cites [6] Luscher, C. & Malenka, R. C. Drug-evoked synaptic plasticity in addiction: from molecular changes to circuit remodeling. Neuron 69, 650–663 (2011). sourceNot checked yet

  Not checked in the free run.

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

Neural activity can induce widespread transcriptional changes in neurons, which in turn can encrypt structural and/or functional changes in them.

“Neural activity can induce widespread transcriptional changes in neurons4, which in turn can encrypt structural and/or functional changes in them4.”

• cites [4] West, A. E. & Greenberg, M. E. Neuronal activity-regulated gene transcription in synapse development and cognitive function. Cold Spring Harb Perspect Biol 3, https://doi.org/10.1101/cshperspect.a005744 (2011). sourceNot checked yet

  Not checked in the free run.

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

The PFC is a major brain region regulating behavior.

“As the site of origin of executive function, the prefrontal cortex (PFC) is a major brain region regulating behavior1.”

• cites [1] Miller, E. K., Freedman, D. J. & Wallis, J. D. The prefrontal cortex: categories, concepts and cognition. Philos Trans. R. Soc. Lond. B Biol. Sci. 357, 1123–1136 (2002). sourceNot checked yet

  Not checked in the free run.

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

Profound sensory experiences even in later life can significantly impact neural circuits, which in turn can recondition behavior.

“However, profound sensory experiences even in later life can significantly impact circuits, which in turn can recondition behavior5,6.”

• cites [5] McEwen, B. S. & Morrison, J. H. The brain on stress: vulnerability and plasticity of the prefrontal cortex over the life course. Neuron 79, 16–29 (2013). sourceNot checked yet

  Not checked in the free run.

• cites [6] Luscher, C. & Malenka, R. C. Drug-evoked synaptic plasticity in addiction: from molecular changes to circuit remodeling. Neuron 69, 650–663 (2011). sourceNot checked yet

  Not checked in the free run.

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

The cocaine intravenous self-administration (IVSA) model is a gold standard in addiction studies.

“In parallel, to understand transcriptional changes during induced plasticity in the mature brain, we analyzed volitional drug-intake using the cocaine intra-venous self-administration (IVSA) model11, a gold standard in addiction studies.”

• cites [11] Kmiotek, E. K., Baimel, C. & Gill, K. J. Methods for intravenous self administration in a mouse model. J. Vis. Exp., e3739, https://doi.org/10.3791/3739 (2012). sourceNot checked yet

  Not checked in the free run.

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

Several genetic aberrations can affect early life PFC development leading to psychiatric disorders such as schizophrenia, bipolar disorder, chronic depression, mania, or personality disorders.

“For example, several genetic aberrations can affect early life PFC development leading to psychiatric disorders such as schizophrenia, bipolar disorder, chronic depression, mania, or personality disorders7.”

• cites [7] Nurnberger, J. I., Berrettini, W. & Niculescu, A. B. in The Medical Basis of Psychiatry (eds S. Hossein Fatemi & Paula J. Clayton) 553–600 (Springer New York, 2016). sourceNot checked yet

  Not checked in the free run.

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

The PFC continues to develop significantly during postnatal life and retains a certain degree of plasticity lifelong.

“To sustain this dynamic experience-dependent neuroplasticity, the PFC, unlike any other brain region, continues to develop significantly during postnatal life, and retains a certain degree of plasticity lifelong3.”

• cites [3] Kolb, B. et al. Experience and the developing prefrontal cortex. Proc. Natl Acad. Sci. USA 109(Suppl 2), 17186–17193 (2012). sourceNot checked yet

  Not checked in the free run.

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

Developmentally, Sst+ and Pavlb+ originate from the medial ganglionic eminence (MGE), while the Vip+ originate from the caudal ganglionic eminence (CGE).

“Developmentally, Sst+and Pavlb+ originate from the medial ganglionic eminence (MGE), while the Vip+ originate from the caudal ganglionic eminence (CGE)15.”

• cites [15] Mayer, C. et al. Developmental diversification of cortical inhibitory interneurons. Nature 555, 457–462 (2018). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Many of these altered genes are members of the 68 known epigenetic regulatory complexes, which have well-defined roles in transcription activation or repression.

“We realized that many of these altered genes are members of the 68 known epigenetic regulatory complexes30, which have well-defined roles in transcription activation or repression.”

• cites [30] Medvedeva, Y. A. et al. EpiFactors: a comprehensive database of human epigenetic factors and complexes. Database 2015, bav067–bav067 (2015). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

All these functions together contribute to long term potentiation or synaptic plasticity in many neuron subtypes, consistent with the hypothesis of increased neuroplasticity during adolescence.

“Not surprisingly, all these functions together contribute to long term potentiation or synaptic plasticity in many neuron subtypes (Fig. 4e), consistent with the hypothesis of increased neuroplasticity during adolescence.”

May combine two claims.

Collectively, the comparative transcriptome analysis suggests that selective transcriptional programs operate in each PFC cell subtype during adolescence, driving some broad and some highly specific pathways that facilitate the adaptive transition of the PFC, which likely contribute to proper behavioral programing.

“Collectively, our comparative transcriptome analysis suggests that selective transcriptional programs operate in each PFC cell subtype during adolescence, driving some broad and some highly specific pathways that facilitate the adaptive transition of the PFC, which likely contribute to proper behavioral programing.”

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

Different PFC layers are involved in different neuronal functions.

“Previous studies have demonstrated that different PFC layers are involved in different neuronal functions13.”

• cites [13] Shipp, S. Structure and function of the cerebral cortex. Curr. Biol.: CB 17, R443–R449 (2007). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

While transcription is significantly altered in all excitatory neurons, different subtypes in each layer are impacted to a different degree.

“Therefore, while transcription is significantly altered in all excitatory neurons, different subtypes in each layer are impacted to a different degree (Fig. 4c, d, Supplementary Fig. 6a).”

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

Adolescence marks one of the most dynamic phases of experience-dependent plasticity in the PFC.

“Adolescence marks one of the most dynamic phases of experience-dependent plasticity in the PFC8.”

• cites [8] Selemon, L. D. A role for synaptic plasticity in the adolescent development of executive function. Transl. Psychiatry 3, e238 (2013). sourceNot checked yet

  Not checked in the free run.

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

Broad expression of Reln in coronal sections within PFC can be confirmed from RNA-FISH.

“Indeed, broad expression of Reln in coronal sections within PFC can be confirmed from RNA-FISH (Supplementary Fig. 3c).”

May combine two claims.

Layer properties confer further identity to each subtype, which provides a foundation for understanding cell cluster-specific function.

“Thus, layer properties confer further identity to each subtype, which provides a foundation for understanding cell cluster-specific function.”

May combine two claims.

These results indicate that PFC has neuron populations that are transcriptionally distinct from its closest (ALM) as well as farthest (VISp) neighbors, which may explain unique functions of the PFC compared to the other cortical regions.

“These results indicate that PFC has neuron populations that are transcriptionally distinct from its closest (ALM), as well as farthest (VISp) neighbors, which may explain unique functions of the PFC compared to the other cortical regions.”

May combine two claims.

Cumulatively, these pathways facilitate functions like membrane protrusion, ruffling, adherence, and neurite/dendrite growth that are pivotal for structural changes in synapses, dendrites and axons, implying extensive structural plasticity in most excitatory populations.

“Cumulatively, these pathways facilitate functions like membrane protrusion, ruffling, adherence, and neurite/dendrite growth that are pivotal for structural changes in synapses, dendrites and axons: implying extensive structural plasticity (or modifications) in most excitatory populations20–24.”

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

Formation and migration of different cell types in cortex is complete by P21.

“consistent with prior knowledge that formation and migration of different cell types in cortex is complete by this time19.”

• cites [19] Loo, L. et al. Single-cell transcriptomic analysis of mouse neocortical development. Nat. Commun. 10, 134 (2019). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Overall, these analyses revealed that PFC neurons have very distinct transcriptional properties relative to other cortical regions, which may underlie their unique functional abilities for dynamic integration of varied convergent signals, or their sustained postnatal neuroplasticity/adaptability.

“Overall, these analyses revealed that PFC neurons have very distinct transcriptional properties relative to other cortical regions, which may underlie their unique functional abilities for dynamic integration of varied convergent signals, or their sustained postnatal neuroplasticity/adaptability.”

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

L2/3 are largely involved in intra-cortical (cortico-cortical) regulations, while L5 sends long distance projections outside cortex and L6 receives subcortical inputs (e.g., from thalamus) and projects to superficial layers.

“For example, L2/3 are largely involved in intra-cortical (cortico-cortical) regulations, while L5 sends long distance projections outside cortex and L6 receives subcortical inputs (e.g., from thalamus) and projects to superficial layers13.”

• cites [13] Shipp, S. Structure and function of the cerebral cortex. Curr. Biol.: CB 17, R443–R449 (2007). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Comparable cell numbers and data quality were obtained from both saline (11,886 cells) and cocaine (12,936 cells) groups.

“Comparable cell numbers and data quality were obtained from both saline (11,886 cells) and cocaine (12,936 cells) groups”

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

Dysfunction of the Chd8 complex has been associated with autism, and H4K16 acetylation by this complex is associated with brain aging.

“Dysfunction of this complex has been associated with autism33, and H4K16 acetylation by this complex is associated with brain aging34.”

• cites [33] Bernier, R. et al. Disruptive CHD8 mutations define a subtype of autism early in development. Cell 158, 263–276 (2014). sourceNot checked yet

  Not checked in the free run.

• cites [34] Jun, G. et al. A novel Alzheimer disease locus located near the gene encoding tau protein. Mol. psychiatry 21, 108–117 (2016). sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim.

Induced plasticity under psychological, emotional or stressful conditions like PTSD, depression, social isolation or drug addiction is believed to elicit significant cellular re-adaptations in PFC leading to long term behavioral changes.

“induced plasticity under psychological, emotional or stressful conditions like PTSD, depression, social isolation or drug addiction is believed to elicit significant cellular re-adaptations in PFC leading to long term behavioral changes5.”

• cites [5] McEwen, B. S. & Morrison, J. H. The brain on stress: vulnerability and plasticity of the prefrontal cortex over the life course. Neuron 79, 16–29 (2013). sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim.

DNA methylation events in cortex occur earlier in development.

“prior reports that DNA methylation events in cortex occur earlier in development40”

• cites [40] Lister, R. et al. Global epigenomic reconfiguration during mammalian brain development. Science 341, 1237905 (2013). sourceNot checked yet

  Not checked in the free run.

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

These observations are limited by the low abundance of the inhibitory cell populations and are consequently difficult to interpret.

“these observations are limited by the low abundance of the inhibitory cell populations and are consequently difficult to interpret.”

May combine two claims.

Oxidative phosphorylation is pivotal in fueling pre- and post-synaptic processing, whose alteration impacts mitochondrial function, which in turn affects the sirtuin pathway.

“Oxidative phosphorylation is pivotal in fueling pre- and post-synaptic processing45, whose alteration impacts mitochondrial function, which in turn affects the sirtuin pathway46.”

• cites [45] Hall, C. N., Klein-Flugge, M. C., Howarth, C. & Attwell, D. Oxidative phosphorylation, not glycolysis, powers presynaptic and postsynaptic mechanisms underlying brain information processing. J. Neurosci. 32, 8940–8951 (2012). sourceNot checked yet

  Not checked in the free run.

• cites [46] Verdin, E., Hirschey, M. D., Finley, L. W. & Haigis, M. C. Sirtuin regulation of mitochondria: energy production, apoptosis, and signaling. Trends Biochem. Sci. 35, 669–675 (2010). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Many relevant genes exhibit cell type-specific expression for each disease, providing higher specificity for both diagnosis and treatment.

“many relevant genes exhibit cell type-specific expression for each disease, providing higher specificity for both diagnosis and treatment.”

May combine two claims.

This is a fundamental difference from adaptive neuroplasticity of adolescence, where structural changes in neurons/synapses appear to play a more predominant role in neuroadaptation with altered oxidative phosphorylation, although affected, play a secondary role.

“This is a fundamental difference from adaptive neuroplasticity of adolescence, where structural changes in neurons/synapses appear to play a more predominant role in neuroadaptation with altered oxidative phosphorylation, although affected, play a secondary role”

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

Schizophrenia and bipolar disorder share highly similar behavioral features and are often difficult to diagnose or treat differentially.

“schizophrenia and bipolar disorder, two major PFC-related disorders, share highly similar behavioral features and are often difficult to diagnose or treat differentially42.”

• cites [42] Altamura, A. C. & Goikolea, J. M. Differential diagnoses and management strategies in patients with schizophrenia and bipolar disorder. Neuropsychiatr. Dis. Treat. 4, 311–317 (2008). sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim.

Transcriptional plasticity during adolescence is believed to play a key role in disease pathogenesis.

“transcriptional plasticity during this period is believed to play a key role in disease pathogenesis10.”

• cites [10] Paus, T., Keshavan, M. & Giedd, J. N. Why do many psychiatric disorders emerge during adolescence? Nat. Rev. Neurosci. 9, 947–957 (2008). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Progressive engagement of the deeper PFC layers (L5-6) upon withdrawal might explain psychomotor aspects of drug withdrawal and would be consistent with symptoms seen in humans.

“progressive engagement of the deeper PFC layers (L5-6) upon withdrawal might explain psychomotor aspects of drug withdrawal and would be consistent with symptoms seen in humans44.”

• cites [44] Donroe, J. H. & Tetrault, J. M. Substance use, intoxication, and withdrawal in the critical care setting. Crit. Care Clin. 33, 543–558 (2017). sourceNot checked yet

  Not checked in the free run.

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

Broadly expressed genes often comprise enzymes, metabolic or cytoskeletal regulators essential for general cellular functions.

“These broadly expressed genes often comprise enzymes, metabolic or cytoskeletal regulators essential for general cellular functions.”

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

Cocaine self-administration is a well-established drug addiction model.

“we chose cocaine self-administration11, a well-established drug addiction model.”

• cites [11] Kmiotek, E. K., Baimel, C. & Gill, K. J. Methods for intravenous self administration in a mouse model. J. Vis. Exp., e3739, https://doi.org/10.3791/3739 (2012). sourceNot checked yet

  Not checked in the free run.

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

Downregulation of Mbd3, a component of the NuRD complex, can prevent deacetylation to promote transcription.

“Likewise, downregulation of Mbd3 (Fig. 4f), a component of the NuRD complex (Supplementary Fig. 6c), can prevent deacetylation to promote transcription36.”

• cites [36] Saito, M. & Ishikawa, F. The mCpG-binding domain of human MBD3 does not bind to mCpG but interacts with NuRD/Mi2 components HDAC1 and MTA2. J. Biol. Chem. 277, 35434–35439 (2002). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Neural regulation (transcriptional) during maintenance is largely operated by the basal ganglia, and only withdrawal induces greater cortical involvement.

“it likely indicates that neural regulation (transcriptional) during maintenance is largely operated by the basal ganglia, and only withdrawal induces greater cortical involvement.”

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

Many neuropsychiatric disorders have been linked to genetic mutations and impaired gene expression.

“Many neuropsychiatric disorders have been linked to genetic mutations and impaired gene expression7.”

• cites [7] Nurnberger, J. I., Berrettini, W. & Niculescu, A. B. in The Medical Basis of Psychiatry (eds S. Hossein Fatemi & Paula J. Clayton) 553–600 (Springer New York, 2016). sourceNot checked yet

  Not checked in the free run.

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

Upregulation of pathways like sirtuin signaling, mitochondrial function and oxidative phosphorylation also contribute to neuronal stress.

“upregulation of pathways like sirtuin signaling, mitochondrial function and oxidative phosphorylation also contribute to neuronal stress.”

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

Our results are consistent with prior observations that inhibitory neurons show uniform molecular features even between distant cortical areas.

“In general, our results are consistent with these observations.”

May combine two claims.

Excitatory neurons show more widespread changes than inhibitory neurons, but the lower abundance of the latter underpowering the differential gene expression analysis cannot be fully omitted as a possibility.

“While excitatory neurons show more widespread changes than inhibitory neurons, the lower abundance of the later underpowering the differential gene expression analysis cannot be fully omitted as a possibility.”

May combine two claims.

One or more of these PFC neuron subtypes (5 out of 13 excitatory) respond to dopamine released from midbrain projections and participate in reward processing in conjunction with regions like the VTA and NAc.

“it is likely that one or more of these PFC neuron subtypes (5 out of 13 excitatory) respond to dopamine released from midbrain projections and participate in reward processing in conjunction with regions like the VTA and NAc.”

May combine two claims.

Cocaine-taking associated transcriptome changes provide supporting evidence that cocaine-taking does have an impact on the transcriptome of the adult PFC and that the impact could be variable across different neuron sub-types.

“these cocaine-taking associated transcriptome changes provide supporting evidence that cocaine-taking does have an impact on the transcriptome of the adult PFC and that the impact could be variable across different neuron sub-types.”

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

A patient with a particular disorder can be diagnosed based on the specific disease-related mutation identified by sequencing of genomic DNA isolated from blood sample or oral swab.

“A patient with a particular disorder can be diagnosed based on the specific disease-related mutation identified by sequencing of genomic DNA isolated from blood sample or oral swab.”

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

Many psychiatric disorders associated with genetic defects surface during adolescence.

“Many psychiatric disorders associated with genetic defects surface during adolescence10.”

• cites [10] Paus, T., Keshavan, M. & Giedd, J. N. Why do many psychiatric disorders emerge during adolescence? Nat. Rev. Neurosci. 9, 947–957 (2008). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

Cocaine alters oxidative phosphorylation and other metabolic parameters in PFC has been observed in prior human studies, and we reveal that this function is altered only in specific cell types.

“That cocaine alters oxidative phosphorylation and other metabolic parameters in PFC has been observed in prior human studies53, we reveal that this function is altered only in specific cell types.”

• cites [53] Lehrmann, E. et al. Transcriptional profiling in the human prefrontal cortex: evidence for two activational states associated with cocaine abuse. Pharm. J. 3, 27–40 (2003).Not checked yet

  Not checked in the free run.

May combine two claims.

Adolescence broadly invoked structural plasticity of axons and synapses, cocaine predominantly altered metabolic parameters to support synaptic potentiation.

“While adolescence broadly invoked structural plasticity of axons and synapses, cocaine predominantly altered metabolic parameters to support synaptic potentiation”

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

This can in turn cause excitotoxicity and deterioration of neural health and excitability that eventually resulting in a long-term deleterious effects of drug addiction.

“This can in turn cause excitotoxicity and deterioration of neural health and excitability that eventually resulting in a long-term deleterious effects of drug addiction.”

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

Prior studies demonstrated prominent transcriptional differences between the two geometric poles of the cortex (anterior and posterior).

“prior studies that demonstrated prominent transcriptional differences between the two geometric poles of the cortex (i.e., anterior and posterior)47”

• cites [47] Saunders, A. et al. Molecular Diversity and Specializations among the Cells of the Adult Mouse Brain. Cell 174, 1015–1030 (2018). e1016. sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim.

We performed gene expression analyses using the 13 "low resolution" clusters for a better power and higher accuracy to determine differential transcription.

“we performed gene expression analyses using the 13 “low resolution” clusters for a better power and higher accuracy to determine differential transcription.”

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

Prior human studies have predicted such possibilities.

“prior human studies have predicted such possibilities52.”

• cites [52] Zhou, Z., Enoch, M. A. & Goldman, D. Gene expression in the addicted brain. Int Rev. Neurobiol. 116, 251–273 (2014). sourceNot checked yet

  Not checked in the free run.

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

The age difference between the two models (adolescence- P21 and cocaine starting at P60) may also have some contribution towards the difference.

“the age difference between the two models (adolescence- P21 and cocaine starting at P60), which can naturally impact the basal adaptability/plasticity of the brain may also have some contribution towards the difference.”

May combine two claims.

The impact of cocaine in PFC is highly cell subtype-specific and temporally regulated.

“our data suggest that the impact of cocaine in PFC is highly cell subtype-specific and temporally regulated.”

May combine two claims.

Our integrative analyses have allowed us to identify disease-relevant cell types for a disorder, identify unique dysfunctions in individual cohorts, and deduce a potential functional basis of a disorder based on the cells affected.

“Our integrative analyses have allowed us to: (a) identify disease-relevant cell types for a disorder; (b) identify unique dysfunctions in individual cohorts (due to impact on very specific subtypes); (c) deduce a potential functional basis of a disorder (such as excitatory/inhibitory balance48,49, lack of myelination50 etc.) based on the cells affected.”

• cites [48] Nelson, S. B. & Valakh, V. Excitatory/Inhibitory Balance and Circuit Homeostasis in Autism Spectrum Disorders. Neuron 87, 684–698 (2015). sourceNot checked yet

  Not checked in the free run.

• cites [49] Yizhar, O. et al. Neocortical excitation/inhibition balance in information processing and social dysfunction. Nature 477, 171–178 (2011). sourceNot checked yet

  Not checked in the free run.

• cites [50] Nave, K. A. & Ehrenreich, H. Myelination and oligodendrocyte functions in psychiatric diseases. JAMA Psychiatry 71, 582–584 (2014). sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S2 (checkable), S3 (load-bearing). May combine two claims.

Psychiatric disorders are complex, poorly understood and mostly incurable.

“Psychiatric disorders are complex, poorly understood and mostly incurable7.”

• cites [7] Nurnberger, J. I., Berrettini, W. & Niculescu, A. B. in The Medical Basis of Psychiatry (eds S. Hossein Fatemi & Paula J. Clayton) 553–600 (Springer New York, 2016). sourceNot checked yet

  Not checked in the free run.

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

Prior studies indicate that inhibitory neurons show uniform molecular features even between distant cortical areas.

“Prior studies indicate that inhibitory neurons show uniform molecular features even between distant cortical areas14.”

• cites [14] Tasic, B. et al. Shared and distinct transcriptomic cell types across neocortical areas. Nature 563, 72–78 (2018). sourceNot checked yet

  Not checked in the free run.

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

These findings align with preexisting notion that exposure to addictive substances during adolescence can impair PFC maturation.

“These findings align with preexisting notion that exposure to addictive substances during adolescence can impair PFC maturation, and open doors for more precise mechanistic and therapeutic studies51.”

• cites [51] Walker, D. M. et al. Cocaine Self-administration Alters Transcriptome-wide Responses in the Brain’s Reward Circuitry. Biol. Psychiatry 84, 867–880 (2018). sourceNot checked yet

  Not checked in the free run.

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

Cortical DNA methylation takes place during postnatal 1-4 weeks.

“Prior studies show that cortical DNA methylation takes place during postnatal 1–4 weeks40.”

• cites [40] Lister, R. et al. Global epigenomic reconfiguration during mammalian brain development. Science 341, 1237905 (2013). sourceNot checked yet

  Not checked in the free run.

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

Acquisition of reward possibly has a greater impact on basal forebrain structures of the reward circuit like the nucleus accumbens, and a craving response upon withdrawal greatly engages the PFC.

“This implies that acquisition of reward possibly has a greater impact on basal forebrain structures of the reward circuit like the nucleus accumbens, and a craving response upon withdrawal greatly engages the PFC.”

Classified as probably not a substantial claim.

GWAS have linked these disorders to a group of genes.

“Genome wide association studies (GWAS) have linked these disorders to a group of genes41.”

• cites [41] Collins, A. L. & Sullivan, P. F. Genome-wide association studies in psychiatry: what have we learned? Br. J. Psychiatry 202, 1–4 (2013). sourceNot checked yet

  Not checked in the free run.

Probably not a substantial claim: may not pass S2 (checkable), S4 (based). May combine two claims.

With knowledge of the cellular basis of pathogenesis, understanding of disease mechanisms/manifestation, progress and targeted cell-type specific therapy can be developed/executed.

“with knowledge of the cellular basis of pathogenesis, understanding of the disease mechanisms/manifestation, progress and targeted cell-type specific therapy can be developed/executed.”

Probably not a substantial claim: may not pass S2 (checkable), S3 (load-bearing). May combine two claims.

There are many factors modulating drug self-administration, and further studies are needed for a complete understanding of neural mechanisms underlying the pathogenesis of voluntary drug taking.

“there are many factors modulating drug self-administration54, and further studies are needed for a complete understanding of neural mechanisms underlying the pathogenesis of voluntary drug taking.”

• cites [54] Ahmed, S. H. & Koob, G. F. Transition from moderate to excessive drug intake: change in hedonic set point. Science 282, 298–300 (1998). sourceNot checked yet

  Not checked in the free run.

May combine two claims.

PFC, unlike other brain regions, retains significant plasticity much later in life, and cell type-specific transcriptional responses can adapt to stimuli such as cocaine withdrawal.

“our study provides evidence that PFC, unlike other brain regions, retains significant plasticity much later in life, and cell type-specific transcriptional responses can adapt to stimuli such as cocaine withdrawal.”

May combine two claims.

While histone modifiers are clearly predominant regulators, a marginal role of DNA methylation in some of the initial impact of P21→P60 cannot be fully ruled out.

“while histone modifiers are clearly predominant regulators, a marginal role of DNA methylation in some of the initial impact of P21→P60 cannot be fully ruled out.”

Classified as probably not a substantial claim.

As an initial quality control, cells with >10% mitochondrial transcripts or likely double-droplets (expressing more than the 99th percentile of the number of genes, 4766 genes) were removed.

“As an initial quality control, cells that: have a potential mitochondria contamination (>10% of their total transcripts from mitochondrial transcriptome) or likely represent double-droplets (cells expressing more than the 99th percentile of the number of genes, 4766 genes) were removed.”

• AssumptionRests on an assumption: Abnormally high mitochondrial mRNA is an indication of dead or dying cells.
• AssumptionRests on an assumption: Incorporation of cells with abnormally high mitochondrial mRNA into the analysis can skew the results and introduce errors in detection of differentially expressed genes.

Classified as probably not a substantial claim.

Cluster markers were calculated using a Bonferroni-corrected negative binomial generalized linear model while controlling for the number of UMI in each cell, the percent of mitochondrial genes, and sample-to-sample variation.

“Specifically, the cluster markers were calculated by using a Bonferroni-corrected negative binomial generalized linear model (FindAllMarkers function of the Seurat package, test.use = “negbinom”) while controlling for the number of UMI in each cell, the percent of mitochondrial genes, and sample-to-sample variation.”

Classified as probably not a substantial claim.

The multiBatchNorm function of the scran R/Bioconductor package was used to rescale the datasets to similar sequencing depth.

“Then, the ‘multiBatchNorm” function of the scran R/Bioconductor package75 was used to rescale the datasets to similar sequencing depth.”

• cites [75] Lun, A. T., McCarthy, D. J. & Marioni, J. C. A step-by-step workflow for low-level analysis of single-cell RNA-seq data with Bioconductor. F1000Research 5, 2122 (2016).Not checked yet

  Not checked in the free run.

• AssumptionRests on an assumption: Canonical correlation analysis (CCA) can only present cells into another projection space that brings similar cells into near proximity; it does not correct the gene expression between two datasets.

May combine two claims.

The P21 cells that did not show high similarity to PFC cells showed a random localization in the t-SNE plot, indicating they couldn't be faithfully classified due to technical issues rather than representing new cell-types.

“When we checked their distribution in the t-SNE they showed a random localization in the t-SNE plot indicating that they couldn’t be faithfully classified due to technical issues rather than representing new cell-types.”

Classified as probably not a substantial claim.

To avoid outlier effect, the mean expression of each cluster was calculated using cells with an expression value <99th percentile for each gene.

“To avoid outlier effect, the mean expression of each cluster was calculated using cells with an expression value <99th percentile for each gene.”

Classified as probably not a substantial claim.

To remove any potential batch effect between saline and cocaine samples, CCA analysis was run.

“To remove any potential batch effect between saline and cocaine samples, we run the CCA analysis17.”

• cites [17] Butler, A., Hoffman, P., Smibert, P., Papalexi, E. & Satija, R. Integrating single-cell transcriptomic data across different conditions, technologies, and species. Nat. Biotechnol. 36, 411–420 (2018). sourceNot checked yet

  Not checked in the free run.

Classified as probably not a substantial claim.

Canonical correlation analysis (CCA) was used to find a projection space that maximizes the correlation between the two samples so that batch effect can be removed.

“We thus first used canonical correlation analysis (CCA)17 to find a projection space that maximize the correlation between the two samples so that batch effect can be removed.”

• cites [17] Butler, A., Hoffman, P., Smibert, P., Papalexi, E. & Satija, R. Integrating single-cell transcriptomic data across different conditions, technologies, and species. Nat. Biotechnol. 36, 411–420 (2018). sourceNot checked yet

  Not checked in the free run.

• AssumptionRests on an assumption: Conventional correlation analysis can lead to inconsistent results when two datasets are sequenced at different depths.