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Cortical fast-spiking parvalbumin interneurons enwrapped in the perineuronal net express the metallopeptidases Adamts8, Adamts15 and Neprilysin.

Rossier J, Bernard A, Cabungcal JH, Perrenoud Q, Savoye A, Gallopin T, Hawrylycz M, Cuénod M, Do K, Urban A, Lein ES - Mol. Psychiatry (2014)

Bottom Line: The pattern of expression of metalloproteases (MPs) was analyzed by single-cell reverse transcriptase multiplex PCR after patch clamp and was compared with the expression of 10 canonical interneurons markers and 12 additional genes from the Allen Atlas.Among these five clusters, two fast-spiking interneuron clusters expressing the calcium-binding protein Pvalb were identified, one co-expressing Pvalb with Sst (PV-Sst) and another co-expressing Pvalb with three metallopeptidases Adamts8, Adamts15 and Mme (PV-MP).By using Wisteria floribunda agglutinin, a specific marker for PNN, PV-MP interneurons were found surrounded by PNN, whereas the ones expressing Sst, PV-Sst, were not.

View Article: PubMed Central - PubMed

Affiliation: Centre de Psychiatrie et Neurosciences, Institut National de la Santé et de la Recherche Médicale CPN INSERM U894, Université Paris Descartes, Hôpital Sainte Anne, Paris, France.

ABSTRACT
The in situ hybridization Allen Mouse Brain Atlas was mined for proteases expressed in the somatosensory cerebral cortex. Among the 480 genes coding for protease/peptidases, only four were found enriched in cortical interneurons: Reln coding for reelin; Adamts8 and Adamts15 belonging to the class of metzincin proteases involved in reshaping the perineuronal net (PNN) and Mme encoding for Neprilysin, the enzyme degrading amyloid β-peptides. The pattern of expression of metalloproteases (MPs) was analyzed by single-cell reverse transcriptase multiplex PCR after patch clamp and was compared with the expression of 10 canonical interneurons markers and 12 additional genes from the Allen Atlas. Clustering of these genes by K-means algorithm displays five distinct clusters. Among these five clusters, two fast-spiking interneuron clusters expressing the calcium-binding protein Pvalb were identified, one co-expressing Pvalb with Sst (PV-Sst) and another co-expressing Pvalb with three metallopeptidases Adamts8, Adamts15 and Mme (PV-MP). By using Wisteria floribunda agglutinin, a specific marker for PNN, PV-MP interneurons were found surrounded by PNN, whereas the ones expressing Sst, PV-Sst, were not.

No MeSH data available.


Related in: MedlinePlus

Clustering of 218 neurons in five (Ward) or six (K-means) classes by gene expression. Each individual cell from the somatosensory cortex of 15–19-day-old mice is represented along the x-axis. In each of the 218 columns, detection of a gene by scRTmPCR in a single neuron is indicated by black or colored squares. The list of 27 genes detected is on the y-axis. Anatomical and electrophysiological profiles of most of these cells have been described previously.11 Complete data are in Supplementary Table S2. (a) Ward's hierarchical and K-means (k=6) clustering for the presence or absence (binarization) of gene expression only. The electrophysiological profiles from Supplementary Table S2 were not used for the clustering. (b) Cluster stability analysis based on principal component analysis (PCA).12 FS-PV, fast-spiking parvalbumin; KS, c-Kit with semaphorin3c; NG, neurogliaform; PV-MP, parvalbumin with metallopeptidases; PV-Sst, parvalbumin with somatostatin; Sst, somatostatin; VGluT1, vesicular glutamate transporter 1; VIP, vasoactive intestinal peptide.
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fig1: Clustering of 218 neurons in five (Ward) or six (K-means) classes by gene expression. Each individual cell from the somatosensory cortex of 15–19-day-old mice is represented along the x-axis. In each of the 218 columns, detection of a gene by scRTmPCR in a single neuron is indicated by black or colored squares. The list of 27 genes detected is on the y-axis. Anatomical and electrophysiological profiles of most of these cells have been described previously.11 Complete data are in Supplementary Table S2. (a) Ward's hierarchical and K-means (k=6) clustering for the presence or absence (binarization) of gene expression only. The electrophysiological profiles from Supplementary Table S2 were not used for the clustering. (b) Cluster stability analysis based on principal component analysis (PCA).12 FS-PV, fast-spiking parvalbumin; KS, c-Kit with semaphorin3c; NG, neurogliaform; PV-MP, parvalbumin with metallopeptidases; PV-Sst, parvalbumin with somatostatin; Sst, somatostatin; VGluT1, vesicular glutamate transporter 1; VIP, vasoactive intestinal peptide.

Mentions: On the basis of binary mRNA expression for the 27 markers, Ward's clustering was performed. Ward's clustering is a hierarchical method that groups cells to maximize the differences between groups and to minimize them within groups. This algorithm identified five groups of neurons (Figure 1a) in agreement with previous findings10, 11 and discriminated excitatory neurons expressing Slc17a7 from inhibitory neurons expressing Gad1 and Gad2.


Cortical fast-spiking parvalbumin interneurons enwrapped in the perineuronal net express the metallopeptidases Adamts8, Adamts15 and Neprilysin.

Rossier J, Bernard A, Cabungcal JH, Perrenoud Q, Savoye A, Gallopin T, Hawrylycz M, Cuénod M, Do K, Urban A, Lein ES - Mol. Psychiatry (2014)

Clustering of 218 neurons in five (Ward) or six (K-means) classes by gene expression. Each individual cell from the somatosensory cortex of 15–19-day-old mice is represented along the x-axis. In each of the 218 columns, detection of a gene by scRTmPCR in a single neuron is indicated by black or colored squares. The list of 27 genes detected is on the y-axis. Anatomical and electrophysiological profiles of most of these cells have been described previously.11 Complete data are in Supplementary Table S2. (a) Ward's hierarchical and K-means (k=6) clustering for the presence or absence (binarization) of gene expression only. The electrophysiological profiles from Supplementary Table S2 were not used for the clustering. (b) Cluster stability analysis based on principal component analysis (PCA).12 FS-PV, fast-spiking parvalbumin; KS, c-Kit with semaphorin3c; NG, neurogliaform; PV-MP, parvalbumin with metallopeptidases; PV-Sst, parvalbumin with somatostatin; Sst, somatostatin; VGluT1, vesicular glutamate transporter 1; VIP, vasoactive intestinal peptide.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4356748&req=5

fig1: Clustering of 218 neurons in five (Ward) or six (K-means) classes by gene expression. Each individual cell from the somatosensory cortex of 15–19-day-old mice is represented along the x-axis. In each of the 218 columns, detection of a gene by scRTmPCR in a single neuron is indicated by black or colored squares. The list of 27 genes detected is on the y-axis. Anatomical and electrophysiological profiles of most of these cells have been described previously.11 Complete data are in Supplementary Table S2. (a) Ward's hierarchical and K-means (k=6) clustering for the presence or absence (binarization) of gene expression only. The electrophysiological profiles from Supplementary Table S2 were not used for the clustering. (b) Cluster stability analysis based on principal component analysis (PCA).12 FS-PV, fast-spiking parvalbumin; KS, c-Kit with semaphorin3c; NG, neurogliaform; PV-MP, parvalbumin with metallopeptidases; PV-Sst, parvalbumin with somatostatin; Sst, somatostatin; VGluT1, vesicular glutamate transporter 1; VIP, vasoactive intestinal peptide.
Mentions: On the basis of binary mRNA expression for the 27 markers, Ward's clustering was performed. Ward's clustering is a hierarchical method that groups cells to maximize the differences between groups and to minimize them within groups. This algorithm identified five groups of neurons (Figure 1a) in agreement with previous findings10, 11 and discriminated excitatory neurons expressing Slc17a7 from inhibitory neurons expressing Gad1 and Gad2.

Bottom Line: The pattern of expression of metalloproteases (MPs) was analyzed by single-cell reverse transcriptase multiplex PCR after patch clamp and was compared with the expression of 10 canonical interneurons markers and 12 additional genes from the Allen Atlas.Among these five clusters, two fast-spiking interneuron clusters expressing the calcium-binding protein Pvalb were identified, one co-expressing Pvalb with Sst (PV-Sst) and another co-expressing Pvalb with three metallopeptidases Adamts8, Adamts15 and Mme (PV-MP).By using Wisteria floribunda agglutinin, a specific marker for PNN, PV-MP interneurons were found surrounded by PNN, whereas the ones expressing Sst, PV-Sst, were not.

View Article: PubMed Central - PubMed

Affiliation: Centre de Psychiatrie et Neurosciences, Institut National de la Santé et de la Recherche Médicale CPN INSERM U894, Université Paris Descartes, Hôpital Sainte Anne, Paris, France.

ABSTRACT
The in situ hybridization Allen Mouse Brain Atlas was mined for proteases expressed in the somatosensory cerebral cortex. Among the 480 genes coding for protease/peptidases, only four were found enriched in cortical interneurons: Reln coding for reelin; Adamts8 and Adamts15 belonging to the class of metzincin proteases involved in reshaping the perineuronal net (PNN) and Mme encoding for Neprilysin, the enzyme degrading amyloid β-peptides. The pattern of expression of metalloproteases (MPs) was analyzed by single-cell reverse transcriptase multiplex PCR after patch clamp and was compared with the expression of 10 canonical interneurons markers and 12 additional genes from the Allen Atlas. Clustering of these genes by K-means algorithm displays five distinct clusters. Among these five clusters, two fast-spiking interneuron clusters expressing the calcium-binding protein Pvalb were identified, one co-expressing Pvalb with Sst (PV-Sst) and another co-expressing Pvalb with three metallopeptidases Adamts8, Adamts15 and Mme (PV-MP). By using Wisteria floribunda agglutinin, a specific marker for PNN, PV-MP interneurons were found surrounded by PNN, whereas the ones expressing Sst, PV-Sst, were not.

No MeSH data available.


Related in: MedlinePlus