rabbit anti mag  (Proteintech)

Journal: bioRxiv

Article Title: Segregated basal ganglia output pathways correspond to genetically divergent neuronal subclasses

doi: 10.1101/2024.08.28.610136

Figure Lengend Snippet: (A) NeuN enrichment strategy for droplet-based single-nucleus RNAseq profiling of SNr neurons. (B) UMAP representation of 1499 SNr GABAergic neurons colored by assigned cluster identity. (C) Heatmap showing relative expression of cluster-enriched genes. (D) Dotplot showing expression of selected marker genes across SNr clusters. (E) SNr cluster identity mapped to Allen Brain Cell Atlas mouse taxonomy with MapMyCells. UMAP representation of SNr GABAergic neurons from 1B colored by ABCA subclass. (F) Number and proportion of neurons per cluster mapping to corresponding ABCA subclass. (G) RNAscope reveals that SNr cluster markers display different topographic distributions within SNr. Scale bars are 250µm, inset panel scalebar is 100µm. (H) Reconstruction of coordinate locations of Foxp2 + and Pou6f2 + neurons labeled by immunohistochemistry within SNr. (I) Topographic organization of ABCA subclasses from ABCA MERFISH of whole coronal mouse brain. See also .

Article Snippet: Primary antibodies used in this study were Rabbit anti-Foxp2 (Abcam, 1:10k), Guinea Pig anti-Pou6f2 (Columbia, 1:2k), Rabbit anti-GFP-488 (Invitrogen, 1:1k), Chicken anti-GFP (Aves labs, 1:2k), Rabbit anti-RFP (Rockland, 1:2k), Rat anti-mCherry (Invitrogen, 1:2k), Mouse anti-TH (Immunostar, 1:10k) and Chicken anti-Myc (Invitrogen, 1:250).

Techniques: Expressing, Marker, RNAscope, Labeling, Immunohistochemistry

Journal: bioRxiv

Article Title: Segregated basal ganglia output pathways correspond to genetically divergent neuronal subclasses

doi: 10.1101/2024.08.28.610136

Figure Lengend Snippet: (A) UMAP representation of 2596 nuclei isolated after SNr microdissection, colored by assigned cluster (top) and cell type label (bottom). (B) SNr cluster identity mapped to Allen Brain Cell Atlas mouse taxonomy with MapMyCells. UMAP representations of SNr GABAergic neurons from 1B colored by ABCA supertype (top left) or ABCA cluster (bottom left). Number and proportion of neurons per cluster mapping to corresponding ABCA label (right). (C) RNAscope reveals marker distribution within SNr. Scale bars are 250µm, inset and rightside panel scalebars are 100µm. (D) Violin plots showing average expression of Gad2 and Pvalb per SNr cluster. (E) Proportion of Foxp2 + , Pou6f2 + , Tmem132c + and Scn5a + neurons labeled by RNAscope that colocalize with Pvalb . N = 3 mice, 6 slides, Foxp2 : 279 neurons, Pou6f2 : 335 neurons, Tmem132c : 150 neurons, Scn5a : 187 neurons. (F) Relative intensity of Pvalb expression in Foxp2 + , Pou6f2 + , Tmem132c + and Scn5a + neurons colocalizing with Pvalb . N = 3 mice, 6 slides, Foxp2 : 225 neurons, Pou6f2 : 153 neurons, Tmem132c : 31 neurons, Scn5a : 71 neurons. Data are represented as mean ± SEM. *: p < 0.0001.

Article Snippet: Primary antibodies used in this study were Rabbit anti-Foxp2 (Abcam, 1:10k), Guinea Pig anti-Pou6f2 (Columbia, 1:2k), Rabbit anti-GFP-488 (Invitrogen, 1:1k), Chicken anti-GFP (Aves labs, 1:2k), Rabbit anti-RFP (Rockland, 1:2k), Rat anti-mCherry (Invitrogen, 1:2k), Mouse anti-TH (Immunostar, 1:10k) and Chicken anti-Myc (Invitrogen, 1:250).

Techniques: Isolation, Laser Capture Microdissection, RNAscope, Marker, Expressing, Labeling

Journal: bioRxiv

Article Title: Segregated basal ganglia output pathways correspond to genetically divergent neuronal subclasses

doi: 10.1101/2024.08.28.610136

Figure Lengend Snippet: (A) Experimental schematic for single-nucleus RNAseq profiling of SNr neurons retrogradely labeled by injection of AAVretro-hSyn-H2B-mCherry in each of six target sites. (B) UMAP representation of 3049 SNr GABAergic neurons from integrated cluster analysis of whole SNr 10x (1507 neurons) and projection-tagged plate-based RNAseq (1542 neurons), colored by mapped ABCA subclass (left), assigned cluster (middle) and projection target (right). (C) Feature plots showing single-nucleus gene expression of Foxp2 , Pou6f2 , Tcf7l2 and Scn5a . (D) Proportion of 10x or retrogradely labeled SNr neurons corresponding to a given cluster or mapped ABCA subclass. (E) Dotplot showing expression of selected marker genes across retrogradely labeled SNr neurons. (F) Immunostaining after retrograde labeling of SNr neurons by injection of AAVretro-hSyn-H2B-GFP in each of six target sites reveals pattern of colocalization of retrogradely labeled SNr neurons with Foxp2 or Pou6f2. The same brain section is shown in the top and bottom panels for each target site. Scalebar is 50µm. (G) Quantification of (F), showing proportion of retrogradely labeled SNr neurons colocalizing with Foxp2 or Pou6f2 immunolabeling. Data are represented as mean ± SEM. See also .

Article Snippet: Primary antibodies used in this study were Rabbit anti-Foxp2 (Abcam, 1:10k), Guinea Pig anti-Pou6f2 (Columbia, 1:2k), Rabbit anti-GFP-488 (Invitrogen, 1:1k), Chicken anti-GFP (Aves labs, 1:2k), Rabbit anti-RFP (Rockland, 1:2k), Rat anti-mCherry (Invitrogen, 1:2k), Mouse anti-TH (Immunostar, 1:10k) and Chicken anti-Myc (Invitrogen, 1:250).

Techniques: Labeling, Injection, Expressing, Marker, Immunostaining, Immunolabeling

Journal: bioRxiv

Article Title: Segregated basal ganglia output pathways correspond to genetically divergent neuronal subclasses

doi: 10.1101/2024.08.28.610136

Figure Lengend Snippet: (A) Experimental schematic and representative images for brain-wide anterograde tracing from three SNr subclasses. Scale bars are 250µm. (B) Quantification of relative axon density across brain regions (see for brain region acronyms). N = 3 mice for Foxp2 and Pou6f2 subclasses and 4 mice for SNL-IC subclass. Data are represented as mean ± SEM. See also .

Article Snippet: Primary antibodies used in this study were Rabbit anti-Foxp2 (Abcam, 1:10k), Guinea Pig anti-Pou6f2 (Columbia, 1:2k), Rabbit anti-GFP-488 (Invitrogen, 1:1k), Chicken anti-GFP (Aves labs, 1:2k), Rabbit anti-RFP (Rockland, 1:2k), Rat anti-mCherry (Invitrogen, 1:2k), Mouse anti-TH (Immunostar, 1:10k) and Chicken anti-Myc (Invitrogen, 1:250).

Techniques: Anterograde Tracing

Journal: bioRxiv

Article Title: Segregated basal ganglia output pathways correspond to genetically divergent neuronal subclasses

doi: 10.1101/2024.08.28.610136

Figure Lengend Snippet: (A) TH + dopamine neurons in the substantia nigra pars compacta (SNc) express Foxp2. SNc adjacent Pou6f2 + SNr neurons do not express TH. Scalebar is 250µm (left) and 100µm (right) (B) Injection of AAV-Con/Fon-YFP into the SNr of Vgat-Flp;Foxp2-Cre mice labels TH - /Foxp2 + cells in SNr but not TH + cells in SNc. Scalebar is 250µm (large panels) and 50µm (small panels). (C) Validation of Pou6f2-Flp mouse line with RNAscope. TdTomato + cells in Pou6f2-Flp;Ai65F mice colocalize with endogenous expression of Pou6f2 but not Foxp2 . Scalebars are 250µm (left and middle panels) and 100µm (right panels). (D) Proportion of SNr neurons expressing endogenous Pou6f2 by RNAscope that are labeled by tdTomato in Pou6f2-Flp;Ai65 mice. N = 3 mice, 6 sections, 3870 Pou6f2 + neurons. Data are represented as mean ± SEM.

Article Snippet: Primary antibodies used in this study were Rabbit anti-Foxp2 (Abcam, 1:10k), Guinea Pig anti-Pou6f2 (Columbia, 1:2k), Rabbit anti-GFP-488 (Invitrogen, 1:1k), Chicken anti-GFP (Aves labs, 1:2k), Rabbit anti-RFP (Rockland, 1:2k), Rat anti-mCherry (Invitrogen, 1:2k), Mouse anti-TH (Immunostar, 1:10k) and Chicken anti-Myc (Invitrogen, 1:250).

Techniques: Injection, RNAscope, Expressing, Labeling

Journal: bioRxiv

Article Title: Segregated basal ganglia output pathways correspond to genetically divergent neuronal subclasses

doi: 10.1101/2024.08.28.610136

Figure Lengend Snippet: (A) Experimental schematic and representative images for rabies virus presynaptic input tracing from three SNr subpopulations. Scale bars are 500 (B) Quantification if number of neurons across striatum subdomains (see for striatum subdomain acronyms). N = 4 mice for Foxp2 subclass, 3 mice for Pou6f2 subclass and 3 mice for SNL-IC subclass. Data are represented as mean ± SEM. See also .

Article Snippet: Primary antibodies used in this study were Rabbit anti-Foxp2 (Abcam, 1:10k), Guinea Pig anti-Pou6f2 (Columbia, 1:2k), Rabbit anti-GFP-488 (Invitrogen, 1:1k), Chicken anti-GFP (Aves labs, 1:2k), Rabbit anti-RFP (Rockland, 1:2k), Rat anti-mCherry (Invitrogen, 1:2k), Mouse anti-TH (Immunostar, 1:10k) and Chicken anti-Myc (Invitrogen, 1:250).

Techniques: Virus

Journal: bioRxiv

Article Title: Segregated basal ganglia output pathways correspond to genetically divergent neuronal subclasses

doi: 10.1101/2024.08.28.610136

Figure Lengend Snippet: (A) UMAP representation of 39110 human non-dopaminergic midbrain neurons from 8 control donors, colored by cluster (left), cell type label (middle) and donor ID (right). (B) UMAP representation of 73889 human non-dopaminergic midbrain neurons from 8 control donors (39110 neurons) and 7 Parkinson’s Disease donors (34779 neurons), colored by cluster (left), cell type label (middle) and disease state (left). (C) UMAP representation of 13910 human GABAergic SNr neurons (9307 control neurons and 4603 PD neurons) subsetted from (B) and feature plots showing relative expression of SIX3 and POU6F2 . (D) Number of neurons in the SIX3 + /FOXP2 + and PAX5 + /POU6F2 + clusters deriving from control or PD donors. Proportion from PD donors shown in parentheses. (E) Average gene expression of control and PD neurons within SIX3 + /FOXP2 + clusters (top) and PAX5 + /POU6F2 + clusters (bottom). (F) Volcano plots of genes up- or downregulated in PD compared to control within SIX3 + /FOXP2 + clusters (top) and PAX5 + /POU6F2 + clusters (bottom). Genes shown in red have fold change > 2 and p < 10e-100.

Article Snippet: Primary antibodies used in this study were Rabbit anti-Foxp2 (Abcam, 1:10k), Guinea Pig anti-Pou6f2 (Columbia, 1:2k), Rabbit anti-GFP-488 (Invitrogen, 1:1k), Chicken anti-GFP (Aves labs, 1:2k), Rabbit anti-RFP (Rockland, 1:2k), Rat anti-mCherry (Invitrogen, 1:2k), Mouse anti-TH (Immunostar, 1:10k) and Chicken anti-Myc (Invitrogen, 1:250).

Techniques: Control, Expressing