Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: CO-EXPRESSION OF SEROTONIN AND NITRIC OXIDE IN THE RAPHE COMPLEX: CORTICAL VS SUBCORTICAL CIRCUIT

doi: 10.1002/ar.21222

Figure Lengend Snippet: Color photographs demonstrating Fluoro-Gold (FG) retrogradely labeled dorsal raphe- (DR-), median raphe- (MR-) and supralemniscal- (SLN-) medial prefrontal cortex (mPFC) projecting neurons (upper row; FG-yellow) co-expressing both tryptophan hydroxylase (TPH; middle row; Cy3-red) and neuronal nitric oxide synthase (nNOS; bottom row; Cy2-green) immunoreactivities. Note that many retrogradely labeled neurons in these three raphe nuclei contained both TPH and nNOS (arrows) immunoreactivities. Scale bar: 50 μm.

Article Snippet: Immunofluorescent Staining In order to visualize TPH or nNOS immunoreactive neurons in the raphe system, every third section through the raphe nuclear complex were incubated in either the monoclonal TPH antiserum (1: 1-2000; Mouse; MAB5278; Millipore), polyclonal nNOS antiserum (1:1000; Rabbit; 24287; Immunostar) or both antisera at 4 °C in PBS with 0.3% Triton and 1% bovine serum albumin (BSA) overnight.

Techniques: Labeling, Expressing

Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: CO-EXPRESSION OF SEROTONIN AND NITRIC OXIDE IN THE RAPHE COMPLEX: CORTICAL VS SUBCORTICAL CIRCUIT

doi: 10.1002/ar.21222

Figure Lengend Snippet: DR-mPFC projecting neurons co-express both TPH and nNOS immunoreactivity.

Article Snippet: Immunofluorescent Staining In order to visualize TPH or nNOS immunoreactive neurons in the raphe system, every third section through the raphe nuclear complex were incubated in either the monoclonal TPH antiserum (1: 1-2000; Mouse; MAB5278; Millipore), polyclonal nNOS antiserum (1:1000; Rabbit; 24287; Immunostar) or both antisera at 4 °C in PBS with 0.3% Triton and 1% bovine serum albumin (BSA) overnight.

Techniques: Labeling

Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: CO-EXPRESSION OF SEROTONIN AND NITRIC OXIDE IN THE RAPHE COMPLEX: CORTICAL VS SUBCORTICAL CIRCUIT

doi: 10.1002/ar.21222

Figure Lengend Snippet: MR-mPFC projecting neurons co-express both TPH and nNOS immunoreactivity.

Article Snippet: Immunofluorescent Staining In order to visualize TPH or nNOS immunoreactive neurons in the raphe system, every third section through the raphe nuclear complex were incubated in either the monoclonal TPH antiserum (1: 1-2000; Mouse; MAB5278; Millipore), polyclonal nNOS antiserum (1:1000; Rabbit; 24287; Immunostar) or both antisera at 4 °C in PBS with 0.3% Triton and 1% bovine serum albumin (BSA) overnight.

Techniques: Labeling

Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: CO-EXPRESSION OF SEROTONIN AND NITRIC OXIDE IN THE RAPHE COMPLEX: CORTICAL VS SUBCORTICAL CIRCUIT

doi: 10.1002/ar.21222

Figure Lengend Snippet: SLN-mPFC projecting neurons co-express both TPH and nNOS immunoreactivity.

Article Snippet: Immunofluorescent Staining In order to visualize TPH or nNOS immunoreactive neurons in the raphe system, every third section through the raphe nuclear complex were incubated in either the monoclonal TPH antiserum (1: 1-2000; Mouse; MAB5278; Millipore), polyclonal nNOS antiserum (1:1000; Rabbit; 24287; Immunostar) or both antisera at 4 °C in PBS with 0.3% Triton and 1% bovine serum albumin (BSA) overnight.

Techniques: Labeling

Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: CO-EXPRESSION OF SEROTONIN AND NITRIC OXIDE IN THE RAPHE COMPLEX: CORTICAL VS SUBCORTICAL CIRCUIT

doi: 10.1002/ar.21222

Figure Lengend Snippet: Color photomicrographs demonstrating serotonin transporter (SERT) positive fibers (A1–B1; Cy3-red) and neuronal nitric oxide synthase (nNOS) positive profiles (A2–B2; Cy2-green) in the medial prefrontal cortex (mPFC). Note that both immunostainings exhibited similar distribution patterns in the mPFC and most fibers were double-labeled with SERT and nNOS immunoreactivities (A3–B3; merged). In addition, nNOS positive neurons and their processes in the cortex were also identified but none of them contained SERT immunoreactivity (arrows in A1–A3). Scale bar: 50 μm.

Article Snippet: Immunofluorescent Staining In order to visualize TPH or nNOS immunoreactive neurons in the raphe system, every third section through the raphe nuclear complex were incubated in either the monoclonal TPH antiserum (1: 1-2000; Mouse; MAB5278; Millipore), polyclonal nNOS antiserum (1:1000; Rabbit; 24287; Immunostar) or both antisera at 4 °C in PBS with 0.3% Triton and 1% bovine serum albumin (BSA) overnight.

Techniques: Labeling

Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: CO-EXPRESSION OF SEROTONIN AND NITRIC OXIDE IN THE RAPHE COMPLEX: CORTICAL VS SUBCORTICAL CIRCUIT

doi: 10.1002/ar.21222

Figure Lengend Snippet: Color photographs demonstrating the relationship between tryptophan hydroxylase (TPH; upper row; Cy3-red) and neuronal nitric oxide synthase (nNOS; middle row; Cy2-green) positive neurons in the nucleus raphe pallidus (B1), raphe obscurus (B2) and raphe magnus (B3). Note that several small nNOS positive neurons were scattered and adjacent to the medium sized TPH positive neurons, but none contained TPH immunoreactivity (bottom row; merged). Scale: 50 μm.

Article Snippet: Immunofluorescent Staining In order to visualize TPH or nNOS immunoreactive neurons in the raphe system, every third section through the raphe nuclear complex were incubated in either the monoclonal TPH antiserum (1: 1-2000; Mouse; MAB5278; Millipore), polyclonal nNOS antiserum (1:1000; Rabbit; 24287; Immunostar) or both antisera at 4 °C in PBS with 0.3% Triton and 1% bovine serum albumin (BSA) overnight.

Techniques:

Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: CO-EXPRESSION OF SEROTONIN AND NITRIC OXIDE IN THE RAPHE COMPLEX: CORTICAL VS SUBCORTICAL CIRCUIT

doi: 10.1002/ar.21222

Figure Lengend Snippet: Color photographs demonstrating the relationship between Fluoro-Glod (FG; A and C) retrogradely labeled neurons in the nucleus raphe magnus (B3) and the corresponding tryptophan hydroxylase (TPH; B; Cy-3) or neuronal nitric oxide synthase (nNOS; D; Cy-3) immunostaining, respectively. Note that a few retrogradely labeled neurons in A contained TPH immunoreactivity (arrows in A and B), but none of retrogradely labeled neurons in C was double-labeled with nNOS immunoreactivity (C and D). Scale: 50 μm.

Article Snippet: Immunofluorescent Staining In order to visualize TPH or nNOS immunoreactive neurons in the raphe system, every third section through the raphe nuclear complex were incubated in either the monoclonal TPH antiserum (1: 1-2000; Mouse; MAB5278; Millipore), polyclonal nNOS antiserum (1:1000; Rabbit; 24287; Immunostar) or both antisera at 4 °C in PBS with 0.3% Triton and 1% bovine serum albumin (BSA) overnight.

Techniques: Labeling, Immunostaining

Journal: Nature Communications

Article Title: The histone demethylase Kdm6b regulates subtype diversification of mouse spinal motor neurons during development

doi: 10.1038/s41467-022-28636-7

Figure Lengend Snippet: a , b Unsupervised clustering analysis of the single-cell transcriptomes of Slc18a3 + cells from E12.5 control spinal cord ( Kdm6b f/+ ;Olig2-Cre;mTmG ). 11 c ontrol c lusters, CC0 to CC10, were visualized using tSNE. Cell types were assigned according to the expression of specific marker genes. Each dot represents a single cell and was color-coded based on the cluster identity. c , d Violin plots for selected marker genes in CC0-CC10. e – j tSNE plots to show the expression of the indicated genes in each plot. The color intensity indicates the expression levels of the genes. The tSNE plots in f , g , and i reveal the co-expression pattern of the two genes. f CC8 (V2IN) and CC9 (V3IN) co-express Slc18a3 and Slc17a6 (cyan, Double+), while the remaining clusters express only Slc18a3 (red, Slc18a3+). g CC1 (nbMN) and CC2 (MMC) co-express Isl1 and Lhx3 (cyan, Double+), whereas CC3 (HMC), CC4 (LMCm), and CC6 (PGC-Isl1 + ) express Isl1 only (red, Isl1+). CC8 (V2IN) express Lhx3 only (green, Lhx3+). i CC4 (LMCm) and CC5 (LMCl) co-express Foxp1 and Aldh1a2 (cyan, Double+), whereas CC6 (PGC-Isl1 + ) and CC7 (PGC-Isl1 − ) express only Foxp1 (red, Foxp1+). k Immunohistochemical analyses with Isl1 and Nos1 antibodies in E13.5 wild-type spinal cords at thoracic levels. Nos1 + PGC cells were divided into Isl1 + and Isl1 − subpopulations. Scale bar, 50 μm. 3–6 sections in 3 embryos were analyzed and the representative images were shown.

Article Snippet: Primary antibodies were listed as follow: goat anti-Olig2 (R&D Systems AF2418, 1:500), rabbit anti-Olig2 (Millipore AB9610, 1:1000), guinea pig anti-Mnx1 (Homemade, 1:500), rabbit anti-Isl1 (Abcam ab109517, 1:250), guinea pig anti-Isl1 (Homemade, 1:2000), rabbit anti-Lhx3 (Abcam ab14555, 1:1000), guinea pig anti-Lhx3 (Homemade, 1:250), guinea pig anti-Vsx2 (Homemade, 1:1000) , guinea pig anti-Sox14 (Homemade, 1:500) , rabbit anti-Nkx2-2 (Abcam ab191077, 1:50), rabbit anti-Foxp1 (Abcam ab16645, 1:2000), rabbit anti-Nos1 (Immunostar 24287, 1:1000), rabbit anti-Kdm6b (Novus Biologicals NBP1-06640, 1:100), guinea pig anti-Klf5 , rabbit anti-Mecom (Cell Signaling Technology #2593, 1:500), sheep anti-Zfhx3 (Novus Biologicals AF7384, 1:200), and rabbit anti-Zfhx4 (Sigma HPA023837, 1:200).

Techniques: Expressing, Marker, Immunohistochemical staining

Journal: Nature Communications

Article Title: The histone demethylase Kdm6b regulates subtype diversification of mouse spinal motor neurons during development

doi: 10.1038/s41467-022-28636-7

Figure Lengend Snippet: a Immunohistochemical analyses for Lhx3, FoxP1, and Nos1 and in situ hybridization analyses for Aldh1a2 in E12.5 spinal cords. Foxp1 + MNs, Aldh1a2 + LMC, and Nos1 + PGC were markedly reduced in Mnx1p::Lhx3 transgenic mice, in which Lhx3 expression was expanded to all MNs. Scale bars, 50 μm. b Immunohistochemical analysis with Kdm6b antibody. Kdm6b protein was detected at E11.5 and substantially removed by E12.5 in Kdm6b -cKO; Isl1-Cre mice. Scale bars, 50 μm. c–g Immunohistochemical analyses for Lhx3, FoxP1, Mnx1, and Isl1, in situ hybridization analyses for Aldh1a2, Slc18a3, Klf5, and Kcnip4 in E13.5 spinal cords, and the quantification of specific marker-labeled MNs per 12 μm thick section. In Kdm6b -cKO; Isl1-Cre mice, Lhx3 + and Mnx1 + MNs were significantly reduced, but neither Foxp1 + nor Isl1 + MNs changed significantly. Scale bars, 50 μm. The error bars represent the standard deviation of the mean. ** p < 0.01; ns non-significant in the two-tailed Student’s t -test. n = 3–6 mice per genotype and 9 slices per genotype. Source data are provided as a Source Data file. The exact p -values are as follows. Lhx3, brachial, 1.44 × 10 −8 ; thoracic, 8.05 × 10 −10 . Foxp1, brachial, 0.68; thoracic, 0.94. Mnx1, brachial, 1.79×10 −9 ; thoracic, 2.36 × 10 −7 . Isl1, brachial, 0.55; thoracic, 0.76.

Article Snippet: Primary antibodies were listed as follow: goat anti-Olig2 (R&D Systems AF2418, 1:500), rabbit anti-Olig2 (Millipore AB9610, 1:1000), guinea pig anti-Mnx1 (Homemade, 1:500), rabbit anti-Isl1 (Abcam ab109517, 1:250), guinea pig anti-Isl1 (Homemade, 1:2000), rabbit anti-Lhx3 (Abcam ab14555, 1:1000), guinea pig anti-Lhx3 (Homemade, 1:250), guinea pig anti-Vsx2 (Homemade, 1:1000) , guinea pig anti-Sox14 (Homemade, 1:500) , rabbit anti-Nkx2-2 (Abcam ab191077, 1:50), rabbit anti-Foxp1 (Abcam ab16645, 1:2000), rabbit anti-Nos1 (Immunostar 24287, 1:1000), rabbit anti-Kdm6b (Novus Biologicals NBP1-06640, 1:100), guinea pig anti-Klf5 , rabbit anti-Mecom (Cell Signaling Technology #2593, 1:500), sheep anti-Zfhx3 (Novus Biologicals AF7384, 1:200), and rabbit anti-Zfhx4 (Sigma HPA023837, 1:200).

Techniques: Immunohistochemical staining, In Situ Hybridization, Transgenic Assay, Expressing, Marker, Labeling, Standard Deviation, Two Tailed Test