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Inhibitory transmitters in neurons of human nucleus raphe interpositus (RIP). (A) Consecutive 5 μm thick coronal paraffin sections through RIP stained with <t>GABA</t> A receptor (GABRA1, left) and glutamate decarboxylase (GAD65/67, right) antibodies. Note that cholinergic non-OPNs (green arrows) express intense GABRA1 immunoreactivity compared to OPNs’ (red arrows) weak labeling on their somatic membrane. This trend is reflected in the dense GABAergic inputs shown by GAD65/67 immunolabeling (right) to the cholinergic non-OPNs (green arrows) and sparse inputs to somatic membranes of OPNs (red arrows). The density difference between somatic and dendritic labelling of OPNs can be seen in the inset on the top-right corner of the image (black arrowheads). The close up (inset) is magnified from the boxed region. (B) Quantitative analysis of the GAD65/67 immunopositive puncta on somatic (dark-colored bars) and dendritic (light-colored bars) membranes of OPNs (red) and cholinergic non-OPNs (green) demonstrating significantly denser GABAergic inputs to non-OPNs. (C) Glycine receptor 1a immunolabeling in neurons of RIP demonstrate a complimentary pattern to GABRA1. Note that cholinergic non-OPNs (green arrows) express weak GlyR1 immunoreactivity compared to OPNs’ (red arrows) intense labeling on their somatic membrane. Scale bars represent 50 μm, in A and C; 10 μm in the inset in right column (GAD) of A. *** p < 0.05, * p < 0.1.
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Inhibitory transmitters in neurons of human nucleus raphe interpositus (RIP). (A) Consecutive 5 μm thick coronal paraffin sections through RIP stained with <t>GABA</t> A receptor (GABRA1, left) and glutamate decarboxylase (GAD65/67, right) antibodies. Note that cholinergic non-OPNs (green arrows) express intense GABRA1 immunoreactivity compared to OPNs’ (red arrows) weak labeling on their somatic membrane. This trend is reflected in the dense GABAergic inputs shown by GAD65/67 immunolabeling (right) to the cholinergic non-OPNs (green arrows) and sparse inputs to somatic membranes of OPNs (red arrows). The density difference between somatic and dendritic labelling of OPNs can be seen in the inset on the top-right corner of the image (black arrowheads). The close up (inset) is magnified from the boxed region. (B) Quantitative analysis of the GAD65/67 immunopositive puncta on somatic (dark-colored bars) and dendritic (light-colored bars) membranes of OPNs (red) and cholinergic non-OPNs (green) demonstrating significantly denser GABAergic inputs to non-OPNs. (C) Glycine receptor 1a immunolabeling in neurons of RIP demonstrate a complimentary pattern to GABRA1. Note that cholinergic non-OPNs (green arrows) express weak GlyR1 immunoreactivity compared to OPNs’ (red arrows) intense labeling on their somatic membrane. Scale bars represent 50 μm, in A and C; 10 μm in the inset in right column (GAD) of A. *** p < 0.05, * p < 0.1.
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Inhibitory transmitters in neurons of human nucleus raphe interpositus (RIP). (A) Consecutive 5 μm thick coronal paraffin sections through RIP stained with <t>GABA</t> A receptor (GABRA1, left) and glutamate decarboxylase (GAD65/67, right) antibodies. Note that cholinergic non-OPNs (green arrows) express intense GABRA1 immunoreactivity compared to OPNs’ (red arrows) weak labeling on their somatic membrane. This trend is reflected in the dense GABAergic inputs shown by GAD65/67 immunolabeling (right) to the cholinergic non-OPNs (green arrows) and sparse inputs to somatic membranes of OPNs (red arrows). The density difference between somatic and dendritic labelling of OPNs can be seen in the inset on the top-right corner of the image (black arrowheads). The close up (inset) is magnified from the boxed region. (B) Quantitative analysis of the GAD65/67 immunopositive puncta on somatic (dark-colored bars) and dendritic (light-colored bars) membranes of OPNs (red) and cholinergic non-OPNs (green) demonstrating significantly denser GABAergic inputs to non-OPNs. (C) Glycine receptor 1a immunolabeling in neurons of RIP demonstrate a complimentary pattern to GABRA1. Note that cholinergic non-OPNs (green arrows) express weak GlyR1 immunoreactivity compared to OPNs’ (red arrows) intense labeling on their somatic membrane. Scale bars represent 50 μm, in A and C; 10 μm in the inset in right column (GAD) of A. *** p < 0.05, * p < 0.1.
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Inhibitory transmitters in neurons of human nucleus raphe interpositus (RIP). (A) Consecutive 5 μm thick coronal paraffin sections through RIP stained with <t>GABA</t> A receptor (GABRA1, left) and glutamate decarboxylase (GAD65/67, right) antibodies. Note that cholinergic non-OPNs (green arrows) express intense GABRA1 immunoreactivity compared to OPNs’ (red arrows) weak labeling on their somatic membrane. This trend is reflected in the dense GABAergic inputs shown by GAD65/67 immunolabeling (right) to the cholinergic non-OPNs (green arrows) and sparse inputs to somatic membranes of OPNs (red arrows). The density difference between somatic and dendritic labelling of OPNs can be seen in the inset on the top-right corner of the image (black arrowheads). The close up (inset) is magnified from the boxed region. (B) Quantitative analysis of the GAD65/67 immunopositive puncta on somatic (dark-colored bars) and dendritic (light-colored bars) membranes of OPNs (red) and cholinergic non-OPNs (green) demonstrating significantly denser GABAergic inputs to non-OPNs. (C) Glycine receptor 1a immunolabeling in neurons of RIP demonstrate a complimentary pattern to GABRA1. Note that cholinergic non-OPNs (green arrows) express weak GlyR1 immunoreactivity compared to OPNs’ (red arrows) intense labeling on their somatic membrane. Scale bars represent 50 μm, in A and C; 10 μm in the inset in right column (GAD) of A. *** p < 0.05, * p < 0.1.
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Inhibitory transmitters in neurons of human nucleus raphe interpositus (RIP). (A) Consecutive 5 μm thick coronal paraffin sections through RIP stained with <t>GABA</t> A receptor (GABRA1, left) and glutamate decarboxylase (GAD65/67, right) antibodies. Note that cholinergic non-OPNs (green arrows) express intense GABRA1 immunoreactivity compared to OPNs’ (red arrows) weak labeling on their somatic membrane. This trend is reflected in the dense GABAergic inputs shown by GAD65/67 immunolabeling (right) to the cholinergic non-OPNs (green arrows) and sparse inputs to somatic membranes of OPNs (red arrows). The density difference between somatic and dendritic labelling of OPNs can be seen in the inset on the top-right corner of the image (black arrowheads). The close up (inset) is magnified from the boxed region. (B) Quantitative analysis of the GAD65/67 immunopositive puncta on somatic (dark-colored bars) and dendritic (light-colored bars) membranes of OPNs (red) and cholinergic non-OPNs (green) demonstrating significantly denser GABAergic inputs to non-OPNs. (C) Glycine receptor 1a immunolabeling in neurons of RIP demonstrate a complimentary pattern to GABRA1. Note that cholinergic non-OPNs (green arrows) express weak GlyR1 immunoreactivity compared to OPNs’ (red arrows) intense labeling on their somatic membrane. Scale bars represent 50 μm, in A and C; 10 μm in the inset in right column (GAD) of A. *** p < 0.05, * p < 0.1.
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Inhibitory transmitters in neurons of human nucleus raphe interpositus (RIP). (A) Consecutive 5 μm thick coronal paraffin sections through RIP stained with <t>GABA</t> A receptor (GABRA1, left) and glutamate decarboxylase (GAD65/67, right) antibodies. Note that cholinergic non-OPNs (green arrows) express intense GABRA1 immunoreactivity compared to OPNs’ (red arrows) weak labeling on their somatic membrane. This trend is reflected in the dense GABAergic inputs shown by GAD65/67 immunolabeling (right) to the cholinergic non-OPNs (green arrows) and sparse inputs to somatic membranes of OPNs (red arrows). The density difference between somatic and dendritic labelling of OPNs can be seen in the inset on the top-right corner of the image (black arrowheads). The close up (inset) is magnified from the boxed region. (B) Quantitative analysis of the GAD65/67 immunopositive puncta on somatic (dark-colored bars) and dendritic (light-colored bars) membranes of OPNs (red) and cholinergic non-OPNs (green) demonstrating significantly denser GABAergic inputs to non-OPNs. (C) Glycine receptor 1a immunolabeling in neurons of RIP demonstrate a complimentary pattern to GABRA1. Note that cholinergic non-OPNs (green arrows) express weak GlyR1 immunoreactivity compared to OPNs’ (red arrows) intense labeling on their somatic membrane. Scale bars represent 50 μm, in A and C; 10 μm in the inset in right column (GAD) of A. *** p < 0.05, * p < 0.1.
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Inhibitory transmitters in neurons of human nucleus raphe interpositus (RIP). (A) Consecutive 5 μm thick coronal paraffin sections through RIP stained with <t>GABA</t> A receptor (GABRA1, left) and glutamate decarboxylase (GAD65/67, right) antibodies. Note that cholinergic non-OPNs (green arrows) express intense GABRA1 immunoreactivity compared to OPNs’ (red arrows) weak labeling on their somatic membrane. This trend is reflected in the dense GABAergic inputs shown by GAD65/67 immunolabeling (right) to the cholinergic non-OPNs (green arrows) and sparse inputs to somatic membranes of OPNs (red arrows). The density difference between somatic and dendritic labelling of OPNs can be seen in the inset on the top-right corner of the image (black arrowheads). The close up (inset) is magnified from the boxed region. (B) Quantitative analysis of the GAD65/67 immunopositive puncta on somatic (dark-colored bars) and dendritic (light-colored bars) membranes of OPNs (red) and cholinergic non-OPNs (green) demonstrating significantly denser GABAergic inputs to non-OPNs. (C) Glycine receptor 1a immunolabeling in neurons of RIP demonstrate a complimentary pattern to GABRA1. Note that cholinergic non-OPNs (green arrows) express weak GlyR1 immunoreactivity compared to OPNs’ (red arrows) intense labeling on their somatic membrane. Scale bars represent 50 μm, in A and C; 10 μm in the inset in right column (GAD) of A. *** p < 0.05, * p < 0.1.
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Inhibitory transmitters in neurons of human nucleus raphe interpositus (RIP). (A) Consecutive 5 μm thick coronal paraffin sections through RIP stained with GABA A receptor (GABRA1, left) and glutamate decarboxylase (GAD65/67, right) antibodies. Note that cholinergic non-OPNs (green arrows) express intense GABRA1 immunoreactivity compared to OPNs’ (red arrows) weak labeling on their somatic membrane. This trend is reflected in the dense GABAergic inputs shown by GAD65/67 immunolabeling (right) to the cholinergic non-OPNs (green arrows) and sparse inputs to somatic membranes of OPNs (red arrows). The density difference between somatic and dendritic labelling of OPNs can be seen in the inset on the top-right corner of the image (black arrowheads). The close up (inset) is magnified from the boxed region. (B) Quantitative analysis of the GAD65/67 immunopositive puncta on somatic (dark-colored bars) and dendritic (light-colored bars) membranes of OPNs (red) and cholinergic non-OPNs (green) demonstrating significantly denser GABAergic inputs to non-OPNs. (C) Glycine receptor 1a immunolabeling in neurons of RIP demonstrate a complimentary pattern to GABRA1. Note that cholinergic non-OPNs (green arrows) express weak GlyR1 immunoreactivity compared to OPNs’ (red arrows) intense labeling on their somatic membrane. Scale bars represent 50 μm, in A and C; 10 μm in the inset in right column (GAD) of A. *** p < 0.05, * p < 0.1.

Journal: Frontiers in Neuroanatomy

Article Title: Transmitter and ion channel profiles of saccadic omnipause neurons and cholinergic non-omnipause neurons in human nucleus raphe interpositus

doi: 10.3389/fnana.2025.1670220

Figure Lengend Snippet: Inhibitory transmitters in neurons of human nucleus raphe interpositus (RIP). (A) Consecutive 5 μm thick coronal paraffin sections through RIP stained with GABA A receptor (GABRA1, left) and glutamate decarboxylase (GAD65/67, right) antibodies. Note that cholinergic non-OPNs (green arrows) express intense GABRA1 immunoreactivity compared to OPNs’ (red arrows) weak labeling on their somatic membrane. This trend is reflected in the dense GABAergic inputs shown by GAD65/67 immunolabeling (right) to the cholinergic non-OPNs (green arrows) and sparse inputs to somatic membranes of OPNs (red arrows). The density difference between somatic and dendritic labelling of OPNs can be seen in the inset on the top-right corner of the image (black arrowheads). The close up (inset) is magnified from the boxed region. (B) Quantitative analysis of the GAD65/67 immunopositive puncta on somatic (dark-colored bars) and dendritic (light-colored bars) membranes of OPNs (red) and cholinergic non-OPNs (green) demonstrating significantly denser GABAergic inputs to non-OPNs. (C) Glycine receptor 1a immunolabeling in neurons of RIP demonstrate a complimentary pattern to GABRA1. Note that cholinergic non-OPNs (green arrows) express weak GlyR1 immunoreactivity compared to OPNs’ (red arrows) intense labeling on their somatic membrane. Scale bars represent 50 μm, in A and C; 10 μm in the inset in right column (GAD) of A. *** p < 0.05, * p < 0.1.

Article Snippet: GABA receptor alpha subunit was detected by a polyclonal rabbit antibody (Cat #: 12410-1-AP; RRID; AB_2108692; Chromotek/Proteintech, Planegg/Martinsried, Bavaria, Germany), which recognizes an immunogen consisting of 260 aminoacids (21–280 aa encoded by BC030696 ).

Techniques: Staining, Labeling, Membrane, Immunolabeling