Journal: Frontiers in Neural Circuits

Article Title: Muscarinic Acetylcholine Receptor Localization on Distinct Excitatory and Inhibitory Neurons Within the ACC and LPFC of the Rhesus Monkey

doi: 10.3389/fncir.2021.795325

Figure Lengend Snippet: Primary antibodies utilized in immunohistochemistry.

Article Snippet: Vesicular GABAergic Transporter (VGAT) , Guinea Pig , 1:400 , Synaptic Systems, 131004 , AB_887873.

Techniques: Immunohistochemistry

Journal: Frontiers in Neural Circuits

Article Title: Muscarinic Acetylcholine Receptor Localization on Distinct Excitatory and Inhibitory Neurons Within the ACC and LPFC of the Rhesus Monkey

doi: 10.3389/fncir.2021.795325

Figure Lengend Snippet: Presynaptic m2 + colocalization with excitatory and inhibitory axon terminals. (A) Representative confocal images of m2 + (magenta), VGLUT1 + (cyan), and VGLUT2 + (green) in the ACC (top) and LPFC (bottom). (B,C) The density (% area labeled) of VGLUT1 + puncta (B) was significantly greater in L1 of LPFC compared to L3 while VGLUT2 + (C) boutons were equivalent across layers and areas. (D,E) Colocalization coefficient (% area colocalized) of VGLUT1 + and VGLUT2 + with m2 + . (D) In ACC, a significantly greater percent of VGLUT1 + boutons expressing m2 + was found in L2 compared to L1. (E) In LPFC, a significantly greater percent of VGLUT2 + boutons expressing m2 + was found in L3 compared to L1. (F) Representative confocal images of m2 + (magenta) and VGAT + (green), and colocalized particles in white. Scale bar 20 μm. (G) % area label showing that the laminar density of VGAT + was significantly greater within the ACC. (H) Colocalization coefficient showing % area of VGAT + colocalized with m2 + : approximately 11% of inhibitory terminals were colocalized with m2 + in both the ACC and LPFC. * p ≤ 0.05, ** p ≤ 0.01.

Article Snippet: Vesicular GABAergic Transporter (VGAT) , Guinea Pig , 1:400 , Synaptic Systems, 131004 , AB_887873.

Techniques: Labeling, Expressing

Journal: Frontiers in Neural Circuits

Article Title: Muscarinic Acetylcholine Receptor Localization on Distinct Excitatory and Inhibitory Neurons Within the ACC and LPFC of the Rhesus Monkey

doi: 10.3389/fncir.2021.795325

Figure Lengend Snippet: Co-localization of m2 + with neurochemically distinct and compartment-specific inhibitory axon terminals. (A,B) Representative confocal images of VGAT + (red), m2 + (magenta) terminals with terminals from either (A) CB (green) in L1 (top) or L3 (bottom) or (B) PV (green) in L3. (C) Box and whisker plots of % area of m2 colocalization with VGAT + /CB + in L1 (light hue) and L3 (dark hue) and with VGAT + /PV + L3 (dark hue). Within the ACC, L3 had significantly greater colocalization of CB + VGAT + terminals with m2 + than PV + VGAT + with m2 + . (D–F) VGAT + /m2 + terminals apposed (putative synapses) to specific somatodendritic MAP2 + compartments. (D) Representative confocal image of L3 MAP2 + neuron (green), VGAT + (red), m2 + (magenta). White outline illustrates the ROIs of the proximal apical dendrite and the soma. Scale bar 10 μm. (E,F) The relative proportion (top) and density (bottom) of colocalized VGAT + m2 − and VGAT + m2 + appositions on the (E) soma and (F) proximal apical dendrite of MAP2 + neurons in the ACC ( n = 10 cells) and LPFC ( n = 10 cells). (G) Representative confocal image of L3 SMI-32 + neuron (green), VGAT + (red), m2 + (magenta) showing proximal dendritic and somatic VGAT + appositions. White arrows denote examples of VGAT + only appositions and yellow arrows denote VGAT + m2 + apposition. Scale bar 20 μm. (H,I) The density of VGAT + m2 + appositions (putative synapses) per cell was significantly greater in the (H) somatic (appositions/μm 2 ) but not in the (I) dendritic (appositions/ μm) compartments of neurons in the ACC compared to the LPFC. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Vesicular GABAergic Transporter (VGAT) , Guinea Pig , 1:400 , Synaptic Systems, 131004 , AB_887873.

Techniques: Whisker Assay

Journal: Frontiers in Neural Circuits

Article Title: Muscarinic Acetylcholine Receptor Localization on Distinct Excitatory and Inhibitory Neurons Within the ACC and LPFC of the Rhesus Monkey

doi: 10.3389/fncir.2021.795325

Figure Lengend Snippet: Differential m2 + expression of VGAT + cartridges in the ACC and LPFC. (A) Representative confocal image of distinct VGAT + cartridges (arrow) in ACC ( left ) and LPFC ( right ). Boxed insets are shown in higher magnification (in the upper left corner of each image) to highlight the difference in length of VGAT + cartridges between the two areas. (B) The ACC revealed a significantly greater cartridge length compared to the LPFC. (C) The density of VGAT + cartridges (number per mm 3 ) was equivalent between the two areas. (D) Representative image of either VGAT + only ( top) , VGAT + with lightly-labeled m2 + ( middle ) or with strongly-labeled m2 + + ( bottom ), the white arrows show the VGAT + cartridges, and examples of VGAT + m2 + boutons along each cartridge are indicated by red arrows. Note that m2 + and m2 + + cartridges are made up of rows of both VGAT + m2 − and VGAT + m2 + boutons distributed along the length of each cartridge. Scale bar 5 μm. (E) The density of VGAT + only cartridges was greater in the LPFC, while the ACC had greater density of m2 + + VGAT cartridges. (F) The relative proportion of VGAT + only cartridges ( white ), lightly-labeled m2 + ( light colored ) or strongly-labeled m2 + + ( dark colored ) VGAT + cartridges in ACC ( left) and LPFC ( right). * p ≤ 0.05. (G) The density of individual VGAT + m2 − (unfilled bars) and VGAT + m2 + (filled bars) boutons along the length of distinct VGAT + cartridge types classified as either lightly-labeled m2 + or strongly-labeled m2 + + in ACC and LPFC. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Vesicular GABAergic Transporter (VGAT) , Guinea Pig , 1:400 , Synaptic Systems, 131004 , AB_887873.

Techniques: Expressing, Labeling

Journal: Frontiers in Neural Circuits

Article Title: Muscarinic Acetylcholine Receptor Localization on Distinct Excitatory and Inhibitory Neurons Within the ACC and LPFC of the Rhesus Monkey

doi: 10.3389/fncir.2021.795325

Figure Lengend Snippet: Summary of ACC and LPFC micro-circuitry influenced by muscarinic receptors. (A) Normalized densities of differentially distributed excitatory and inhibitory neuronal populations expressing mAChRs in ACC and LPFC. LPFC had a greater density of m1 + /m2 + SMI-32 + while ACC had a greater density of m1 + /m2 + SMI-32 + CB + pyramidal neurons. The two areas differed with regards to m2 + inhibitory neurons, with inhibitory neurons in the ACC having a greater extent of m2 + expression than in the LPFC. (B) Micro-circuit schematic based on the main findings of the subcellular distribution of postsynaptic m1 + on MAP2 + dendrites and m2 + inhibitory terminals on specific somatodendritic and axonal compartments. The number of connections and line thickness represents the relative strength of connection, while the dotted line indicates m2 mediated pre-synaptic suppression. Compared to LPFC, L3 ACC pyramidal neurons had a greater density of m1 + on the dendritic and somatic compartments. The dendrites of these ACC pyramidal neurons had a greater density of total VGAT + , including m2 + VGAT + , inhibitory inputs likely from CB + (yellow) neurons. The LPFC pyramidal neurons received a lower density of perisomatic input, mostly belonging to PV + interneurons (Medalla et al., ), and only a small subset expressed m2 + (pink). In contrast, the ACC had a greater density of VGAT + m2 + and VGAT + m2 − inhibitory inputs, presumably from non-PV basket cells (great) with a subset likely from CCK + interneurons (Medalla et al., ). Note that the m2 localization on CR and CCK terminations (grey) remains unknown in the present study. Finally, compared to LPFC, the ACC had a greater density of axonal targeting VGAT + m2 + cartridges, presumably from PV + chandelier cells.

Article Snippet: Vesicular GABAergic Transporter (VGAT) , Guinea Pig , 1:400 , Synaptic Systems, 131004 , AB_887873.

Techniques: Expressing

Journal: Neural Regeneration Research

Article Title: Mice lacking perforin have improved regeneration of the injured femoral nerve

doi: 10.4103/1673-5374.332152

Figure Lengend Snippet: Soma size of motoneurons and nerve terminals around retrogradely labeled motoneurons. Representative images of choline-acetyl transferase (ChAT) (A, B, green) and vesicular inhibitory transmitter transporter (VGAT) (C, D, green) immunostained motoneurons of perforin-deficient (Pfp –/– ) (B, D) and wild-type (WT; A, C) mice, 2 months after injury. Asterisks mark retrogradely labeled motoneurons which were analyzed. Scale bars: 15 μm. Cell body area (E) and linear densities (number of puncta per mm) of ChAT + (F) and VGAT + (G) perisomatic terminals in non-injured and injured motoneurons, 2 months after injury. Graphs represent group mean values ± SEM calculated from > 300 cells taken from 6 animals per group. * P < 0.05, vs . WT mice; # P < 0.05, vs . non-injured (one-way analysis of variance with Tukey’s post hoc test).

Article Snippet: The slices were then incubated at 4°C for 3 days with primary antibody against goat choline-acetyl transferase (ChAT), for cholinergic synapses (1:100; Millipore; product number AB144P; RRID: AB_2079751), or anti-mouse vesicular inhibitory transmitter transporter (VGAT), for inhibitory synapses (1:1000; Synaptic Systems, Göttingen, Germany; product number 131011; RRID: AB_887872) diluted in PBS containing 0.5% lambda-carrageenan and 0.02% w/v sodium azide in PBS.

Techniques: Labeling