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. 2016 Nov 18;10:92. doi: 10.3389/fncir.2016.00092

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Copyright © 2016 Hernández, Hernández, Perez de la Mora, Gómora, Fuxe, Eiden and Zhang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution and reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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AVP receptor V1a mRNA expression in GABAergic neurons in the CeA. Panel (A’s) sequential coronal sections were hybridized with antisense radioactive AVP V1a riboprobe. Autoradiographs with overnight-exposure were read by phosphorimager. Rostro-caudal coordinates were indicated according to Rat Brain Atlas (Paxinos and Watson, 2006). AA, Anterior amygdala; CeA, central amygdala. (B–D) In situ hybridization using RNAscope-multiplex method targeting the Gad-1, Gad-2 and V1a mRNAs in CeA. V1a mRNA is particularly expressed by Gad-1 and Gad-2 positive neurons. (B) Low magnification showing the selective location of Gad1 + Gad2 expressing cell population in the CeA (dash-line circumscribed region). (C) Further magnifications showing the exclusive expression of V1a mRNA in Gad1 and Gad2 positive cells (100%, n = 98 neurons analyzed). Optical section thickness 1 μm. (D’s) Confocal 1 μm optical slice of two cells in CeA expressing Gad1 (red), Gad2 (green) and V1a receptor (white) mRNA. Using the same method for V1b mRNA, we did not find any positive labeling. This data is not shown. The positive control for this latter experiment was done using adenohypophysis tissue. Scale bars: (B) 100 mm; (C) 20 μm; (D) 10 μm.