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. 2020 Jul 16;14:43. doi: 10.3389/fncir.2020.00043

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Copyright © 2020 Noftz, Beebe, Mellott and Schofield.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) 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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Injection of viral vector into the PPT of ChAT-Cre transgenic rats labeled neurons at the site of injection. Photomicrographs show virally-expressed fluorescent protein in PPT neurons. (A) A representative example of mCherry expression in neurons of the left PPT following injection of rAAV2/EF1a-DIO-mCherry. Transverse section; lateral is left, dorsal is up. Scale bar = 500 μm. (B) A representative example of EYFP expression in neurons of the right PPT following injection of rAAV2/EF1a-DIO-EYFP. Transverse section; lateral is right, dorsal is up. Scale bar = 500 μm. (C) A representative three-dimensional reconstruction showing the extent of labeled neurons in the PPT through six transverse sections (spacing: 240 μm between sections). The image on the left shows the six sections stacked, with each EYFP-labeled cell indicated by a green circle. The image on the right shows the stack rotated for a lateral view. The “cloud” of labeled cells shifts ventrally moving from caudal to rostral, reflecting the orientation of the PPT. Aq, cerebral aqueduct; D, dorsal; IC, inferior colliculus; LL, lateral lemniscus; M, medial; PAG, periaqueductal gray; PPT, pedunculopontine tegmental nucleus; R, rostral, scp, superior cerebellar peduncle.