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Published February 2013 | Accepted Version + Supplemental Material
Journal Article Open

New transgenic reporters identify somatosensory neuron subtypes in larval zebrafish

Abstract

To analyze somatosensory neuron diversity in larval zebrafish, we identified several enhancers from the zebrafish and pufferfish genomes and used them to create five new reporter transgenes. Sequential deletions of three of these enhancers identified small sequence elements sufficient to drive expression in zebrafish trigeminal and Rohon-Beard (RB) neurons. One of these reporters, using the Fru.p2x3-2 enhancer, highlighted a somatosensory neuron subtype that expressed both the p2rx3a and pkcα genes. Comparison with a previously described trpA1b reporter revealed that it highlighted the same neurons as the Fru.p2x3-2 reporter. To determine whether neurons of this subtype possess characteristic peripheral branching morphologies or central axon projection patterns, we analyzed the morphology of single neurons. Surprisingly, although these analyses revealed diversity in peripheral axon branching and central axon projection, PKCα/p2rx3a/trpA1b-expressing RB cells did not possess obvious characteristic morphological features, suggesting that even within this molecularly defined subtype, individual neurons may possess distinct properties. The new transgenes created in this study will be powerful tools for further characterizing the molecular, morphological, and developmental diversity of larval somatosensory neurons.

Additional Information

© 2012 Wiley Periodicals, Inc. Received 2 July 2012; accepted 26 July 2012. Article first published online: 27 Sep. 2012. We thank Lindsey Mork for initial cloning of zebrafish enhancers, Holly Vu for tracing neurons, Hillary McGraw for advice on trkA in situ hybridization, Michael Granato for the Gt(T2KSAG)j1229a line, Albert Pan and Alex Schier for the Tg(trpA1b:EGFP) line, Mark Voigt for the Tg(p2rx3b:EGFP) line, Matt Veldman, Ann Cavanaugh, and Kevin Mouillesseaux for technical advice and members of the Sagasti Lab for comments on the manuscript. Funds were provided by an NRSA (5F31NS064817) to AMSP from the NINDS and grants from the NSF (RIG:0819010) and NIDCR (5R01DE018496) to AS.

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Accepted Version - nihms410643.pdf

Supplemental Material - DNEU_22049_sm_SuppInfo.pdf

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