Elsevier

Gene Expression Patterns

Volume 11, Issue 7, October 2011, Pages 409-414
Gene Expression Patterns

A zebrafish SKIV2L2-enhancer trap line provides a useful tool for the study of peripheral sensory circuit development

https://doi.org/10.1016/j.gep.2011.06.002Get rights and content

Abstract

The zebrafish is an ideal model for elucidating the cellular and molecular mechanisms that underlie development of the peripheral nervous system. A transgenic line that selectively labels all the sensory circuits would be a valuable tool for such investigations. In this study, we describe such a line: the enhancer trap zebrafish line Tg(SKIV2L2:gfp)j1775 which expresses green fluorescent protein (gfp) in the peripheral sensory ganglia. We show that this transgene marks all peripheral ganglia and sensory nerves, beginning at the time when the neurons are first extending their processes, but does not label the efferent nerves. The trapped reporter is inserted just upstream of a previously poorly described gene: lhfpl4 on LG6. The expression pattern of this gene by in situ hybridization reveals a different, but overlapping, pattern of expression compared to that of the transgene. This pattern also does not mimic that of the gene (skiv2l2), which provided the promoter element in the construct. These findings indicate that reporter expression is not dictated by an endogenous enhancer element, but instead arises through an unknown mechanism. Regardless, this reporter line should prove to be a valuable tool in the investigation of peripheral nervous system formation in the zebrafish.

Highlights

► Tg(SKIV2L2:gfp)j1775 is a zebrafish enhancer trap line that labels sensory neurons. ► GFP expression starts soon after neurogenesis and is maintained into adulthood. ► There is no labeling of efferent axons in the periphery. ► The transgene is inserted in the 5′ end of the lhfpl4 gene. ► The expression pattern is novel and does not arise solely from the trapped gene nor the basal promoter in the construct.

Section snippets

Results and discussion

The sensory peripheral nervous system carries information from the internal organs and the external environment to the CNS. In vertebrates, the nerves that carry such signals originate in ganglia lying outside the CNS (Butler and Hodos, 2005). Although the anatomy of these nerves has been known for over a century, the cellular and molecular mechanisms that underlie the establishment of their connections are poorly understood. The zebrafish is an ideal vertebrate model for studying the

Zebrafish

Zebrafish and embryos were maintained as previously described (Kucenas et al., 2006). Embryos were treated with 0.003% phenylthiourea (PTU) to prevent pigmentation. The described line was constructed by a Tol2 transposon-mediated enhancer trap method (Fisher et al., 2006), using a fragment containing the eGFP gene and a 243 bp fragment of promoter and 5′-untranslated region from the human SKIV2L2 gene (superkiller viralicidic activity 2-like protein 2). Tg(SKIV2L2:gfp) was identified from a pool

Acknowledgements

We would like to thank Rachael Sheridan for help with fish husbandry, and Linda Cox and Dr. J. Chrivia for help with the qPCR experiments. This work was supported by grants from the NIH to A. S. McC. (GM71648), S.L.J. (GM56988) and M.M.V. (NS060074) and from the Saint Louis University School of Medicine to M.M.V.

References (15)

There are more references available in the full text version of this article.
View full text
Copyright © 2011 Elsevier B.V. All rights reserved.