Dopamine plays key roles in many basic functions in the central nervous system. In order to study developmental and functional roles of dopaminergic cells in the zebrafish, we have examined a transgenic line of zebrafish expressing green fluorescent protein (GFP) under the control of the tyrosine hydroxylase (TH) promoter. TH-driven GFP was expressed in cells located in the inner nuclear layer. Immunocytochemistry with antibodies for GFP and TH showed that 29 ± 2% of GFP-labeled cells also expressed TH. Loose-patch voltage-clamp recording from GFPlabeled cells revealed that these dopaminergic neurons are spontaneously active in darkness. This transgenic line provides a useful tool to target retinal dopaminergic cells in vivo and in situ.
The vertebrate retina is profoundly influenced by circadian rhythmicity, yet little is known about the mechanisms of the zebrafish retinal circadian clock. To further the study of the zebrafish retinal clock, we have constructed a recombinant BAC in which short-half life GFP is under the control of the zebrafish circadian gene Per3 promoter. Expression of the modified BAC clone was observed in injected zebrafish embryos. Further intercrossing of injected fish and screening of their progeny may identify transgenic Per3::d2GFP fish. Meanwhile, by using transgenic Per3::LUC fish (generated by G. Cahill, University of Houston), we are able to examine the effect of light stimuli on bioluminescence rhythm at different times of a circadian cycle. Our results demonstrate that cultured zebrafish retina shows large phase shifts with phaseresponse curve close to type 0.
| Identifer | oai:union.ndltd.org:VANDERBILT/oai:VANDERBILTETD:etd-03132008-112959 |
| Date | 13 March 2008 |
| Creators | Meng, Shi |
| Contributors | Bruce H. Appel, Douglas G. McMahon |
| Publisher | VANDERBILT |
| Source Sets | Vanderbilt University Theses |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.vanderbilt.edu/available/etd-03132008-112959/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to Vanderbilt University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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