title: Simple, economical heat-shock devices for zebrafish housing racks.
abstract: One reason for the popularity of the zebrafish (Danio rerio) as a model vertebrate is the ability to manipulate gene expression in this organism. A common method is to induce gene expression transiently under control of a heat-shock promoter (e.g., hsp70l). By making simple mechanical adjustments to small aquarium heaters (25-50W), we were able to produce consistent and reliable heat-shock conditions within a conventional zebrafish housing system. Up to two heat-shock intervals per day (>37°C) could be maintained under conditions of continuous flow (5-25 mL/min). Temperature logging every 30 s indicated rapid warm up times, consistent heat-shock lengths, and accurate and precise peak water temperatures (mean±SD=38°C±0.2°C). The biological effects of these heat-shock treatments were confirmed by observing inducible expression of enhanced green fluorescent protein (EGFP) and inhibition of caudal fin regeneration in a transgenic fish line expressing a dominant negative fibroblast growth factor receptor (Tg(hsp70l:dnfgfr1-EGFP)(pd1)). These devices are inexpensive, easily modified, and can be calibrated to accommodate a variety of experimental designs. After setup on a programmable timer, the heaters require no intervention to produce consistent daily heat shocks, and all other standard care protocols can be followed in the fish facility. The simplicity and stability of these devices make them suitable for long-term heat shocks at any stage of the zebrafish lifecycle (>7 days postfertilization), and useful for both laboratory and classroom experiments on transgenic zebrafish.
authors.affiliation: Department of Biological Sciences, DePaul University, Chicago, Illinois 60614, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Duszynski
authors.forename: Jacek
authors.initials: J
authors.lastname: Topczewski
authors.forename: Elizabeth E
authors.initials: EE
authors.lastname: LeClair
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018869:Heat-Shock Response / Q000235:genetics / Q000502:physiology*; D006361:Heating / Q000191:economics / Q000295:instrumentation* / Q000379:methods; D006358:Hot Temperature; D006799:Housing, Animal; D023421:Models, Animal; D011897:Random Allocation; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics / Q000378:metabolism; D012038:Regeneration; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D017468:Receptors, Fibroblast Growth Factor; D029961:Zebrafish Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins