Identification of Germ-Line Stem Cells in Zebrafish
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Both male and female zebrafish have a population of germ-line stem cells that produce gametes throughout the life of the fish. These cells localize to specific regions in the gonads and can be identified because they uniquely express the nanos2 gene, which encodes a conserved regulator of translation. A method is presented here for identifying germ-line stem cells in the ovary and testis using a combined protocol for whole-mount fluorescent RNA in situ hybridization to detect nanos2 mRNA and immunofluorescence to detect the pan-germ cell marker Vasa.
Key wordsZebrafish germ-line stem cells Oogonial stem cell Spermatogonial stem cell Zebrafish ovary Zebrafish testis nanos2 Vasa Fluorescent RNA in situ hybridization Confocal immunofluorescence
The author would like to thank Holger Knaut for the anti-Vasa antibodies. This work was supported by a NIH grant R01-HD081551.
- 5.Thisse B, Thisse C (2014) In situ hybridization on whole-mount zebrafish embryos and young larvae. In: Nielsen BS (ed) In situ hybridization protocols, vol 1211, Methods in molecular biology. Springer Science + Business Media, New York, NY, pp 53–67Google Scholar