Effect of valproic acid on zebrafish somitogenesis

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Experimental Design

Danio embryos were collected at the 16 or 32 cell stage, prior to the mid-blastula transition. The embryos were stored in Zebrafish embryo media. The embryos were divided into five separate groups. Each group of embryos contained approximately 15-30 Danio embryos. Different concentrations of valproic acid and Zebrafish Embryonic Solution (ZES) were prepared. A control of only Zebrafish Embryonic Solution and four mixtures of valproic acid and Zebrafish embryonic solution were created. The solutions contained 0.025M valproic acid, 0.05M valproic acid, 0.1M valproic acid, and 0.2M valproic acid. These five solutions were poured into five dishes, and each group of 15-30 embryos were placed into each of the five dishes containing the solution approximately 7 hours after fertilization. All embryos were incubated at 18-20°C for 20 hours. The embryos were fixed and stained with antibodies against somite boundaries and motor neurons in order to observe the patterning during late gastrulation and organogenesis. The staining protocol was followed except HRP substrate was used in the place of fluorescent stain. Embryos were photographed at a set time interval to record any changes in somite development among the different groups.

© Cebra-Thomas, 2001
Last Modified: 31 May 2001

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		Experimental Design
		Danio embryos were collected at the 16 or 32 cell stage, prior to the mid-blastula transition. The embryos were stored in Zebrafish embryo media. The embryos were divided into five separate groups. Each group of embryos contained approximately 15-30 Danio embryos. Different concentrations of valproic acid and Zebrafish Embryonic Solution (ZES) were prepared. A control of only Zebrafish Embryonic Solution and four mixtures of valproic acid and Zebrafish embryonic solution were created. The solutions contained 0.025M valproic acid, 0.05M valproic acid, 0.1M valproic acid, and 0.2M valproic acid. These five solutions were poured into five dishes, and each group of 15-30 embryos were placed into each of the five dishes containing the solution approximately 7 hours after fertilization. All embryos were incubated at 18-20°C for 20 hours. The embryos were fixed and stained with antibodies against somite boundaries and motor neurons in order to observe the patterning during late gastrulation and organogenesis. The staining protocol was followed except HRP substrate was used in the place of fluorescent stain. Embryos were photographed at a set time interval to record any changes in somite development among the different groups. © Cebra-Thomas, 2001 Last Modified: 31 May 2001 [Lab Protocols \| Students \| Cebra-Thomas \| Course \| Links ]