Abstract
Successful studies about implementation of electroporation method which sperm becomes gene transfer medium for fish transgenesis have reported. Motility and viability are two major factors in successful of electroporation. The objective of the study was to determine optimal pulse length of electroporation towards the motility and viability of Koi fish sperm. The study was conducted at the Central Laboratory of Biological Sciences and Reproduction of Brawijaya University. The data analysis was conducted using completely randomized design. The electroporation method was carried out using voltage of 30 volt with pulse number of 4 times. The pulse length of the electroporation (based on treatment) was 0.5, 1 and 1.5 ms. The result showed the highest percentage of motility was 3.67% in treatment A (pulse length 0.5 ms) with coefficient correlation R 2 = 0.9643 and the best percentage of viability was 79.67% in treatment A (pulse length 0.5 ms) with coefficient correlation R 2 = 1. Correlation between the pulse length treatment (electroporation) toward motility and viability of Koi fish sperm, in which longer duration of pulse length treatment (electroporation) would decrease the motility and viability percentage of the fish sperm. Keywords: Electroporation, Motility, Pulse length, Sperm, Viability.
Highlights
Electroporation is one of the effective and efficient gen transfer method widely applied in aquaculture besides microinjection
The success of gene transfer method by electroporation method using sperm as the gene transfer media depends on the voltage level and the concentration of DNA vector during electroporation [4]
Previous research has shown that Chinook Salmon (Oncorhynchus tshawytscha) sperm activity decreased from 82% to 2% post electroporation [5], it is important to find out how to use electroporation method optimally
Summary
Electroporation is one of the effective and efficient gen transfer method widely applied in aquaculture besides microinjection. Electroporation method is mostly conducted using sperm as vector that carries a foreign gene to be introduced (sperm-mediated gene transfer). The main factor of success in fish sperm transgenesis is to succeed in maintaining the quality of sperm which includes motility and viability for fertilization. The study of optimum pulse length in Ornamental Japanese carp (Cyprinus carpio Var. Koi) fish transgenesis using electroporation method has not been studied and it is necessary to conduct experiment on influence of different pulse length towards motility and viability of Ornamental Japanese carp fish sperm to get optimum result. The purpose of this research is to describe the influence of electroporation method using voltage level (30 volt) and different pulse length towards motility and viability of Ornamental Japanese carp fish (Cyprinus carpio Var.Koi) sperm
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