Abstract
Spermatozoa are male gametes that have an advantage in acting as gene transfer media (SMGT), because in transferring genetic material, sperm uses relatively natural vectors. This study was conducted to know sperm quality of Koi after short electroporation with different voltages using Gene Pulser. The study was conducted using a complete randomized design (RAL) with three (3) replications and five (5) treatments, namely: 10 V.cm -1 , 20 V.cm -1 , 30 V.cm -1 , 40 V.cm -1 , and 1 control. Electroporation was done with square wave, 1 ms duration of electroporation, 4 times of electroporation shocks with 0.1 s duration. The results showed that electroporation with different voltage levels significantly affected the motility and viability of sperm. Based on orthogonal polynomial analysis, it was known that the treatment affected motility and viability of sperm with a linear equation y = -0.9x + 5, 4 R² = 0.920 and r = 0.96 for motility and y = -11.26x + 91.61 R² = 0.823 and r = 0.91 for viability, meaning that the higher the level of electroporation the lower the viability and sperm motility produced. From the results of this study, it can be suggested that the electroporation of Koi sperm as a gene transfer medium should be performed at a voltage of 10 V.cm -1 . Keywords: electroporation, Koi, sperm quality, transgenesis.
Highlights
Spermatozoa are male gametes that have an advantage in acting as gene transfer media, because in transferring genetic material, sperm uses relatively natural vectors [1]
Electroporation is a method of gene transfer by using a series of short electrical vibrations to temporarily dissolve the cell membrane, thereby the foreign DNA molecule can enter the cell and interact to integrate it with the genome of the test cell and can be expressed on the organism [3,4,5]
The application of transgenesis technology to Koi using electroporation method in fish sperm as the gene transfer medium has not been done in Indonesia
Summary
Spermatozoa are male gametes that have an advantage in acting as gene transfer media, because in transferring genetic material, sperm uses relatively natural vectors [1]. Sperm cells have been used as gene transfer vectors in fish where the entry of gene construction into sperm can be facilitated by the use of electroporator [2]. The application of transgenesis technology to Koi using electroporation method in fish sperm as the gene transfer medium has not been done in Indonesia. To know the succesful of using gene transfer method with sperm electroporation especially for Koi, it is necessary to do the experiment about preliminary study on sperm quality after giving electric shock. A preliminary study on the sperm quality of Koi fish after electroporation is important to determine the ability of sperm to fertilize egg. So, it is necessary to experiment with different stress levels. The purpose of this study was to determine the effect of stress on sperm quality
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