การนำส่งสารฮอร์โมนเพศผู้ 17 -Methyltestosterone เข้าสู่เอ็มบริโอด้วยสนามไฟฟ้าพัลส์เพื่อการแปลงเพศปลานิลเชิงพาณิชย์

Sakshin   Bunthawin; Thanate   Juntadech; Pantima   Ngimkoh; Kata  Jaruwongrungsee; Adisorn   Tuantranont

Authors

Sakshin Bunthawin Biotechnology of Electromechanics, Science of Physics, Faculty of Technology and Environment, Prince of Songkla University
Thanate Juntadech Biotechnology of Electromechanics, Science of Physics, Faculty of Technology and Environment, Prince of Songkla University
Pantima Ngimkoh Biotechnology of Electromechanics, Science of Physics, Faculty of Technology and Environment, Prince of Songkla University
Kata Jaruwongrungsee National Electronics and Computer Technology Center, National Science and Technology Development Agency
Adisorn Tuantranont National Electronics and Computer Technology Center, National Science and Technology Development Agency

Keywords:

tilapia , sex reversal rate , pulse electric fields , 17 -Methyltestosterone, gene

Abstract

The present study proposes an alternative technique of electrical sex reversal to enhance all-male sex reversal (MSR) of Nile tilapia using transient square pulse-electric fields for commercial benefits focusing on the study of mechanisms of electro-permibilization of 17 -Methyltestosterone (MT) androgen hormones through induced membrane of tilapia’s embryo (Oreochromis niloticus L., Chitralada), during segmentation-pharyngula (2-3 days post fertilization). The technique were carried out for large-scale inductions of embryos suspensions with MT concentrations of 1,000 – 100,000 µg l^-1using square-pulse electric fields of amplitudes 100-500 volts (electric field intensities of 25-125 kV m^-1), 100 µs pulse-period (50:50 mark space ratio) and 3 square pulses, respectively. It was found that the most suitable conditions of 350 V with MT concentration of 5,000 µg l^-1 gave the maximum rate of male-sex reversal rate (MSR) of 89.42%±2.28%, hatching rate (HR) of 93.33%±0.58% and survival rate (SR) of 87.49%±1.71%, respectively. The off-spring rate (OSR) of male and female in the control group were 59.71%±3.00% and 46.33%%±1.53%, respectively, with hatching rate (HR) of 91.33%±1.53% and survival rate (SR) of 85.08% ± 4.16% (increasing MSR of 29.71%). Verifications of MT delivery were carried out through LC-MS/MS technique and RNA expression profiles by extraction of total RNA from induced MT embryos. Genes of male-sex development (up-regulated genes) of amh, ar, dmrt1, bmp15, gfd9 and down-regulated genes of gsdf (where house keeping genes of rpl13a) were investigated by Quantitative real-time PCR. The induced MT embryos with transient square pulse-electric fields showed expression of male-sex development genes.

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