Journal of Aquaculture (한국양식학회지)

The Korean Society of Fisheries and Aquatic Science (한국양식학회)
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Effects of Diluents and Cryoprotectants on Sperm Cryopreservation in Starry Flounder (Platichthys stellatus)

강도다리(Platichthys stellatus) 정자의 냉동보존에 미치는 희석액과 동해방지제의 영향

  • Lim, Han-Kyu (Aquaculture Management Team, National Fisheries Research and Development Institute) ;
  • An, Cheul-Min (Fisheries Resources Restoration Development and Management Center, National Fisheries Research and Development Institute) ;
  • Noh, Gyong-Ane (Department of Fisheries Biology, Graduate School, Pukyong National University) ;
  • Min, Byung-Hwa (Aquaculture Management Team, National Fisheries Research and Development Institute)
  • 임한규 (국립수산과학원 양식관리팀) ;
  • 안철민 (국립수산과학원 자원회복사업단) ;
  • 노경언 (부경대학교 수산생물학과) ;
  • 민병화 (국립수산과학원 양식관리팀)
  • Published : 2007.08.31
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Abstract

An experiment was performed to obtain cryopreservation techniques of starry flounder (Platichthys stellatus) sperm. Milt obtained from 24 males were cryopreserved using two diluents, artificial seminal plasma (ASP) and Stein's solution (SS) with three cryoprotectants, dimethyl sulfoxide (DMSO), methanol, and glycerol concentrated from 5% to 20%. Post-thaw sperm activity (motility and/or speed) revealed the highest in 10% DMSO and 15% methanol in ASP and SS as diluent. Motility and speed of cryopreserved sperm were decreased according to increase of glycerol concentration. To conclude, DMSO was a better cryoprotectant than methanol or glycerol for cryopreservation of starry flounder sperm. Glycerol was incongruent cryoprotectant because of toxic to starry flounder sperm. Most cryopreserved spermatozoa without cryoprotectant showed the enlarged head with granulated chromatin and ruptured plasma membrane by freezing and thawing injuries compared with unfrozen normal spermatozoa.

희석액과 동해방지제로 각각 ASP와 DMSO를 사용하여 1년 동안 냉동보존한 후 해동한 강도다리(Platichthys stellatus) 정자의 운동속도는 10% 농도에서 가장 빨랐다. Methanol의 경우 $10{\sim}20%$ 범위에서 유의한 차이를 보이지 않았지만 15%에서 가장 높은 값을 보였다. Glycerol을 동해방지제로 사용하였을 때는 첨가농도가 높아질수록 정자의 운동성과 운동속도가 낮아졌다. 희석액으로 SS를 사용한 경우도 ASP와 비슷한 결과를 보였다. 투과형전자현미경으로 관찰한 냉동하지 않은 신선한 강도다리 정자는 머리, 중편부 및 꼬리로 구성되어 있으며, 치밀한 핵질로 충만한 구형의 머리에는 첨체구조가 관찰되지 않았다. 동해방지제 없이 냉동보존한 경우, 대부분의 정자들이 머리가 찌그러지거나 부분적으로 수축되는 경우가 많았으며 머리의 원형질막이 이탈되고 염색질이 과립상으로 변하거나 균질화되지 않았다. 반면에 냉동보존을 위해 동해방지제를 사용한 경우 부분적으로 정자의 구조적 손상이 관찰되었으나 대부분의 정자는 냉동과 해동과정에서 손상을 입지 않았다.

Keywords

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