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109.精子流式細胞儀應用於種公豬精子品質之評估

郭廷雍、林秀蓮、詹明展、李三玉、顏念慈、吳明哲

行政院農業委員會畜產試驗所遺傳育種組

精子之細微結構如精子細胞膜、頭帽細胞膜、粒線體膜及染色體的完整性及細胞質內鈣離子濃度均與受精力有密切的關連性。而流式細胞儀可觀測細胞細微結構及各類狀態之儀器，是未來可能成為檢測精子品質之利器。本研究目的乃藉由流式細胞儀針對台灣區種 豬發展基金會出場及未出場兩大類年輕種公豬精子之細微結構及組成分加以測定，並建立各項分析值之資料庫以作為未來評估選留產精品質或能力強的種公 豬之應用。本試驗採集之新鮮種豬精液儲存於17℃溫度並攜帶回實驗室，分別予以稀釋成0.5×106 /ml濃度，加入所需染劑後於37℃培養數分鐘後上機分析。初步結果顯示在出場（n=205）與未出場（n=85）公豬部分其各項分析項目之初步結果以平均值±標準偏差表示，分別為精子膜完整性 (75±23%; 69±27%)、頭帽及精子膜完整性 (40±22%與36±25%)、粒腺體不完整性 (28±18%與31±20 %)、細胞內游離鈣水平的檢測 (443±98與443±89)、高鈣精子之比率 (83±20%與83±17 %)、DNA染色質結構完整性 (94±7%與92±11%)、精子細胞內自由基程度(71±17%與67±21%)。

關鍵語：種豬、精子品質、流式細胞儀

ASSESSMENT OF BOAR SPERM QUALITY BY FLOW CYTOMETER

T.Y. Kuo, H. L. Lin, M. C. Chan, S. Y. Lee, N.T. Yen and M.C. Wu.

Livestock Research Institute, Council of Agriculture

The fertility of sperm is related to its structures such as the integrity of sperm membrane, acrosomal membrane, mitochondrial membrane, chromatin and the calcium reservation inside the cytoplasm. Flow cytometer can be used to determine cell structure and the composition of cytoplasm and could be a very useful equipment to determine the semen quality. The purpose of this study was to develop an evaluation system of sperm quality for establishing the database of selection standards for boars. For determination of sperm quality of the boars from Formosan Farmers Association, the semen of each boar was collected and stored at 17℃ and diluted to 0.5×106 /mL. After mixing with specific dyes, the semen was incubated at 37℃ for couple minutes before analyzing. The results showed that the sperm characteristics of boars for auction (n=205) and not auction (n=85), respectively, were as below (%): sperm membrane integrity (75±23%; 69±27%), intact acrosome (40±22%; 36±25% ), depolarized mitochondria (28±18%; 31±20 % ), calcium mean level (443±98; 443±89 ), high calcium level sperm (83±20%; 83±17 % ), intact chromatin structure (94±7%; 92±11% ), oxidation degree (71±17%; 67±21% ).

Key words: Breeding pigs, Sperm Quality, Flow Cytometer