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اثر افزودن ال-گلوتامین به رقیق‌کنندة تریس بر انجماد اسپرم پوشش‌دارشدة قوچ تالشی

نوع مقاله: مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد گروه علوم دامی دانشکدة علوم کشاورزی دانشگاه گیلان

2 دانشیار، گروه علوم دامی دانشکدة علوم کشاورزی دانشگاه گیلان

چکیده

آزمایش به‌منظور بررسی اثر ال-گلوتامین بر انجماد اسپرم پوشش‌دارشده با استفاده از چهار رأس قوچ انجام گرفت. منی با استفاده از واژن مصنوعی متصل به لولة حاوی تریس-فروکتوز- 15 درصد زردة تخم‌مرغ جمع‌آوری شد. نمونه‌ها تجمیع و سانتریفیوژ شدند. پس از حذف مایع فوقانی و رقیق‌سازی رسوب، نمونه‌ها به پنج قسمت تقسیم شدند و به آن‌ها مقدار صفر، 40، 80، 120 و 160 میلی‌مول ال-گلوتامین اضافه شد. نمونه‌ها با استفاده از نیتروژن مایع منجمد و بعد از دو هفته یخ‌گشایی شدند. بعد از یخ‌گشایی و ذخیره‌سازی در 37 درجة سانتی‌گراد، تحرک پیش‌رونده، سلامت غشای پلاسمایی، زنده‌مانی (هوخست بیس بنزامید 33258) و سلامت غشای آکروزومی (آلکسافلور-488PNA-) پس از صفر، سه، شش و نه ساعت بررسی شدند. اثر مستقل ال-گلوتامین نشان داد زنده‌مانی اسپرم در مقدار صفر (95/35 درصد)، 40 (65/40 درصد) و 80 (95/45 درصد) میلی‌مول بیشتر از160 میلی‌مول ال-گلوتامین بود (40/30 درصد، 05/0>P). اثر متقابل ال-گلوتامین و زمان ذخیره‌سازی بر تحرک پیش‌رونده و سلامت غشای پلاسمایی اسپرم معنادار بود (05/0P<). سه ساعت پس از یخ‌گشایی، بیشترین سلامت غشای پلاسمایی (به ترتیب 25/49 و 34/47 درصد) در مقدار 80 میلی‌مول ال-گلوتامین مشاهده شد (05/0P<). نه ساعت پس از یخ‌گشایی، بیشترین تحرک پیش‌روندۀ اسپرم (4/26 درصد) در مقدار 80 میلی‌مول ال-گلوتامین مشاهده شد (05/0P<). بنابراین افزودن 80 میلی‌مول ال-گلوتامین به رقیق‌کنندة تریس- گلیسرول (5 درصد) سبب بهبود ماندگاری اسپرم پوشش‌دار شد.

کلیدواژه‌ها


عنوان مقاله [English]

Effect of adding L-glutamine to Tris diluent on cryopreservation of coated sperm of Taleshi ram

نویسندگان [English]

  • Sanaz Hasanpoor 1
  • Mohammad Roostaei-Ali Mehr 2
  • Mehrdad Mohammadi 2
1 M.Sc. Student, Department of Animal Science, Faculty of Agricultural Sciences, University of Guilan
2 Associate professor, Department of Animal Science, Faculty of Agricultural Sciences, University of Guilan, Rasht
چکیده [English]

Experiment was conducted to evaluate the effect of L-glutamine on coated sperm freezing by using four rams. Semen was collected by artificial vagina contact with a tube containing Tris-fructose-egg yolk 15%. Samples were pooled and centrifuged. After removing supernatant and diluting pellet, samples were split into 5 equal part and added 0, 40, 80, 120 and 160 mM L-glutamine. Samples were frozen by using liquid nitrogen After two weeks, samples were thawed and incubated at 37 °C for nine h. Sperm motility, plasma membrane integrity, viability (Hoechst 33258) and acrosome reaction (PNA-Alexa -flur-488) were investigated at 0, 3, 6 and 9 h. The main effect of L-Glutamine showed that sperm viability was higher in 0 (35.95%), 40 (40.65%) and 80 (45.95%) mM L-Glutamine than 160 mM L-Glutamine (30.40%; P<0.05). There was an interaction between incubation time and L-Glutamine on sperm motility and plasma membrane integrity (P<0.05). After thawing and 3 h later, the highest of plasma membrane integrity was observed in 80 mM L-Glutamine (49.25% and 47.34% respectively; P<0.05). Nine hours after thawing, the highest of sperm motility (26.4%) was observed in 80 mM L-Glutamine (P<0.05). Therefore, the addition of 80 mM L-glutamine to Tris-glycerol diluent (5%) improves coated sperm survival.

کلیدواژه‌ها [English]

  • L-Glutamine
  • Taleshi ram
  • Coated spermatozoa
  • Freezing
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