Stallion Sperm Selection by Density Gradient CentrifugationInvolving a Double Layer Colloid: Effects on Sperm Subpopulation Dynamics in Fresh and Stored Semen
محورهای موضوعی : Camel
جی. میرو
1
(Departmentof Animal Medicine and Surgery, Faculty of Veterinary, University of Autónoma de Barcelona, 08193 Bellaterra, Spain)
ف. رکوئنا
2
(Department of Celular Biology, Phisiology and Immunology, Faculty of Veterinary, University of Córdoba, 14071 Córdoba, Spain)
اچ. مارین
3
(Departmentof Animal Medicine and Surgery, Faculty of Veterinary, University of Autónoma de Barcelona, 08193 Bellaterra, Spain)
جی. جردنا
4
(Departmentof Animal Medicine and Surgery, Faculty of Veterinary, University of Autónoma de Barcelona, 08193 Bellaterra, Spain)
ای. آگوئرا
5
(Department of Celular Biology, Phisiology and Immunology, Faculty of Veterinary, University of Córdoba, 14071 Córdoba, Spain)
کلید واژه: density gradient centrifugation, EquiPureTM, sperm selection, sperm subpopulations, subfertile stallions,
چکیده مقاله :
Sperm selection techniques have became an important tool to improve sub fertile ejaculates. The main aim of this study was to assess the dynamics of motile sperm subpopulations of ejaculates from subfertile stallions subjected to density gradient centrifugation (DGC) involving a double layer colloid [DGC2], before and after storage for 24 and 48 hours. Ejaculates from eight stallions with fertility problems were subjected to the different treatments: 1) control [C]; 2) centrifugation [CT]; 3) density gradient centrifugation involving a double layer colloid [DGC2] and 4) post storage density gradient centrifugation involving a double layer colloid DGC2 post]. In the C, CT and DGC2 treatments, sperm variables were analysed at 0 h and after 24 and 48 h of storage at 4-7 ˚C. In the DGC2 post treatment, the semen was stored at 4-7˚Cand sperm samples selected by DGC2 at 24 and 48 h. Viability, sperm abnormalities and motility were then examined using a computerized system. Four motile sperm subpopulations with different motility characteristics were observed in the all treatments. The DGC2 sperm had faster, straighter moving and more active cells at 0 h. At 24 and 48 h, however, the motility variable values fell and the slower spermatozoa subpopulations increased in size. The DGC2 post treatment returned better overall sperm motility variable values at 24 and 48 h; a larger percentage of spermatozoa fell into the faster subpopulations. In conclusion, DGC2 could be used to improve semen from subfertile stallions as well as semen showing poor fertility after storage.
تکنیکهای انتخاب اسپرم یکی از ابزارهای مهم در بهبود انزالهای کم بارور است. هدف اصلی در این مطالعه، دستیابی به دینامیک زیرجمعیتهای اسپرم متحرک از انزالهای اخذ شده از نریانهای کم بارور با استفاده از سانتریفیوژ گرادیان چگالی (DGC) و یک کلویید دو لایه (DGC2)، پیش و پس از 24 ساعت و 48 ساعت ذخیره است. انزالهای اخذ شده از 8 نریان دارای مشکلات باروری در تیمارهای مختلف مورد استفاده قرار گرفتند. تیمارها شامل کنترل (C)، سانتریفیوژ (CT)، سانتریفیوژ گرادیان چگالی همراه با کلویید دولایه (DGC2) و سانتیرفیوژ گرادیان چگالی همراه با کلویید دو لایه پس از چگالی (DGC post) بودند. در تیمارهای C، CT و DGC2، متغیرهای اسپرم در 0، 24 و 48 ساعت پس از ذخیره در 4 تا 7 درجه سلسیوس مورد آنالیز قرار گرفت. در تیمار DGC2 post منی در 4 تا 7 درجه سلسیوس ذخیره، و نمونههای اسپرم با کمک DGC2 در 24 و 48 ساعت انتخاب شدند. سپس زندهمانی، نقایص و تحرک اسپرم با کمک یک سامانه رایانهای مورد سنجش قرار گرفت. 4 زیرجمعیت اسپرم متحرک با خصوصیات تحرک متفاوت در کلیه تیمارها مشاهده شدند. اسپرم DGC2 سریعتر بوده و مستقیمتر حرکت کرده و دارای سلولهای فعالتر در ساعت صفر بودند. ولی در ساعت 24 و 48، مقادیر متغیر تحرک کاهش یافته و اندازه زیرجمعیتهای اسپرم کُندتر افزایش یافتند. تیمار DGC2 post مقادیر متغیر تحرک بهتری را در ساعت 24 و 48 نشان داده و درصد بیشتری از اسپرمها در زیرجمعیتهای سریعتر قرار گرفتند. به طور کلی می‌توان نتیجه گرفت که DGC2 میتواند برای بهبود منی اخذ شده از نریانهای کم بارور و منیهایی که باروری پایینی پس از ذخیرهسازی نشان میدهند، بهکار گرفته شود.
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