نوع مقاله : مقاله پژوهشی
نویسندگان
1 کارشناسی ارشددانش آموخته ژنتیک و اصلاح دام گروه علوم دامی ;دانشکده علوم دامی و صنایع غذایی -دانشگاه علوم کشاورزی و منابع طبیعی خوزستان-اهواز-ایران
2 استاد گروه علوم دامی ;دانشکده علوم دامی و صنایع غذایی -دانشگاه علوم کشاورزی و منابع طبیعی خوزستان-اهواز-ایران
3 استادیار گروه علوم دامی ;دانشکده علوم دامی و صنایع غذایی -دانشگاه کشاورزی و منابع طبیعی خوزستان-اهواز-ایران
4 دانشیار گروه علوم دامی ;دانشکده علوم دامی و صنایع غذایی -دانشگاه کشاورزی و منابع طبیعی خوزستان-اهواز-ایران
چکیده
اثر دو رقیق‌کننده مختلف تریس- زرده تخم‌مرغ و آندرومد بر کیفیت منی و نسبت جنسیت اسپرم گاو هلشتاین در طول فرآیند رقیق‌سازی و انجماد با استفاده از تکنیک real- time qPCR بررسی شد. این آزمایش با استفاده از چهار گاو نر نژاد هلشتاین در سن چهار سالگی و به‌مدت چهار هفته در قالب طرح کاملاً تصادفی انجام شد .ژنهای PLP و SRY به ترتیب برای جداسازی قطعات خاصی از توالی های کروموزوم X- و Y- تکثیر شدند. استفاده از رقیق‌کننده آندرومد در مقایسه با تریس- زرده تخم‌مرغ، کیفیت منی در انجماد و یخ‌گشایی را بهبود داد. استفاده از رقیق‌کننده آندرومد موجب افزایش معنی‌دار بیان ژن (PLP( 0/16±1/43، و( p=0/01) و کاهش معنی‌دار بیان ژن (SRY(0/11±0/64 و ( p=0/009) در حالت مایع و فرایند انجماد شد. استفاده از رقیق کننده تریس- زرده تخم مرغ باعث کاهش معنی‌دار بیان ژن (PLP (0/06±0/3)، (p<0/0001 و افزایش معنی‌دار بیان ژن (SRY (1/19±0/05)، (p=0/001 در حالت انجماد شد. براساس نتایج، استفاده از منی رقیق کننده آندرومد و منی منجمد شده با تریس- زرده تخم مرغ به ترتیب سبب افزایش ماده زایی و نرزایی در گله گاو شیری می شود.
کلیدواژه‌ها
عنوان مقاله [English]
Effect of freezing with different extenders on the semen quality and sex ratio of Holstien bull sperm by Real-time qPCR technique
نویسندگان [English]
- Sepideh Rostami 1
- Mohammad taghi Beigi nassiri 2
- Mahmood Nazari 3
- Saleh Tabatabaei Vakili 4
1 MS Student of Animal genetic and Breeding, Faculty of Animal Science and Food Technology, Agricultural science and Natural Resources University of Khuzestan, Iran
2 Professor, Department of Animal Science, Faculty of Animal science and Food Technology, Agricultural Science and Natural Resources University of Khuzestan, Ahvaz, Iran
3 Assistant Professor, Department of Animal Science, Faculty of Animal science and Food Technology, Agricultural Science and Natural Resources University of Khuzestan, Ahvaz, Iran
4 Associate Professor, Department of Animal Science, Faculty of Animal science and Food Technology, Agricultural Science and Natural Resources University of Khuzestan, Ahvaz, Iran
چکیده [English]
The effects of two different extenders were investigated (Tris- egg yolk and Andromed extenders) on semen quality and sex ratio during the dilution and freezing process in Holstien bull by using the real-time qPCR technique. The experiment was conducted using four 4-years old Holstien bulls for a period of four weeks in a completely randomized design. The PLP and SRY genes were amplified to isolate the specific fragments of X- and Y- chromosome sequences, respectively. Using Andromed diluent in comparison with Tris- eggs yolk improved semen quality during freezing and thawing. Using Andromed extender significantly increased the PLP gene expression (1.43±0.16), (P= 0/01) and reduced SRY gene expression (0.64±0.11), (P= 0/009). Using Tris- egg yolk extender led to a significant reduced in expression of PLP gene (0.3±0.06), (P< 0/0001) and s increased SRY gene expression during freezing (1.19±0.05), (P= 0/001). Based on the results, sperm dilution with Andromed extender and frozen semen with Tris-egg yolk increase birth of female and male in dairy cow herd, respectively.
کلیدواژه‌ها [English]
- Andromed
- PLP gene
- SRY gene
- Y- chromosome
- Yolk egg
4. Barbas JP and Mascarenhas RD (2009) Cryopreservation of domestic animal sperm cells. Cell and tissue banking 10(1): 49-62.
5. Delgado PA, Lester TD, and Rorie RW (2010) Variation in the Ratio of X-to Y-Chromosome Bearing Spermatozoa in Ejaculates of Semen Collected Weekly from Beef Bulls. Animal Science Arkansas Animal Science. 28-30.
7. Fukui Y, Kohno H, Togari T, Hiwasa M, and Okabe K (2008) Fertility after artificial insemination using a soybean-based semen extender in sheep. Journal of Reproduction and Development 54(4): 286-289.
8. Gil J, Lundeheim N, Soderquist L, and Rodriguez-Martinez H- 2003) Influence of extender, temperature, and addition of glycerol on post-thaw sperm parameters in ram semen. Theriogenology 59(5-6): 1241-1255.
9. Hansen PJ (2013) Alternative Sperm Purification Proce. University of Florida. Animal Science.
10. Hassan BJ, Hossain SO, and Jebur MSh (2016) Evaluation of semen sex ratio in cooled and frozen semen straws by real -time PCR. Journal of Veterinary Research 15(3): 225-235.
11. Hu JH, Li QW, Jiang ZL, and Li WY (2008) Effects of different extenders on DNA integrity of boar spermatozoa following freezing–thawing. Cryobiology 57(3): 257-262.
12. Ijaz A, Hussain A, Aleem M, Yousaf MS, and Rehman H (2009) Butylated hydroxytoluene inclusion in semen extender improves the postthawed semen quality of Nili-Ravi buffalo (Bubalus bubalis). Theriogenology 71(8): 1326-1329.
13. Larionov A, Krause A, and Miller W (2005) A standard curve based method for relative real time PCR data processing. BMC bioinformatics 6(1): 62.
14. Miller SA, Dykes DD, and Polesky, HFRN (1988) A simple salting out procedure formextracting DNA from human ucleated cells. ucleic acids research 16(3): 12-15.
15. Najafi A, Asadi E, Moawad AR, Mikaeili S, Amidi F, Adutwum E, and Sobhani AG (2016) Supplementation of freezing and thawing media with brain-derived neurotrophic factor protectsm human sperm from freeze-thaw-induced damage. Fertility and sterility 106(7): 1658-1665.
16. Papa F, Henrion R, and Breart G (1983) Preconceptional selection of sex using the ionic method. Dietary regime. Results of a 2 years' prospective clinical study. Journal de gynecologie, obstetrique et biologie de la reproduction 12(4): 415-422.
17. Parati K, Bongioni G, Aleandri R, and Galli A (2006) Sex ratio determination in bovine semen: A new approach by quantitative real time PCR. Theriogenology 66(9): 2202-2209.
18. Sharma R, Kattoor AJ, Ghulmiyyah J, and Agarwal A (2015) Effect of sperm storage and selection techniques on sperm parameters. System Biology Reproduction Medicen 61(1): 1-12.
19. Skoff RP, Bessert D A, Cerghet M, Franklin MJ, Rout UK, Nave KA, and Armant DR (2004) The myelin proteolipid protein gene modulates apoptosis in neural and non-neural tissues. Cell death and differentiation 11(12): 1247.
20. Sumner AT, Robinson JA and Evans HJ (1971) Distinguishing between XY and YY-bearing human spermatozoa by fluorescence and DNA content. Nature new biology 229: 231-233.
21. Tavalaee M, Abbasi H, Deemeh MR, Fotohi F, Gilani, MAS and Esfahani M HN (2012) Semen parameters and chromatin packaging in microsurgical varicocelectomy patients. International journal of fertility & sterility 6(3): 165.
22. Ward WS (2009) Function of sperm chromatin structural elements in fertilization and development. MHR: Basic science of reproductive medicine 16(1): 30-36.
23. Xu ZZ, Johnson DL, and Burton LJ (2000) Factors affecting the sex ratio in dairy cattle in New Zealand. Proceedings-new zealand society of animal production. New Zealand Society of Animal Production 60: 301-302.
24. Yeste M (2016) Sperm cryopreservation update: Cryodamage, markers, and factors affecting the sperm freezability in pigs. Theriogenology 85(1): 47-64.
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