آنالیز تحرک اسپرم آواسی در دو محیط در سطوح مختلف دما، pH و اسمولاریته

Subject Areas : Camel

م. آلومار ¹ , م. زرکاوی ² , م.آ. آلزوآبی ³

1 - Department of Agriculture, Atomic Energy Commission, Damascus, Syria
2 - Department of Agriculture, Atomic Energy Commission, Damascus, Syria
3 - Department of Agriculture, Atomic Energy Commission, Damascus, Syria

Received: 2017-11-02 Accepted : 2017-12-31 Published : 2018-09-01

Keywords: دما, قوچ, تحلیل‌گر اسپرم مبتنی بر کامپیوتر, اسمولاریته,

Abstract :

اثرات پنج سطح اسمولاریته و pH همراه با سه دمای مختلف روی پارامترهای تحرک اسپرم قوچ­های آواسی با استفاده از تحلیل­گر اسپرم مبتنی بر کامپیوتر (CASA) در محیط کشت­های تیرود آلبومین لاکتات پیرووات (TALP) و تریس زرده تخم مرغ (YET) مطالعه گردید. اسپرم حاصل از 5 قوچ آواسی بالغ با هم مخلوط گردیده و روی دو محیط با میلی­اسمولاریته 100، 200، 300، 400 و 500 به ازای هر کیلوگرم در دمای 4، 20 و 37 درجه سلسیوس و نیز در سطح pH 5، 6، 7، 8 و 9 در دماهای یکسان رقیق گردیدند. اسمولاریته، pH و دما اثر معنی­داری (05/0>P) روی درصد تحرک (MOT %) و درصد اسپرم با حرکت پیش­رونده (PMOT %) داشتند. غیر از ناهنجاری سر جانبی، همه پارامترهای دیگر تحرک CASA نیز به طور معنی­داری تحت تأثیر قرار گرفتند (05/0>P). اسپرم انکوباسیون شده در YET قادر بود در اسمولاریته­های بین 200 تا 400 میلی­اسمول به ازای هر کیلوگرم دامنه برای MOT % مقاومت کند. شرایط بازی در 9 =pH اثر منفی بزرگتری در مقایسه با شرایط اسیدی در 5 =pH داشت. در مقایسه با 20 و 37 درجه سلسیوس، دمای 4 درجه سلسیوس تأثیر منفی بر MOT % و PMOT % داشته و این امر در محیط TALP با وضوح بیشتری دیده می­شد. به عنوان نتیجه­گیری، نتایج ما نشان داد که: 1) اثرات اسمولاریته، pH و دما بر اسپرم آواسی انکوباسیون شده در هر دو محیط TALP و YET معنی‌دار بود و 2) لازم است که دما برای اسپرمی که در تلقیح مصنوعی و آزمایشات باروری خارج رحمی در این دو محیط مورد استفاده قرار می­گیرند، به دقت انتخاب شوند.

References:

Ak K., Cirit U., Nur Z., Bacinoglu S., Pabuccuoglu S., Ozdas O.B. and Birler S. (2010).Effects of extender osmolarity, cooling rate, dilution rate and glycerol addition time on post-thaw ram semen characteristics and fertilization. J. Fac. Vet. Med. Istanbul Univ. 36, 33-46.

Alomar M., Mahieu J., Verhaeghe B., Defoin L. and Donnay I. (2006). Assessment of sperm quality parameters of six bulls showing different abilities to promote embryo development. Reprod. Fertil. Dev. 18, 395-402.

Amann R.P. and Waberski D. (2014). Computer-assisted sperm analysis (CASA): Capabilities and potential developments. Theriogenology. 81, 5-17.

Bag S., Joshi A., Rawat P.S. and Mittal J.P. (2002a).Effect of initial freezing temperature on semen characteristics of frozen-thawed ram spermatozoa in a semi-arid tropical environment. Small Rumin. Res. 43, 23-29.

Bag S., Joshi A., Naqvi S.M.K., Rawat P.S. and Mittal J.P. (2002b). Effect of freezing temperature, at which straws were plunged into liquid nitrogen, on the post-thaw motility and acrosomal status of ram spermatozoa. Anim. Reprod. Sci. 72, 175-183.

Carr D.W. and Acott T.S. (1989). Intracellular pH regulates bovine sperm motility and protein phosphorylation. Biol. Reprod. 41, 907-920. 

Comhaire F.H., Huysse S., Hinting A., Vermeulen L. and Schoonjans F. (1992). Objective semen analysis: Has the target been reached? Hum. Reprod. 7, 237-241.

Contri A., Gloria A., Robbe D., Valorz, C., Wegher L. and Carluccio A. (2013). Kinematic study on the effect of pH on bull sperm function. Anim. Reprod. Sci. 136, 252-259.

Curry M.R., Millar J.D. and Watson P.F. (1994).Calculated optimal cooling rates for ram and human sperm cryopreservation fail to conform with empirical observations. Biol. Reprod. 51, 1014-1021.

Drobnis E.Z., Crowe L.M., Berger T., Anchordoguy T.J., Overstreet J.W. and Crowe J.H. (1993).Cold shock damage is due to lipid phase-transitions in cell-membranes a demonstration using sperm as a model. J. Exp. Zool. 265, 432-437.

Eppleston J., Salamon S., Moore N.W. and Evans G. (1994).The depth of cervical insemination and site of intrauterine insemination and their relationship to the fertility of frozen-thawed ram semen. Anim. Reprod. Sci. 36, 211-225.

Farrell P.B., Foote R.H., Mcardle M.M., Trouern-Trend V.L. and Tardif A.L. (1996).Media and dilution procedures tested to minimize handling effects on human, rabbit and bull sperm for computer-assisted sperm analysis (CASA). J. Androl. 17, 293-300.

Farrell P.B., Presicce G.A., Brockett C.C. and Foote R.H. (1998). Quantification of bull sperm characteristics measured by computer-assisted sperm analysis (CASA) and the relationship to fertility. Theriogenology. 49, 871-879.

Galal S., Gursoy O. and Shaat I. (2008). Awassi sheep as a genetic resource and efforts for their genetic improvement: A review. Small Rumin. Res. 79, 99-108.

Herrara C., Brogliatti G., Cavia R., Conde P., Revora M. and Pasqualini R.S. (2005). CASA sperm parameters and their relation with in vitro fertilization. Pp. 411-415 in Proc. 15^th Int. Congr. Anim. Reprod., Porto Seguro, Brazil.

Holt W.V. (2000). Basic aspects of frozen storage of semen. Anim. Reprod. Sci. 62, 3-22.

Holt W.V. and Palomo M.J. (1996).Optimization of a continuous real-time computerized semen analysis for ram sperm motility assessment, and evaluation of four methods of semen preparation. Reprod. Fertil. Dev. 8, 219-230.

Joshi A., Mathur A.K., Naqvi S.M.K. and Mittal J.P. (2006).Influence of osmolality of complete semen extender on motion characteristics of frozen-thawed ram spermatozoa. Asian-Australasian J. Anim. Sci. 19, 716-1721.

Langlais J. and Roberts K.D. (1985).A molecular membrane model of sperm capacitation and the acrosome reaction of mammalian spermatozoa. Gamete Res. 12, 183-224.

Makler A. and Jakobi P. (1981). Factors affecting sperm motility. V. Washing and resuspension of human spermatozoa in various artificial media. Fertil. Steril. 35, 442-446.

Makler A., David R., Blumenfeld Z. and Better O.S. (1981). Factors affecting sperm motility. VII sperm viability as affected by changes of pH and osmolarity of semen and urine specimens. Fertil. Steril. 36, 507-511.

Mortimer S.T. (1997). A critical review of the physiological importance and analysis of sperm movement in mammals. Hum. Reprod. Update. 3, 403-439.

Mortimer S.T. (2000). CASA-Practical aspects. J. Androl. 21, 515-524.

Mortimer D. and Mortimer S.T. (2013). Computer-aided sperm analysis (CASA) of sperm motility and hyperactivation. Methods Mol. Biol. 927, 77-87.

Nur Z., Zik B., Ustuner B., Sagirkaya H. and Ozguden C.G. (2010). Effects of different cryoprotective agents on ram sperm morphology and DNA integrity. Theriogenology. 73, 1267-1275.

Rijsselaere T., Mase D., Hoflack G., de Kruif A. and Van Soom A. (2007). Effect of body weight, age and breeding history on canine sperm quality parameters measured by the Hamilton-Thorne Analyser. Reprod. Domest. Anim. 42, 143-148.

Robayo I., Montenegro V., Valdes C. and Cox J.F. (2008). CASA assessment of kinematic parameters of ram spermatozoa and their relationship to migration efficiency in ruminant cervical mucus. Reprod. Domest. Anim. 43, 393-399. 

Salamon S. and Maxwell W.M.C. (1995a). Frozen storage of ram semen. I. Processing, freezing, thawing and fertility after cervical insemination (Review). Anim. Reprod. Sci. 37, 185-249.

Salamon S. and Maxwell W.M.C. (1995b). Frozen storage of ram semen. II. Causes of low fertility after cervical insemination and methods of improvement. Anim. Reprod. Sci. 38, 1-36.

Sleiman F.T. and AbiSaab S. (1995). Influence of environment on respiration, heart rate and body temperature of filial crosses compared to local Awassi sheep. Small Rumin. Res. 16, 49-53.

Vantman D., Bands S.M., Koukoulis G., Dennison L. and Sherins R.J. (1989).Assessment of sperm motion characteristics from fertile and infertile men using a fully automated computer-assisted semen analyzer. Fertil. Steril. 51, 156-161.   

Varisli O., Uguz C., Agca C. and Agca Y. (2009). Motility and acrosomal integrity comparisons between electro-ejaculated and epididymal ram sperm after exposure to a range of anisosmotic solutions, cryoprotective agents and low temperatures. Anim. Reprod. Sci. 110, 256-268.

Visconti P.E., Westbrook V.A., Chertihin O., Demarco I., Sleight S. and Diekman A.B. (2002).Novel signaling pathways involved in sperm acquisition of fertilizing capacity. J. Reprod. Immunol. 53, 133-150.

Watson P.F. (1975). The interaction of egg yolk and ram spermatozoa studied with a fluorescent probe. J. Reprod. Fertil. 42, 105-111.