(PhD) Musabekov A.T.

The South-Kazakhstan state university him M.Auezov

Change decrease membranes structures spermatozoa

a male pig at freezing

Summary

Established that in the process of freezing and thawing in spermatozoa undergoing considerable destructive changes in membrane structures, resulting in rapid hydrolysis of ATP under the action contained in the cells and seminal plasma enzymes. These changes can to some extent, reduce the dilution method using dialysis sperm before freezing.

It is known, that in an estimation quality of sperm of agricultural animals the important criterion is synthesis macro ergs. Level АТF and a functional condition of power system as it was shown, in a number works have high communication with mobility and impregnating ability spermatozoa. The high level of oxidizing processes and the big sensitivity of mechanisms of interface of oxidation with syntheses АТF, low ability adenilatsiklaz systems in sperm of male pigs are the reasons of fast infringement of an oxidizing exchange and disbalans in work of fermentable systems, infringements of permeability and transport of ions in membranes at dilution, fast cooling and freezing of sperm.

Doubtless practical interest represents to communications by it and perfection of methods cry preservation sperms of male pigs studying of dynamics of maintenance АТF in spermatozoa under influence of deep freezing depending on methods cry preservation. The purpose of our experiments was studying maintenance АТF in natives and subjected to freezing by various ways spermatozoa male pigs.

Experiments are carried out on sperm of male pigs of large white breed in pig-breeding facilities "Dostik" Sairams area of the South-Kazakhstan area. In connection with absence АТF in seed plasma, the preliminary branch spermatozoa from plasma was not carried out. Extraction АТF from spermatozoa it was carried out on method Ugarovs with authors (1991). For this purpose from each sample of sperm selected test in volume of 0,05 ml and added in it of 0,45 ml dimethilsulfhydryls. After 20 minute extraction at room temperature defined maintenance АТF a bioluminescent method with the help portable luminometers ЕМILIТЕ-1003А (Russia).

Results of experiment with studying maintenance АТF in spermatozoa male pigs are submitted in table 1.

From table 1 it is visible, that during freezing - thawing there is a sharp decrease of level АТF in spermatozoa male pigs. In 5 minutes after thawing level АТФ has decreased on 38 % in comparison with native samples. Thus on 1 % mobility spermatozoa also has decreased and the number spermatozoa with damaged acrosome has increased on 40 %.

Table 1.

Influence of deep freezing on maintenance АТF in spermatozoa a male pig

Time of research	Mobility spermatozoa, %	Number spermatozoa with intact acrosome,	Maintenance АТF in spermatozoa, nmole/108кл
Before freezing	83,4±5,0	93,9±4,2	28,1±0,7
Right after thawing	37,1±1,9	53,2±2,6	16,3±1,1
Through 1 c after thawing	21,8±0,9	45,3±2,7	6,2±0,6
Through 2 c after thawing	18,7±0,7	37,6±1,7	4,9±0,3

The further incubation оттаянных samples at 37°С promoted significant decrease of mobility and maintenance АТF in spermatozoa. In 2 hours after thawing maintenance АТF in spermatozoa has decreased in 5,7 times in comparison with an initial level (Р < 0,01). Hence, during freezing - thawing in spermatozoa male pigs occur significant destructive changes membranes structures therefore there is fast hydrolysis АТF under action contained in cells and seed plasma of enzymes. Studying of influence of various ways dilution sperms of male pigs before freezing on level АТФ in spermatozoa is submitted in table 2. Table 2. Influence of a way dilution sperms before freezing on maintenance АТF in spermatozoa

Time of research АТF	Usual dilution		Dialysis processing
	Mobility spermatozoa, %	Maintenance АТФ, nmole/108кл	Mobility spermatozoa, %	Maintenance АТF, nmole/108кл
Before freezing	78,8±4,3	28,2±2,4	80,1±6,8	32,8±2,1
Right after thawing	31,0±1,2	15,8±1,7	34,1±2,9	19,8±1,2
Through 1 c after thawing	18,6±0,6	5,9±0,3	22,1±1,7	7,7±0,6
Through 2 c after thawing	14,6±1,3	4,7±0,8	22,0±1,8	6,6±0,7

From table 2 it is visible, that the way dilution sperms before freezing renders the certain influence on maintenance АТF in spermatozoa after freezing - thawing and on their mobility. The best results are received at dilution dialysis by a way. Thus in 2 hours equilibration sperms in group with dialysis processing higher deduction АТФ was revealed in comparison with usual dilution: 32,8 nmole/108 kl. Against 28,2 nmole/108 kl. Accordingly. Thus in dialysis to group maintenance АТФ in spermatozoa at once, 1 and 2 hours after thawing has decreased on 60,3%, 4,2 and 4,9 times and against 78,4%, 4,7 and 6,0 times at usual dilution sperms. Apparently, also in dialysis to group less sharp decrease of mobility thawed spermatozoa also is marked. So after 2 ч thawing at usual dilution mobility spermatozoa has made 14,6%, and in dialysis to group of 22,0 % or on 7,4% is higher. These data testify that dialysis the method разбавления sperms of male pigs promotes more effective protection spermatozoa during them cryopreservation. Probably, it speaks the best conditions of stabilization membranes structures spermatozoa in process equilibration. Besides thus from structure of sperm the low-molecular components rendering biodestructive influence on membranes сперматозоидов can leave.

Thus, the carried out laboratory and industrial experiments testify, that during freezing-thawing in spermatozoa male pigs occur significant destructive changes membranes structures therefore there is fast hydrolysis АТF under action contained in cells and seed plasma of enzymes. These changes can be lowered in the certain degree applicable dialysis a method dilution sperms before freezing which renders cryoprotectixe action on spermatozoa.