Expressão de proteínas do plasma seminal em carneiros das raças santa ines e morada nova: associaçãoes com parâmetros seminais , influencia de fontes de proteínas e de nitrogênio não proteico nas dietas.

Abstract

The Brazilian Northeast has about almost half of Brazilian sheep livestock, estimated at twenty million animals. The races created here, Santa Ines and Morada Nova stand out because they have good physical development, weight gain and good adaptation to the tropical climate. Recently showed the protein profile of mature rams through the techniques of two-dimensional electrophoresis associated with mass spectrometry, which several proteins have been identified, including those RSVPs 14 and 22 kDa, belonging to the family of BSPs and bodesinas - 1 and 2, that are part of the family of espermadesinas as well as the changes suffered by the protein profile of seminal plasma of animals during reproductive development, while changes in the parameters of sperm concentration and motility, although the association between the protein expression of these proteins with reproductive parameters testicular and measures have not yet been made in northeastern Brazil. Semen samples were collected from twenty-one adult sheep Santa Ines and eleven Morada Nova with normal reproductive and mean age of two years and seminal plasma was obtained by centrifugation and subjected to two-dimensional electrophoresis, and the spots that differed significantly between the groups of animals from high (G1) and low motility (G2) were digested with trypsin and subjected to mass spectrometry and research database for identification. The gels were stained with colloidal Cromassie, scanned and analyzed with PDQuest application and quantity of the spots quantitatively estimated. Evaluated, then, statistical associations between these variables and the reproductive parameters scrotal circumference (EC), percentage of motile cells (PEM) and total sperm defects (MDD) of the animals Santa Inês and scrotal circumference (EC) motility individual rogressive (MIP), percentage of motile cells (PEM), Sperm concentration (CONC) and Massal motility (MM) for the animails Morada Nova. Were detected on average 236 ± 7.65 spots per gel, according to the pairing generated by PDQuest application to the Santa Ines and 261 ± 15.09 spots per gel in the seminal plasma of animals Morada Nova, ranging between 182 and 375 spots gel. For Santa Inês a total of 63 spots were identified consistently in all gels, which represented 41.5% of all spots detected intensity. For Morada Nova animails, a total of 98 spots were detected consistently in all gels. The intensity of these spots amounted to 60.82% of the intensity of all spots shown on the master gel was observed a great expression of proteins of low molecular weight (14.5 to 18.4 kDa) and pI ranging from 4.2 to 5, 9. to Santa Ines. The average total of 236 spots found thirteen spots showed significant difference (p <0.05) between animals of high and low motility, being more intense ten spots in animals with high motility and three in animals with low motility. After digestion of spots and identification by mass spectrometry these spots were identified as proteins Arylsulfatase A and zinc alpha 2 glycoprotein more intense in G1and RSVP-22 and Bodesina with higher expression in G2. For the Morada Nova race, four spots showed significant difference (p <0.05) for the parameter percentage of motile cells (PEM). After digestion of spots and identification by MALDI-TOF-TOF, we identified as proteins, RSVP-14, RSVP-22, a protein α-1 of t-complex and aldose reductase, which are all more intense in animals with high motility (G1). In a second study, evaluated the influence of different sources of dietary protein (soybean meal, leucaena leaf hay and cottonseed meal) and non-protein nitrogen (urea) on body weight, testicular development and sperm quality of the animals studied and protein expression in the seminal plasma of the animals potentially related to the diets. We used 20 Morada Nova lambs aged between 24 and 39 weeks. For the testicular development criteria (cirrcunferência scrotal, testicular thickness, width and length testicular testicular) no significant difference (p <0.05) was observed at the end of experiment. Animals fed cottonseed meal had a significant increase in body weight (p <0.05) compared with Leucaena hay. In the same period, the body weight of animals fed leucaena hay, soybeans and urea was similar. Differences between treatments (p <0.05) were observed only for individual progressive motility and total defects. The semen of animals fed diets containing leucaena showed low motility individual when compared with those fed diets containing soybean meal. However, animals fed diets containing cottonseed meal and urea had similar values for progressive motility. When the total defects were analyzed, there was a significant difference between groups of leucaena hay and urea. Detected an amount of 246.6 ± 13.9, 236.0 ± 12.4, 261.2 ± 20.2 and 243.8 ± 20.5 spots per gel in groups of soybean meal, leucaena leaf hay, cottonseed meal and urea, respectively. No significant differences in the number of spots per gel, in both groups. Significant differences (p <0.05) were observed in the intensities of twelve spots in the proteic maps . An increased expression of spot S8707 (62.8 kDa, pI 6.9) in animals fed with hay and leaves of leucaena compared to those fed soybean meal, and a negative correlation with individual motility (MPI .) These observations indicate that factors present in hay sheets leucaena promote an increase in protein expression (S8707), the likely cause deleterious changes in the mechanism controlling the motility of sperm cells, as observed in this group lowest for MPI.