Fertility-associated proteins in male and female reproductive fluids of cattle (original) (raw)
Related papers
2006
We evaluated the expression of proteins in the accessory sex gland fluid (AGF) and their relationships with fertility indexes of dairy bulls. Fertility was normalized as the percentage point deviation of their nonreturn rates (PD) from the average fertility of all bulls from a given artificial insemination center. Services associated with each sire ranged from 269 to 77 321 and PD values from ϩ7.7% to Ϫ18.1%. AGF, from 37 bulls, was obtained with an artificial vagina after cannulation of the vasa deferentia. Proteins from AGF were separated by 2-dimensional SDS-PAGE followed by staining with Coomassie blue and analysis of polypeptide maps using PDQuest software. Bulls were divided in groups based on PD values and the optical density of spots in the AGF gels used as independent variables to predict bull fertility. Proteins were identified by capillary liquid chromatography nanoelectrospray ionization tandem mass spectrometry (CapLC-MS/MS). An average of 52 Ϯ 5 spots was detected in the AGF gels, but there were no spots unique to groups of either high-(PD Ն 0) or low-(PD Ͻ 0) fertility sires. The former were neither less nor more homogeneous than the latter based on correlations of all matched spots between pairs of AGF maps. However, high fertility of dairy bulls was significantly associated with lower expression of 14-kDa spermadhesin Z13 isoforms and higher amounts of 55-kDa osteopontin and 58-kDa phospholipase A 2 (PLA 2 ) isoforms. The average intensity of 5 spots identified as BSP 30 kDa in the AGF gels had a quadratic association with fertility indexes (R 2 ϭ .18; P ϭ .03). PD values of bulls were related (R 2 ϭ .56) to the quantity of spermadhesin, osteopontin, and BSP 30 kDa in the AGF polypeptide maps. Bull fertility was also determined by another equation (R 2 ϭ .53) with spermadhesin, BSP 30 kDa, and PLA 2 as independent variables. We conclude that interactions among several proteins in accessory sex gland fluid explain a significant proportion of the variation in fertility scores of mature dairy sires.
Proteins of the Cauda Epididymal Fluid Associated With Fertility of Mature Dairy Bulls
Journal of Andrology, 2006
We evaluated the relationships between proteins in cauda epididymis fluid (CEF) and fertility scores of dairy bulls. Fertility was expressed as the percentage point deviation (PD) of bull nonreturn rate from the average fertility of all bulls at an artificial insemination center. The number of services for each bull ranged from 1074 to 52 820, and PD values ranged from +7.7% to 26.6%. CEF from 20 bulls was obtained from vasa deferentia cannulae and was separated from sperm by centrifugation immediately after collection. Samples were evaluated by 2-dimensional (2-D) sodium dodecyl sulfate polyacrylamide gel electrophoresis gels stained with Coomassie blue, and polypeptide maps were analyzed by PDQuest software. Protein quantities, defined as the total integrated optical density of the spots, were compared between groups of high-fertility sires (n 5 12; PD $ 0) and low-fertility sires (n 5 8; PD , 0) and were also used as independent variables in regression analysis. Proteins were identified by capillary liquid chromatography-nanoe-lectrospray ionization-tandem mass spectrometry. An average of 118 spots was detected in 2-D maps of the CEF, but we were unable to distinguish any protein that was expressed only in high-fertility or in low-fertility bulls. However, the amount of a-L-fucosidase 2 and cathepsin D was 2.3-and 2.4-fold greater (P , .05) in high-fertility than in low-fertility bulls, respectively. Conversely, the intensities of 3 isoforms (24-27 kd; pI 6.3-5.8) of prostaglandin D-synthase (PGDS) were from 3.2-to 2.2-fold greater in low-fertility sires (P , .05). An empirical regression model established that a significant proportion (R 2 5 0.72; P , .0001) of the variation in fertility scores (PD values) was explained by the intensities of cathepsin D and 1 isoform of PGDS (24 kd; pI 6.3). Thus, multiple proteins present in the CEF are potential biomarkers of fertility in high-use, mature Holstein bulls.
Fertility-associated proteins in Holstein bull seminal plasma
Biology of Reproduction, 1993
This study was undertaken to determine whether bovine seminal plasma contained protein markers associated with bull fertility, and whether these markers were of value in predicting bull fertility. Seminal plasma was obtained from 35 Holstein bulls of known fertility. Two-dimensional PAGE of seminal plasma samples indicated that two proteins (26 kDa, pi 6.2; 55 kDa, pI 4.5) predominated in higher-fertility bulls, and two proteins (16 kDa, pi 4.1; 16 kDa, pi 6.7) predominated in lower-fertility bulls. Densitometry data for these proteins in individual samples were combined for bulls grouped by fertility level. Average density of the 26-kDa protein was significantly greater n seminal plasma of high-fertility bulls, and high-fertility seminal plasma also contained more of the 55-kDa protein than that of average-and below average-fertility bulls. Below average-and low-fertility bull seminal plasma had significantly more of both 16-kDa proteins than that of average-and high-fertility bulls. A regression model was developed to predict bull fertility using the four fertility-associated protein densities. A plot of actual bull fertility versus that calculated by this model was linear and positively correlated (r = 0.89). These findings indicate that bull seminal plasma contains fertility-associated proteins that are predictive of bull fertility.
Evaluation of bull semen for fertility-associated protein, in vitro characters and fertility
Proteins present in the seminal plasma and sperm influence sperm function and fertilization. The present study was carried out to screen breeding bull semen samples for the presence of fertilityassociated 28–30 kDa heparin-binding protein (HPB) and its effect on in vitro sperm characters and fertility. Semen samples were collected from 22 breeding bulls and the sperm proteins were extracted by Triton X detergent extraction method. HBPs were eluted, and the molecular weight of the proteins was assessed by discontinuous sodium dodecyl sulphate polyacrylamide gel elecrophoresis. Based on the presence/absence 28 kDa HBPs, bulls were categorized into group I and group II. Frozen semen samples were evaluated for in vitro sperm characters at immediate post thaw, 60, 120 and 180 min post-thaw incubation. To assess the field fertility of the bulls, 50 frozen semen straws/bull were used for insemination. Results indicated that only 50% of the bulls screened had 28–30 kDa HBPs in their sperm. Bulls positive for fertility-associated protein had better in vitro sperm characters, better protection against oxidative stress, readily underwent capacitation induction by heparin and had 13% higher conception than the bulls lacking the protein. So, it can be concluded that the bulls positive for of 28–30 kDa HBPs in sperm had higher chance of fertility and screening for its presence can be included in the regular breeding soundness examination for selection of bulls.
Fertility-associated proteins in Nelore bull sperm membranes
Animal Reproduction Science, 2006
The present study was undertaken to evaluate the protein composition of the sperm membranes (SM) of Nelore bulls, assessing protein markers associated with bull fertility, and whether these markers can be used for predicting bull fertility. Samples were obtained of 20 Nelore bulls, with fertility ranked and divided into three groups (greater, normal and least). To rank the bull's fertility weighted classification was used (according to the number of pregnant cows, number of AI cows and number of herds, considering three different breeding seasons), using the PROC GENMOD as a statistical model, with 99% significance. A total of 7897 Nelore cows, randomly distributed among 28 different farms, were considered in the statistical analyses. The bulls were divided into three fertility groups (pregnancy rates): greater (%F > 80), normal (79 < %F > 71) and least (<68%F) with 3, 13 and 4 bulls, respectively. Two-dimensional gel electrophoresis (2DE) of sperm membranes indicated in 27 spots (SM40,
Bull sperm and seminal plasma proteins and their relationship with fertility: a review
Online Journal of Animal and Feed Research
The efficiency of artificial insemination (AI) is greatly influenced by the quality of semen. Spermatozoa and seminal plasma are found in semen, which play a role in the reproductive process and its ability to fertilize an egg and maintain the development of an embryo. Various factors will determine the fertility capacity of a sperm, both from the intrinsic factors of the sperm and the plasma component of the semen. Seminal plasma proteins are crucial for maintaining the stability of the membrane, viability, motility of spermatozoa, acrosome reactions, maintaining osmotic pressure and helping the fertilization process. Good quality semen will support the fertilization process. The purpose of this scoping review is to increase our understanding of protein from sperm and seminal plasma of bulls and their relationship with fertility. The sperm proteins that were significantly correlated with fertility were Outer Dense Fiber protein 2 (ODF2), Protamine (PRM), Testis specific histine 2B ...
A comprehensive proteomic analysis of the accessory sex gland fluid from mature Holstein bulls
Animal Reproduction Science, 2007
The expression of proteins in accessory sex gland fluid (AGF) of proven, high use mature Holstein bulls was evaluated. Thirty-seven bulls with documented fertility based on their non-return rates were studied. AGF was obtained by artificial vagina after bulls were surgically equipped with cannulae in the vasa deferentia. Samples of AGF were evaluated by two-dimensional SDS-PAGE, gels stained with Coomassie blue and polypeptide maps analyzed by PDQuest software. A master gel generated by the software representing the best pattern of spots in the AGF polypeptide maps was used as a reference for protein identification. Proteins were identified by Western blots and capillary liquid chromatography-nanoelectrospray ionization tandem-mass spectrometry (CapLC-MS/MS). The product ion spectra were processed using Protein Lynx Global Server 2.1 prior to database search with both PLGS and MASCOT (Matrix Science) software. The entire NCBI database was considered for mass fingerprint matching. An average of 52 ± 5 spots was detected in the AGF 2D gels, which corresponded to proteins potentially involved in capacitation (bovine seminal plasma protein-BSP-A1/A2 and A3, BSP 30 kDa, albumin); sperm membrane protection, prevention of oxidative stress, complement-mediated sperm destruction and anti-microbial activity (albumin, clusterin, acidic seminal fluid protein-aSFP, 5 -nucleotidase-5 -NT, phospholipase A 2 -PLA 2 ); acrosome reaction and sperm-oocyte interaction (PLA 2 , osteopontin); interaction with the extracellular matrix (tissue inhibitor of metalloproteinase 2, clusterin) and sperm motility (aSFP, spermadhesin Z13, 5 -NT). The 20 spots distinguished in all gels were matched to proteins associated with these functions. Proteins identified by tandem mass spectrometry as ecto-ADP-ribosyltransferase 5 and nucleobindin, never described before in the accessory sex gland secretions, were also detected. In summary, we identified a diverse range of components in the * Corresponding author. Tel.: +1 814 863 7434; fax: +1 814 863 0833.
Stable bull fertility protein markers in seminal plasma
Journal of Proteomics, 2021
Bull fertility is an important trait in breeding as the semen of one bull can, potentially, be used to perform thousands of inseminations. The high number of inseminations needed to obtain reliable measures from Non-Return Rates to oestrus creates difficulties in assessing fertility accurately. Improving molecular knowledge of seminal properties may provide ways to facilitate selection of bulls with good semen quality. In this study, liquid chromatography mass spectrometry (LC-MS/MS) was used to analyze the protein content from the seminal plasma of 20 bulls with Non-Return Rates between 35 and 60%, sampled across three seasons. Overall, 1343 proteins were identified and proteins with consistent correlation to fertility across multiple seasons found. From these, nine protein groups had a significant Pearson correlation (p < 0.1) with fertility in all three seasons and 34 protein groups had a similar correlation in at least two seasons. Among notable proteins showing a high and consistent correlation across seasons were Osteopontin, a lipase (LIPA) and N-acetylglucosamine-1phosphotransferase subunit gamma. Three proteins were combined in a multiple linear regression to predict fertility (r = 0.81). These sets of proteins represent potential markers, which could be used by the breeding industry to phenotype bull fertility. Significance: The ability of bull spermatozoa to fertilize oocytes is crucial for breeding efficiency. However, the reliability of this trait from field measures is relatively low and the prediction of fertility given by conventional methods to evaluate sperm quality is currently not very accurate. In this work, we identify sets of proteins in bull seminal plasma from repeated samples collected at different times of the year that correlate to fertility in a consistent way. We combined these individual proteins to build a molecular signature predictive of fertility. This study provides an overview of proteins linked to fertility in seminal plasma, thereby increasing knowledge of the bull seminal plasma proteome. Protein signatures from the latter, potentially related to fertility, may be of use to predict fertility for individual bulls.