Bull sperm and seminal plasma proteins and their relationship with fertility: a review (original) (raw)
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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.
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.
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.
Protein profile and functionality of spermatozoa from two semen collection methods in Bali bulls
The aim of this study was to evaluate the effect of two semen collection methods (electroejaculation [EE] and transrectal massage [RM]) on in vitro sperm functionality and protein composition of seminal plasma in Bali bulls. Ten untrained Bali bulls were selected for semen collection by EE and RM. Parameters analysed were acrosome and plasma membrane integrity, sperm motility (by CASA), normal morphology, functionality (sperm penetration assay), acrosome reaction, total protein content and protein profiles (by 2DPAGE). Bulls collected by RM had a higher (po0.05) percentage of spermatozoa with intact acrosome and plasma membrane, functionality and individual motility, and a lower proportion of seminal plasma, total protein content and lower ratio of low molecular weight proteins than those collected by EE. Analysis of 2D-PAGE gel detected about 116 spots in the range of 10–250 kDa and isoelectric points (pI) ranging from 3 to 10. Approximately 52% of seminal plasma protein spots were represented by four major protein fractions with molecular weights around 37–45 kDa (15.66%), 25–30 kDa (12.46%), 14–16 kDa (11.73%) and 12–15 kDa (11.52%). Ten of the seminal plasma proteins identified by mass spectrometry belonged to major bovine seminal plasma proteins. A very significant finding in this study was related to the two proteins identified, PGK and PLA2, with MW of approximately 37–40 and 50–55 kDa and pI of 8.5–8.8 and 5.2–6.0, respectively. These two protein spots can only be detected in the seminal plasma of ejaculates obtained through RM. In conclusion, semen quality as examined by in vitro sperm functionality was found to be better in RM than EE samples after treatment with heparin and calcium ionophore A-23187. In addition some low molecular weight proteins were up-regulated in the seminal plasma obtained from the EE method
Veterinary World, 2020
Background and Aim: Holstein cows and heifers are widely bred in Indonesia by artificial insemination (AI) to increase population and milk production. Sperm fertility is modulated by genetic factors, but the analysis of sperm quality is still based on macro- and microscopic characteristics. This study aimed to analyze both sperm quality and proteins of Holstein bulls at different fertility levels. Materials and Methods: The frozen semen samples were collected from the Indonesia National AI Center. They were classified based on the reproductive efficiency data and were grouped into high fertile (HF) and low fertile (LF). Sperm qualities were evaluated by microscopic evaluation. The Holstein sperm proteins were extracted using phenylmethanesulfonyl fluoride as a protease inhibitor and the benzidine detergent extraction method. Discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was conducted to analyze the molecular weights (MWs) of the sperm proteins. The data obtained were analyzed by a t-test using the one-factor bull fertility level, and Spearman's correlation analysis was used to identify the correlation between the sperm microscopic evaluation parameters and protein bands. Results: The sperm motility post-freeze thawing was not significantly different between the HF and LF (p>0.05). The HF level had a higher percentage of viability, intact plasma membrane integrity, and intact acrosomes than the LF (p<0.05). Five protein bands were found in the SDS-PAGE of sperm proteins of Holstein bulls with different concentrations. Sperm proteins with MWs of 17.51 kDa, 14.87 kDa, 33.71 kDa, and 41.97 kDa were abundant in the Holstein bulls with an HF level, while 55 kDa proteins were abundant in the LF level of Holstein bulls. The sperm of Holstein bulls in the HF level contained proteins of about 33.71 kDa that were not detected in the LF. Conclusion: The sperm protein with a molecular weight of 33.71 kDa was predicted to be a specific protein biomarker that influences bull fertility. Sperm fertilization abilities were also determined by the sperm proteins, the morphology of sperm acrosomes, and the quality of plasma membranes. This method can be used to select bulls with high fertility to increase the population of Holstein bulls.
Protamine and other proteins in sperm and seminal plasma as molecular markers of bull fertility
Veterinary World, 2020
Fertility is the most important aspect in the efforts to increase livestock populations. Protamine and various proteins in sperm and seminal plasma are the results of the molecular analysis which can be used as a marker of fertility. Each of the proteins plays an important role in the normal function of sperm, starting from the formation of sperm structure, motility, capacitation, cell protection, acrosome reactions, successful fertilization, egg activation, and embryonic development. Finally, these molecular components can be a marker of fertility and can help to diagnose the cases of infertility/subfertility in livestock in the field.
Protein profile of seminal plasma of Murrah buffalo bulls
Journal of biotechnology & biomaterials, 2015
A rtificial insemination in cattle and buffalo is the common practice and to keep the success at high, the quality of the semen must be good. Seminal plasma is a complex secretion composed by fluids from accessory sex glands, epididymis and testicles and its molecular composition is capable of modulating sperm function. Proteins are the most abundant organic compounds in seminal plasma and play a crucial role in processes related to fertilizing capacity of sperm and can be considered as potential molecular markers of fertility. This study was carried out to assess the protein profile of the Murrah buffalo seminal plasma by using SDS-PAGE and to correlate them with the semen characteristics. Semen samples were collected by a bovine artificial vagina from 8 Murrah buffalo bulls maintained at
Spermatozoa Molecules in Relation to Bulls Fertility
Iranian Journal of Applied Animal Science, 2017
Bull fertility may be defined as the process by which spermatozoa fertilize and activate the ovum and then support embryonic development. Bull fertility is a complex trait having relatively low heritability and plays a vital role for efficient production and reproduction of bovine. Various mechanisms involved in regulating bull fertility associated phenotype and reliable biomarkers are poorly defined. Primary spermatozoa physiology indicators namely sperm molecules and epigenetic factors may play a vital role in predicting sperm physiology and fertility. Spermatozoa and their fingerprints or patterns can play a pivotal role in prediction of fertility in bovine. No reliable tests exist despite genomic selection for evaluating quality of semen and fertility of bull. This review focuses on various molecules related to bull fertility viz., sperm RNAs, seminal plasma proteins, sperm proteins and sperm epigenome. In combination with single-nucleotide polymorphism (SNP), microarrays or spe...
Veterinary World, 2021
Background and Aim: Protamine (PRM) is the major protein in the sperm nucleus and plays an essential role in its normal function. Moreover, PRM has great potential as a protein marker of semen production and quality. This study aimed to assess the potential of sperm bovine PRM as a protein marker of semen production and quality in bulls at the National Artificial Insemination (AI) Center of Indonesia. Materials and Methods: The semen production capacity of each bull was collected from frozen semen production data at the Singosari AI Center for 6 months, and was then divided into two groups (high and low). A total of 440 frozen semen straws from six Limousin (LIM), six Friesian Holstein (FH), six Peranakan Ongole (PO), and four Aceh bulls aged 4-5 years were used in the study. The frozen semen was used to measure the concentration of PRM1, PRM2, and PRM3 using the enzyme immunoassay method. The frozen semen was also used to assess the quality of the semen, including progressive motility (PM) through computer-assisted semen analysis, sperm viability through eosin–nigrosin analysis, and the DNA fragmentation index through Acridine Orange staining. Results: PRM1 was significantly higher in all bull breeds included in the study (p<0.00), followed by PRM2 (p<0.00) and PRM3 (p<0.00). PRM1 significantly affected semen production in LIM, FH, PO, and Aceh bulls (p<0.05). Moreover, PRM2 significantly affected semen production only in FH and Aceh bulls (p<0.05), whereas PRM3 affected this parameter in PO and Aceh bulls exclusively (p<0.05). Consistently and significantly, PRM1 was positively correlated with the PM and viability of sperm and negatively associated with its DNA fragmentation in LIM, FH, PO, and Aceh bulls (p<0.05; p<0.01). The correlation analysis between PRM2 and PRM3 and semen quality parameters varied across all bull breeds; some were positively and negatively correlated (p<0.05; p<0.01), and some were not correlated at all. Conclusion: PRM1 has excellent potential as a protein marker of semen production and quality in bulls at the National AI Center of Indonesia.
Biomarkers of in vivo fertility in sperm and seminal plasma of fertile stallions
Theriogenology, 2010
The global proteome of sperm and seminal plasma of fertile stallions was investigated to determine whether associations with relative in vivo fertility exist. Seven stallions at stud in a commercial breeding station were collected throughout the breeding season and bred to a total of 164 mares to determine conception rates. On three occasions during the breeding season, raw semen was obtained from a regular collection for proteomic analysis using two-dimensional electrophoresis and also assessed for routine semen quality end points. First cycle conception rate was negatively related to ejaculate volume (r ϭ Ϫ0.43, P ϭ 0.05) and total IGF1 content (ng) per ejaculate (r ϭ Ϫ0.58, P ϭ 0.006), whereas overall pregnancy rate was positively related to sperm concentration (r ϭ 0.56, P ϭ 0.01). The abundance of three proteins known to be involved in carbohydrate metabolism in sperm was positively related to fertility. Furthermore, the abundance of four seminal plasma proteins were identified as being negatively related to fertility; these were identified as kallikrein-1E2 (KLK2), clusterin, and seminal plasma proteins 1 (SP1) and 2 (SP2). Abundance of cysteine-rich secretory protein 3 (CRISP3) was positively related to first cycle conception rate (r ϭ 0.495, P ϭ 0.027) and may provide a good marker of fertility. Based on stepwise regression analysis, clusterin and SP1 in seminal plasma together with sperm citrate synthase were predictive of fertility (r ϭ 0.77, P Ͻ 0.0001). This study identified proteins within sperm and seminal plasma that could serve as biomarkers of semen quality and fertility in stallions. Theriogenology 74 (2010) 956 -967 www.theriojournal.com 0093-691X/$ -see front matter