Osteopontin (OPN) is a protein found at higher concentrations in the seminal plasma of bulls with above average fertility. Polymorphisms have been reported within the OPN gene promoter that can affect production of th...Osteopontin (OPN) is a protein found at higher concentrations in the seminal plasma of bulls with above average fertility. Polymorphisms have been reported within the OPN gene promoter that can affect production of this protein and thus, affect fertility. Therefore, Angus (n = 5) and Angus x Gelbvieh (Balancer, n = 14) and Angus x Brahman (n = 15) bulls were evaluated for presence of single nucleotide polymorphisms (SNP) in the Bos taurus OPN gene (GenBank: AY878328.1) promoter region, and their possible effects on bull semen quality as evaluated by computer-assisted semen analysis (CASA). Semen was collected by electroejaculation 6 to 9 times from each bull, and each semen collection was evaluated by CASA for motile, progressive and rapid sperm within 5 mins of ejaculation. The bulls were genotyped for reported single nucleotide polymorphisms (SNP) in the promoter region of the OPN gene through amplification of two 700 base pair (bp) DNA fragments and sequencing of the resulting PCR products. Seven SNP sites were identified, at bp 3379, 3490, 3492, 5075, 5205, 5209, and 5263 of the OPN gene. The SNP identified at bp 5205, 5209 and 5263 had not been previously reported. Individual SNP sites were evaluated as the main effect on CASA sperm motility variables in a SAS MIXED model for repeated measures. A thymine to guanine substitution at bp 3379 was associated with increased (P ≤ 0.02) percentage of motile, progressive and rapid sperm in Angus x Brahman bulls, and tended (P ≤ 0.10) to increase the same sperm motility parameters in Angus, and Angus x Gelbvieh bulls. The percentages of motile, progressive and rapid sperm were similar (P ≥ 0.05) among genotypes for the other 6 SNP identified. These results suggest that identification and genotyping of polymorphisms within the promoter region of the bovine OPN gene may be useful for selecting bulls with improved sperm motility parameters.展开更多
This study evaluated sperm viability over time, after dilution and refrigerator storage of fresh semen extended in either synthetic cauda epididymal plasma (CEP2), or in a low sodium medium (CJ2) supplemented with eit...This study evaluated sperm viability over time, after dilution and refrigerator storage of fresh semen extended in either synthetic cauda epididymal plasma (CEP2), or in a low sodium medium (CJ2) supplemented with either AlbuMAX or egg yolk. Semen collected weekly for 4 weeks from 4 bulls and assigned within bulls, across treatments. After extension in either CEP2, or CJ2 containing either egg yolk or AlbuMAX, semen was cooled to 4°C, and evaluated for 7 days. A computer assisted sperm analysis (CASA) system was used for sperm evaluation. Particular emphasis was placed on sperm motility since it is the single most important sperm parameter influencing bull fertility. Total and progressive motility of sperm in CEP2 and CJ2-AlbuMAX were similar (P = 0.85 and P = 0.23, respectively), but both were lower (P < 0.01) when compared to CJ2-yolk. Fewer sperm had rapid motility in CEP2 and CJ2-AlbuMAX compared to CJ2-yolk (P < 0.01). Sperm straightness and linearity were greater in CJ2-AlbuMAX and CJ2-yolk than in CEP2 (P < 0.01). Mean velocity (VAP) and linear velocity (VSL) were greater (P < 0.01) in CJ2-AlbuMAX than either CEP2 or CJ2-yolk. The calculated curvilinear velocity (VCL) of spermatozoa in CEP2 was lower than CJ2-AlbuMAX (P = 0.01), but similar with CJ2-yolk (P = 0.54). Overall, every sperm parameter measured by the CASA system was equal to or higher for sperm stored 7 days in CJ2 medium as compared with CEP2. The CJ2 extender supplemented with egg yolk is a viable alternative for storing fresh bovine semen.展开更多
A study was conducted to evaluate variation in the ratio of X- to Y-bearing sperm of individual ejaculates and to determine any relationship between skewed sex ratio and either routine morphological evaluation or comp...A study was conducted to evaluate variation in the ratio of X- to Y-bearing sperm of individual ejaculates and to determine any relationship between skewed sex ratio and either routine morphological evaluation or computer assisted sperm analysis (CASA). Semen was collected from bulls weekly for 6 consecutive weeks, sperm DNA was recovered and quantitative, and real-time PCR was used to determine the ratio of X- to Y-bearing sperm in each ejaculate. The overall mean of X-bearing sperm within ejaculates was 54.7% over the 6 weeks of semen collections. The percentages of X-bearing sperm were similar (P > 0.5) across all collections. Between bulls, there were differences (P < 0.05) in the mean percentage of X-bearing sperm. No significant correlations were found between CASA parameters and percentage of X-bearing sperm across bulls, so analysis was done within each bull. Different combinations of CASA and/or morphological parameters were found to correlate with the percentage of X-bearing sperm but the prediction equations were specific for individual bulls and unlikely to be of use across bulls. These results confirm that the ratio of X- to Y-bearing sperm may be skewed in some ejaculates of bull semen. Some sperm parameters measured by CASA or routine morphological evaluation were associated with semen sex ratio.展开更多
文摘Osteopontin (OPN) is a protein found at higher concentrations in the seminal plasma of bulls with above average fertility. Polymorphisms have been reported within the OPN gene promoter that can affect production of this protein and thus, affect fertility. Therefore, Angus (n = 5) and Angus x Gelbvieh (Balancer, n = 14) and Angus x Brahman (n = 15) bulls were evaluated for presence of single nucleotide polymorphisms (SNP) in the Bos taurus OPN gene (GenBank: AY878328.1) promoter region, and their possible effects on bull semen quality as evaluated by computer-assisted semen analysis (CASA). Semen was collected by electroejaculation 6 to 9 times from each bull, and each semen collection was evaluated by CASA for motile, progressive and rapid sperm within 5 mins of ejaculation. The bulls were genotyped for reported single nucleotide polymorphisms (SNP) in the promoter region of the OPN gene through amplification of two 700 base pair (bp) DNA fragments and sequencing of the resulting PCR products. Seven SNP sites were identified, at bp 3379, 3490, 3492, 5075, 5205, 5209, and 5263 of the OPN gene. The SNP identified at bp 5205, 5209 and 5263 had not been previously reported. Individual SNP sites were evaluated as the main effect on CASA sperm motility variables in a SAS MIXED model for repeated measures. A thymine to guanine substitution at bp 3379 was associated with increased (P ≤ 0.02) percentage of motile, progressive and rapid sperm in Angus x Brahman bulls, and tended (P ≤ 0.10) to increase the same sperm motility parameters in Angus, and Angus x Gelbvieh bulls. The percentages of motile, progressive and rapid sperm were similar (P ≥ 0.05) among genotypes for the other 6 SNP identified. These results suggest that identification and genotyping of polymorphisms within the promoter region of the bovine OPN gene may be useful for selecting bulls with improved sperm motility parameters.
文摘This study evaluated sperm viability over time, after dilution and refrigerator storage of fresh semen extended in either synthetic cauda epididymal plasma (CEP2), or in a low sodium medium (CJ2) supplemented with either AlbuMAX or egg yolk. Semen collected weekly for 4 weeks from 4 bulls and assigned within bulls, across treatments. After extension in either CEP2, or CJ2 containing either egg yolk or AlbuMAX, semen was cooled to 4°C, and evaluated for 7 days. A computer assisted sperm analysis (CASA) system was used for sperm evaluation. Particular emphasis was placed on sperm motility since it is the single most important sperm parameter influencing bull fertility. Total and progressive motility of sperm in CEP2 and CJ2-AlbuMAX were similar (P = 0.85 and P = 0.23, respectively), but both were lower (P < 0.01) when compared to CJ2-yolk. Fewer sperm had rapid motility in CEP2 and CJ2-AlbuMAX compared to CJ2-yolk (P < 0.01). Sperm straightness and linearity were greater in CJ2-AlbuMAX and CJ2-yolk than in CEP2 (P < 0.01). Mean velocity (VAP) and linear velocity (VSL) were greater (P < 0.01) in CJ2-AlbuMAX than either CEP2 or CJ2-yolk. The calculated curvilinear velocity (VCL) of spermatozoa in CEP2 was lower than CJ2-AlbuMAX (P = 0.01), but similar with CJ2-yolk (P = 0.54). Overall, every sperm parameter measured by the CASA system was equal to or higher for sperm stored 7 days in CJ2 medium as compared with CEP2. The CJ2 extender supplemented with egg yolk is a viable alternative for storing fresh bovine semen.
文摘A study was conducted to evaluate variation in the ratio of X- to Y-bearing sperm of individual ejaculates and to determine any relationship between skewed sex ratio and either routine morphological evaluation or computer assisted sperm analysis (CASA). Semen was collected from bulls weekly for 6 consecutive weeks, sperm DNA was recovered and quantitative, and real-time PCR was used to determine the ratio of X- to Y-bearing sperm in each ejaculate. The overall mean of X-bearing sperm within ejaculates was 54.7% over the 6 weeks of semen collections. The percentages of X-bearing sperm were similar (P > 0.5) across all collections. Between bulls, there were differences (P < 0.05) in the mean percentage of X-bearing sperm. No significant correlations were found between CASA parameters and percentage of X-bearing sperm across bulls, so analysis was done within each bull. Different combinations of CASA and/or morphological parameters were found to correlate with the percentage of X-bearing sperm but the prediction equations were specific for individual bulls and unlikely to be of use across bulls. These results confirm that the ratio of X- to Y-bearing sperm may be skewed in some ejaculates of bull semen. Some sperm parameters measured by CASA or routine morphological evaluation were associated with semen sex ratio.
