Lipoxygenases (LOXs) are a family of enzymes capable of peroxidizing phospholipids. A member of the LOX family of enzymes, 15-LOX, participates in the degradation of mitochondria and other organelles within differentiating red blood cells, the reticulocytes. The present study provides biochemical and immunocytochemical evidence for the presence of 15-LOX in the sperm cytoplasmic droplet (CD). Testicular, epididymal and ejaculated spermatozoa were evaluated for the presence of 15-LOX using an affinity-purified immune serum raised against a synthetic peptide corresponding to the C-terminal sequence of rabbit reticulocyte 15-LOX. Western blotting revealed an appropriate single band of ,81 kDa in boar spermatozoa but not in boar seminal plasma. When ejaculated boar spermatozoa were subjected to separation on a 45/90% Percoll gradient, 15-LOX co-migrated with the immotile sperm and cellular debris/CD fractions, but not with the motile sperm fraction containing morphologically normal spermatozoa without CDs. Varied levels of 15-LOX were expressed in ejaculated sperm samples from boars with varied semen quality. By immunofluorescence, prominent 15-LOX immunoreactivity was found within the residual body in the testis and within the CDs from caput, corpus and cauda epididymal and ejaculated spermatozoa. Components of the ubiquitin-dependent proteolytic pathway, which is thought to facilitate both spermiogenesis and reticulocyte organelle degradation, were also detected in the sperm CD. These included ubiquitin, the ubiquitin-conjugating enzyme E2, the ubiquitin C-terminal hydrolase PGP 9.5, and various 20S proteasomal core subunits of the a-and b-type. The 15-LOX and various components of the ubiquitin-proteasome pathway were also detected in sperm CDs of other mammalian species, including the human, mouse, stallion and wild babirusa boar. We conclude that 15-LOX is prominently present in the mammalian sperm CD and thus may contribute to spermiogenesis, CD function or CD removal. Reproduction (2005) 130 213-222
The objectives of this study were to estimate response to divergent selection for an index of placental efficiency in swine, and to evaluate the effect of placental efficiency on litter size. The selection index (SI) included total born (TB), birth weight (BRWT), and placental weight (PW), and was designed to increase in the high line (H) or decrease in the low line (L) the efficiency of the placental function (PE), defined as the ratio BRWT:PW. (Co)variance components were estimated for direct and maternal additive effects by using an animal model with MTDFREML procedures. Estimated breeding values were calculated by using records on individual BRWT (n = 2,111), PW (n = 2,006), PE (n = 1,677), and SI (n = 1,677). Litter traits were evaluated using records on 193 litters. The model included the fixed effects of contemporary group for all traits, with the addition of sex for individual traits and parity for litter traits. Litter was fitted as an uncorrelated random effect for all traits, and TB was used as a linear and quadratic covariate for BRWT, PW, and PE. Direct heritability estimates from single-trait models were 0.03, 0.25, 0.18, 0.11, and 0.08 for BRWT, PW, PE, SI, and TB, respectively. Estimated breeding values were compared between lines by using a model including generation, line within generation, and replicate within line as the error term. Estimates of genetic divergence were 20.7 +/- 2.7 g, 0.24 +/- 0.03, 0.11 +/- 0.02, and 0.07 +/- 0.02 per generation for PW, PE, SI, and TB, respectively (P < 0.01), but divergence was not significant for BRWT. At Generation 4, direct EBV was higher in L than in H for PW (55.9 +/- 8.7 vs. -24.2 +/- 9.5 g, respectively; P < 0.01) and higher in H than in L for PE (0.58 +/- 0.10 vs. -0.35 +/- 0.09 g, respectively; P < 0.01). However, EBV was not different for BRWT, SI, or TB. These results indicate that PW and PE are susceptible to change by genetic selection; however, the correlated response in TB was an unexpected genetic trend toward a higher TB in L of 0.05 +/- 0.01 piglets per generation (P < 0.01).
The purpose of this study was to investigate the relationship between fertility and quantitative measures of boar semen quality, including various patterns of sperm cytoplasmic droplet (CD) retention, as determined by high power differential interference contrast (DIC) microscopy. A total of 116 ejaculates were collected from a nucleus herd of 18 Large White boars over an eight month period. Semen quality parameters were analyzed for each ejaculate by calculating the percentage of normal spermatozoa, spermatozoa possessing a CD in the proximal, distal, or distal midpiece reflex position, total spermatozoa with an attached cytoplasmic droplet, spermatozoa with non-CD related aberrations and total spermatozoa with abnormalities. Of the 116 ejaculates received, 71 ejaculates from 13 boars had corresponding fertility data from single-sire inseminations of multiparous sows. The fertility data included farrowing rate (FR) and total number born (TNB). The monthly FR encompassed one month before and one month after the date of semen collection. Detection of differences for fertility and semen quality parameters was performed by separating the boars into either an above-average or below-average group based on the mean FR (74.01 AE 1.43%) or TNB (12.34 AE 0.17) for the study. For FR, the boars in the below-average group had a significantly lower percentage of normal spermatozoa and significantly higher percentage of spermatozoa possessing distal CDs, total attached CDs and total abnormalities compared to the boars in the above-average group. Conversely, for TNB there were no significant differences between the above-and below-average groups for the semen quality parameters. These data suggest that the attached CD may negatively affect FR, but not TNB. The detection of relationships between the boar fertility parameters and the retention of the sperm CD after ejaculation, document the advantage of high power DIC microscopy in conventional semen evaluation.
High numbers of live born and vital piglets are required for an effective piglet production. It has been reported that embryonic losses in swine can be as high as 20 to 50%. Generally, up to 30% of embryonic losses are considered within the normal biological range. The aim of our study was first to determine embryonic and fetal losses in sows of the German Landrace breed, and second to identify sows, which are able to realize high numbers of intact embryos and fetuses in combination with low rates of embryonic and fetal losses. This study was conducted on a commercial farrow-finish operation and involved 64 gilts. Gilts were synchronized for ovulation and inseminated artificially (AI) twice at fixed times. Pregnant gilts were slaughtered on Day 30 (n=34) and Day 80 (n=30) after second AI. Corpora lutea (CL) and embryos/fetuses (E/F) were counted. The length of uterine horns was measured. Based on the difference between the number of CL and the number of E/F, embryonic and fetal losses were 36.9 and 37.9%, respectively. The results obtained regarding the number of CL, the number E/F (intact and total), the rate of embryonic or fetal losses and uterine space per E/F underwent a cluster analysis. The results show that several gilts are able to realize high numbers of intact E/F in combination with low rates of losses and limited uterine space per E/F. Numbers of 15 to 20 intact fetuses on day 80 of pregnancy are considered as a minimum for an adequate litter size at the end of pregnancy. In this study only 35% of gilts at Day 30 and 37% of gilts at Day 80 demonstrate this potential. Cluster analysis is a useful mathematical tool which helps to assess results of embryonic and fetal losses and to group gilts according to their performance. As a consequence cluster analysis provides clues which gilts or family structures should be analyzed more intensively.
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