White Composite Boars Research Articles

Relationships of testicular iron and ferritin concentrations with testicular weight and sperm production in boars12

The inverse relationship of testicular size and circulating follicle-stimulating hormone (FSH) concentrations has been documented, and accompanying this relationship is the change in color of the parenchymal tissue of the testes. Large testes (300 to 400 g) are pink to light red and small testes (100 g) are dark maroon with color gradations for weights in between. It was hypothesized that this color most likely represented an iron protein. Chromatographic analysis of testicular tissue indicated that the Fe was associated primarily with ferritin, and immunohistochemistry showed that Leydig cells were the primary location of ferritin storage within the testes. Concentrations of Fe and ferritin were higher in small testes and decreased as testes weight increased (P < 0.05). As testicular Fe concentrations increased, daily sperm production (DSP) and total DSP declined (P < 0.05). Genotyping six generations of Meishan x White composite boars (n = 288) for a quantitative trait locus that is indicative of elevated FSH and small testes in boars indicated that the Meishan genotype had elevated testicular iron concentrations and darker color in conjunction with reduced total DSP (P < 0.01). It is not thought the elevated iron concentrations affect testicular weights but are probably a result of elevated FSH and FSH inducement of Fe transport. The storage of Fe in Leydig cells may provide a reservoir of Fe for easy access by Sertoli and germ cells, but still provide a degree of protection to germ cells from ionic iron.

Specific Staining of Sertoli Cell Nuclei and Evaluation of Sertoli Cell Number and Proliferative Activity in Meishan and White Composite Boars During the Neonatal Period1

The positive relationship between Sertoli cell number and testicular size emphasizes the importance of determining factors involved in the regulation of the Sertoli cell population. Based on data from other species and indirect evidence in the boar, it is generally accepted that porcine Sertoli cells proliferate rapidly throughout the early postnatal period. However, direct evaluation of Sertoli cell number and the proliferative activity of Sertoli cells during the early postnatal period in boars have not been reported. Stereological enumeration of Sertoli cells is a labor-intensive process and would be greatly facilitated by a marker for these cells especially in the sexually mature male. Thus, the first objective of this study was to determine if expression of the transcription factor GATA-4 is an effective marker for fetal, postnatal, and adult Sertoli cells to facilitate enumeration procedures. The second objective was to evaluate the proliferative activity and growth of the Sertoli cell population in neonatal White Composite and Meishan boars, known to differ in mature testis size and Sertoli cell number, to determine the importance of this developmental period for the adult Sertoli cell population. GATA-4 was abundantly expressed by Sertoli cells throughout fetal and prepubertal stages of development and specifically stained both type A and B Sertoli cell nuclei in the sexually mature boar. Immunoreactivity was never observed in the germ cells regardless of their stage of development, illustrating that GATA-4 is a useful marker for both developing and adult Sertoli cells in the boar. Testicular size did not differ between breeds on Day 1 postpartum, but by 14 days postpartum White Composite boars had significantly larger testes compared to Meishan boars (P: < 0.001). Similarly, Sertoli cell number did not differ between breeds at 1 day postpartum; however, at 14 days postpartum White Composite boars had a significantly larger Sertoli cell population compared to Meishan boars (P: < 0.05). Surprisingly, despite having more Sertoli cells than Meishan boars at 14 days postpartum, the proportion of actively proliferating Sertoli cells in the White Composite boars was almost 50% less than the Meishan boars. This result illustrates that rapid rates of Sertoli cell proliferation probably occurred prior to 14 days postpartum in the White Composite boars. Collectively, these results illustrate that the relationship between testicular size and Sertoli cell number is manifested very early in the postnatal period for these two breeds. The substantial difference in the size of the Sertoli cell population and their proliferative activity between Meishan and White Composite boars during the early postnatal period emphasizes the importance of this early period for the establishment of the Sertoli cell population and subsequent adult testicular size.

Relationships of gonadotropins, testosterone, and cortisol in response to GnRH and GnRH antagonist in boars selected for high and low follicle-stimulating hormone levels.

Considerable variation exists in the serum levels of gonadotropins in boars; this results in differential testicular function. Boars (Chinese Meishan, European White composite, and crosses of the two breeds) selected for high and low circulating FSH concentrations were used to define possible differences in pituitary sensitivity to GnRH and GnRH antagonist and gonadal and adrenal responses. After a 2-h pretreatment sampling period, boars were injected with GnRH or GnRH antagonist and repetitively sampled via jugular cannula for changes in serum concentrations of FSH, LH, testosterone, and cortisol. In response to varying doses of GnRH or GnRH antagonist, FSH, LH, or testosterone changes were not different in high- or low-FSH boars. Declines in LH after GnRH stimulation were consistently faster in boars selected for high FSH. Chinese Meishan boars had considerably higher cortisol concentrations than White composite boars (132.2 +/- 28.5 vs 67.4 +/- 26.8 ng/mL, respectively; P < .01). When select high- and low-gonadotropin Meishan:White composite crossbreds were sampled, cortisol levels were elevated but comparable between the two groups (126.5 +/- 13.7 vs 131.4 +/- 13.4 ng/mL, respectively). After GnRH antagonist lowered LH concentrations, administration of hCG resulted in increased testosterone and cortisol concentrations. Although testosterone concentrations remained high for 30 h, cortisol concentrations returned to normal levels within 10 h after hCG injection. The mechanism by which boars selected for high gonadotropins achieve increased levels of LH and FSH may not be due to differences in pituitary sensitivity to GnRH but to differences in clearance from the circulation.

Effects of the thymic peptide thymulin on in vitro and in vivo testicular steroid concentrations in white composite and Meishan boars.

Immuno-peptides may have positive or negative effects on gonadal steroidogenesis, but few have been tested outside of rodent species or in vivo. In Exp. 1, thymulin, a secreted nonapeptide of the thymus, was incubated (1, 10, 100, or 1,000 ng/mL) with testicular minces (sampled at 3, 6, or 12 h) from Chinese Meishan boars of high gonadotropin/testicular steroidogenic function (n = 8) and White composite boars of European origin (n = 8 ) to test the hypothesis that thymulin could augment hCG stimulation of testicular androgen concentrations. Thymulin alone had few effects on androgen concentrations (testosterone, dehydroepiandrosterone+dehydroepiandrosterone sulfate [DHEA+DHEASO4]) in Meishan boar testicular incubates. In minces from White composites incubated with thymulin, testosterone concentrations were generally below control values (P<.05), but DHEA+DHEASO4 concentrations were unaffected. Thymulin had no effect on estrone concentrations in testicular incubates of White composite boars but stimulated estrone concentrations in Meishan testicular incubates. Thymulin plus hCG increased testosterone (3 and 6 h of incubation; P<.05) but not DHEA+DHEASO4 concentrations in White composite testicular incubates. Thymulin plus hCG did not alter androgen or estrogen concentrations from control values in Meishan testicular incubates. In Exp.2 with a protocol similar to that of Exp. 1 for testicular minces from White composite boars (n = 30), thymulin increased testosterone concentrations during the early incubation period (1 to 3 h; P<.05) and depressed testosterone concentrations at later times (6 h; P<.05). Thymulin synergized with hCG in stimulating increases in testosterone and DHEA+DHEASO4 concentrations (P<.05) but had no effect on estrone concentrations in vitro. Thymulin was tested in vivo in boars from three genetic lines selected for high, medium, or normal circulating LH concentrations (Meishan, select White composites, and control White composites, respectively). Injection of thymulin i.v. (4.4, 44.4, or 444.4 ng/kg BW) generally increased circulating testosterone concentrations (2 to 3 h later; P<.01), but the response was dependent on the boar's general circulating LH concentrations and dose of thymulin. Overall results from these studies support the hypothesis of a thymulin augmentation of LH stimulation of androgen increases in vitro and in vivo in the testis of boars.

Identification of quantitative trait loci affecting female reproductive traits in a multigeneration Meishan-White composite swine population.

A multigeneration crossbred Meishan-White composite resource population was used to identify quantitative trait loci (QTL) for age at first estrus (AP) and the components of litter size: ovulation rate (OR; number of ova released in an estrous period) and uterine capacity (UC). The population was established by reciprocally mating Meishan (ME) and White composite (WC) pigs. Resultant F1 females were mated to either ME or WC boars to produce backcross progeny (BC) of either 3/4 WC 1/4 ME or 1/4 WC 3/4 ME. To produce the next generation (F3), 3/4 WC 1/4 ME animals were mated to 1/4 WC 3/4 ME animals yielding half-blood (1/2 WC 1/2 ME) progeny. A final generation (F4) was produced by inter se mating F3 animals. Measurements for AP and OR were recorded on 101 BC, 389 F3, and 110 F4 gilts, and UC data were from 101 BC and 110 F4 first parity litters. A genomic scan was conducted with markers (n = 157) spaced approximately 20 cM apart. All parental, F1, BC, and F4 animals but only 84 F3 animals were genotyped and included in this study. The QTL analysis fitted a QTL at 1-cM intervals throughout the genome, and QTL effects were tested using approximate genome-wide significance levels. For OR, a significant (E[false positive] < .05) QTL was detected on chromosome 8, suggestive (E[false positive] < 1.0) QTL were detected on chromosomes 3 and 10, and two additional regions were detected that may possess a QTL (E[false positive] < 2.0) on chromosomes 9 and 15. Two regions possessed suggestive evidence for QTL affecting AP on chromosomes 1 and 10, and one suggestive region on chromosome 8 was identified for UC. Further analyses of other populations of swine are necessary to determine the extent of allelic variation at the identified QTL.

Positive association between expression of follicle-stimulating hormone beta and activin betaB-subunit genes in boars.

This study tested our hypothesis that inhibin/activin (I/A) betaB subunit and not follistatin (FS) gene expression relates positively to plasma FSH concentrations in the anterior pituitary gland of boars. Mature crossbred boars (n = 12) were selected for divergence in plasma FSH concentrations, and their anterior pituitary glands were evaluated for expression of the FSHbeta, I/A ssB, FS, calmodulin, and GnRH receptor (GnRH-R) genes by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and/or RNase protection assays (RPAs). Expression of I/A ssB was greater (p < 0. 01) in the six boars with high FSH than in the six with low FSH; expression of the I/A betaB-subunit gene was positively correlated to that of the FSHbeta gene (RT-PCR: r = 0.96; p < 0.01; RPA: r = 0.68; p < 0.05). In contrast, expression of the FS (p > 0.10), GnRH-R (p > 0. 08), and calmodulin (p > 0.10) genes was similar in the two groups of boars. Additionally, expression of the FSHbeta gene was correlated positively with pituitary and plasma FSH concentrations (r = 0.69 and 0.88, respectively; p < 0.05). These results support the hypothesis that activin B is partially responsible for elevated FSH concentrations in boars. Furthermore, the expression difference of the calmodulin gene observed previously between Meishan and White Composite boars represents a breed difference unrelated to FSH.

Identification of quantitative trait loci affecting carcass composition in swine: I. Fat deposition traits.

One of the major determining factors in the price of market hogs today is backfat depth. Therefore, identification of regions of the genome affecting this trait could be very useful to the swine industry. A large resource population (n = 540) was developed by backcrossing F1 Meishan-White composite females to either Meishan or White composite boars. A genomic scan was conducted by genotyping all animals with microsatellite markers spaced at approximately 20-cM intervals across the entire porcine genome. Breed of origin for all chromosomal segments was determined using multipoint linkage procedures, and a least squares regression analysis was conducted. Nominal P-values were converted to a genome-wide level of significance to adjust for the numerous tests conducted. Traits analyzed were backfat depths at the first rib (FRIB); 10th rib (10th); last rib (LRIB); last lumbar (LLUM) vertebra; the average of FRIB, LRIB, and LLUM (AVBF); and weight of leaf fat (LEAF). Significant quantitative trait loci (QTL) were detected on chromosomes 1, 7, and X. Suggestive evidence for QTL was present on chromosomes 5, 8, 9, 10, 13, and 14. Genotypic means for the loci detected indicated a predominantly additive mode of inheritance. Meishan alleles produced fatter pigs for all loci except those on chromosomes 7 and 10. Additional research should be conducted to refine the estimated position of each QTL and its effect and determine epistatic interactions. These loci should be evaluated in other germplasms to determine whether allelic variation at the QTL exists in other breeds.

Survival, body weights, feed efficiency, and carcass traits of 7/8 White Composite and 1/8 Duroc, 1/8 Meishan, 1/8 Fengjing, or 1/8 Minzhu pigs.

Pigs were the progeny of White Composite boars mated to gilts that were either 1/4 Duroc, 1/4 Meishan, 1/4 Fengjing, or 1/4 Minzhu and the remainder 3/4 White Composite. One-eighth Meishan and 1/8 Fengjing pigs averaged approximately .5 more nipples than 1/8 Duroc pigs and .2 more nipples than 1/8 Minzhu pigs (P < .05), respectively. Duroc, Meishan, and Minzhu crosses did not differ (P > .05) for survival at birth or at 14 and 28 d. Fengjing crosses had a lower survival rate (P < .05) at all three ages than Duroc and Meishan crosses. Duroc crosses were heavier (P < .05) than Chinese crosses at birth. At 56 d, Duroc and Meishan crosses did not differ (P > .05) for BW, but both were heavier (P < .05) than Minzhu crosses. Body weight at 70 d did not differ significantly (P > .05) among breed types. Duroc crosses were heavier (P < .05) than any of the Chinese crosses at 98, 126, and 154 d of age. At 154 d, Fengjing crosses were lighter (P < .05) than Meishan or Minzhu crosses. The effect of breed type was not detected for average probe backfat thickness of gilts at 99.7 kg. Duroc crosses consumed the most feed, and Fengjing crosses consumed the least during each interval and during the total period (P < .05). Feed consumption of Meishan and Minzhu crosses was intermediate to and significantly (P < .05) different from that of Duroc and Fengjing crosses. At 184 d of age, breed types did not differ (P > .16) for measures of carcass fat thickness, marbling score, color score, and firmness score. At 184 d of age, weights of all carcass cuts were heavier (P < .05) for Duroc than for Chinese crosses with the exception of untrimmed loin weight of Minzhu crosses. There were few significant differences among breed types when carcass traits were compared at a constant carcass weight, although Duroc crosses generally were superior. Relative to similar Duroc crosses, these results suggest that 1/8 Chinese pigs would grow less rapidly (P < .05) and produce less weight of trimmed lean cuts at a constant carcass weight.

Breed Differences in Expression of Inhibin/Activin Subunits in Porcine Anterior Pituitary Glands

Chinese Meishan (MS) boars have greater plasma FSH concentrations than European White Composite boars, but this difference does not occur in females of these breeds. To understand this disparity, we studied expression of the follistatin gene and of genes for the inhibin/activinα -, βA-, and βB-subunits in porcine anterior pituitary glands using quantitative reverse transcription-PCR and ribonuclease protection techniques. We found that 1) the inhibin/activin βA- and βB-subunits and follistatin were expressed in porcine pituitary; 2) the α-subunit was not detected in the porcine pituitary, but was highly expressed in porcine follicles; and 3) the βB-subunit gene is more abundantly expressed (2-fold greater) in MS boar pituitaries than in pituitaries of White Composite boars. We conclude that this is not due to a breed difference, because the expression levels of this gene were similar in pituitaries of females of these breeds. No breed differences were detected for other genes screened in this study. From these observations, we propose that activin B, a dimer ofβ B-subunits and a stimulator of FSH secretion, may be partially responsible for the elevated plasma FSH concentrations in MS boars, and intrapituitary inhibin plays no or a very minimal role.

Negative relationship between blood concentrations of follicle-stimulating hormone and testicular size in mature boars.

Relationships between blood concentration of FSH and testicular size and daily sperm production were evaluated with data obtained from five studies originally designed to investigate regulation of FSH secretion in Meishan (MS), White composite (WC), and crossbreds of these. A minimum of three blood samples/boar were obtained at greater than 4-d intervals for determination of FSH, and testes were obtained at castration or slaughter. In a random sample of boars, FSH was fivefold greater (P < .01) in MS than in WC boars (n = 22/group). Daily sperm production (DSP)/gram of testis (estimated by counting elongated spermatid nuclei in testicular homogenates) was similar in the groups, but testicular weight (TWT), adjusted for body weight, was less (P < .01) in MS than in WC, yielding lower total daily sperm production (TDSP; P < .05) in MS boars. In four populations (one with MS, one with WC, and two with crossbreds; n = 34 males), boars were selected for extremes in FSH concentrations from larger groups. Across all populations, a threefold greater plasma FSH concentration was associated with a 32% smaller TWT (P < .01). Coincident with increased FSH, TDSP was 33% less (P < .05). In 48 MS x WC boars that were selected for divergence in plasma FSH during pubertal development (4 to 6 mo of age), this divergence was retained at 1 yr (P < .01). Retrospectively, the divergence in FSH was also apparent at 2 and 8 wk of age (P < .05), and the boars with elevated FSH had smaller testicles, lower DSP, and lower TDSP (P < .01). These studies document a negative relationship in mature boars between FSH secretion and testicular size accompanied with decreased TDSP.

Effects of porcine somatotropin on circulating testosterone concentrations in boars and mechanism of action.

Four experiments were conducted to elucidate the effects of porcine somatotropin (pST, growth hormone) on circulating testosterone concentrations in boars. In Exp. 1, jugular-cannulated obese boars were administered 4 mg recombinant (r) pST/d for 24 d before collection of samples at 15-min intervals over 6 h for measurement of pST, testosterone, and LH. Somatotropin treatment decreased plasma concentrations of testosterone and LH (P < .05). In Exp. 2, White composite boars were administered rpST with implants (4 mg/d) for 10 to 28, 16 to 28, or 22 to 28 wk of age. Reductions in testosterone concentrations were proportional to the length of time rpST was administered; boars treated for 18 wk had the lowest testosterone concentrations. Concentrations of LH declined throughout the study in rpST-implanted boars but remained static in untreated, control boars. In vitro production of testosterone and dehydroepiandrosterone/dehydroepiandrosterone sulfate was unaffected by rpST treatment, and hCG stimulation of in vitro androgen secretion was similar in rpST and control treatments. In Exp. 3, i.v. injection of pituitary pST (USDA-B1; 5 micrograms/kg BW) into jugular-cannulated White composite and Meishan boars resulted in an acute increase in circulating LH followed by an increase in testosterone concentrations, which then declined to below preinjection levels at 6 to 7 h after pST injection. Multiple injections, two or four per day, of 5 micrograms pST/kg BW resulted in decreased testosterone concentrations in White composite boars, whereas in Meishan boars testosterone concentrations were unaffected (Exp. 4). Concentrations of LH were not different from control values in either breed, but in both breeds, four injections of pST per day produced lower LH concentrations than did two injections per day (P < .01). Depression of circulating concentrations of androgens in boars requires extended periods of pST treatment.

Survival, body weights, feed efficiency, and carcass traits of 3/4 white composite and 1/4 Duroc, 1/4 Meishan, 1/4 Fengjing, or 1/4 Minzhu pigs.

Pigs were the progeny of White Composite (WC) boars mated to F1 Duroc x WC, Meishan x WC, Fengjing x WC, and Minzhu x WC gilts. Meishan and Fengjing crosses had more (P < .05) nipples than Duroc and Minzhu crosses. Meishan and Minzhu crosses had a higher survival rate at birth (P < .05) than Duroc and Fengjing crosses, but breed types did not differ significantly (P approximately .29) for survival to 14 or 28 d of age. Duroc crosses were heavier (P < .05) than Fengjing and Minzhu crosses at 0, 28, 56, 70, 98, 126, and 154 d of age; they were heavier (P < .05) than Meishan crosses at 0, 28, 98, 126, and 154 d of age. Chinese crosses had similar weights at most ages, although Meishan crosses were heavier (P < .05) than Fengjing and Minzhu crosses at 126 and 154 d of age. Breed types did not differ significantly (P approximately .27) for feed efficiency during the nursery period. Over the entire finishing period, Duroc-cross gilts (.3310) were less efficient (P < .05) than Meishan (.3436), Fengjing (.3454), or Minzhu crosses (.3466); among barrows Meishan crosses (.3176) were least efficient (P < .05), Fengjing crosses (.3263) were most efficient (P < .05), and Duroc (.3211) and Minzhu (.3209) were intermediate but not significantly different from the Meishan or Fengjing crosses. At a constant age, Duroc crosses had longer carcasses, greater longissimus muscle area, and heavier slaughter weight, carcass weight, and weight of untrimmed cuts and trimmed cuts than the Chinese crosses (P < .05). There were few significant differences among breed types for carcass traits at a constant carcass weight. Results show that, relative to one-half Duroc females, incorporation of one-half Chinese females into a crossbreeding program will result in progeny with a significant decrease in rate of growth (approximately 9% for weight at 154 d of age) and a small, nonsignificant decrease in yield of trimmed lean cuts (approximately 5%) at a constant carcass weight.