Leydig Cells in Immunocastrated Polish Landrace Pig Testis: Differentiation Status and Steroid Enzyme Expression Status.

Porker immunocastration against gonadoliberin (GnRH) secretion has been utilized since 2009; however, consumers are still skeptical of it. This is due to not having full information available on the problem of a boar taint, as well as a lack of research on morphological and molecular changes that may occur in the animal reproductive system and other body systems. The present study aimed to explore the functional status of steroidogenic Leydig cells of the testicular interstitial tissue in immunocastrated Polish Landrace pigs. Analyses were performed using Western blot, immunohistochemistry for relaxin (RLN), insulin-like 3 protein (INSL3), pelleted growth factor receptor α (PDGFRα), cytochrome P450scc, 3ß- and 17ß-hydroxysteroid dehydrogenases (3ß-HSD, 17ß-HSD), cytochrome P450arom, and 5α-reductase (5α-RED). Immunoassay ELISA was used to measure the androstenone, testosterone, and estradiol levels in the testis and serum of immunocastrates. We revealed disturbances in the distribution and expression of (i) RLN, indicating an inflammatory reaction in the interstitial tissue; (ii) INSL3 and PDGFRα, indicating alterations in the differentiation and function of fetal, perinatal, or adult Leydig cell populations; (iii) P450scc, 3ß-HSD, 17ß-HSD, P450arom, and 5α-RED, indicating disturbances in the sex steroid hormone production and disturbed functional status of Leydig cells; as well as (iv) decreased levels of androstenone, testosterone, and estradiol in testicular tissue and serum, indicating the dedicated action of Improvac to reduce boar taint at both the hypothalamic-hypophysis-gonadal axis and local level (Leydig cells). In summary, our study provides a significant portion of knowledge on the function of Leydig cells after immunocastration, which is also important for the diagnosis and therapy of testis dysfunction due to GnRH action failure and/or Leydig cell differentiational-functional alterations.

The roles and expression of HOXA/Hoxa10 gene: A prospective marker of mammalian female fertility?

This review addresses the influence of homebox A10/a10 (HOXA/Hoxa10) gene on reproductive tract anatomy and functional fertility in mammalian species, and discusses major endocrine and environmental regulators of HOXA/Hoxa10 expression. Female reproductive efficiency or success is a function of several factors including the ovulation and fertilization rate, and uterine receptivity. A family of HOX/Hox genes establishes the segmental identity of the reproductive tract during embryogenesis and retains its physiological plasticity in sexually mature animals and humans. In particular, the HOXA/Hoxa10 gene is an intrinsic component of implantation, decidualization, and immunomodulation in the adult uterus. It was, therefore, suggested that knowledge of HOXA/Hoxa10 regulation might be essential in navigating molecular mechanisms with the aim of enhancing female reproductive potential. However, a recent study in pigs revealed a lack of associations between endometrial HOXA10 expression and reproductive tract morphology, and very poor correlations with sows' fertility metrics. Retinoic acid mainly regulates 3' HOX/Hox paralogs but may also modify the expression of downstream HOX/Hox genes, including HOXA/Hoxa10. Sex steroids directly regulate HOXA/Hoxa10 expression. The vitamin D receptor pathway modulates HOXA/Hoxa10 expression in the adult reproductive tract. Lastly, endocrine disruptors such as diethylstilbestrol, methoxychlor, bisphenol A, and isoflavones were shown to alter HOXA/Hoxa10 expression, thus affecting reproductive competence of the female.

Peroxisome Proliferator-Activated Receptor γ, but Not α or G-Protein Coupled Estrogen Receptor Drives Functioning of Postnatal Boar Testis-Next Generation Sequencing Analysis.

Porcine tissue gene expression is highly similar to the expression of homologous genes in humans. Based on this fact, the studies on porcine tissues can be employed to understand human physiology and to predict or treat diseases. Our prior studies clearly showed that there was a regulatory partnership of the peroxisome proliferator-activated receptor (PPAR) and the G-protein coupled membrane estrogen receptor (GPER) that relied upon the tumorigenesis of human and mouse testicular interstitial cells, as well as the PPAR-estrogen related receptor and GPER-xenoestrogen relationships which affected the functional status of immature boar testes. The main objective of this study was to identify the biological processes and signaling pathways governed by PPARα, PPARγ and GPER in the immature testes of seven-day-old boars after pharmacological receptor ligand treatment. Boar testicular tissues were cultured in an organotypic system with the respective PPARα, PPARγ or GPER antagonists. To evaluate the effect of the individual receptor deprivation in testicular tissue on global gene expression, Next Generation Sequencing was performed. Bioinformatic analysis revealed 382 transcripts with altered expression. While tissues treated with PPARα or GPER antagonists showed little significance in the enrichment analysis, the antagonists challenged with the PPARγ antagonist displayed significant alterations in biological processes such as: drug metabolism, adhesion and tubule development. Diverse disruption in the Notch signaling pathway was also observed. The findings of our study proposed that neither PPARα nor GPER, but PPARγ alone seemed to be the main player in the regulation of boar testes functioning during early the postnatal developmental window.

Correlates of reproductive tract anatomy and uterine histomorphometrics with fertility in swine.

Economic potential of the swine industry hinges upon the reproductive performance of sows, which may be enhanced by improving uterine capacity, a component trait of litter size and piglet productivity. Previous attempts at characterizing morphological traits indicative of high uterine volume have not been completely successful, resulting in the continued need for a reliable method of predicting reproductive value to improve production efficiency of the sow. Hence, the main objective of this study was to scrutinize macro- and micro-morphology of the sow's reproductive tract for quantitative correlations with fertility indices. Reproductive records from Polish Landrace × Polish Large White sows were used to examine the associations between fertility and ovarian/uterine morphology (n = 34) or uterine histomorphometry (n = 10). Several measures related to the ovary, including right and left ovarian weight (r = 0.50, p = 0.005 and r = 0.49, p = 0.006, respectively), were positively correlated with the litter size, while left ovarian number of corpora lutea (r = -0.38, p = 0.04) was negatively correlated with the mean litter size. Analysis of histomorphological characteristics of the uterine wall collected during the luteal phase of the estrous cycle revealed correlations between mean litter size and myometrial vascular content (r = 0.75, p = 0.03), the proportion of myometrial stroma (r = -0.68, p = 0.03), and the variability of endometrial thickness (r = -0.72, p = 0.02) in sows. Eight ovarian, vaginal and uterine characteristics were significantly correlated with mean lifetime numbers of live born and stillborn piglets/litter or the last litter size before slaughter. In conclusion, several anatomical and histomorphological metrics that relate to reproductive performance of swine may be used to inform production protocols and as a tool for selection of elite breeding sows, warranting future research into non-invasive or minimally invasive techniques for obtaining such measures.

Senescent cells in rabbit, nutria and chinchilla testes-Results from histochemical and immunohistochemical studies.

Rabbit, nutria and chinchilla testes were evaluated to compare testicular cellular senescence. There were no major species-specific differences in structure of either seminiferous tubules or interstitial tissue. There, however, were occasional abnormalities in seminiferous tubule structure with there being multinucleated and exfoliated cells present in rabbit testes. Furthermore, there were seminiferous tubules without a lumen that were filled with premeiotic/meiotic cells in nutria; and tubules with vacuolization with there being no post-meiotic cells in chinchillas. There were no differences in distribution or content of acids, total proteins and polysaccharides in the testis of any of the three species. Results using comparative immunohistochemistry procedures indicated the testes contained a few senescent cells in seminiferous tubules with typical morphology and there was a large number of senescent cells in seminiferous tubules of nutrias and chinchillas that had an abnormal structure (P <0.001). Compared to rabbit testes, in which there was the least number of senescent cells in seminiferous tubules, there was a greater abundance of senescence markers in both nutria and chinchilla testes (P < 0.05; P < 0.001, respectively). Furthermore, there were small abundances of caspase 3 and LC3 in the testes of all species. In chinchilla testes, there was a lesser concentration of cholesterol (P < 0.001) and testosterone compared with the other species. Cellular senescence in testes, therefore, can be assessed by detection of morpho-functional disorders of the testis of the three species evaluated in the present study.

Implication of Membrane Androgen Receptor (ZIP9) in Cell Senescence in Regressed Testes of the Bank Vole.

Here, we studied the impact of exposure to short daylight conditions on the expression of senescence marker (p16), membrane androgen receptor (ZIP9) and extracellular signal-regulated kinase (ERK 1/2), as well as cyclic AMP (cAMP) and testosterone levels in the testes of mature bank voles. Animals were assigned to groups based on an analysis of testis diameter, weight, seminiferous tubule diameter and the interstitial tissue area: group 1, not fully regressed (the highest parameters); group 2 (medium parameters); or group 3, regressed (the lowest parameters). Cells positive for p16 were observed only in the seminiferous tubule epithelium. However, in groups 1 and 2, these were mostly cells sloughed into the tubule lumen. In group 3, senescent cells resided in between cells of the seminiferous epithelium. Staining for ZIP9 was found in Sertoli cells. Western blot analysis showed a trend towards a decreased expression of p16 and ZIP9 in the testes of the voles in groups 2 and 3, compared to group 1. In addition, a trend towards an increased expression of ERK, as well as an increase of cAMP and testosterone levels, was revealed in group 2. In the regressed testes, a functional link exists between senescence and androgen levels with implication of ZIP9 and cAMP/ERK signaling pathways.

The Use of Vagina⁻Cervix Length Measurement in Evaluation of Future Reproductive Performance of Sows: A Preliminary Study under Commercial Conditions.

The length of the distal part of the internal reproductive tract seems to be related to the length and capacity of uterine horns, which is the most important anatomical property influencing litter size in sows. The aim of this study was to evaluate variation in vagina-cervix length (VCL) in gilts and differences in reproductive performance of sows according to VCL. The study was performed in a commercial farm using 221 gilts introduced into the breeding herd. Females were divided into three groups: (S) short (26.0 ± 2.0 cm, n = 36), (M) medium (31.3 ± 1.46 cm; n = 121), and (L) long VCL (36.0 ± 1.4 cm; n = 42) (p < 0.01). Mean live weight of gilts did not differ significantly among groups. Mean first litter size significantly varied between groups S (10.47 ± 3.01) and L (11.98 ± 2.32) (p = 0.0075) and M (10.67 ± 2.98) and L (p = 0.0054), while there was no significant difference between group S and M. Significant advantage (p = 0.023) was noted in the number of litters obtained from sows in groups L (4.69 ± 3.14), M (3.67 ± 2.71), and S (3.36 ± 2.40), and thus in total life production of sows (p = 0.0054), i.e., the number of piglets born alive. To conclude, the differences in vagina-cervix length in gilts during the first service was associated with significant variability in litter size during the first reproductive cycle, giving an advantage to females with longer VCL. Gilts with longer VCL were culled later and gave significantly more litters. Consequently, their lifetime piglet production was greater than gilts with shorter VCL.

Storage of boar semen at 16-18 °C in the long-term commercial extender prepared with deionized water or nanowater.

The aim of this study was to assess the usefulness of nanowater (NW; water declusterized using cold plasma treatment) as a diluent for a commercial boar semen extender during the 15-day storage (Days 1 to 15) at 16-18 °C. Ejaculates collected from 8 boars were subjected to the standard evaluation and then diluted in the extender prepared with deionized water (DW) or NW to a final concentration of 3×109 spermatozoa/ml. The proportion of defective spermatozoa increased (P<0.05) from Day 10 to Day 15 of storage (22.8±16.6% to 41.8±26.4% in DW group and 18.6±11.7% to 34.8±25.4% in NW group) and it was significantly greater in DW group compared with NW group on Days 5 and 10 due mainly to a greater (P<0.05) number of mid-piece defects in semen stored in the DW-containing extender. Sperm progressive motility decreased (P<0.05) in both groups between Days 2 and 6, Days 6 and 10, and Days 10 and 12, whereas the percentage of motile spermatozoa declined (P<0.05) to Day 14 only in NW group. Sperm motility was greater (P<0.05) in NW group compared with DW group from Day 5 to Day 13. A decline in sperm progressive motility below 40% in all semen samples occurred by Day 11 in DW group and by Day 12 in NW group. The mean survival time of sperm at 37 °C ex situ was greater in NW group than in DW group on Day 5 (314±87 min compared with 284±87 min) and Day 10 (223±34 min compared with 182±27 min; NW group compared with DW group, respectively). There were no differences (P>0.05) between the two groups in the concentrations of alkaline phosphatase and aspartate aminotransferase in semen extender. To summarize, the use of NW as an extender diluent exerts cytoprotective effects on boar spermatozoa and delays a decline in sperm progressive motility.

Laparoscopically implanted system for stimulation of the hypogastric plexus induces colonic motility, defecation, and micturition: experimental study.

BACKGROUND: Modulation of the enteric nervous system seems to be promising in several functional colorectal disorders for which targeted, causal treatment methods do not exist. However, sacral nerve stimulation can induce undesirable muscle contraction or paresthesia. Therefore, we have developed a laparoscopic technique for implanting a neural electrode, placed directly over the pelvic autonomic nerve plexus. The aim of this experimental study was to evaluate the effect of stimulating the hypogastric plexus and pelvic nerves on inducing distal colon contraction, defecation, and micturition. METHOD: A total of 10 white, male healthy pigs (25-30 kg) were subjected to the laparoscopic implantation of the electrode and the stimulator. In the third and fourth weeks postimplantation, the efficacy of the acute and chronic stimulation to induce defecation was evaluated. RESULTS: The average operative time was 105 minutes (85-150 minutes). In all pigs, acute stimulation activated induced defecation, every second day, every time on demand, with an average delay of 139.7 s. Micturition was induced incidentally. Acute or chronic stimulation did not cause any harm, pain, or suffering to the animals. No adverse effects of the stimulation were observed, and no septic complications or macroscopic fibrosis around the electrodes were found on autopsy. CONCLUSION: Hypogastric plexus stimulation can be a useful and safe option of distal colon contraction, defecation, and micturition. However, the efficacy of the stimulation was observed for a relatively short period of time, and it is not known if it will be sustained for a longer duration.

Morphofunctional alterations in testicular cells of deslorelin-treated boars: an immunohistochemical study.

In this study we thoroughly scrutinized testes morphology and investigated whether treatment of recipient boars with gonadotropin-releasing hormone (GnRH)-agonist deslorelin could alter the expression of 3beta-hydroxysteroid dehydrogenase (3beta-HSD), luteinizing hormone receptors (LHRs), and androgen receptors (ARs) in testicular cells. An implant containing 4.7 mg of the GnRH-agonist deslorelin was subcutaneously inserted into crossbred male pigs at 91 and 147 days of age. Testicular traits, morphology of the testes, the proteins' expression, and testosterone concentration in blood plasma were analyzed in all boars after slaughter at 175 days of age. Histological analysis revealed significant alterations in both the interstitial tissue and seminiferous tubules of experimental animals after 28 and 84 days of deslorelin treatment. The intensity of the AR immunostaining within the testis appeared as a function of the severity of testicular dysgenesis. Time-dependent action of deslorelin on the expression of LHR and 3beta-HSD in Leydig cells was also detected. Staining for LHR and 3beta-HSD was very weak or the Leydig cells were immunonegative. Concomitantly, a significant decrease in plasma testosterone level was found in both groups of deslorelin-treated boars when compared with the control group. This is the first report showing the cellular distribution of AR, LHR, and 3beta-HSD in testicular cells of deslorelin-treated boars. It is concluded that morphological and immunohistochemical studies are important for the evaluation of testicular histoarchitecture and steroidogenic function. Subsequently, the endocrine control of reproduction in the GnRH-agonist deslorelin-treated males will be better understood.

Effects of vagal pacing on food intake and body mass in pigs.

UNLABELLED: In obesity patients inexpensive, non-invasive, low risk treatment remains a holy grail. The purpose of this study was to evaluate effect of long term and low frequency vagal pacing on feeding behavior in pigs. METHODS: Two groups of animals were investigated, first control with sham operation and second group with microchip (MC) on both vagal nerves placed laparoscopically. In both groups EGG was performed before and after MC implantation. Parameters of stimulation were stable (amplitude 170 mV, frequency 1 Hz, impulse duration 170 ms). RESULTS: MC group demonstrated continuous decrease in body weight gain during 8 weeks of experiment at average of 3.73 +/- 0.5 vs. 4.83 +/- 1.1 (p < 0.05). Food intake also decreased in MC group and was 16.38 +/- 1.3 vs. control 17.5 +/- 1.7 kg/w (ns). EGG recording showed decreased percent normogastria in MC group 26.66 +/- 10 vs. 76.2 +/- 16 (p < 0.05) mostly at cost of tachygastria 62.3 +/- 15 vs. 35.4 +/- 16% of the recording time. CONCLUSION: MC vagal pacing mostly decreased body weight being without significant influence on food intake causing gastric dysrhythmia.