Mature Gilts Research Articles

Seasonal ovarian activity and oocyte size in ovarian follicles of sexually mature gilts

Seasonal ovarian activity and oocyte size in ovarian follicles of sexually mature gilts

Expression of visfatin in the ovarian follicles of prepubertal and mature gilts and in vitro effect of gonadotropins, insulin, steroids, and prostaglandins on visfatin levels

Recent studies have demonstrated that visfatin participates in the regulation of female reproduction. Due to the lack of data concerning the level of visfatin in the ovarian follicles of pigs, one of the most economically important livestock species, the aim of this study was to investigate the expression and localisation of visfatin and the follicular fluid concentration in the ovarian follicles of prepubertal and mature gilts. We also aimed to examine the in vitro effects of gonadotropins (LH, FSH), insulin, progesterone (P4), oestradiol (E2), prostaglandin E2 (PGE2) and F2α (PGF2α) on visfatin levels. In the present study, we have demonstrated that visfatin expression is dependent on the maturity of the animals and the stage of ovarian follicle development. Visfatin signal was detected in individual follicular compartments from primordial to antral follicles and even in atretic follicles. We have shown that the expression of visfatin in granulosa cells was higher than in theca cells. The level of visfatin is upregulated by LH, FSH, E2, and P4 and downregulated by insulin, while prostaglandins have modulatory effects, dependent on the dose and type of ovarian follicular cells. To summarise, our research has shown that visfatin is widely expressed in the ovarian follicles of prepubertal and mature pigs, and its expression is regulated by gonadotropins, insulin, steroids, and prostaglandins, suggesting that visfatin appears to be an important intra‐ovarian factor that could regulate porcine ovarian follicular function.

Attainment of Sexual Maturity and Gonadotropin Priming in Gilts Determine Follicular Development, Endocrine Milieu and Response to Ovulatory Triggers.

The routine procedure of estrous cycle synchronization in pigs allows for the use of gonadotropins to stimulate ovarian activity. The applied protocols of eCG and hFSH priming similarly affected development of ovarian follicles in two classes 3–6 mm and >6 mm of diameter, however, the number of small follicles (<3 mm) was 2-fold higher in hFSH- than in eCG-primed prepubertal gilts. The attainment of sexual maturity increased concentration of estradiol, testosterone and androstenedione in the follicular fluid of hFSH/eCG-primed gilts, however, prostaglandin E2 and F2α metabolite increased in mature hFSH- and eCG-primed gilts, respectively. The maturity increased mRNA and/or protein expression of key steroidogenic enzymes, prostaglandin synthases or luteinizing hormone receptors in follicular walls. Both hormonal primers played a moderate role in affecting expression of steroidogenic enzymes in follicular walls. In vitro studies showed higher estradiol production in r-hLH (p = 0.04)- and r-hCG (p = 0.049)-stimulated follicular walls of mature gilts than in prepubertal hFSH-primed gilts. Both ovulatory triggers decreased the abundance of LHCG/FSH mRNA receptors in follicular walls, which mimic downregulation of these receptors by a preovulatory LH surge, confirmed in vivo. These data revealed the importance of sexual maturity in the protection of the estrogenic environment, and the selective, moderate role of eCG and FSH in the activation of steroidogenic enzymes in preovulatory follicles.

Uterine Inflammation Changes the Expression of Cholinergic Neurotransmitters and Decreases the Population of AChE-Positive, Uterus-Innervating Neurons in the Paracervical Ganglion of Sexually Mature Gilts.

Simple SummaryEndometritis, both with non-infectious and infectious backgrounds, is one of the most prevalent pathological states among domestic animals. In animals, it generates severe economic problems, including lowered reproductive indices and rising medical treatment costs, and in women, it might lead to severe fertility impairment. In order to determine how the autonomic nervous system responds to such a pathological state, an experimental group of pigs were treated with Escherichia coli injection into the uterine horns, and several ganglions responsible for innervation of this organ were examined, including the paracervical ganglion located on both sides of the broad ligament of the uterus. The results clearly showed a strong impact of the inflammation on the chemical coding of neurons, some even synthesizing neurotransmitters de novo such as the GAL-expressing perikarya. Additionally, applied injections decreased the number of parasympathetic, acetylcholinesterase-expressing neurons implying the importance of the cholinergic population to keep the inflammation under control. The obtained data serve as a basis for the future implementation of modern treatment and enhancements in animal breeding.The focus of this study was based on examining the impact of endometritis on the chemical coding of the paracervical ganglion (PCG) perikaryal populations supplying pig uterus. Four weeks after the injection of Fast Blue retrograde tracer into uterine horns, either the Escherichia coli (E. coli) suspension or saline solution was applied to both horns. Laparotomy treatment was performed for the control group. Uterine cervices containing PCG were extracted on the eighth day after previous treatments. Subsequent macroscopic and histopathologic examinations acknowledged the severe form of acute endometritis in the E. coli-treated gilts, whereas double-labeling immunofluorescence procedures allowed changes to be analyzed in the PCG perikaryal populations coded with vesicular acetylcholine transporter (VAChT) and/or somatostatin (SOM), vasoactive intestinal polypeptide (VIP), a neuronal isoform of nitric oxide synthase (nNOS), galanin (GAL). The acetylcholinesterase (AChE) detection method was used to check for the presence and changes in the expression of this enzyme and further confirm the presence of cholinergic perikarya in PCG. Treatment with E. coli resulted in an increase in VAChT+/VIP+, VAChT+/VIP−, VAChT+/SOM+, VAChT+/SOM−, VAChT+/GAL− and VAChT+/nNOS− PCG uterine perikarya. An additional increase was noted in the non-cholinergic VIP-, SOM- and nNOS-immunopositive populations, as well as a decrease in the number of cholinergic nNOS-positive perikarya. Moreover, the population of cholinergic GAL-expressing perikarya that appeared in the E. coli-injected gilts and E. coli injections lowered the number of AChE-positive perikarya. The neurochemical characteristics of the cholinergic uterine perikarya of the PCG were altered and influenced by the pathological state (inflammation of the uterus). These results may indicate the additional influence of such a state on the functioning of this organ.

Long-Term Changes in Ovarian Follicles of Gilts Exposed Neonatally to Methoxychlor: Effects on Oocyte-Derived Factors, Anti-Müllerian Hormone, Follicle-Stimulating Hormone, and Cognate Receptors.

In this paper, we investigated the effects of neonatal exposure to methoxychlor (MXC), a synthetic organochlorine used as an insecticide with estrogenic, antiestrogenic, and antiandrogenic activities on ovarian follicles of adult pigs. Piglets were injected with MXC (20 μg/kg body weight) or corn oil (controls) from postnatal Day 1 to Day 10 (n = 5 per group). Then, mRNA expression, protein abundance and immunolocalization of growth and differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), anti-Müllerian hormone (AMH) and cognate receptors (ACVR1, BMPR1A, BMPR1B, TGFBR1, BMPR2, and AMHR2), as well as FSH receptor (FSHR) were examined in preantral and small antral ovarian follicles of sexually mature gilts. The plasma AMH and FSH levels were also assessed. In preantral follicles, neonatal exposure to MXC increased GDF9, BMPR1B, TGFBR1, and BMPR2 mRNAs, while the levels of AMH and BMP15 mRNAs decreased. In addition, MXC also decreased BMP15 and BMPR1B protein abundance. Regarding small antral follicles, neonatal exposure to MXC upregulated mRNAs for BMPR1B, BMPR2, and AMHR2 and downregulated mRNAs for AMH, BMPR1A, and FSHR. MXC decreased the protein abundance of AMH, and all examined receptors in small antral follicles. GDF9 and BMP15 were immunolocalized in oocytes and granulosa cells of preantral follicles of control and treated ovaries. All analyzed receptors were detected in the oocytes and granulosa cells of preantral follicles, and in the granulosa and theca cells of small antral follicles. The exception, however, was FSHR, which was detected only in the granulosa cells of small antral follicles. In addition, MXC decreased the plasma AMH and FSH concentrations. In conclusion, the present study may indicate long-term effects of neonatal MXC exposure on GDF9, BMP15, AMH, and FSH signaling in ovaries of adult pigs. However, the MXC effects varied at different stages of follicular development. It seems that neonatal MXC exposure may result in accelerated initial recruitment of ovarian follicles and impaired cyclic recruitment of antral follicles.

Endometritis decreases the population of uterine neurons in the paracervical ganglion and changes the expression of sympathetic neurotransmitters in sexually mature gilts

BackgroundThe focus of the study was to examine the impact of the inflamed uterus on the population of the paracervical ganglion (PCG) uterus-innervating perikarya and their chemical coding. Fast Blue retrograde tracer was injected into the wall of uterine horns on the 17th day of the first studied estrous cycle. After 28 days, either Escherichia coli suspension or saline was applied to the horns of the uterus, whereas the control group received laparotomy only. Eight days after the above-mentioned procedures, uterine cervices with PCG were collected. Both macroscopic and histopathologic examinations confirmed severe acute endometritis in the Escherichia coli-injected uteri. The double immunofluorescence method was used to analyze changes in the PCG populations coded with dopamine-β‐hydroxylase (DβH) and/or neuropeptide Y (NPY), somatostatin (SOM), vasoactive intestinal polypeptide (VIP) and neuronal isoform of nitric oxide synthase (nNOS).ResultsThe use of Escherichia coli lowered the total number of Fast Blue-positive neurons. Moreover, an increase in DβH+/VIP+, DβH+/NPY+, DβH+/SOM + and DβH+/nNOS + expressing perikarya was noted. A rise in non-noradrenergic VIP-, SOM- and nNOS-immunopositive populations was also recorded, as well as a drop in DβH-positive neurotransmitter-negative neurons.ConclusionsTo sum up, inflammation of the uterus has an impact on the neurochemical properties of the uterine perikarya in PCG, possibly affecting the functions of the organ.

Endocrine and molecular milieus of ovarian follicles are diversely affected by human chorionic gonadotropin and gonadotropin-releasing hormone in prepubertal and mature gilts

Different strategies are used to meet optimal reproductive performance or manage reproductive health. Although exogenous human chorionic gonadotropin (hCG) and gonadotropin-releasing hormone (GnRH) agonists (A) are commonly used to trigger ovulation in estrous cycle synchronization, little is known about their effect on the ovarian follicle. Here, we explored whether hCG- and GnRH-A-induced native luteinizing hormone (LH) can affect the endocrine and molecular milieus of ovarian preovulatory follicles in pigs at different stages of sexual development. We collected ovaries 30 h after hCG/GnRH-A administration from altrenogest and pregnant mare serum gonadotropin (eCG)-primed prepubertal and sexually mature gilts. Several endocrine and molecular alternations were indicated, including broad hormonal trigger-induced changes in follicular fluid steroid hormones and prostaglandin levels. However, sexual maturity affected only estradiol levels. Trigger- and/or maturity-dependent changes in the abundance of hormone receptors (FSHR and LHCGR) and proteins associated with lipid metabolism and steroidogenesis (e.g., STAR, HSD3B1, and CYP11A1), prostaglandin synthesis (PTGS2 and PTGFS), extracellular matrix remodeling (MMP1 and TIMP1), protein folding (HSPs), molecular transport (TF), and cell function and survival (e.g., VIM) were observed. These data revealed different endocrine properties of exogenous and endogenous gonadotropins, with a potent progestational/androgenic role of hCG and estrogenic/pro-developmental function of LH.

Effects of neonatal exposure to methoxychlor on corpus luteum in gilts: A transcriptomic analysis.

This study investigated the effects of neonatal exposure to methoxychlor (MXC), a synthetic organochlorine used as an insecticide with estrogenic, antiestrogenic, and antiandrogenic activities, on luteal function in pigs. Piglets were injected subcutaneously with MXC (20 μg/kg body weight) or corn oil (control) between postnatal Days 1 and 10 (N = 5/group). Corpora lutea from sexually mature gilts were examined for luteal steroid and prostaglandin concentrations and processed for total RNA isolation and subsequent RNA sequencing. Intra-luteal concentrations of androstenedione and prostaglandin E2 were greater, while that of estrone was lower when compared to control. Fifty-three differentially expressed (DE) microRNAS (miRNAs) (p-adjusted <.05 and log2(fold change) ≥.5) and 359 DE genes (p-adjusted <.05 and log2(fold change) ≥1) were identified in luteal tissue in response to neonatal MXC treatment. MXC was found to affect the expression of genes related to lipogenesis, steroidogenesis, membrane transport, immune response, cell signaling and adhesion. These results suggest an earlier onset of structural luteolysis in pigs caused by MXC actions in neonates. Since negative correlation analysis showed the potential interactions of miRNAs with specific messenger RNAs, we propose that these miRNAs are potential mediators of the long-term MXC effect on the CL function in pigs.

Improving ovulation in gilts using anti-inhibin serum treatment combined with fixed-time artificial insemination.

For successful batch farrowing, porcine oestrus and ovulation must be synchronized using fixed-time artificial insemination (FTAI). However, exogenous gonadotropins, which are currently used in FTAI, negatively affect gilt ovulation. Here, we aimed to improve sexually mature gilt superovulation efficiency using passive immunization against inhibin during FTAI. Altrenogest-treated gilts were challenged with 10ml anti-inhibin serum (AIS group, n=6), 1,000IU pregnant mare serum gonadotropin (PMSG group, n=6), or 10ml goat serum (control group, n=6). Gilts in the AIS and PMSG groups were inseminated according to the FTAI protocol, and gilts in the control group were inseminated during natural oestrus. When PMSG was replaced by AIS during FTAI of gilts, ovulation rate and embryos recovered were significantly greater in the AIS group as compared to the other two groups (p<.05). Especially the average number of 6-8-cell embryos in the AIS group was significantly higher than that in the PMSG group (p<.01). Moreover, the blastocyst number in the AIS group was significantly higher than that in the PMSG group and the control group (p<.05). But there was no significant difference in the blastocyst number between the PMSG group and the control group (p>.05). Besides, plasma levels of estradiol-β (E2) and progesterone (P4) were significantly greater in the AIS group as compared to the other two groups on Day 23 and D 27, respectively (p<.01). In summary, we devised an improved high-yield FTAI protocol for sexually mature gilts using AIS; this protocol had a greater superovulation efficiency than the FTAI using PMSG.

Vitamin D3 in ovarian antral follicles of mature gilts: Expression of its receptors and metabolic enzymes, concentration in follicular fluid and effect on steroid secretion in vitro

Recent studies have reported that vitamin D3 (VD) regulates ovarian function under physiological and pathological conditions. Due to a lack of information concerning the expression of VD-related molecules (receptors: VDR, PDIA3, and metabolic enzymes: CYP27B1, CYP24A1) in the porcine ovary, this research aimed to determine the mRNA expression, protein abundance and localization of VDR, PDIA3, CYP27B1 and CYP24A1 in small (SFs), medium (MFs) and large (LFs) antral follicles of sexually mature gilts. We also examined the concentration of active VD in the follicular fluid of SFs, MFs and LFs, and the effect of 1,25(OH)2D3 on their steroidogenic activity in vitro. In the present study, we have demonstrated for the first time the patterns of VDR, PDIA3, CYP27B1 and CYP24A1 immunolocalization in porcine antral follicles of different sizes. Furthermore, the expression of VD-related molecules was influenced by follicle developmental stage. VDR and PDIA3 mRNA expression and protein abundance decreased with the follicle size: they were the greatest in SFs, and the lowest in LFs. CYP27B1 mRNA expression was the highest in MFs and differed from that in SFs, whereas protein abundance was greater in MFs and SFs than in LFs. The expression of mRNA for CYP24A1 was higher in MFs than in SFs and LFs, while protein abundance did not differ between follicle classes. We have also described changes in the concentration of 1,25(OH)2D3 in the follicular fluid of antral follicles with its highest level in MFs. These findings show that the porcine antral follicle is a target tissue for direct VD action and is a local site of VD metabolism. Furthermore, we found that 1,25(OH)2D3 increased the secretion of progesterone and estradiol-17β by SFs and MFs in vitro, implying a crucial role of VD in the regulation of ovarian steroidogenesis in mature gilts. Therefore, VD appears to be an important intraovarian factor that could regulate follicular development and function in pigs.

Effect of neonatal exposure to endocrine-active compounds on epigenetic regulation of gene expression in corpus luteum of gilts

Recently, we have demonstrated that neonatal exposure to environmental endocrine-active compounds (EACs) with androgenic/antiandrogenic and estrogenic/antiestrogenic activities led to morphological and functional changes in the porcine corpus luteum (CL). To gain insight into the regulatory mechanisms of the long-term effects of EACs, we analyzed the impact of neonatal exposure of such compounds on global DNA methylation and the expression of miRNA biogenesis components in the porcine CL. Piglets were injected subcutaneously with testosterone propionate (TP, an androgen), flutamide (FLU, an antiandrogen), 4-tert-octylphenol (OP, an estrogenic compound), ICI 182,780 (ICI, an antiestrogen), methoxychlor (MXC, a compound with mixed activities) or corn oil (control) between postnatal days 1 and 10 (n = 5/group). The CLs from sexually mature gilts were examined for global DNA methylation and for the abundance of proteins related to DNA methylation (DNMT1, DNMT3A and DNMT3B) and miRNA biogenesis (DROSHA, XPO5, DICER1, AGO2) using an enzyme-linked immunosorbent assay, Western blotting and immunohistochemical staining. ICI and MXC increased the global DNA methylation levels and DNMT1 protein abundance in the luteal tissue. OP treatment led to a lower DROSHA protein abundance, while ICI treatment resulted in a greater DROSHA protein abundance. Both FLU and ICI increased DICER1 protein abundance in the luteal tissue. In addition, XPO5 showed immunolocalization exclusively in small luteal cells in the OP-treated pigs, in contrast to localization in both small and large luteal cells in the controls. In conclusion, the changes in DNA methylation, as well as the altered miRNA biogenesis components, seem to be a part of the regulatory network that mediates the long-term effects of EACs on CL function in pigs.

Altrenogest affects the development and endocrine milieu of ovarian follicles in prepubertal and mature gilts†.

Altrenogest with gonadotropins is commonly used to synchronize the estrous cycle, but it can also lead to follicular cyst formation, especially in prepubertal gilts. Here, we aimed to investigate how maturity and altrenogest treatment affect the development, endocrine milieu, and molecular control of ovarian follicles. Crossbred prepubertal and mature gilts were challenged or not (control) with altrenogest, and ovaries were collected in the morning on the first day of behavioral estrus. In prepubertal gilts, altrenogest decreased the percentage of primordial and atretic small follicles, but increased large antral follicles when compared with controls. In mature gilts, altrenogest reduced the percentage of primary follicles and elevated the total number of antral follicles. Maturity affected the estradiol level in the follicular fluid of preovulatory follicles, luteinizing hormone (LH)-stimulated cyclic adenosine monophosphate (cAMP) generation, and LH receptor messenger RNA (mRNA) expression in granulosa. Moreover, cytochrome P45017A1 (CYP17A1) mRNA levels in the theca layer were affected and correlated with follicular androstendione and estradiol concentration. Altrenogest negatively affected follicular fluid progesterone concentration and decreased levels of prostaglandin (PG) E2 in prepubertal gilts and PGF2alpha metabolite in mature gilts. LH-stimulated cAMP release in granulosa cells of mature gilts as well as human chorionic gonadotropin- and forskolin-induced cAMP were also affected. In addition, altrenogest downregulated CYP17A1 mRNA in the prepubertal theca layer and PGF2alpha synthase expression in the granulosa and theca layer of mature gilts. To the best of our knowledge, this is the first study to report multiple effects of maturity and altrenogest on the endocrine milieu and molecular regulations governing ovarian follicle development in gilts.

Expression of chemerin and its receptors in the ovaries of prepubertal and mature gilts.

Recent studies have demonstrated that chemerin participates in the regulation of female reproductive function at the level of the ovaries. Due to the lack of data concerning the presence of the chemerin system (chemerin and its receptors: CMKLR1, GPR1, CCRL2) in the ovaries of pigs, one of the most economically important livestock species, the aim of this study was to investigate the expression and localization of chemerin and its receptors in the ovaries of prepubertal and mature gilts. We also aimed to examine the concentrations of chemerin in the follicular fluid of prepubertal and mature animals. In the present study, we have demonstrated the expression patterns of chemerin system components in the porcine follicles of different sizes of prepubertal and mature animals, as well as in corpora lutea of mature gilts during the estrous cycle and early pregnancy. The obtained results suggest that the expression of chemerin system components is influenced by the reproductive stage, cell type, and the hormonal status of gilts (the estrous cycle/pregnancy). We have also presented the localization of the chemerin system components in various ovarian structures, and also showed changes in the concentration of chemerin in the follicular fluid of pigs. The presented findings not only confirm that chemerin is produced locally in the porcine ovarybut they also demonstrate that chemerin directly affects ovarian cells, as confirmed by the presence of chemerin receptors in all ovarian structures. Therefore, chemerin appears to be an important intra-ovarian factor that could regulate ovary function in pigs.

Transcription Analysis of the Chemerin Impact on Gene Expression Profile in the Luteal Cells of Gilts.

Chemerin is a recently discovered adipokine that participates in the regulation of many physiological and disorder-related processes in mammals, including metabolism, inflammatory reactions, obesity, and reproduction. We investigated how chemerin affects the transcriptome profile of porcine luteal cells. The luteal cells were acquired from mature gilts. After the in vitro culturing with and without chemerin, the total RNAs were isolated and high-throughput sequencing was performed. Obtained datasets were processed using bioinformatic tools. The study revealed 509 differentially expressed genes under the chemerin influence. Their products take part in many processes, important for the functions of the corpus luteum, such as steroids and prostaglandins synthesis, NF-κB and JAK/STAT signal transducing pathways, and apoptosis. The expression of the CASP3, HSD3B7, IL1B, and PTGS2 genes, due to their important role in the physiology of the corpus luteum, was validated using the quantitative real-time polymerase chain reaction (qPCR) method. The qPCR confirmed the changes of gene expression. Chemerin in physiological concentrations significantly affects the expression of many genes in luteal cells of pigs, which is likely to result in modification of physiological processes related to reproduction.

A transcriptome approach evaluating effects of neonatal androgen and anti-androgen treatments on regulation of luteal function in sexually mature pigs

The current study was designed to gain insights into regulatory mechanisms mediating long-term effects of androgen excess or deficiency on corpus luteum function in pigs. Piglets were injected subcutaneously with testosterone propionate (TP, an androgen), flutamide (FLU, an anti-androgen) or corn oil (control) between postnatal Days 1 and 10. Corpora lutea from sexually mature gilts were examined for luteal steroid concentrations and processed for total RNA isolation and subsequent RNA sequencing to determine abundances of mRNA transcripts and microRNAs (miRNAs). Potential miRNA-mRNA interactions were explored in silico. Androstenedione, testosterone and estrone concentrations in corpora lutea were altered due to the disrupted androgen action in neonates. The luteal tissue had 465 and 353 genes for which there were differential mRNA abundances as compared with the control group (P-adjusted < 0.05; log2FC ≥ 1.0) in response to neonatal TP and FLU piglet treatments, respectively. Disruption of androgen signalling in neonates affected mRNA transcript abundance, as compared with the control group, for genes associated with apoptosis, angiogenesis and immune functions in the corpora lutea. Furthermore, there was a differential abundance of a group of miRNAs in the treatment groups compared with the control group. These results indicate the neonatal androgenic milieu affects the onset of luteolysis when these animals are sexually mature, although mechanisms for responses to TP or FLU likely differ. It is proposed that changes in specific miRNAs and mRNAs may, in part, account for long-term effects of androgen excess or androgen deficiency on corpus luteum function in pigs.

Effects of feeding melatonin during proestrus and early gestation to gilts and parity 1 sows to minimize effects of seasonal infertility1.

This study tested whether supplemental melatonin given to mimic the extended nighttime melatonin pattern observed in the higher fertility winter season could minimize infertility during summer and fall in swine. Exogenous melatonin was fed during periods coinciding with follicle selection, corpus luteum formation, pregnancy recognition, and early embryo survival. Experiments were conducted at a commercial farm in 12 sequential replicates. In Exp. 1a, mature gilts (n = 420) that had expressed a second estrus were assigned by weight to receive once daily oral Melatonin (MEL, 3 mg) or Control (CON, placebo) at 1400 h for 3 wk starting before insemination at third estrus. In Exp. 1b, parity 1 sows (n = 470) were randomly assigned by lactation length to receive MEL or CON for 3 wk, starting 2 d before weaning. Follicles, estrus, pregnancy, and farrowing data were analyzed for the main effects of treatment, season (4-wk periods), and their interaction. Environmental measures were also analyzed for reproductive responses. In Exp. 1a, there was no effect (P > 0.10) of MEL on age at third estrus (203 d), follicle size after 7 d of treatment (5.0 mm), estrous cycle length (22.6 d), return to service (9.2%), farrowing rate (FR, 80.0%), or total born pigs (TB, 13.6). However, there was an effect of season (P = 0.03) on number of follicles and on gilts expressing estrus within 23 d of the previous estrus (P < 0.005). In Exp. 1b, there was no effect of MEL (P > 0.10) on follicle measures, wean to estrous interval, FR (84.0%), or TB (13.0). But MEL (73.5%) reduced (P = 0.03) estrous expression within 7 d of weaning compared with CON (82.0%) and season (P = 0.001) decreased FR by ~14.0% during mid summer. Also, gilts and parity 1 sows exposed to low light intensity (<45 lx) during breeding had reduced conception (-8%) and farrowing (-15%) rates, compared with higher light intensity. Similarly, high temperatures (>25 °C) during breeding also reduced gilt conception rates by 7%. Although there was clear evidence of seasonal fertility failures in gilts and sows, MEL treatment did not improve fertility in gilts and reduced estrus in parity 1 sows. It is possible that differences in lighting and thermal environments before breeding could explain the differential response to MEL in sows and gilts.

Ovarian Activity and the Size of Oocytes in Ovarian Follicles in Sexually Mature Gilts in Individual Seasons

Abstract The aim of this study was to evaluate ovarian activity and the size of oocytes in ovarian follicles in sexually mature Landrace-Yorkshire gilts in relation to the individual seasons of the year. The study was carried out on 240 gilts slaughtered at an abattoir during the four yearly seasons. The size and weight of the ovaries, the number of follicles and corpora lutea (CL) according to individual size categories were evaluated. The oocytes were aspirated from follicles and their sizes were measured. Our evaluation of the size of the ovaries showed that they were the largest in autumn, when their mean length reached 25.8 ± 3.4 mm, while in winter their mean length was 24.2 ± 2.9 mm. The smallest weight of the ovaries was determined in autumn (mean 5.7 ± 1.4 g) and the highest in spring (mean 6.2 ± 2.2 g). The largest number of follicles in the ovaries of the gilts was recorded during the autumn months, with a predominance of follicles up to 3 mm (mean number 17.9 ± 7.5). The largest number of corpora lutea was observed in spring (mean number 12.1 ± 2.6) and the smallest in winter (mean number 6.1 ± 1.1). The oocytes from follicles of up to 3 mm size, were the smallest in spring (mean size 16.99 × 103 ± 3.42 × 103 µm2) and the largest in winter (mean size 18.90 × 103 ± 2.99 × 103 µm2). In total, the largest oocytes were aspirated from 4—6 mm follicles in autumn (mean size 19.60 × 103 ± 5.37 × 103 ± µm2). The values recorded indicated that the seasons affected the ovarian activity and the growth of oocytes in gilts.

Effect of neonatal exposure to endocrine-active compounds on plasma lipid and steroid concentrations, and morphology of luteal tissue in the adult pig

To study the long-term impact of neonatal exposure to endocrine-active compounds (EACs) on plasma lipid profiles, steroid concentrations, and morphology of porcine luteal tissue, piglets were injected with testosterone propionate (TP), flutamide (FLU), 4-tert-octylphenol, ICI 182,780 (ICI), methoxychlor, or corn oil (controls) between postnatal days 1 and 10 (n = 5/group). Blood samples and corpora lutea were obtained from sexually mature gilts. The investigated compounds differentially affected plasma lipid and steroid concentrations. Moreover, we demonstrated hypertrophy of luteal cells after neonatal EAC administration. In addition, a predominant abundance of lipid droplets was found in luteal cells of TP-, FLU-, and ICI-treated animals. It seems that the pathways leading to changes in the plasma lipid profile may contribute to the development of long-term alterations that have the potential to affect luteal steroidogenic capability in pigs.

Endometritis affects chemical coding of the dorsal root ganglia neurons supplying uterus in the sexually mature gilts

This study reports on the influence of experimentally-induced uterine inflammation on chemical phenotypes, number and distribution of neurons in the dorsal root ganglia (DRGs) innervating the uterus in sexually mature gilts. On day 17 of the first studied estrous cycle, the uterine horns were injected with retrograde tracer Fast Blue (FB). After 28 days (on an expected day 3 of third studied estrous cycle), 50 ml of either saline (group SAL) or Escherichia coli (E. coli) suspension (109 colony-forming units/ml, group E. coli) were injected into each uterine horn. In the control pigs (group CON), only laparotomy was performed. Eight days later DRGs and uteri were collected. All infected gilts developed severe form of acute endometritis. By use of double immunofluorescence labelling the numbers of uterine perikarya expressing substance P (SP), calcitonin gene-related peptide (CGRP), neurokinin A (NKA), galanin (GAL) and pituitary adenylate cyclase-activating polypeptide (PACAP) were analyzed. Injection of E. coli decreased the total number of the FB positive perykaria in the Th10-S4 DRGs. We revealed an increase in the populations of uterine perikarya coded SP+/CGRP−, SP+/NKA−, SP−/NKA+, SP+/GAL+, SP+/GAL−, SP−/GAL+, SP+/PACAP+ and SP−/PACAP+. Our results suggest that uterine inflammation affects both the spatial and neurochemical organization pattern of uterine sensory innervation. Additionally, the inflammation may affect the transmission of sensory information from uterus to spinal cord.

Presence of trophoblast in the uterine lumen affects VEGF-C expression in porcine endometrium

Early pregnancy is associated with morphological and functional changes within the uterus, accompanied by angiogenesis, increased vascular permeability and activation of immune tolerance. Intensive angiogenesis leads to accelerated vascular leakage and accumulation of interstitial fluid in endometrium. To protect the trophoblast from the harmful effect of extracellular fluid, process known as lymphangiogenesis is crucial. These studies are focused on VEGF-C, factor responsible of lymphatic vessels creating, and its receptors: Flk1 (VEGFR2) and Flt4 (VEGFR3) during the time of implantation as well as the effect of trophoblast signals (IFNG and E2) on VEGF-C production. Endometrial samples were collected from mature gilts from days 8, 10, 12, 14 of estrous cycle and pregnancy. Real-Time PCR analysis revealed increased mRNA expression of VEGF-C on days 10, 12, 14 of pregnancy compared to corresponding days of estrous cycle. The highest VEGF-C mRNA expression was observed on 14 day of pregnancy (p < 0.05). Increased mRNA expression of Flk1 and Flt4 was noticed on day 14 of pregnancy in comparison to day 10. Enhanced Flk1 mRNA expression during 14 day of pregnancy was observed compared to corresponding day of estrous cycle (p < 0.05). No significant difference on the protein level was revealed. VEGF-C and its receptors were localized mainly in luminal and glandular epithelial cells, but their presence were confirmed also in endothelial cells of blood and lymphatic vessels and 14 d trophoblasts. In vitro studies revealed positive effect of IFNG on VEGF-C mRNA expression in stromal cells and protein content in medium after stromal cells culture (p < 0.05). Our studies demonstrated the presence of VEGF-C system in porcine endometrium and indicated its possible important role during the time of implantation.