Estrous Cycle In Pigs Research Articles

Transcriptome profiling of different developmental stages of corpus luteum during the estrous cycle in pigs

To better understand the molecular basis of corpus luteum (CL) development and function RNA-Seq was utilized to identify differentially expressed genes (DEGs) in porcine CL during different physiological stages of the estrous cycle viz. early (EL), mid (ML), late (LL) and regressed (R) luteal. Stage wise comparisons obtained 717 (EL vs. ML), 568 (EL vs. LL), 527 (EL vs. R), 786 (ML vs. LL), 474 (ML vs. R) and 534 (LL vs. R) DEGs with log2(FC) ≥1 and p < 0.05. The process of angiogenesis, steroidogenesis, signal transduction, translation, cell proliferation and tissue remodelling were significantly (p < 0.05) enriched in EL, ML and LL stages, where as apoptosis was most active in regressed stage. Pathway analysis revealed that most annotated genes were associated with lipid metabolism, translation, immune and endocrine system pathways depicting intra-luteal control of diverse CL function. The network analysis identified genes AR, FOS, CDKN1A, which were likely the novel hub genes regulating CL physiology.

Progesterone inhibits cytokine/TNF-α production by porcine CL macrophages via the genomic progesterone receptor

In pigs, luteolytic sensitivity to PGF-2α (=LS) is delayed until d 13 of the estrous cycle. While the control of LS is unknown, it is temporally associated with macrophage (MAC; which secretes tumor necrosis factor [TNF]-α) infiltration into the corpora lutea (CL), and previous studies have shown that TNF-α induces LS in porcine luteal cells (LCs) in culture. This study was designed to explore the control of LS by CL macrophage (CL MAC)/TNF-α by progesterone (P4), and to examine the hypothesis that P4 acting via the genomic P4 receptor (PGR) inhibits CL MAC TNF-α and thus plays a key role in regulating LS during the pig estrous cycle. In experiment 1, the effects of LCs on CL MAC cytokine/TNF-α mRNA expression in co-culture were examined (MID cycle; ~d 7–12; no LS); results showed that LC was inhibitory to cytokine/TNF-α. In experiment 2, the effects of P4 or R5020 (PGR-agonist) on CL MAC cytokine/TNF-α mRNA expression were examined (MID cycle; ~d 7–12; no LS); results showed that both P4 and R5020 dose-dependently inhibited TNF-α. In experiment 3, CL MACs were isolated from CL at MID (~d 7–12; no LS) and LATE (~d 13–18; + LS) cycle, and TNF-α/PGR mRNA measured. Results indicated that while TNF-α mRNA was 4.2-fold greater in CL MACs from LATE vs MID cycle, PGR mRNA was 4.5-fold greater in CL MACs from MID vs LATE cycle. These data support our hypothesis and suggest that progesterone, acting via PGR, plays a critical physiological role in the control of TNF-α production by CL MACs and LS during the pig estrous cycle.

Conjugated estrogens in the endometrium during the estrous cycle in pigs

Estrogen metabolism results in the formation of inactive estrogen sulphates and glucuronides. Despite the lack of receptor binding, circulating conjugated estrogens might serve as a reservoir for the active form through the involvement of specific cleaving enzymes. In order to elucidate the potential role that estrogen conjugates play in the regulation of the estrous cycle, we determined the concentration of progesterone, estrogen and estrogen conjugates in serum and endometrial homogenates of cycling gilts. In addition, we determined the mRNA expression changes of enzymes (UDP glucuronosyltransferase (UGT), β-glucuronidase (GUSB), sulphotransferases (SULT) and steroid sulphatase (STS)) and transporters (multidrug resistance-associated protein (MRP), organic anion-transporting polypeptide (OATPs)) involved in the estrogen metabolism in the endometrium across the estrous cycle. GUSB displayed highest expression at estrous (day 0), decreasing expression during metestrus (day 3 and 6), minimal expression on day 10 and 12, and increasing expression towards proestrus (day 18), suggesting either a stimulation by estrogens or a negative impact of progesterone. The mRNA expression of the influx-transporter OATP1A2 significantly increased from day 0 to 6 and decreased again by day 10, while the efflux-transporters (MRP1, MRP2, and MDR1) displayed minimal expression at day 3 and 6. The mRNA expression of the UDP-glucuronsyltransferases followed a similar pattern, with minimal expression found at day 6. The analyses of the concentration of local and circulating steroid hormones points towards an interaction of the analyzed transporters and enzymes with steroid hormones, thereby possibly regulating the reservoir of active steroids contributing to the endometrial function.

The expression of VEGF, myoglobin and CRP2 proteins regulating endometrial remodeling in the porcine endometrial tissues during follicular and luteal phase.

Endometrial remodeling is important for successful embryo development and implantation in pigs. Therefore, this study investigated change of proteins regulating endometrial remodeling on follicular and luteal phase in porcine endometrial tissues. The endometrial tissue samples were collected from porcine uterus during follicular and luteal phase, vascular endothelial growth factor (VEGF), myoglobin and cysteine-rich protein 2 (CRP2) proteins were expressed by immnofluorescence, immunoblotting, and determined by 2-DE and MALDI-TOF/MS. We found that VEGF, myoglobin and CRP2 were strongly localized in endometrial tissues during luteal phase, but not follicular phase. The protein levels of VEGF, myoglobin and CRP2 in endometrial tissues were higher than luteal phase (P<0.05). These results may provide understanding of intrauterine environment during estrous cycle in pigs, and will be used in animal reproduction for developing specific biomarkers in the future.

Pro-inflammatory Cytokines and Their Receptors: Expression and Regulation in the Uterine Endometrium during the Estrous Cycle in Pigs

Pro-inflammatory Cytokines and Their Receptors: Expression and Regulation in the Uterine Endometrium during the Estrous Cycle in Pigs

The effect of interleukin-1β, interleukin-6, and tumor necrosis factor-α on estradiol-17β release in the myometrium: The in vitro study on the pig model

Estradiol-17β (E2) is a potent regulator of early pregnancy and the estrous cycle in pigs. Production of E2 occurs in the porcine myometrium, but the factors involved in its regulation are unknown. In this in vitro study, it was investigated whether interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α affect the release of E2 from the porcine myometrium on Days 10 to 11, 12 to 13, and 15 to 16 of pregnancy and the estrous cycle. The expression of the cytochrome P450 family 19 (CYP19) gene and the presence of the aromatase cytochrome P450 protein in the myometrium confirmed the ability of the tissue to produce E2. In gravid pigs, the expression of IL1RI mRNA and IL6R mRNA was markedly increased on Days 15 to 16 of gestation, whereas TNFRI mRNA was increased on Days 10 to 11 of gestation. In cyclic pigs, the expression of myometrial IL1RI mRNA did not differ among the studied days, although the expression of IL6R and TNFRI mRNAs was increased on Days 15 to 16. In gravid pigs, IL-1β, IL-6, and TNF-α increased myometrial E2 secretion on Days 15 to 16 but did not affect E2 release on Days 10 to 11 and 12 to 13 of pregnancy. In cyclic pigs, IL-1β, IL-6, and TNF-α did not increase myometrial E2 release. In conclusion, IL-1β, IL-6, and TNF-α affected myometrial E2 release in a manner that is dependent on the physiologic status of the female. The porcine myometrium expresses IL1RI, IL6R, and TNFRI genes and is the target tissue for IL-1β, IL-6, and TNF-α. In gravid pigs, IL-1β, IL-6, and TNF-α may increase myometrial release of E2in vitro specifically on Days 15 to 16 of pregnancy. These findings may be of interest to researchers using pigs as an animal model for fetal programming.

The Activity and Localization of 3β-hydroxysteroid Dehydrogenase/Δ&lt;sup&gt;5&lt;/sup&gt;-Δ&lt;sup&gt;4&lt;/sup&gt; Isomerase and Release of Androstenedione and Progesterone by Uterine Tissues During Early Pregnancy and the Estrous Cycle in Pigs

Steroid hormones are produced by the porcine uterus. We hypothesized that the uterus in pigs possesses active 3β-hydroxysteroid dehydrogenase/Δ5-Δ4 isomerase (3β-HSD) responsible for progesterone and androstenedione production, that uterine steroids may supplement the amount of steroid hormones produced by embryos and corpus luteum and that these steroids are necessary for maintenance of pregnancy. In this study, we examined 1) endometrial and myometrial expression of 3β-HSD mRNA, 2) uterine 3β-HSD protein activity and 3) in vitro production of A4 and P4 by uterine slices harvested from pigs on days 10 to 11, 12 to 13 and 15 to 16 of pregnancy and the estrous cycle. The expression of 3β-HSD and the presence and activity of 3β-HSD protein were different in the endometrium and the myometrium during the examined periods of pregnancy and the estrous cycle. Production of A4 by the endometrium and myometrium was highest on days 12 to 13 of pregnancy and the estrous cycle. Endometrial secretion of P4 did not differ in the course of early pregnancy and on the respective days of the estrous cycle. The gravid myometrium was the highest source of P4 in pregnant pigs on days 12 to 13. The release of P4 by the cyclic myometrium rose during the examined days of the estrous cycle. The steroidogenic activity of the uterus, as described in this study, may support early pregnancy or the luteal phase of the estrous cycle in pigs.

Expression of aquaporin 1 in the pig peri-ovarian vascular complex during the estrous cycle and early pregnancy

Aquaporin 1 (AQP1) is a water channel protein expressed in endothelial and epithelial cells of many tissues, including the vasculature, where it serves to increase water permeability of the cell membrane. The aim of this study was to investigate the expression and distribution of AQP1 in porcine peri-ovarian vascular complex (PVC) during the estrous cycle and early pregnancy. Immunohistochemistry and semi-quantitative immunoblotting techniques were used. We have demonstrated the presence of AQP1 protein in the endothelial cells of the lymphatic and vascular endothelium of the PVC during the pig estrous cycle and early pregnancy. The expression of AQP1 protein in the PVC did not change significantly between Days 10-12 and 14-16, but increased on Days 2-4 and 18-20 when compared with Days 10-12 and 14-16 of the estrous cycle. In pregnant gilts, the expression of AQP1 did not differ significantly during the onset and the end of the implantation process and also when compared to the mid- and late-luteal phases of the estrous cycle. In conclusion, AQP1 is expressed in the endothelial cells of PVC and may modulate hormonal regulation of reproductive organs.

396 CHANGES OF PLASMINOGEN ACTIVATOR ACTIVITY IN OVIDUCTAL EPITHELIAL CELLS DURING THE ESTROUS CYCLE IN PIGS

Plasminogen activator (PA) activity in rat, porcine, and bovine embryos and in oocytes have been demonstrated, but reports of PA activity in the uterus and oviduct are limited. The present study was undertaken to identify changes of PAs in porcine oviductal epithelial cells (pOECs) during the estrous cycle in pigs. pOECs obtained from pigs in post-ovulatory (Post-Ov), early-to-mid luteal (Early-mid L), and pre-ovulatory (Pre-Ov) stages were washed twice with Hank&amp;apos;s balanced salt solution (HBSS); the oviductal lumen was flushed with 10 mL HBSS, and centrifuged at 300g for 10 min at 4�C. The collected pOECs were washed twice with DMEM containing 0.1% BSA and centrifuged at 300g for 10 min at 4�C. To examine the changes in activity of PAs in pOEC, 2.0 � 105 fresh cells were cultured in a 4-well dish with DMEM/Ham&amp;apos;s F-12 medium containing inactivated 10% FBS and amphotericin under a humidified atmosphere of 5% CO2 in air at 38�C for 1, 3, 5, and 7 days. The culture medium was replaced every 48 h with fresh medium during culture for 7 days, after which cells and conditioned medium were stored separately at -20�C until they were used for zymographic analysis. PA activities of pOECs were measured using SDS-PAGE, casein-agar zymography, and densitometry. The urokinase-type PA (uPA) was observed at 5–7 days of culture in pOECs of different stages of the estrous cycle in pigs. On the other hand, uPA, tissue-type (tPA), and tPA-PA inhibitor (tPA-PAI) activities were observed at days 1, 3, 5, and 7 of culture in medium conditioned with pOEC from different stages of the estrous cycle. uPA activity was decreased during prolonged culture of Post-Ov pOECs. At this time, uPA activity was significantly (P &amp;lt; 0.05) higher at 1 day than at 3–7 days of culture in Post-Ov pOEC-conditioned medium. tPA activity was significantly (P &amp;lt; 0.05) higher at 5 days than at 1–3 days or 7 days of culture in Post-Ov pOEC-conditioned medium. tPA-PAI activity was significantly (P &amp;lt; 0.05) higher at 7 days than at 5 days of culture in Pre-Ov pOEC-conditioned medium, and tPA-PAI activity was significantly (P &amp;lt; 0.05) higher at Post-Ov than Pre-Ov or Early-mid L in 7 days of culture in pOEC-conditioned medium of different stages of the estrous cycle. All tPA activities were increased until Day 5 and were decreased at Day 7 of culture in pOEC-conditioned medium. uPA activity was observed in pOEC-conditioned medium throughout the estrous cycle. uPA activity was significantly (P &amp;lt; 0.05) different during culture of Post-Ov in pOECs-conditioned medium. tPA-PAI activity was observed at all stages, and tPA-PAI activity was significantly (P &amp;lt; 0.05) different among different stages of the estrous cycle. These results suggest that uPA, tPA, and tPA-PAI are produced by pOEC, and that variations of PA activity are found at different stages of the estrous cycle in porcine oviductal epithelial cells. This work was supported by grant No. R01-2003-000-10500-0 from the Basic Research Program of Korea Science &amp;amp; Engineering Foundation.

PGF&lt;SUB&gt;2&amp;alpha;&lt;/SUB&gt;の反復投与による豚の発情周期の短縮とその繁殖成績

PGF&lt;SUB&gt;2&amp;alpha;&lt;/SUB&gt;の反復投与による豚の発情周期の短縮とその繁殖成績

Cyclic Changes in Serum Follicle-Stimulating Hormone, Luteinizing Hormone and Inhibin During the Guinea Pig Estrous Cycle.

We proposed to describe the cyclic changes in serum follicle-stimulating hormone (FSH), luteinizing hormone (LH) and inhibin during the guinea pig estrous cycle. Serum gonadotropin concentrations had not been determined routinely because antibodies to guinea pig gonadotropins were rarely available. Serum gonadotropin concentrations provide important information about the stimuli received by the ovary. Inhibin plays a major role in modulating FSH synthesis and secretion at the anterior pituitary; and inhibin concentrations can be used as a marker for follicular development. We used only sexually mature guinea pigs that showed at least two consecutive normal 15-17 day cycles. The day of maximal vaginal cornification, the day of ovulation, was designated as day 0. Blood samples were taken every other day at 0900 hours throughout one estrous cycle. Serum was analyzed for FSH, LH, and inhibin by radioimmunoassay. Cyclic changes in peripheral FSH, LH, and inhibin concentrations were observed. Serum inhibin rose to a peak on day 7 of the guinea pig estrous cycle. Inhibin increased again, although not significantly, on day 13. Thus, this study confirms the biphasic phenomenon of ovarian follicle development in guinea pigs. Since serum FSH concentrations declined as serum inhibin concentrations reached a peak, it is also suggested that inhibin regulates FSH secretion in this species. This is the first study in which all three hormones were measured during the estrous cycle of the guinea pig.

Coordinate developmental expression of genes regulating sterol economy and cholesterol side-chain cleavage in the porcine ovary.

To investigate the coordinate developmental expression of low-density lipoprotein (LDL) receptor, 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, sterol carrier protein 2 (SCP2), steroidogenic acute regulatory protein (StAR), and cytochrome P450 side-chain cleavage (P450scc) enzyme messages throughout the pig estrous cycle, RNase protection analysis was performed using homologous (partially cloned) porcine sequences. Total RNA was isolated from ovarian tissues from unstimulated prepubertal gilts and gilts stimulated with eCG (Day -3) and hCG (Day 0) to induce follicular growth and ovulation. Specific transcripts (relative to 18S rRNA) were quantified in immature ovaries, preovulatory follicles (> or = 5 mm), corpora lutea (CL), and corpora albicantia. As an index of steroidogenesis, tissue progesterone content (per microgram protein) was low in the unstimulated ovary and preovulatory follicles, and it began to increase 4 days post-hCG, peaked at 12 days, and returned to preovulatory concentrations by 20 days post-hCG. HMG-CoA reductase mRNA was expressed at low levels and did not change significantly throughout the estrous cycle. The amount of LDL receptor mRNA increased approximately 6-fold after eCG stimulation and was expressed at similar concentrations in both preovulatory follicles and functional CL. Expression of SCP2 mRNA did not differ among the four tissue types but tended to be highest in midcycle (Day 12) CL compared other stages of CL (p = 0.007). StAR mRNA expression was minimal in unstimulated ovaries, was higher in preovulatory follicles (p = 0.014), and then rose again in CL (p = 0.009 compared with unstimulated ovary). P450scc mRNA concentrations were low in unstimulated ovaries, increased in preovulatory follicles (p = 0.044), and increased further in CL (p = 0.001 compared with preovulatory follicles). P450scc and StAR mRNA levels correlated with progesterone levels (r = +0.37, p = 0.025, and r = +0.71, p < 0.001, respectively). The expression of LDL receptor, StAR, and P450scc messages showed a dramatic decline by Day 20 post-hCG (p = 0.002, p = 0.003, p = 0.006, respectively, compared with CL) corresponding with functional regression of the CL. In summary, P450scc and StAR message expression are coordinately amplified during the pig follicular and luteal phase, whereas LDL receptor message after an initial increase is expressed at constitutively high levels, thus indicating a differential regulation of ovarian sterol-metabolizing genes during the steroidogenic life of the follicle and CL.

Pharmacological characterization of α1-adrenoceptors in porcine uterine artery

This study was undertaken to ascertain if changes in the affinity to α 1-adrenoceptor agonists and antagonists could explain the increase in uterine artery vasoconstriction to adrenergic stimuli during the luteal phase of the estrous cycle in pigs. We also sought to determine the subtype(s) of adrenoceptor ( α 1A and α 1B) in the porcine uterine artery. When phenylephrine was the agonist, uterine artery pA 2 values for prazosin were 8.98, 9.04 and 9.10 in the luteal and follicular phases and in early pregnancy, respectively. The K A (dissociation constant) values for phenylephrine were 6.5, 3.7 and 4.4 (μM) in the luteal and follicular phases and in early pregnancy, respectively. The use of the putative α 1A-adrenoceptor WB4101 (2-[[[2-(2,6-dimethoxyphenoxy)ethyl]amino]methyl]-1,4-benzodioxane) and the α 1B-adrenoceptor antagonist chloroethylclonidine indicated that both α 1A- and α 1B-adrenoceptors are present in the porcine uterine artery and that a similar magnitude of inhibition of responses to noradrenaline by each of the antagonists occurred in arteries from the luteal and follicular phases and in early pregnancy. This suggests that the α 1A : α 1B adrenoceptor ratio does not vary significantly during the estrous cycle and in early pregnancy. This study indicates that porcine uterine artery α 1-adrenoceptor affinity is similar in the luteal and follicular phases of the estrous cycle and in early pregnancy.

Follicular Atresia, Follicular Fluid Hormones, and Circulating Hormones During the Midluteal Phase of the Estrous Cycle in Pigs1

The incidence of atresia, concentrations of follicular fluid steroids, and circulating concentrations of estradiol, FSH, and progesterone were investigated to determine whether there was any evidence for a wave of follicular activity or atresia between Days 7 and 15 of the estrous cycle in pigs, Cyclic pigs (gilts) were slaughtered on Days 7, 9, 11, 13, and 15 (4, 4, 4, 4, and 3 pigs per day), and 287 follicles > 2 mm were dissected from ovaries to recover follicular fluid and granulosa cells. Apoptotic (Ao) granulosa cells were those containing sub-diploid DNA fluorescence as determined by flow cytometry. follicles with > or = 10% Ao granulosa cells were classified as atretic, and those with < 10% Ao granulosa cells were designated nonatretic. The percentage of atretic follicles (AF) per pig was 35.5 +/- 4.7 (mean +/- SEM), and while the percentage of AF ranged from 12% to 73%, means did not differ significantly among days. Mean follicular estradiol concentration was higher (p < 0.05) in nonatretic follicles (NAF) than in AF; in NAF, the concentration decreased (p < 0.05) from 15 ng/ml on Day 7 to 2.5 ng/ml on Day 11 and then increased (p < 0.05) to 10 ng/ml on Day 15 of the cycle. Mean follicular progesterone concentration was higher (p < 0.05) in AF than in NAF, while follicular androstenedione concentration (20 +/- 1.5 ng/ml, overall mean +/- SEM) did not differ between AF and NAF. Mean plasma estradiol, FSH, and progesterone concentrations did not differ among days of the cycle or differ significantly during the last 42 h before slaughter. Plasma estradiol and FSH concentrations were not significantly correlated with percentage of AF or with follicular steroid concentrations. These results support the notion of continuous growth and atresia of ovarian follicles in pigs independent of changes in plasma FSH and follicular steroid concentration, without evidence for a dominant cohort of follicles during the luteal phase of the estrous cycle.

Immunohistochemical localization of 3 beta-hydroxysteroid dehydrogenase and P450 17 alpha-hydroxylase during follicular and luteal development in pigs, sheep, and cows.

Follicular and luteal morphology and steroidogenic function were investigated by immunohistochemistry for cytochrome P450 17 alpha-hydroxylase (P450c17) and 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) during the estrous cycle in pigs, sheep, and cows. The theca interna of all species expressed P450c17 during follicular development. In the pig, this constituted a continuous layer of cells around the follicle, but a sheath of cells lining the basement membrane appeared not to express P450c17. Neither was expression of P450c17 in ovine and bovine follicles uniform throughout the theca interna. In these two species, a beaded appearance was given by P450c17, since it was expressed in some regions but not in others. Therefore, staining for P450c17 defined functional sub-populations of cells within the theca interna of pigs, sheep, and cows. Ovulation was associated with a decrease in P450c17 in all species, but some expression persisted in theca-derived cells of developing and mature porcine CL. Expression of 3 beta-HSD in the preovulatory follicle was confined to the theca of the pig and sheep; in contrast, in the cow, it was highest in the granulosa. In general, 3 beta-HSD expression appeared to be greater in porcine than ovine or bovine follicles, the physiological relevance of which is discussed. Porcine and ovine theca continued to express 3 beta-HSD after ovulation, and granulosa-derived cells increased their 3 beta-HSD expression markedly as they luteinized in all three species. During early luteal development in pigs and sheep, theca-derived cells with high 3 beta-HSD encircled luteal lobules, but these cells appeared throughout the parenchyma of the mature CL. Luteal regression in sheep and cows was typified by the loss of many cells expressing 3 beta-HSD, whereas others, adjacent to them, appeared to be intact without loss of enzyme expression. These data further define differences in steroidogenesis during follicular and luteal development among the pig, sheep, and cow.

Effects of progesterone on prolactin, hypothalamic beta-endorphin, hypothalamic substance P, and midbrain serotonin in guinea pigs.

Unlike rats, but similar to primates, guinea pigs exhibit prolonged function of the corpus luteum and elevated progesterone secretion after ovulation. The gonadotropins, estrogen (E) and progesterone (P) have been examined throughout the guinea pig estrous cycle. However, neither prolactin secretion nor its regulation by steroid hormones has been characterized, perhaps due to the lack of a specific radioimmunoassay. beta-Endorphin (BE), substance P (SP), and serotonin (5-HT) increase prolactin secretion in rats and monkeys. BE and SP neurons in guinea pigs and 5-HT neurons in monkeys contain progestin receptors which could mediate neuroendocrine effects of steroid hormones. Therefore, the effects of E and P on prolactin, BE, SP, and 5-HT and its metabolite 5-HIAA were examined in guinea pigs which were ovariectomized, E treated (28 days), and E+P treated (14 days E+14 days E+P). The rat NB2 lymphoma cell line was used as a bioassay for serum prolactin. BE and SP levels were measured by radioimmunoassay in four hypothalamic areas: the preoptic region (POA), the mediobasal hypothalamus (MBH), the dorsomedial hypothalamus (DMH), and the mamillary bodies (MB). 5-HT and 5-HIAA were measured in the midbrain raphe area by high-pressure liquid chromatography. E alone had little effect on serum prolactin levels, but E+P significantly increased prolactin as compared with ovariectomized controls. The BE levels increased with E treatment and remained elevated with E+P treatment in MBH and POA. The BE content was stimulated in DMH and MB by E+P treatment and not with E alone. The SP content in MBH, DMH, and MB increased in E-treated guinea pigs.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in Follicular Populations, in Serum Estrogen and Progesterone, and in Ovarian Steroid Secretion in Vitro during the Guinea Pig Estrous Cycle12

It has been proposed that the guinea pig estrous cycle manifests biphasic follicular development. The follicles of one cohort apparently achieve their greatest diameter by approximately Day 10 of the cycle and then undergo atresia while the second cohort ovulates; this constitutes an uninvestigated and novel model for the evaluation of atresia. In this study, follicular development was evaluated in vivo and in vitro to confirm this pattern. On cycle Day 3, 5, 7, 10, 12, 15 or 0 (ovulation), guinea pigs were killed and trunk blood was collected; ovaries were excised, weighed, and measured, and size and number of large (greater than 400 microns) follicles were determined. Ovaries were quartered and placed into culture dishes for incubation. Culture variables were presence or absence of human follicle-stimulating hormone (100 ng/ml) and time. Ovarian fragments were processed for histology. Estrogen (E) and progesterone (P) in sera and culture media were analyzed by radioimmunoassay. The abundance of large follicles on both the ovarian surface (greater than 635 microns) and in histologic section (greater than 500 microns) relative to all follicles observed was high on Day 7 and Day 10, respectively; lower on Day 10 and Day 12; and high again at Day 12 and Day 15. Mean secretion of E in culture was elevated on Day 10 (253.0 +/- 60.3 pg/ml/mg ovary), low on Day 12 (67.9 +/- 13.0), and high again on Day 0 (185.8 +/- 56.8). Peripheral P reached a maximum of 2.93 +/- 0.09 ng/ml (Day 5), and then declined to Day 15.(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of antral follicle populations during the estrous cycle in pigs selected for ovulation rate.

The objectives of this study were to characterize and compare ovarian follicular populations in Gene Pool Control (GPC, randomly selected) and Relax Select line (RS, nine generations of selection for high ovulation rate followed by six generations of random selection) gilts during different stages of the estrous cycle. Thirty-five RS and 23 GPC gilts were allotted randomly within litter for ovary recovery on either d 3, 15 or 19 of the estrous cycle. Surface follicles on the ovaries were classified by size (small, less than 3 mm; medium, 3 to 6.9 mm; large, 7 to 12 mm), and counts were recorded for each ovary. Ovarian weight (OW), number of corpora lutea (CL), follicular fluid volume (FFV) from small, medium and large follicles, residual ovarian weight and follicular fluid weight (FFW) also were recorded. Total numbers of small and medium follicles were greatest on d 15, whereas total number of large follicles and FFW were greatest on d 19. The OW, FFW and follicle numbers of all classes were lowest on d 3. The RS gilts expressed longer interestrous intervals (21.9 vs 20.4 d, P less than .05) and higher ovulation rates (18.5 vs 15.3 CL, P less than .01) than GPC gilts. The left ovary of RS gilts was responsible for most of the ovulation rate advantage (10.3 vs 7.4 CL, P less than .01) Overall, GPC gilts had more total small follicles than RS gilts (P less than .01). The advantage was due primarily to higher numbers of small follicles at d 15.(ABSTRACT TRUNCATED AT 250 WORDS)

Catecholamines in sow graafian follicles at proestrus and at diestrus.

Endogenous dopamine, noradrenaline, and adrenaline were detected in the sow graafian follicular wall and in the follicular fluid. Noradrenaline represented the highest level and adrenaline the lowest. Dopamine and noradrenaline concentrations found in the follicular fluid were lower at early proestrus than at mid-diestrus, whereas adrenaline levels in the fluid did not differ at either stage of the estrous cycle. The sow follicular wall contained less dopamine, noradrenaline and adrenaline at early proestrus than at mid-diestrus. Concomitantly, a decrement of [3H]-dopamine and [3H]-noradrenaline uptake, and of dopamine-beta-hydroxylase activity was detected at early proestrus compared to levels detected at mid-diestrus. The findings in sow graafian follicles show the existence of relationships between hormonal status, dopamine, noradrenaline and adrenaline endogenous levels and uptake, and dopamine-beta-hydroxylase activity. Possible links between estradiol, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) levels during the pig estrous cycle and ovarian catecholamines are discussed, as is a plausible involvement of these neurotransmitters in the contractile activity of the theca layer and the processes of follicular rupture and ovulation.

Immunocytochemical Localization of Relaxin in Corpora Lutea of Sows Throughout the Estrous Cycle1

Porcine relaxin has been sought by localization in the corpus luteum of sows on Days 3, 7, 9, 11, 12, 15, 18, 19, and 21 of the estrous cycle, using the avidin-biotin immunoperoxidase method and an antiserum to purified porcine relaxin. Simultaneous localization of relaxin in corpora lutea from sows on Days 108 and 113 of pregnancy was used to compare the intensity of immunostaining with that of corpora lutea of cyclic animals. However, the antiserum dilution necessary for optimal localization differed considerably in these two states (1:10,000 in pregnancy and 1:750 in the cycle), suggesting that lower levels of antigen are present in the luteal cells of the cycle. Relaxin immunostaining was undetectable on Day 3 of the cycle but became evident by Days 7 and 9. At Day 11 staining intensity increased and persisted through Day 15. On Day 18 some stain was still evident, but by Days 19, 20, and 21 there was complete absence of immunostain. Relaxin immunostaining appeared to be located throughout the cytoplasm of the luteal cell, as clear areas in the nuclear region were often observed. The results suggest that relaxin is produced in low amounts by the luteal cells of the cyclic sow and that the levels fluctuate with stage of the cycle. Lack of evidence from radioimmunoassay for a surge of relaxin secretion into the systemic circulation prior to luteolysis in the pig estrous cycle suggests that the relaxin localized in the luteal cells of the cycle may have an intraovarian function.