Nitric oxide stimulates progesterone and prostaglandin E2 secretion as well as angiogenic activity in the equine corpus luteum

Cytokines and nitric oxide (NO) are potential mediators of luteal development and maintenance, angiogenesis, and blood flow. The aim of this study was to evaluate (i) the localization and protein expression of endothelial and inducible nitric oxide synthases (eNOS and iNOS) in equine corpora lutea (CL) throughout the luteal phase and (ii) the effect of a nitric oxide donor (spermine NONOate, NONOate) on the production of progesterone (P4) and prostaglandin (PG) E(2) and factor(s) that stimulate endothelial cell proliferation using equine luteal explants. Luteal tissue was classified as corpora hemorrhagica (CH; n = 5), midluteal phase CL (mid-CL; n = 5) or late luteal phase CL (late CL; n = 5). Both eNOS and iNOS were localized in large luteal cells and endothelial cells throughout the luteal phase. The expression of eNOS was the lowest in mid-CL (P < 0.05) and the highest in late CL (P < 0.05). However, no change was found for iNOS expression. Luteal explants were cultured with no hormone added or with NONOate (10(-5) M), tumor necrosis factor-α (TNFα; 10 ng/mL; positive control), or equine LH (100 ng/mL; positive control). Conditioned media by luteal tissues were assayed for P4 and PGE(2) and for their ability to stimulate proliferation of bovine aortic endothelial cells (BAEC). All treatments stimulated release of P4 in CH, but not in mid-CL. TNFα and NONOate treatments also increased PGE(2) levels and BAEC proliferation in CH (P < 0.05). However, in mid-CL, no changes were observed, regardless of the treatments used. These data suggest that NO and TNFα stimulate equine CH secretory functions and the production of angiogenic factor(s). Furthermore, in mares, NO may play a role in CL growth during early luteal development, when vascular development is more intense.     

In vitro effect of leptin on growth hormone (GH)- and insulin-like growth factor-I (IGF-I)-stimulated progesterone secretion and apoptosis in developing and mature corpora lutea of pig ovaries

This study was designed to determine whether leptin modulates growth hormone (GH)- and insulin like growth factor-I (IGF-I)-stimulated progesterone (P4) production by corpora lutea (CL). Luteal cells were recovered from early developing (ELP) and mature (MLP) corpora lutea and cultured in defined medium with various combinations of GH, IGF-I, and leptin (0-200 ng/ml). P4 concentrations in the media were determined after 48 h of culture. During the early luteal phase, leptin at all used doses had no effect on basal P4 secretion, but it did suppress caspase-3 activity. When added in combination with GH, it had no effect on either GH-stimulated P4 secretion or apoptosis. Concomitant treatment with IGF-I and leptin decreased P4 secretion and parallelly increased the apoptosis rate. In mature corpora lutea of full secreting capacity, leptin at all doses had no effect on basal and GH-stimulated P4 secretion and caspase-3 activity. Only at the highest dose (200 ng/ml) when leptin was added with IGF-I did P4 secretion decrease with no effect on the caspase-3 activity. We conclude that the role of leptin is to restrict the stage of CL formation. During this luteal phase, leptin acts as an antiapoptotic factor and, at the same time, reverses antiapoptotic action of IGF-I, thereby protecting cells from excessive apoptosis and supporting retention of appropriate cell numbers, which is necessary for maintenance of homeostasis in developing CL.     

Relationships between ultrasonographic image attributes, histomorphology and proliferating cell nuclear antigen expression of bovine antral follicles and corpora lutea ex situ.

The aim of this study was to examine the relationships between quantitative ultrasonographic image characteristics, histological attributes and cell proliferating ability of bovine antral follicles and corpora lutea (CL) ex situ. Bovine ovaries (n = 30) from animals at various reproductive states (metoestrus-early dioestrus, n = 8; mid-dioestrus, n = 12; oestrous phase of peripubertal heifers, n = 6; and pregnancy, n = 4) were collected at the slaughterhouse. High-resolution ultrasonographic images of the ovaries were obtained in the water bath, digitized and subjected to computerized image analyses. The analyses utilized line and spot techniques designed to determine pixel values of the follicular wall (the largest follicles >2 mm in diameter in each ovary) and CL, respectively. Individual ovarian structures were dissected and processed for histology and immunohistochemical detection of proliferating cell nuclear antigen (PCNA). The mean follicular diameter was negatively correlated with total cell density (r = -0.45, p < 0.05), granulosa layer thickness (r = -0.67, p < 0.001) and the percentage of PCNA-positive cells (r = -0.57, p < 0.001). Numerical pixel values and heterogeneity of the follicular wall were positively correlated with total cell density (r = 0.42, p < 0.05 and r = 0.62, p < 0.05; respectively), granulosa layer thickness (both r = 0.39, p < 0.05), and the percentage of PCNA-positive cells (r = 0.54, p < 0.01 and r = 0.69, p < 0.001, respectively). Estimates of cell density and proliferating cell index were not correlated with the ultrasonographic image attributes of CL. We conclude that follicular size and echotextural variables, as determined by computer-assisted image analysis of ovaries ex situ, are reliable markers of the histophysiological properties of bovine antral follicles, but the ultrasonographic characteristics are not indicative of cell density and proliferation in the bovine CL.     

Luteal toxicity evaluation in rats

The corpora lutea (CL) are endocrine glands that form in the ovary after ovulation and secrete the steroid hormone, progesterone (P4). P4 plays a critical role in estrous and menstrual cycles, implantation, and pregnancy. The incomplete rodent estrous cycle stably lasts 4–5 days and its morphological features can be distinguished during each estrous cycle stage. In rat ovaries, there are two main types of CL: newly formed ones due to the current ovulation (new CL), and CL remaining from prior estrous cycles (old CL). In the luteal regression process, CL were almost fully regressed after four estrous cycles in Sprague-Dawley rats. P4 secretion from CL in rodents is regulated by the balance between synthesis and catabolism. In general, luteal toxicity should be evaluated by considering antemortem and postmortem data. Daily vaginal smear observations provided useful information on luteal toxicity. In histopathological examinations, not only the ovaries and CL but also other related tissues and organs including the uterus, vagina, mammary gland, and adrenal glands, must be carefully examined for exploring luteal changes. In this review, histological and functional characteristics of CL in rats are summarized, and representative luteal toxicity changes are presented for improved luteal toxicity evaluation in preclinical toxicity research.     

Tumour Necrosis factor-α Receptors are Present in the Corpus Luteum Throughout the Oestrous Cycle and During the Early Gestation Period in Pigs

To determine the physiological significance of tumour necrosis factor‐α (TNFα) in the regulation of luteal functions in pig, this study was conducted to identify the presence of functional TNFα receptors in porcine corpora lutea (CL) throughout the oestrous cycle and the early gestation. The CL were isolated from pigs on days 4, 6, 8, 12 or 15 of the oestrous cycle (n=3; day 0 = oestrus) and days 15, 20 or 25 of gestation (n=3; day 0 = mating). A Scatchard analysis revealed the presence of a high‐affinity binding site for TNFα in all samples (dissociation constant; 2.7 ± 0.51 to 5.8 ± 0.50 nM ). The concentration of TNFα receptors was higher on day 15 of the oestrous cycle than on days 4 and 8 of the oestrous cycle (p < 0.05). Furthermore, TNFα receptor concentrations in the CL on days 15, 20 and 25 of gestation were significantly lower than on day 15 of the oestrous cycle (p < 0.05). On day 9 of the oestrous cycle, exposure of cultured luteal cells to 0.06–60 nM TNFα stimulated prostaglandin (PG) F_2α and PGE₂ secretion in a dose‐dependent manner (p < 0.05). These results indicate that functional TNFα receptors are present in the porcine CL throughout the oestrous cycle and early gestation, and suggest that TNFα plays one or more physiological roles in regulating CL function throughout the oestrous cycle and the early gestation period. In addition, TNFα receptor concentration in the CL of the late luteal stage (day 15) of the oestrous cycle was higher than on the respective day in the early pregnant pig, suggesting that TNFα plays a role in accomplishing luteolysis in the porcine CL.     

Fasting influences steroidogenesis, vascular endothelial growth factor (VEGF) levels and mRNAs expression for VEGF, VEGF receptor type 2 (VEGFR-2), endothelin-1 (ET-1), endothelin receptor type A (ET-A) and endothelin converting enzyme-1 (ECE-1) in newly formed pig corpora lutea

This study was designed to verify whether fasting influences vascular endothelial growth factor (VEGF) production and VEGF, VEGF receptor-2 (VEGFR-2) as well as endothelin (ET) system members (endothelin converting enzyme-1, ECE-1; ET-1; endothelin receptor type A, ET-A) mRNA expression in pig corpora lutea; furthermore, we wanted to assess whether fasting affects steroidogenesis in luteal cells. Eight prepubertal gilts were induced to ovulate and were randomly assigned to two groups: (A) n = 4, normally fed; and (B) n = 4, fasted for 72 h starting 3 days after ovulation. At the end of fasting, ovaries were removed from all the animals and corpora lutea (CLs) were collected. VEGF and steroid levels in luteal tissue were determined by ELISA and RIA, respectively; VEGF, VEGFR-2, ET-1, ET-A and ECE-1 mRNAs expression was measured by real-time PCR. VEGF protein levels were similar in the two groups, while all steroid (progesterone, testosterone, estradiol 17beta) concentrations were significantly (P < 0.001) higher in CLs collected from fasted animals compared with those from normally fed gilts. VEGF, VEGFR-2, ET-1 and ECE-1 (but not ET-A) mRNA expression was significantly lower (P < 0.05) in fasted versus normally fed animals. The overall conclusion is that all the parameters studied are affected by feed restriction, but the mechanisms activated at luteal level are possibly not fully adequate to compensate for nutrient shortage.     

Uterine influences on the formation of subnormal corpora lutea in seasonally anestrous ewes

The hypothesis that subnormal luteal function after induced ovulation in anestrous ewes was the result of uterine influences exerted during the periovulatory period was tested. Crossbred ewes (n = 27) in seasonal anestrus were induced to ovulate by administration of 12 doses of 250 ng of LHRH at 2-h intervals, followed immediately by a bolus injection of LHRH (250 micrograms; d 0). Ewes were unilaterally hysterectomized on either d -3 (PRELHRH) or 2 (POSTLHRH). Daily blood samples were collected and assayed for progesterone (P4) and 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM). All ewes were slaughtered on d 10, and corpora lutea (CL) were collected, weighed, and assayed for concentration of P4. All ewes that ovulated exclusively in the ovary ipsilateral to the remaining uterine horn had a transient increase in plasma P4 of 2 to 3 d (short luteal phase). In ewes with at least one CL in the isolated ovary, elevated plasma P4 was maintained after hysterectomy but was consistently lower (P less than .05) in POSTLHRH ewes than in PRELHRH ewes. Concentrations of PGFM did not differ between treatments. The CL ipsilateral to the remaining uterine horn weighted less (P less than .01) and contained less P4 (P less than .01) than contralateral CL. These data confirm the hypothesis that premature regression of subnormal CL is uterine-dependent in a local fashion. Presence of the uterus during the follicular and(or) early luteal phase inhibited subsequent luteal function in seasonally anestrous ewes.     

Cystic Degeneration in Porcine Ovaries - First Communication: Morphology of Cystic Ovaries, Interpretation of the Results

Contents: The morphological appearance of ovarian structures in 79 sows with ovarian cysts was examined. Sixty sows had been culled due to sterility; 19 sows had been repeatedly investigated clinically before slaughter. It was found that cystic ovaries without corpora lutea (CL) had the largest diameter and volume (151.3 cm³), in comparison to both cystic ovaries with CL (85.7 cm³) and normalovaries (16.9 cm³). The number of cysts on ovaries without corpora lutea was approximately twice as high as on the ovaries with CL (23.3 vs 12.1 cystslsow; p < 0.001). A combined form of degeneration consisting of large (> 1.5 cm in diameter) and small (1.5 em in diameter) cysts was found in 63% of the animals, whereas degeneration with only small cysts occured in 8% of the cases. Dark CL, interpreted as being functional, were mainly present in animals with up to 10 cysts per sow (24 out of 27 sows, p < 0.001); pale CL, interpreted as being weakly functional, were mainly found on ovaries with more than 10 cysts (19 out of 23 sows; p < 0.001). Based on macroscopical differentiation cyclic activity was found to be dependent to the number of cysts. Regular or irregular estrous cycles were observed in oligocystic animals (10 cysts/animal) with functional CL (58%). Up to 75% of the animals with pale or absent CL showed no estrous activity.     

Follicle Development during Luteal Phase and Altrenogest Treatment in Pigs

Synchronization of the oestrous cycle of gilts using altrenogest treatment has been found to increase ovulation rate. The current experiment investigated if the increase in ovulation rate after altrenogest treatment is related to increased follicle size at the end of altrenogest treatment compared with late luteal phase follicles. Crossbred gilts (n = 15) received altrenogest during 18 days [20 mg Regumate (Janssen Animal Health, Beerse, Belgium)], starting 5-7 days after onset of first oestrus. Control gilts (n = 15) did not receive altrenogest. At days 10-12 of the oestrous cycle [i.e. in the presence of corpora lutea (CL)], average follicle development was 2.51 +/- 0.20 mm (assessed with ultrasound) in altrenogest-treated gilts and 2.58 +/- 0.16 mm in control gilts (p > 0.10). During the last days of altrenogest treatment (i.e. when CL had gone into regression), average follicle size had increased to 3.01 +/- 0.31 mm (p < 0.05). Subsequent ovulation rate was 16.6 +/- 1.7 in altrenogest treated gilts and 15.1 +/- 1.2 in control gilts (p < 0.05). Altrenogest treatment resulted in increased follicle size after regression of the CL, showing that suppression of follicle growth by altrenogest alone is less severe than suppression by endogenous progesterone (either with or without altrenogest). Altrenogest treatment also resulted in a higher ovulation rate. However, it is unclear if the increased follicle size and higher ovulation rate after altrenogest treatment are causally related, as the relation between the two on an animal level was not significant.     

Ultrasonography and progesterone concentration at a single time point for the detection of puberty in dairy heifers

Data were available from 1657 heifers across 48 dairy farms which were visited once, on average 9 days (± 5.2) prior to the mating start date (23 April, ± 12.6). Blood samples were collected via coccygeal venepuncture for progesterone (P4) analysis, and animals were scanned for the presence or absence of corpora lutea (CL), to determine the luteal status. A comparison of luteal status between ultrasound (CL identified) and P4 (≥ 1 ng/ml), based on a single measurement point, showed poor agreement (K = 0.32). The majority of animals were classified as luteal (76%) using both ultrasonography and P4. There was excellent agreement between luteal status detected by ultrasonography alone and luteal status assigned by a combination of ultrasonography and P4 (K = 0.93). The agreement between luteal status assigned by P4 and luteal status assigned by the combination of ultrasonography and P4 was poor (K = 0.37). These results indicate that at a single examination, ultrasonography is the preferred modality to determine the luteal status of maiden heifers.     

Effect of the gonadotrophins treatment on morphological alterations in ovary and peripheral plasma concentrations of steroid hormones in gilts

The present study was designed to examine the influence of gonadotrophins treatment on the ovarian morphology changes and plasma concentrations of steroid hormones in peripheral blood. The experiment was performed on sexually pubertal gilts (Large White x Landrace) of similar age (7-8 months) and body mass (100-110 kg) with two controlled subsequent estrous cycles. The animals were randomly divided into four groups: two control consisting of pigs with the luteal phase (n = 9, the 10th day of the estrous cycle) and the follicular phase (n = 6, the 20th day of the estrous cycle) and two experimental ones consisting of animals with both mentioned periods (n = 7 and n = 9) treated with gonadotrophins (PMSG and hCG). The gilts in the luteal phase were injected (s.c.) with gonadotrophins at a daily dose of PMSG 400 and hCG 200 IU from the 16th to the 27th day (the 6th day of the next estrous cycle). The gilts in the follicular phase, were injected with the same dose of gonadotrophins but from the 8th to the 19th day of the estrous cycle. Plasma concentrations of P4, A4, T, E1, E2 and metabolite of PGF2 alpha-PGFM were determined by radioimmunoassay (RIA) method. Injections of PMSG and hCG in both experimental groups produced several times enlarged: weight, size and volume of ovaries and alterations in a number of structural elements as compared with those found in the control animals. The morphological elements presented in ovaries: corpora haemorrhagica, corpora lutea, regular and atretic follicles and first of all cysts by distinctly differentiation thickness of the walls are characteristic for cystic ovarian degeneration. Plasma concentrations all determined hormones after gonadotrophins treatment in experimental groups were increased except E1 (insignificant decrease) in luteal phase as compared with those found in the control groups. Statistically significant increase (p < 0.001) in plasma concentrations of P4, A4, and T in both experimental groups and E2 (p < 0.001) in luteal phase were noted. In peripheral plasma concentrations increase of E1 and E2 in follicular phase of the estrous cycle were insignificant.     

The induction of a delayed post-ovulatory progesterone rise in dairy cows: a novel model

A delayed rise in post-ovulatory progesterone is associated with poor embryo development in the cow, although the underlying cause of this aberrant luteal function is poorly understood. The objective of this study was to develop a novel model, in which a delayed progesterone rise could be induced by manipulating the dynamics of the follicular phase. Luteolysis was induced in 20 dairy cows in the presence of either a larger follicle > 10 mm (LF, n = 11) or a smaller follicle < 10 mm (SF, n = 9) and transrectal ultrasonography was performed to determine follicle and CL growth and timing of ovulation. Plasma progesterone and oestradiol were analysed 3x daily. Cows were slaughtered on either day 4 (n = 4 per group) or day 7 (SF, n = 5; LF, n = 7) after ovulation. The pre-ovulatory follicle was larger in the LF group than the SF group at luteolysis (13.5 +/- 0.4 mm versus 6.7 +/- 0.7 mm, P < 0.001) and ovulation (16.7 +/- 0.3 mm versus 13.6 +/- 0.6 mm, P < 0.001). The LF group experienced a shorter follicular phase and ovulated 36 h earlier than the SF group (P < 0.001). At luteolysis, plasma oestradiol concentrations were greater in the LF group (P < 0.001), although peak concentrations were not different (P > 0.05). Moreover, higher progesterone concentrations were observed in the LF group during the early luteal phase (P < 0.05). Luteal weights were positively correlated with plasma progesterone concentrations on day 5 (P < 0.05) but not day 8. In conclusion, a model has been developed which has shown that the dynamics of follicle development during the pre-ovulatory period is an important determinant of subsequent CL development and function.     

Immunohistochemical localization of steroidogenic enzymes in corpus luteum of wild sika deer during early mating season

We analyzed the localization of steroidogenic enzymes (P450 scc, 3 beta HSD, P450 arom and P450 c17) in the corpora lutea of two Hokkaido sika deer (Cervus nippon yesoensis) during the early mating season. Two corpora lutea were found in each female and the timing of formation of the corpora lutea seemed different. P450 scc, and 3 beta HSD, positive luteal cells were found in both corpora lutea. The existence of two functional corpora lutea from the early mating season through pregnancy suggests that progesterone secreted by two or more corpora lutea is necessary for maintenance of pregnancy in sika deer.     

Insulin-like growth factor (IGF)-I and IGF binding proteins-2, -3, -4, -5 in porcine corpora lutea during the estrous cycle; evidence for inhibitory actions of IGFBP-3

In this study we measured protein concentrations of insulin-like growth factor (IGF)-I and IGF binding proteins (IGFBPs) 2-5 in porcine corpora lutea (CLs) throughout the estrous cycle (Experiment 1), and examined the effects of IGFBP-3 and IGFBP-3 antibody (AB) on luteal progesterone (P4) secretion in vitro (Experiment 2). For Experiment 1, (CLs) and serum were collected on days (D) 4, 7, 10, 13, 15 and 16 of the estrous cycle (n = 5 animals per day). IGF-I was extracted from CLs and sera, and measured by radioimmunoassay (RIA). IGFBPs were measured in CLs by ligand blots. For Experiment 2, CLs (from Experiment 1) were enzyme dissociated and luteal cells cultured (24 h) in Medium 199 (M199) containing (0-500 ng/ml) IGFBP-3 (+/-IGF-I; 100 ng/ml), or (0-10 microg/ml) IGFBP-3 AB. P4 in media was measured by RIA. In Experiment 1, luteal IGF-I concentrations (ng/g tissue) were maximal on day 4 and gradually decreased thereafter. Serum IGF-I concentrations (ng/ml) were highest on days 4 and 7, compared with days 10-15. Peak levels of luteal IGFBP-3 were also seen on days 4 and 7 of the cycle. Luteal IGFBP-2 concentrations showed a tendency to increase on day 16 (P < 0.05 versus day 10), but no significant changes in IGFBP-4 or -5 were seen. In Experiment 2, IGFBP-3 (w IGF) inhibited the steroidogenic actions of IGF-I, but had no significant actions alone (IGFBP-3 w/o IGF). Finally, IGFBP-3 AB stimulated P4 secretion on days 4 and 7, but not on days 10-16. We conclude that IGFBP-3 inhibits IGF-I actions in the porcine CL.     

Responsiveness of bovine corpora lutea to prostaglandin F2 alpha: comparison of corpora lutea anticipated to have short or normal lifespans

The objective of this study was to determine if corpora lutea anticipated to have short lifespans were more responsive to the luteolytic action of prostaglandin F2 alpha (PGF2 alpha) than corpora lutea anticipated to have normal lifespans. Sixteen cows were allotted randomly to a hysterectomized-control (HC) or hysterectomized-progestogen (norgestomet) implant (HN) group. To verify that progestogen treatment of postpartum cows prior to induction of ovulation with gonadotropin-releasing hormone (GnRH) results in an increased number of cows exhibiting normal-length luteal phases, 21 additional cows were allotted randomly to a uterine intact-control (IC) or a uterine intact-progestogen implant (IN) group. Cows allotted to the HN and IN groups received norgestomet ear implants for 9 d beginning 17 to 21 d postcalving. All cows were injected (i.m.) with 100 micrograms GnRH 28 to 32 d postcalving (48 h after implant removal in the HN and IN groups) to induce ovulation. Two or 3 d after GnRH injection (d 0), cows in the HC (n = 8) and HN (n = 8) groups were hysterectomized to remove the major endogenous source of PGF2 alpha, and on d 7 cows were injected (i.m.) with 10 mg PGF2 alpha to assess luteal sensitivity. The proportion of corpora lutea having normal lifespans was greater (P less than .1) for the IN than for the IC group. In HC and HN groups, concentration of progesterone (P) increased similarly from d 0 to 6. Injection of PGF2 alpha in HC and HN groups on d 7 decreased (P less than .01) concentration of P approximately 50% by 6 h after injection (similar for both groups). Complete luteolysis was induced by PGF2 alpha in none of eight and two of eight cows in the HC and HN groups, respectively. In remaining cows (HC and HN groups) concentration of P increased (P less than .01; similar for HC and HN groups) beginning 24 h after PGF2 alpha and remained elevated through d 30 to 34 (end of experimental-period). In summary, corpora lutea anticipated to be short-lived were not more responsive to PGF2 alpha than corpora lutea anticipated to have normal lifespans.     

Possible roles of intracellular cyclic AMP, protein kinase C and calcium ion in the apoptotic signaling pathway in bovine luteal cells

Structural luteolysis occurs by apoptosis of luteal cells. The present study examined the effects of activators of well-characterized second messengers on Fas and caspase-3 mRNA expression and on P4 production in luteal cells in order to trace the pro- and anti-apoptotic factors in the bovine corpus luteum (CL). Cultured bovine mid luteal cells were treated for 24 h with a cyclic AMP analogue (8-bromo cyclic AMP; 8br-cAMP; 2.5 mM), a protein kinase C (PKC) activator (phorbol 12-myristate 13-acetate; PMA; 10 microM), or calcium ionophore (A23187; 10 microM). Fas and caspase-3 mRNA expression was inhibited by 8br-cAMP and PMA but was increased by A23187 (P<0.05). In addition, P4 production by bovine luteal cells was stimulated by 8br-cAMP and PMA, whereas it was inhibited by A23187, compared with untreated controls (P<0.05). The overall results suggest that cAMP and PKC suppress apoptosis in bovine luteal cells through inhibition of Fas and caspase-3 mRNA expression and through stimulation of P4 production. Therefore, substances that activate cAMP or PKC may act as survival factors in the bovine CL. Furthermore, substances that mobilize Ca2+ may act as apoptotic factors by stimulating Fas and caspase-3 expression in the bovine luteal cells.     

Impacts on conceptus survival in a commercial swine herd

An estimated 30 to 40% of potential piglets are lost before farrowing in U.S. or European pig breeds. Because these studies were conducted in limited numbers of university research herds, we decided to characterize the timing, pattern, and extent of conceptus loss in a commercial swine herd in Iowa (Pig Improvement Company; Camborough Line). Sows (parities 2 to 14) were slaughtered on d 25 (n = 83), 36 (n = 78), or 44 (n = 83) of gestation. These days coincide with periods before, during, and after uterine capacity becomes limiting to conceptus survival. At slaughter, numbers of corpora lutea (CL) and uterine horn length were determined, and conceptuses were removed and evaluated. Uterine horn length and CL number did not differ among these days of gestation, averaging 217 cm and 26.6 CL, respectively. In contrast, numbers of conceptuses decreased (P < 0.05) from 15.8 on d 25 to 12.9 on d 36, then remained relatively constant through d 44 (12.1). Thus, conceptus survival averaged 60.2% on d 25, 50.1% on d 36, and 46.3% on d 44, based on numbers of CL present. There was a positive correlation (P < 0.001; r = 0.50) between numbers of viable conceptuses on d 25 and ovulation rate, but this association was completely lost by d 36 (P > 0.10) when uterine capacity becomes limiting. In agreement with this premise, uterine horn length and conceptus number were not associated on d 25 but exhibited positive correlations (P < 0.05) on d 36 (r = 0.36) and d 44 (r = 0.40). On all 3 d examined, the numbers of viable conceptuses were not associated with fetal weight but were negatively correlated (P < 0.05) with placental weight. Compared with the commonly reported values for ovulation rate and percentage conceptus loss in university research herds, values from these production animals were extremely high. Data suggest that throughout this period, larger litters were associated with conceptuses exhibiting small placentae. These data lend support to the concept that increased placental efficiency (fetal weight/placental weight) may contribute to increased litter size in the pig.     

The influence of the ovulation rate on ultrasonically determined ovine corpus luteum morphometry and progesterone concentrations in cyclic and early pregnant sheep

The objective of the present study was to describe morphology and function of the Corpora lutea (CL) during the oestrous cycle and early pregnancy in sheep with different ovulation rates. In total 40 Booroola. Mutton Merino crosses [heterozygous carriers (FecBFec+) and non-carriers (Fec+Fec+) of the Booroola-fecundity gene (FecB)] with ovulation rates 1 to 4 were examined. During the oestrous cycle (n = 20) and the first month of pregnancy (n = 20) blood samples were taken daily (radioimmunoassay of progesterone) and an ultrasonic ovary diagnosis was conducted. The ewes were scanned transrectally with a 7.5 MHz linear probe lying in a dorsal position. During every examination the CL could be detected. The number and the diameter of the CL were documented and the total volume of luteal tissue per ewe was calculated. The effect of the ovulation rate on CL-morphology (diameter and total volume of luteal tissue per ewe) and peripheral progesterone concentrations were assessed by one-way ANOVA. On day 6 and 7 post ovulationem in cyclic and early pregnant sheep 42% of the diagnosed CL had a cavity. On day 11 (cyclic sheep) and day 10 post ovulationem (early pregnant sheep) this number decreased to 22% (p < 0.05). Both conditions of the CL (compact or with a central cavity) are similar in function and should be regarded as appearances of the same basic process. From the third day onwards the ovulation rate influenced significantly (p < 0.05) the development of the outside diameters of the CL. However, the ovulation rate had no effect on the total volume of the luteal tissue per sheep and on the progesterone concentrations. Yet, in sheep with the ovulation rate 1 significantly lower progesterone concentrations were determined than in sheep with the ovulation rates 2 to 4. In sheep with the ovulation rates 2 to 4 the peripheral progesterone concentrations did not differ significantly. In cyclic and pregnant sheep there is a positive correlation (r = 0.75, p < 0.05) between the progesterone concentration and the total volume of luteal tissue. Considering the smaller diameters of the preovulatory follicles it seems that the development of the CL continues until a threshold-value of progesterone and/or of the total luteal tissue is exceeded. Ewes with low ovulation rates reach this threshold-value with only a few but large CL. With increasing ovulation rate the CL tend to have smaller diameters.     

Priming with progestin, but not GnRH antagonist, induces a consistent endocrine response to exogenous gonadotropins in induced and spontaneously ovulating cats

Ovarian sensitivity to exogenous gonadotropin stimulation (equine chorionic gonadotropin [eCG] and human chorionic gonadotropin [hCG]) following pre-treatment with a progestin (levonorgestrel) versus GnRH antagonist (antide) was studied in cats known to be induced versus spontaneous ovulators. Queens were assigned to one of three treatments: (1) levonorgestrel implants+eCG/hCG (n=7 cats); (2) antide injections+eCG/hCG (n=7) or (3) eCG/hCG alone (control; n=7). Hormonal metabolites were assessed in fecal samples collected daily for 60 days before and during the 37 days inhibitory pre-treatment and for more than 60 days after eCG/hCG. Fecal metabolites of estradiol and progesterone were measured by radioimmunoassay. Females that maintained baseline progesterone were considered induced ovulators, whereas cats that exhibited a luteal phase before inhibition treatment were classified as spontaneous ovulators. Based on fecal hormone profiles, levonorgestrel thoroughly inhibited ovarian activity, whereas antide synchronized follicular phases but did not induce complete ovarian down-regulation. Both treatments prevented ovulation in spontaneous ovulators, but neither caused regression of existing corpora lutea (CL). Levonorgestrel, but not antide, pre-treatment resulted in a quiescent ovary at the time of eCG injection, yet endocrine responses to eCG/hCG were not different among treatments. Interestingly, spontaneously ovulating females exhibited a prolonged estradiol response to gonadotropin stimulation compared to induced ovulators, and this prolonged estradiol surge was replicated by levonorgestrel pre-treatment. Thus, the progestin levonorgestrel effectively suppresses follicular and luteal activity in the cat, resulting in a more consistent response to gonadotropin stimulation, even in females prone to spontaneous ovulation.