Expression of the orexigenic peptide ghrelin in the sheep ovary

Ghrelin has been implicated in the control of cell proliferation in reproductive tissue. Here, we provide evidence that both ghrelin mRNA and protein are present in ovarian follicles. Persistent expression of ghrelin was also demonstrated in sheep ovary throughout the estrous cycle and pregnancy. In fact, the relative mRNA levels of ghrelin varied depending on the stage of the cycle, with the highest expression during the development of the corpora lutea (CL) and minimal expression in the regressing CL. A similar pattern was seen during pregnancy. Dynamic changes in the profile of ghrelin expression during the estrous cycle and throughout pregnancy suggest a precise regulation of ovarian expression of ghrelin, which could represent a potential role for ghrelin in the regulation of luteal development. In conclusion, the presence of the ghrelin signaling system within the sheep ovary especially in the oocytes opens up the possibility of a potential regulatory role of this novel molecule in reproductive function.     

Observations on Ovarian Structure and Development of the Southern Giraffe,Giraffa Camelopardalis Giraffa

The occurrence of vesicular follicles and corpora lutea (CL), without evidence of ovulation, in late fetuses and during prepubertal life was confirmed in the southern giraffe. In this, and other respects, the pattern of ovarian development is similar in the southern giraffe to the northern subspecies. This early ovarian activity recedes during late prepubertal life. Numerous accessory CL were present in pregnant and non-pregnant giraffe but they did not attain the size of the true CL of pregnancy. The latter undergo an initial phase of rapid enlargement lasting about 100 days, followed by a period of recession till mid-pregnancy and a second growth phase which persists until term (457 days). The CL of pregnancy rapidly regresses after parturition. The accessory CL regress slowly and form normal looking corpora albicantia (CA). These CA are most abundant during early adult life as also are vesicular follicles.     

Suitability of sonographic evaluation of ovarian dynamics and uterine involution for prediction of postpartum fertility in the cow

To investigate the suitability of sonography for prediction of puerperal fertility, we used 34 cows with normal puerperium. For this purpose, the animals were examined gynecologically and by transrectal sonography at 10-day intervals between day 15 and day 45 p.p. as well as at oestrus, when they were inseminated. We judged uterine involution by diameter and sonographic structure of the uterine cross-section and ovarian activity by the presence of follicles and corpora lutea. Cows, which became pregnant after first insemination p.p., were assigned to group G1 (n = 16) and animals, which failed to conceive, to group G2 (n = 18). The cows of G1 obtained significantly better results concerning both uterine involution and ovarian activity than cows of G2. Thus, diameter of the uterine horn decreased faster and sonographic structure of the uterine cross section returned markedly earlier to the nonpregnant appearance in G1 than in G2. Group G1 also showed a significantly higher percentage of dominant follicles at all examinations. The results of the study show a distinct relationship between uterine involution and ovarian activity and puerperal fertility. But it has also to be resumed that the prediction of fertility for individuals will always be restricted by other reasons causing failure of conception.     

Studies on Reproduction in the Goat: III. The Functional Activity of the Ovaries of the Goat

1. A study of the functional activity in the ovaries was undertaken on 366 organs from pregnant goats and 171 organs from nonpregnant goats. The ovulations were recorded by the presence of corpora lutea in the ovaries.
2. The right ovary appears to be the most active in the goat, since 57 % of the ovulations occurred in the right ovary. In cases of multiple ovulations there appears to be a greater tendency for the first of these ovulations to occur in the right ovary (62.34 %).
3. In almost 50 % of twin pregnancies both ova will be shed from the same ovary.
4. In six cases, five twin pregnancies and one triplet pregnancy, only one, respectively two corpora lutea were found. This can in all probability be taken as an indication of the occurrence of identical twins.
5. The incidence of “ovum loss” or early embryonic mortality was found to be 19.6 %.

     

Ovariectomy by colpotomy in cows.

For the purpose of collecting active ovarian structures for cell culture, unilateral ovariectomy (n = 34 ovaries) was performed per vagina on 17 dairy cows having normal estrous cycles, bilateral ovariectomy was performed on 9 (n = 18 ovaries) dairy cows, and corpora lutea (n = 13) were removed from 11 beef cows having normal estrous cycles. None of the cows was clinically ill after the operation. Nine of 37 cows developed adhesions of both uterine horns and the body of the uterus. Three instruments were used to perform colpotomy. The described surgical technique for removal of the ovaries or corpora lutea is practical and inexpensive, and has low morbidity associated with it.     

Unusual phenomena during pregnancy in the cat and cow.

Unusual situations were observed during examination of pregnant uteri from a cat and a cow. The feline specimen revealed three corpora lutea at the surface of each ovary with three fetuses developing normally in the left and two in the right uterine horn. The right horn had been ruptured between two of the locular enlargements but the damage was sealed by the omentum. Three conceptuses were present in the cow's uterus. Two of these embryos, each with its individual allantois, were developing within a single chorion within one uterine horn. The adjacent tips of these allantoic vesicles were entangled into a complex knot which could provide opportunity for vascular anastomosis.     

The Immunohistochemical Localization of Desmin and Smooth Muscle Actin in the Ovary of the African Giant Rat (Cricetomys gambianus) During the Oestrous Cycle

The aim of this study was to describe the distribution of smooth muscle actin and desmin immunopositive cells in the ovary of the giant rat. In addition, the study describes the morphological changes in the ovary of this species during the oestrous cycle. Healthy secondary and tertiary follicles dominated the ovary during pro-oestrus and oestrus. The theca externa of the tertiary follicles was immunopositive for smooth muscle actin, but immunonegative for desmin. Oestrus was also characterized by the presence of corpora haemorrhagica, which had an outer layer of smooth muscle actin immunopositive cells. Differentiating corpora lutea were observed during metoestrus. A further notable feature of the ovary during metoestrus was the presence of numerous atretic secondary and tertiary follicles. In the later stages of atresia, the follicles were infiltrated by desmin and smooth muscle actin immunopositive cells. Dioestrus was characterized by the presence of non-regressing and regressing corpora lutea. Immunostaining for smooth muscle actin was demonstrated in the enclosing layer of the corpora lutea, as well as in the tunica media of blood vessels within the corpora lutea. The results of this study have shown that morphological changes in the ovary of the giant rat during the oestrus cycle are similar to those of laboratory rodents. Furthermore, the results of the immunohistochemical study indicate that the perifollicular distribution of desmin and smooth muscle actin cells changes during follicular development and atresia.     

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.     

Expression of nerve growth factor and its receptors, TrkA and p75, in porcine ovaries

The cellular localization of nerve growth factor (NGF) and its receptors (TrkA, p75) was investigated during the estrous cycle in gilts. Also, the levels of expression of these factors in walls of tertiary follicles and corpora lutea (CLs) were determined using Western blot. The ovaries from days 3, 7, 16 and 20 of the cycle revealed the presence of NGF and its receptors in oocytes of secondary and tertiary follicles, follicular cells of primary and secondary follicles, thecal and granulosa cells of tertiary follicles and steroidogenic cells of CLs. In wall cells of primary follicles, NGF, TrkA and p75 staining was strongest on day 16, while in secondary follicles, only p75 was more intensely stained on day 16 and 20. In walls of small (to 3 mm in diameter) and medium (4-6 mm in diameter) follicles, NGF staining was lower on day 16, and the p75 reaction was strongest on day 20. On day 20, NGF staining in large follicles (7-10 mm in diameter) was higher than in smaller follicles. The levels of NGF and p75 in small and medium follicles were highest on day 20. The contents of NGF and TrkA in large follicles on day 20 were higher than in smaller follicles. NGF and TrkA contents in CLs were highest on day 7. Our study demonstrates that NGF, TrkA and p75 are expressed in the ovary during the estrous cycle in gilts. These results suggest that NGF and its receptors may be important for ovarian function in cycling gilts.     

Uterine and ovarian blood flow in a Holstein Friesian cow with aplasia of one uterine horn

The ovarian dynamics and uterine and ovarian blood flows of a 6-year-old (2 parturitions) Holstein Friesian cow with right uterine horn aplasia were observed during two estrous cycles. In one estrous cycle, a corpus luteum (CL) formed in the right ovary, but regression of the CL and subsequent ovulation were not observed. In the other estrous cycle, a CL formed in the left ovary and delayed regression of the CL and subsequent ovulation were observed. The blood velocity of the right uterine artery was lower than that of the left uterine artery throughout both estrous cycles when a CL formed in either the right and left ovary. The blood velocities of the right and left ovarian arteries were unaffected by right uterine horn aplasia and changed depending on the presence of a CL. These results indicated that the blood flow of the right uterine artery was very weak and that aplasia of one uterine horn affects the estrous cycle, especially CL regression.     

Dynamics of ovarian follicular development in cattle following hysterectomy and during early pregnancy

Three experiments were conducted to evaluate ovarian follicular dynamics and functional activity during pregnancy in cattle. In 11 pregnant Charolais cows of Experiment I, size of largest follicle, number of follicles and accumulated follicle size were reduced by day 27 of pregnancy on the ovary bearing the corpus luteum (CL) but not on the non-CL bearing ovary. In experiment II, local attenuation of ovarian follicular development on the CL bearing ovary of seven pregnant heifers was evident compared to the contralateral ovary without the CL. However, in four hysterectomized heifers, follicular development was sustained on both the CL- and non-CL bearing ovaries when CL maintenance was achieved without presence of the uterus or conceptus. In Experiment III, steroidogenic characteristics of the largest and second largest follicles at 17 d postestrus were evaluated for seven pregnant and six cyclic cattle. Follicle by physiological status interactions were detected for both aromatase activity of the follicle and follicular fluid concentrations of estradiol and progesterone. In cyclic cows, the largest follicle had appreciably more aromatase activity than did the second largest follicle; whereas, aromatase activity of the largest follicle from pregnant cows was less than that of cyclic cows. However, in pregnant cows the second largest follicle became the estrogen-active follicle, and this follicle occurred with a higher frequency on the ovary contralateral to the CL-bearing ovary. These changes in aromatase activity were reflected by parallel changes in estrogen concentrations of follicular fluid. The higher progesterone concentration in follicular fluid of the largest follicle in pregnant cows provided further confirmation of their atretic status. In conclusion, during early pregnancy the conceptus and/or uterus ipsilateral to the conceptus appear to secrete compounds which alter local follicular steroidogenic activity and attenuate subsequent follicular growth between 17 to 34 d of pregnancy on the CL-bearing ovary. This local mechanism acting within the ovary may contribute to the antiluteolytic effects of early pregnancy in cattle.     

Ovarian function and hormone secretion of gilts actively immunized against androstenedione

Fifty crossbred gilts immunized against bovine serum albumin (BSA) or androstenedione conjugated to BSA (AD) were used in three experiments. Primary immunizations were given at 120 d of age and boosters at 148 and 176 d. Gilts were moved to pens containing four to five animals each and exposed to boars beginning at 180 d of age. Immunization against AD did not affect age at puberty, percentage of gilts exhibiting estrus or duration of first estrous cycle. Over the three experiments, ovulation rate was 24% greater for AD-immunized gilts than for controls, and the number of corpora lutea was related positively (r = .82) to the log of the antibody titer. Number of ovulations decreased as interval from booster immunization to onset of estrus increased. During diestrus of the first estrous cycle, gilts immunized against AD had more follicles 5 to 10 mm in diameter, more total ovarian follicles and more total ovarian structures (corpora lutea plus follicles) than controls. Immunization against AD increased the frequency of LH pulses on d 16 but not on d 17 or 18, of the estrous cycle. However, average serum concentrations of LH, FSH and estradiol from 5 d before until 2 d after expected estrus were not different between treatment groups. Concentrations of AD in follicles 4 to 6 and greater than 7 mm in diameter were greater in gilts immunized against AD. Mean serum progesterone was higher on d 9 and 12 after mating in AD immunized gilts than in controls. Immunization against AD had no effect on maintenance of pregnancy or embryo survival rate.     

Ovarian remnant syndrome: revascularization of free-floating ovarian tissue in the feline abdominal cavity

Nine, healthy, intact female domestic shorthair cats were ovariohysterectomized. At the time of surgery and following removal, the major portion of one ovary was loosely sutured to the mesentery and replaced in the abdominal cavity. Six months later, an abdominal laparotomy was performed in order to retrieve the ovarian remnants. Histopathological examination of the remnants showed viable tissue and evidence of ovarian follicles or corpora lutea in eight of nine (88.9%) cats. The ninth ovarian remnant was atrophied and fibrotic. Measurement of serum estradiol and progesterone, vaginal cytology, and stimulation of estrus and ovulation with a protocol using pregnant mare serum gonadotropin (PMSG) followed by human chorionic gonadotropin (hCG) were unreliable indicators of ovarian activity in this study. Revitalization of an ovarian remnant was shown to occur in the absence of surgical implantation.     

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)     

Effect of ovarian follicles on luteal regression in heifers

Our objectives were to determine whether or not ovarian follicles contribute to spontaneous luteal regression in heifers and, if so, when during diestrus do follicles exert their effect. Thirty-one Holstein heifers having displayed at least one estrous cycle (19 to 21 d) were assigned, as available, to randomized blocks for a factorial experiment. Reproductive organs were exposed through a midventral incision on d 9, 12 or 15 postestrus (estrus = d 0). Visible follicles were electrocauterized and both ovaries were x-irradiated (1,500 rads) in treated heifers, whereas ovaries of controls were exteriorized but follicles were not destroyed and ovaries were not x-irradiated. In two additional heifers, the ovary containing the corpus luteum was exteriorized and x-irradiated on d 15 postestrus, but follicles were not electrocauterized. Jugular blood was collected before and every 8 h after surgery until d 24 postestrus. All heifers were ovariectomized on d 24 postestrus to inventory follicles and to weigh corpora lutea. No follicles (greater than or equal to 1 mm diameter) were observed in ovaries from treated animals and concentrations of estradiol-17 beta did not change over time, whereas different numbers of follicles were observed in ovaries from controls and concentrations of estradiol-17 beta increased (P less than .05) during proestrus. Hence, treatment destroyed follicles and prevented follicular development. On d 24 postestrus, corpora lutea from treated heifers (5.5 +/- .5 g) were heavier (P less than .001) than corpora lutea from controls (1.1 +/- .1 g), independent of day when follicles were destroyed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Expression of estrogen receptor alpha (ERalpha) and estrogen receptor beta (ERbeta) in the ovarian follicles and corpora lutea of pregnant swine

The objective of the study was to demonstrate the presence of estrogen receptor alpha (ERalpha) and beta (ERbeta) protein and corresponding mRNA in porcine ovarian follicles and corpora lutea obtained on day 10, 18, 32, 50, 71 and 90 post coitum (p.c.) using immunohistochemistry, Western blot, and RT-PCR analysis. Immunohistochemistry showed that ERalpha protein was located in the granulosa cells of ovarian follicles and the strongest immunoreaction was observed on days 32 and 50 p.c. The ERbeta protein was found mainly in theca cells of follicles as well as in luteal cells. The most intense immunoreaction was observed on day 18 p.c. within theca cells, while in the corpus luteum (CL) the intensity of ERbeta staining gradually increased and remained elevated at mid and late pregnancy. In CL by day 50 p.c. immunoreaction for ERbeta was present only in small luteal cells, but starting from day 71 to 90 p.c. it was observed in both small and large luteal cells. Western blot analysis was performed and validated data obtained from immunohistochemistry. RT-PCR results indicated that ERalpha mRNA was expressed only in ovarian follicles of the pregnant swine, while that of ERbeta in both follicles and CL. The results suggest an autocrine/paracrine role of estrogens acting via both ERalpha and ERbeta in the regulation of the ovarian function during pregnancy and for the process of successful reproduction.     

Localization of oestrogen receptors within various bovine ovarian cell types at different stages of the oestrous cycle

In the present study oestrogen receptor alpha(ERalpha) and oestrogen receptor beta (ERbeta) mRNA were localized in various ovarian cell types of 23 cows at different stages of the oestrous cycle. ERalpha was detected by immunohistochemistry and the localization of ERbeta mRNA was examined using in situ hybridization. The immunostaining of ERalpha was low in the ovarian follicles, tunica albuginea and surface epithelium, but high in cells of the deep stroma and superficial stroma, which indicates a functional role of ERalpha in the cells surrounding the follicles. In contrast, ERbeta mRNA scores were low to moderate in primordial and primary follicles, and increased with the development of the follicle. ERbeta mRNA scores were higher in cystic follicles than in obliterative follicles. In the corpora lutea and corpora albicantia the scores for ERbeta mRNA were moderate. Furthermore, in the corpora lutea, ERbeta mRNA levels showed cyclic variations and were low during early dioestrus. The correlation between plasma progesterone levels and the score for ER was low and negative in all ovarian cell types. This study demonstrates the predominant role of ERbeta over ERalpha in bovine ovarian structures. Furthermore, the colocalization of both ERbeta mRNA and ERalpha in most cell types suggests possible interactions between both ER subtypes.     

Relationships among the corpus luteum,follicles and conceptus in sheep

This study aimed to investigate the intra- and interovarian relationships among the corpus luteum (CL), the largest follicle (LF) and follicular population in non-pregnant and between the conceptus and ovarian structures in pregnant ewes. In experiment 1, the follicular and luteal structures were examined in 538 reproductive systems of non-pregnant Awassi ewes. The follicular population was categorised into small (SF), medium (MF) and large (LF) groups. Inter-relationships between CL and follicular population and between LF and subordinate follicles were determined. In experiment 2, the location and number of conceptuses were identified and correlated with the ovarian structures in 58 reproductive systems of pregnant ewes. Effects of pregnancy status, stage of pregnancy, pregnancy side and conceptual number on follicular population were determined. The results showed that the right ovary was more active than the left ovary. CL had intraovarian positive effect on the number of medium and large follicles. LF had no local suppressive effect on the subordinate follicles. Side and stage of pregnancy and the conceptual number did not affect the follicular population. Accordingly, it can be concluded that the LF has no local suppressive effect on the subordinate follicles. The CL has intraovarian positive effect on the follicular population. Follicular population does not show remarkable changes during the first term of pregnancy. The present study probably provides information which may help in the understanding of the ovarian dynamics during pregnancy in sheep.     

Old,new and the newest concepts of inhibition of luteolysis during early pregnancy in pig

In 1977 Bazer and Thatcher proposed that maternal recognition of pregnancy in the pig involves the secretion of PGF(2alpha) towards the uterine lumen (exocrine) rather than towards the uterine venous drainage (endocrine) as occurs in the non-pregnant pig during the mid to late stages of the estrous cycle. The retrograde transfer of PGF(2alpha) from the venous blood and uterine lymph into the uterus and the ability of the uterine vein and artery wall to accumulate PGF(2alpha) could constitute a part of putative mechanism of corpus luteum protection during early pregnancy. A luteotropic/anti-luteolytic effect of PGE(2) in the pig also has been frequently demonstrated and it seems that the most effective agent in changing PGE(2):PGF(2alpha) secretion is estradiol. The role for oxytocin during luteolysis and early pregnancy is controversial. It appears, however, that the main function of this hormone is autocrine and/or paracrine stimulation of PGF(2alpha) secretion. Pig trophoblastic interferons, unlike those of ruminants, do not themselves exert an anti-luteolytic effect in pigs. It is likely, that cytokines and angiogenic growth factors are involved in the initiation of luteolysis and/or maintenance of corpora lutea (CL).A discovery of functional LH receptors in porcine endometrium opened a new possibility for this hormone in luteolysis and perhaps in recognition of pregnancy in pigs. The endogenous LH pulses can provoke prostaglandin secretion from endometrium in pigs. On the other hand prolongation of up-regulation of LH receptors in endometrium of early pregnant gilts can additionally increase angiogenic factor production before the process of implantation is completed. Finally new integrated concepts of luteolysis and inhibition of luteolysis in pigs based on selectively reviewed information are presented.