Changes in serum glucose and lipids, and liver glycogen and phosphorylase during vitellogenesis in nature in the flounder (Platichtys flesus, L.).

1. During vitellogenesis in nature the concentration of phospholipid in serum of female flounders is correlated to the concentration of vitellogenin. 2. High levels of glucose and total lipid in blood occur before the onset of vitellogenesis. During early vitellogenesis the values decrease. At spawning they reach maximum levels. After spawning they decrease to low levels. 3. The concentration of glycogen in the liver oscillates during the period August to May. The phosphorylase activity shows peak activities with high concentrations of glycogen in the liver. There is no simple correlation between glycogen concentration in the liver and glucose concentration in the blood.     

Changes in activity levels of glutamate dehydrogenase and two transaminases of maternal liver during reproduction and after administration of oestradiol and progesterone in Zoarces viviparus (L.)

The enzyme activity per unit liver wet weight, the specific activity and the total liver activity level of glutamate dehydrogenase (GDH) and two transaminases (GOT and GPT) were studied during vitellogenesis and pregnancy. A steady level of activity was observed during the short summer vitellogenesis and during early pregnancy. During mid-term pregnancy in October the activity level significantly increased.
Administration of 5 and 100 μg oestradiol increased serum vitellogenin, total liver protein and the liver somatic index during late pregnancy. Oestradiol decreased activity and specific activity of the enzymes. However, expressing activity as total hepatic enzyme activity units in fish of standard body weight oestradiol had no effect.     

Antioxidant status of ewe's blood during pregnancy and lactation

Antioxidant status of ewe's blood during pregnancy and lactation was studied. It is revealed that the lipid peroxidation in erythrocytes is decreased, whereas in blood plasma is increased in the middle pregnancy, reaching the maximum shortly before peak of progesterone production. The activities of antioxidant enzymes are increased at the end of pregnancy, when the level of progesterone in blood is maximal. The positive correlation between plasma progesterone level and activity of erythrocyte superoxide dismutase (p < 0.01), between plasma cortisol concentration and total antioxidant capacity of blood plasma (p < 0.05) are revealed. It is shown, that the activities of antioxidant enzymes reach a maximum in the critical periods of pregnancy, providing the protection of the maternal organism and the fetus against negative influence of free radicals. The positive correlation between a level of hormones and separate parameters of antioxidant system in blood allows to assume, that progesterone participates in the regulation of enzymatic part of the antioxidant system, while cortisol is implicated in the regulation of the non-enzymatic component of this system in ewes.     

Plasma progesterone in alpaca (Lama pacos) during pregnancy, parturition and early postpartum

Plasma progesterone concentration during pregnancy and the early postpartum period was measured by radioimmunoassay in five alpacas (Lama pacos), of the Huacaya breed, whose pregnancy length had a mean of 344.8 +/- 4.4 days. Concentration of progesterone increased (P < 0.001) from low premating values (0.11 +/- 0.06 nmol/l) to greater values at 30 days of pregnancy (8.05 +/- 1.13 nmol/l) and remained high up to 2 months of pregnancy. A slight transitory decline was observed between 3 to 7 months. Concentration of plasma progesterone dropped markedly during the 72 h before parturition especially, at the day of parturition.     

Lipid dynamics in the viperine snake, Natrix maura, from the Ebro Delta (NE Spain)

Body reserves play a major role in several aspects of vertebrate biology. Accurate identification and quantification of body reserves constitute a useful contribution to the better understanding of the energetic costs of reproduction and the implication of food availability in life history traits of organisms. In this study, lipid content in fat bodies, liver and muscle of the viperine snake (Natrix maura) were measured along an active season. Samples were collected monthly from a natural population of the Ebro Delta Natural Park (NE Spain). This methodology pointed out that lipids stored in fat bodies were the main energetic source during reproduction. In addition, lipids stored in liver appeared to be critical for vitellogenesis, while lipids stored in muscle turned out to be a supplementary energetic resource to fuel reproductive effort. Relationships between changes in body reserves and prey availability in canals of the Ebro Delta were also considered. In males, lipid reserves presented a positive correlation with food availability. On the contrary, lipid reserves in female viperine snakes decreased during vitellogenesis even though food availability increased in this period, which suggests a quick transfer of body lipids to clutch. In April, when rice fields of the Ebro Delta were dry and aquatic prey was scarce for viperine snakes, males and females presented a lower lipid content in fat bodies, liver and muscle than they did in other months, showing a clear link between prey availability and body reserves during food shortage. Thus, patterns of variation in fat levels indicated that Natrix maura is a capital breeder since it acquires resources in advance and stores them until they are invested during the reproductive period. Nevertheless, the shortage in April forces Natrix maura to turn into a facultative income breeder to fuel vitellogenesis. Finally, fat reserves in body components were compared with an estimate of body condition calculated by the residuals from the regression of body mass on body length. In male viperine snakes, the estimate of body condition was correlated with fat levels, revealing that this index is useful to measure condition in living individuals. On the contrary, body condition in females was not correlated with fat levels, which suggests that it is not appropriate to infer condition in female viperine snakes since it depends on the reproductive status of the individuals.     

Liver Changes Related to Oocyte Growth in Roach,a Single Spawner Fish,and in Bleak and White Bream,Two Multiple Spawner Fish

The objectives of this study were to compare the seasonal hepatic activity and to evaluate whether this activity was related to the dynamics of oocyte recruitment in three cyprinids from the River Meuse (Belgium): the roach Rutilus rutilus as a single spawner, and the bleak Alburnus alburnus and the white bream Blicca bjoerkna as multiple spawners. In roach, hepatosomatic index (HSI) was low during exogenous vitellogenesis. However, the strong development of the rough endoplasmic reticulum (RER), the presence of large mitonchondria with many inner folds, and the depletion of glycogen, lipid droplets and phospholipids indicated intense hepatic activity during that period. Prior to spawning, hepatic activity decreased as indicated by a decrease of the HSI and in the hepatocytes a regression of the RER and the disappearance of the lipid droplets and glycogen. In bleak, the oocyte recruitment occurred continuously throughout the spawning season and the vitellogenic activity of the liver remained intense throughout that period as suggested by high HSI and the ultrastructure of the hepatocytes. In contrast in white bream the vitellogenic activity of the liver decreased during the spawning season as the completion of the vitellogenesis took place prior to spawnings. We concluded that the vitellogenic activity of the liver is strongly related to oogenesis.     

Progress report: Results of a long-range captive breeding program for the bottlenose dolphin,Tursiops truncatus and Tursiops truncatus gilli

There have been 36 bottlenose dolphins born in breeding colonies at Sea World, California, and Sea World, Florida, from 1978–1985. Significant features of this successful reproductive program are construction of a compatible breeding colony, early hormonal detection of pregnancy, pre- and postnatal association of inexperienced mothers with experienced females and their calves, and minimized handling of the mothers and calves until the calves are over 1 year of age. Inclusion in the breeding colonies of males old enough to be effective breeders is stressed. Females in the colonies have successfully bred from 8–9 years of age to 23–24 years of age. Calving intervals in the colonies vary from 2 to over 3 years. Calves are born all year round, with some peaking in calving activity in the spring and fall months. Serum progesterone levels greater than 6,000 pg/ml, maintained over a 4–6-week period, are considered indicative of pregnancy. Progesterone levels vary from less than 10,000 to over 50,000 pg/ml during gestation, averaging 25,000 pg/ml. The need for further study of variation in the pattern of progesterone levels during pregnancy is emphasized. A gestation period for Tursiops of 11.5 to 12 months is consistent with our cumulative progesterone data.     

Pregnancy effects on distribution of progesterone receptors, oestrogen receptor alpha, glucocorticoid receptors, Ki-67 antigen and apoptosis in the bovine interplacentomal uterine wall and foetal membranes

Until recently, studies dealing with the uterus of the pregnant cow focus primarily on the placentome or on early and late pregnancy. Thus, there is a paucity of information about many aspects of the interplacentomal uterine wall including adherent foetal membranes. Corresponding tissue specimens were collected at the slaughterhouse and in animals undergoing premature caesarean section. Two specimens per month of pregnancy were assessed immunohistochemically for progesterone receptors, oestrogen receptor alpha and glucocorticoid receptors, Ki-67 protein and TUNEL procedure was performed. The latter two methods were employed in three animals each per months 1 and 2, 3 and 4, 7 and 8 and in six animals undergoing caesarean section at days 274 and 275 post insemination or during spontaneous labour. Results indicate that proliferation and apoptosis are of minor importance for tissue homeostasis since both can histochemically be detected only sporadically. Thus, at the sites investigated here, cellular hypertrophy plays an important role for tissue growth during pregnancy. Progesterone receptors, oestrogen receptor alpha and glucocorticoid receptors, however, exhibit cell type and pregnancy stage specific distribution patterns within the tissues assessed. Progesterone receptor immunoreactive scores remained fairly unchanged during pregnancy. Oestrogen receptor alpha scores, however, generally decreased and glucocorticoid receptors increased with ongoing gestation. Progesterone receptors and oestrogen receptor alpha were present in endometrial stroma and in myometrial smooth muscle cells during whole pregnancy. Oestrogen receptor alpha was detectable during whole pregnancy also in uterine glands. Progesterone receptors were, however, present at a very low level at the latter site only during months 1-3 and 6-9. Oestrogen receptor alpha and glucocorticoid receptors may also mediate uterine blood flow since they were present in the tunica media of uterine blood vessels. Results of the present study indicate, that progesterone and its receptor play an important role during whole gestation, mainly for uterine quiescence. Glucocorticoids and their receptors - possibly in cooperation with oestrogens and decreasing amounts of the oestrogen receptor alpha - should trigger processes initiating parturition, such as endometrial prostaglandin production. Further studies - including the periparturient period - should help to understand the exact role of the extraplacental compartment of the uterine wall for the initiation and progress of parturition.     

Relationship between progesterone, oestrone sulphate and cortisol and the components of renin angiotensin aldosterone system in Spanish purebred broodmares during pregnancy

The coordinated interaction of the components of the renin angiotensin aldosterone system (RAAS) with reproductive hormones such as progesterone, oestrogens and cortisol during pregnancy has been widely reported to play a vital role in foetal and placental development in various species, significantly influencing the proper achievement of pregnancy and foetal viability at birth. These interactions have not yet been clarified in mares. Thus, the purpose of the present research was to analyse the relationship between cortisol (CORT), progesterone (P4) and oestrone sulphate (OESTRONE), and the components of the RAAS, renin (REN), angiotensin II (ANG-II) and aldosterone (ALD) concentrations in Spanish broodmares during pregnancy.Venous blood samples were obtained monthly from a total of 31 Purebred Spanish broodmares aged between 5 and 15 years during the 11 months of pregnancy. Plasma and serum REN, ANG-II, ALD, P4, OESTRONE and CORT concentrations were analysed by competitive immunoassay. Pregnancy in Purebred Spanish broodmares is characterised by a progressive increase in REN and ALD, a decrease in CORT levels, variable fluctuations in P4 and OESTRONE and no variations in ANG-II concentrations (P < 0.05). Serum P4 was not correlated with either ALD or CORT. The OESTRONE and REN levels were not correlated, while OESTRONE and ALD showed a positive correlation (r = 0.16; P < 0.05). These results suggest that the sustained stimulation of the RAAS in normal pregnancy in Spanish broodmares is not totally dependent on the changes in P4 and CORT concentrations, despite the involvement of OESTRONE in the secretion of ALD. This brings into question the possible involvement of oestrogen in the secretion of ALD by a mechanism which is not exclusively dependent on REN. Consequently, at physiological levels, OESTRONE is not the only stimulus for REN synthesis, and the mineralocorticoids ALD and CORT do not show a competitive mechanism with P4 during pregnancy in mares. Other mechanisms which do not depend on these hormones should be considered in the modification of the RAAS during pregnancy in Spanish mares.     

The role of prolactin and progesterone in the regulation of lipogenesis in maternal and foetal rat liver in vivo and in isolated hepatocytes during the last day of gestation.

The administration of progesterone on day 21 of gestation increases the rates of lipogenesis in the liver in vivo and in hepatocytes isolated from rats on day 22 of pregnancy. Bromocriptine administration increases the rates of hepatic lipogenesis in vivo, but has no effect on lipid synthesis in hepatocytes under the same treatment conditions. Concurrently, the administration of progesterone or bromocriptine on day 21 to the mother increases the rates of lipogenesis in the foetal liver in vivo on day 22. The rates of lipid synthesis in foetal isolated hepatocytes are increased by progesterone administration, but remain unchanged by bromocriptine.     

Serum and urinary hormones during pregnancy and the peri- and postpartum period in an Asian elephant (Elephas,maximus)

Blood and urine samples were collected weekly from an Asian elephant (Elephas maximus) for 10 months before conception, throughout pregnancy, and for 10 months after parturition. Additional daily samples were collected for 41 days before through 10 days after parturition to define endocrine events during the peripartum period. During gestation, serum progesterone concentrations increased gradually and, after ～13 weeks, were higher (P < 0.05) than those observed during the nonpregnant luteal phase. Concentrations peaked at ～12 months of gestation, gradually declined during the last month, and then decreased sharply to nondetectable levels 2 days before parturition. A 12 week lactational anestrus was observed before cyclicity resumed. The urinary profile of progestagen excretion paralleled that of circulating progesterone (r = 0.79; P < 0.05); however, radioimmunoassay of HPLC-separated fractions of urinary eluates indicated that this immunoactivity was not associated with native progesterone. After remaining basal through the first 16 weeks of gestation, serum prolactin concentrations increased to 100-fold about midterm and remained elevated until after parturition. Neither serum nor urinary cortisol concentrations were altered during pregnancy, but both increased markedly the day after parturition and remained elevated above prepartum levels for several weeks thereafter. These data indicate that analysis of serum prolactin can confirm pregnancy in the Asian elephant after ～4 months of gestation and that daily monitoring of serum or urinary progestagens is useful for predicting parturition. © 1995 Wiley-Liss, Inc.

1 This article is a US Government work and, as such, is in the public domain in the United States of America.

     

Evidence of a progesterone receptor in the liver of the green frog Rana esculenta and its down-regulation by 17 beta estradiol and progesterone

Progesterone is a versatile hormone showing an ample variety of effects. One of the numerous functions attributed to progesterone is the modulation of vitellogenesis in oviparous vertebrates. As a prerequisite for the possible involvement of progesterone in vitellogenesis modulation, we investigated the presence of a progesterone receptor (PR) in the liver of the female green frog Rana esculenta. 3H-Progesterone (3H-P) binding activity was found in both cytosol and nuclear extract of the liver of Rana esculenta. The progesterone-binding moiety showed the typical characteristics of a true receptor, such as high affinity, low capacity, and specificity for progesterone. It also bound to DNA-cellulose and was eluted with a linear salt gradient at a concentration of 0.05 M of NaCl. The progesterone-binding moiety was down regulated by steroid hormones, in that ovariectomy resulted in a significant increase, in both cytosol and nuclear extract, of 3H-P binding activity with respect to intact females. On the contrary, 3H-P binding activity was almost undetectable after estradiol and/or progesterone treatment. The progesterone binding moiety of Rana esculenta was analyzed by Western blotting with the aid of a monoclonal antibody raised against the subunits A and B of the chicken PR. An immunoreactive band of about 67 kDa was observed in the liver of both intact and treated females. The 67 kDa band showed an increased intensity in ovariectomized animals, while it was faint following treatment with estradiol and/or progesterone. This is the first report on the presence of a progesterone receptor (PR) in the liver of an amphibian. PR of Rana esculenta is down regulated by estradiol and/or progesterone and shows peculiar immunological and biochemical characteristics, which make it rather different from the PR of other vertebrates.     

Peripartal changes of estrone, progesterone and prostaglandin in the water buffalo

The peripheral blood plasma concentration of estrone, progesterone and 15-keto-13, 14-dihydroprostaglandin F2alpha (PGF2alpha metabolite) were determined by radioimmunoassay techniques during the peripartal period in 5 buffalo cows belonging to a river type breed. Estrone levels started to increase from below 200 pg/ml about 15 days prior to parturition, and reached high concentrations (400-750 pg/ml) during the last 5 days of pregnancy. The estrone concentration decreased to baseline levels after delivery. The concentration of progesterone fluctuated between 800 and 2000 pg/ml until 15 days before calving and showed a gradual increase during the last 15 days of pregnancy. The progesterone levels declined abruptly on the day of calving and remained below 100 pg/ml for up to 60 days post-partum. Increased levels of the prostaglandin metabolite were recorded from 15 days prior to parturition with further increases occurring during the last 3 days of pregnancy. PGF2alpha metabolite levels declined gradually after parturition, reaching base line levels 15-20 days after calving.     

Changes in progesterone levels and distribution of progesterone receptor during vitellogenesis in the female mud crab (Scylla paramamosain)

Vertebrate-type steroids, such as progesterone, have been identified in crustaceans. The physiological activity of progesterone during vitellogenesis is still not well understood. In this study, progesterone levels in the female mud crab, Scylla paramamosain, were determined by enzyme-linked immunosorbent assay. Peak levels of progesterone were detected during the previtellogenic stage in the hemolymph, ovary, and hepatopancreas, whereas the progesterone level decreased significantly in vitellogenic stage I. During vitellogenic stage II, progesterone levels rose again in the hemolymph and ovary, but continued to decrease in the hepatopancreas. By using western blotting, progesterone receptor (PR), with an apparent molecular weight of 70 kDa, was identified in the ovary during both vitellogenic stages I and II. By means of immunohistochemistry, PR was detected mainly in the follicle cells during vitellogenic stage I and in the nuclei of oocytes in vitellogenic stage II. Our results strongly suggest that progesterone promotes vitellogenesis in the mud crab, S. paramamosain via a classical genomic mechanism.     

Effect of progesterone,mifepristone, and estrogen treatment during early pregnancy on conceptus development and uterine capacity in Swine

A series of experiments was performed to investigate the influence of progesterone at Days 2 and 3 of pregnancy on conceptus development and uterine capacity. In experiment 1, unilaterally hysterectomized-ovariectomized (UHO) white crossbred gilts were given no treatment, estradiol valerate (5 mg given on Days 11 and 12), or progesterone (200 mg/day on Days 2 and 3 after mating). On Day 105 of pregnancy, each fetus and its associated placenta were weighed, and the number of live and dead fetuses was recorded for each litter. Early progesterone treatment reduced (P < 0.05) litter size (a measure of uterine capacity in UHO gilts). In experiment 2, intact white crossbred gilts were mated, given no treatment or progesterone treatment on Days 2 and 3 of pregnancy, and farrowed. Progesterone treatment decreased (P < 0.05) pregnancy rates. In pregnant gilts, progesterone had no effect on the number of live or stillborn piglets at birth, and gestation length was decreased (P < 0.05). Progesterone treatment did not affect the number of large or small piglets. In experiment 3, intact gilts were mated at estrus and then received 1). no treatment or treatment with 2). 100 mg, 3). 200 mg, or 4). 400 mg mifepristone (also known as RU486) on Day 2 of pregnancy. On Day 11 of pregnancy, both uterine horns were flushed, the number and diameter of each conceptus was recorded, and the flushed material was assayed for total protein and acid phosphatase. The 400 mg mifepristone treatment decreased conceptus diameter (P < 0.05) and total protein (P = 0.06) in the uterine flushings. In experiment 4, UHO gilts were mated at estrus, injected with either corn oil (control) or mifepristone (400 mg) on Day 2 of pregnancy, and killed on Day 105 of pregnancy, and the number and weight of live fetuses and placentas was recorded. In contrast to the effect of progesterone treatment, mifepristone decreased uterine capacity by decreasing the number of small conceptuses. These data suggest that progesterone concentrations on Days 2 and 3 of pregnancy in swine influence the rate of conceptus development during early pregnancy and uterine capacity during later pregnancy.     

Progesterone concentrations and contents in the plasma, ovary, adrenal gland and placenta of the pregnant Natal clinging bat Miniopterus schreibersii natalensis

Reproduction in the Natal clinging bat in South Africa at 25°S is characterized by fertilization in April, implantation during July and parturition in November. In the present study plasma progesterone concentrations peaked two months before implantation, this coinciding with significant peaks in ovarian progesterone content and concentration. Plasma progesterone concentrations remained relatively low during and after implantation, but increased significantly during the limb bud stage to peak levels one month before parturition. Adrenal progesterone content and concentration did not follow a specific trend; post-implantation ovarian progesterone content and concentration remained at baseline values. Placental progesterone content increased significantly with foetal weight (age). The present study showed that pregnancy in these bats is characterized by two major sources of progesterone, with the ovary as the primary source before implantation and the placenta the primary source during embryonic and foetal development.     

Reproductive cycles of male and female tuatara (Sphenodon punctatus) on Stephens Island, New Zealand

Cycles in gonadal activity and plasma sex steroid concentrations were investigated in wild female and male tuatara on Stephens Island, New Zealand. Females nest once every four years on average. Vitellogenesis is spread over the first three years, and mating, ovulation and nesting occur in the fourth. Oviducal eggs are carried for6–8 months before nesting. Although the length of this ovarian cycle is unparalleled among oviparous reptiles, the associated cycles in plasma concentrations of sex steroids are similar to those in other reptiles. Mean plasma concentrations of oestradiol increase during vitellogenesis, peak at mating, fall rapidly before or around ovulation, are low during most of gravidity, and rise slightly during late gravidity-nesting. Plasma concentrations of testosterone show a similarcycle. Mean plasma concentrations of progesterone are low during vitellogenesis and peak around ovulation. This periovulatory surge falls within1–2 months, and mean concentrations are low during the final 5–6 months of gravidity. Male tuatara show an annual, pre-nuptial reproductive cycle. Mean plasma concentrations of testosterone are low during winter, rise during spring, peak during midsummer-early autumn, when pre-nuptial displays and mating occur, and fall in mid autumn. Limited histological data indicate that spermiogenesis occurs during midsummer-early autumn, and also support a previous study suggesting that there may be no period of complete testicular and epididymal regression. In comparison with the pre-nuptial cycles of other temperate-zone reptiles, the cycle in male tuatara shows a more prolonged duration of testicular activity and of rising or elevated plasma testosterone concentrations. The prolonged reproductive cycles of male and female tuatara may be adaptations to a temperate environment with cool summers and cool, but not freezing, winters.     

The effects of progesterone on follicular growth in the rabbit ovary

Studies were undertaken to measure the growth of follicles in the rabbit ovary during periods of elevated blood levels of progesterone. The progestin was increased in the blood by pregnancy or by implantation of progesterone pellets, which raised blood progesterone to near the levels measured during pregnancy. After 1, 2, 3, or 4 weeks of pregnancy or progesterone-pellet treatment, follicles of 1.0 mm external diameter or greater were dissected out of the ovaries and their external diameters were measured; then, each follicle was extracted for measurement of estradiol content. Blood levels of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were measured in these animals as well. Follicles up to 2.5 mm in diameter were found in the ovaries of nonpregnant and untreated animals while 1.8 mm was the maximal size found during pregnancy or progesterone-pellet treatment. Furthermore, both in pregnant and in progesterone-treated rabbits, the follicular estradiol content and concentration were significantly suppressed compared to follicles from untreated rabbits. The progesterone pellets had no major effect on the levels of LH and FSH in the blood; the concentration of these gonadotropins in the progesterone-treated rabbits was virtually identical to levels previously measured in the blood of pregnant animals. The results of these studies indicate that progesterone exerts an inhibitory action on follicular development and steroidogenic function in the rabbit ovary. The progesterone action appears to be exerted directly on the ovary and is not indirect, by way of an inhibition of gonadotropin secretion.     

Patterns of lipid storage and mobilisation in the female green sea turtle ( Chelonia mydas)

Reproductive data from southern Queensland indicate that vitellogenesis in female Chelonia mydas takes approximately 8 months and is followed by a migration to a breeding area. At Heron Island, females lay multiple clutches over approximately 3 months. To investigate how females mobilise and store lipid during the breeding season we collected plasma, yolk, and fat tissue samples from females at a variety of stages during the nesting season. In breeding females, concentrations of plasma triglyceride increased seasonally. They reached peak concentrations during vitellogenesis and courtship, remained high throughout the nesting season, and then declined to a nadir after the last clutch. Plasma protein concentration increased throughout the breeding season, peaking following the last clutch for the season. Yolk lipids were highest during courtship and were similar throughout the nesting season, suggesting that uptake of lipid by ovarian follicles is completed prior to the beginning of the nesting season. Plasma triglyceride decreases in females with prolonged periods of unsuccessful nesting, and total lipid levels in adipose tissue and follicle yolks were significantly lower in atretic females. It appears that: (1) endogenous energy reserves can be reduced by stochastic environmental events (such as those reducing nesting success), and (2) a metabolic shift signalling the end of the nesting season is characterised by a drop in plasma triglycerides and slight increase in total plasma protein.