Prolactin receptors in the mammary gland, corpus luteum and other tissues of the tammar wallaby, Macropus eugenii.

Specific binding of radio-iodinated ovine prolactin to subcellular tissue fractions of the tammar wallaby (Macropus eugenii) was investigated. Specific binding was found, in order of decreasing binding activity, in the lactating mammary gland, corpus luteum, corpus albicans, adrenal gland and ovary. Specific binding was absent in kidney, liver, brain and inactive mammary gland. The mean association constant (Ka at 23 degrees C) was determined as 0.90 x 10(9), 2.20 x 10(9), 2.44 x 10(9), 3.38 x 10(9) and 10.98 x 10(9) l/mol for mammary gland, adrenal, corpus albicans, corpus luteum and ovary respectively. The mean receptor concentration (N) varied from 92.87 x 10(-14) mol/mg protein for the mammary gland to 1.03 x 10(-14) mol/mg protein for the ovary. The concentration in the corpus luteum varied between tissue pools collected at different times of the annual breeding cycle. The specificity for prolactin was shown in the mammary gland and corpus luteum by the failure of ovine FSH, LH, GH and TSH to displace 125I-labelled ovine prolactin, whereas it was displaced readily by both ovine and bovine prolactin.     

Effect of post-ovulatory administered oestrogens on corpus luteum function.

The effect of orally given diethystilboestroldiphophate (DES) and 17alpha-ethinyl-oestradiol-3-methylether (EEM) on plasma progesterone levels was studied. Both compounds were administered for 5 days to 5 women in daily doses of 60 mg (DES) and 30 mg (EEM). The fully informed volunteers were found to have a normal menstrual cycle before the study. The mean corpus luteum phase (corpus luteum phase = days between LH surge and onset of menstruation) of all control cycles lasted 12.8 days. Daily plasma samples were collected for radioimmunoassay (RIA) of progesterone, immunoreactive oestrogens and LH. After a control cycle the first treatment was carried out with DES. The third and the fifth cycle were control cycles again. The EEM-treatment was done in the fourth cycle. Although the effect of the two compounds was different, a dependence of the age of the corpus luteum (CL) could be demonstrated for both. DES-treatment lowered plasma progesterone levels during administration. This effect was only demonstrable if the treatment was begun on the day of the LH-peak. The length of the CL-phase remained unaltered. EEM-treatment if started on the day of the LH surge, suppressed corpus luteum function in the late luteal phase. If the treatment was started later, the effect was less pronounced. The administration of both compounds did not shorten the time between ovulation and the next bleeding. After DES-treatment this interval was not altered. After EEM-treatment the subsequent bleeding was even delayed depending on slowly decreasing levels of plasma oestrogens.     

Steroid metabolism in the yolk sac placenta and endometrium of the tammar wallaby, Macropus eugenii

Yolk sac and endometrial tissue were obtained from tammar wallabies between 11 and 25 days after the removal of pouch young. Tissues were examined histologically and steroid-metabolizing enzymes were identified by incubation for 3 h at 37 degrees C in Medium 199 containing labelled steroid precursors. Yolk sac membrane (YSM) incubated with labelled pregnenolone produced a small amount of progesterone and pregnanediols; 80.5 +/- 8.4 (S.E.M.) % of the original substrate remained unmetabolized. Labelled androstenedione was metabolized to 5 alpha-androstane-3,17-dione and androsterone, and only 5.8 +/- 3.8% of the original substrate remained at the end of incubation. Incorporation of androstenedione or dehydroepiandrosterone (DHA) into phenolic compounds was low (0.5 +/- 0.1%). There was no evidence for the enzymes, arylsulphatase or sulphotransferase, in YSM. Endometrial tissue from the same animals metabolized pregnenolone, DHA and androstenedione, converted progesterone to androstenedione, and produced aqueous-soluble steroid conjugates. The results demonstrated that YSM contains enzymes associated predominantly with steroid catabolism and with incipient progesterone synthesis. The findings are discussed in relation to the histological appearance of the tissues and compared with placental steroid synthesis in eutherian mammals.     

Ovarian (corpus luteum) hemorrhage during anticoagulation therapy

Corpus luteum hemorrhage is a complication of long-term anticoagulant therapy that has rarely been reported in the literature. Within 16 months, we saw six cases in which young women taking anticoagulants to prevent clotting of prosthetic heart valves suffered corpus luteum hemorrhages. Diagnosis was difficult and delayed. Medication stopped the bleeding in three patients; bilateral salpingo-oophorectomy was necessary in the remaining three. Our patients had great difficulty controlling their anticoagulant therapy and had histories of previous bleeding episodes. We believe corpus luteum hemorrhage may become more common as more premenopausal women undergo cardiac valvular surgery.     

Angiogenesis in the corpus luteum

The ovarian corpus luteum plays a critical role in reproduction because it is the primary source of circulating progesterone. After ovulation, as the corpus luteum forms from the wall of the ruptured follicle, it grows and vascularizes extremely rapidly. In fact, the rates of tissue growth and angiogenesis in the corpus luteum rival those of even the fastest growing tumors. Thus, the corpus luteum provides an outstanding model for studying the factors that regulate the angiogenic process, which is critical for normal tissue growth, development, and function. In agreement with data from other tissues, vascular endothelial growth factors (VEGF) seem to be a major angiogenic factor responsible for vascularization of the developing corpus luteum. Recent data suggest that luteal expression of VEGF occurs primarily in specific perivascular cells, including arteriolar smooth muscle and capillary pericytes, and is regulated primarily by oxygen levels. In addition, soon after ovulation, pericytes derived from the thecal compartment appear to be the first vascular cells to invade the developing luteal parenchyma. The granulosa-derived cells produce a factor that stimulates pericyte migration. Moreover, nitric oxide (NO), which is a potent vasodilator and can stimulate VEGF production and angiogenesis, is expressed in endothelial cells of luteal arterioles and capillaries, often in association with expression of VEGF by luteal perivascular cells. Thus, we have proposed a model for the initial process of luteal vascularization in which hypoxia plays a major role. In this model, which we believe will apply to other tissues as well, a paracrine loop exists between the vascular endothelial cells, which produce NO, and the peri-endothelial cells (vascular smooth muscle and pericytes), which produce VEGF, to ensure coordinate regulation of luteal vasodilation and angiogenesis.     

Oxygen transport and acid-base balance during exercise in the tammar wallaby.

Rates of oxygen consumption (V(O)2), body temperatures and pulmonary blood temperatures, blood gases and blood pH were measured for seven 4.9+/-0.8 (SE) kg tammar wallabies (Macropus eugenii) during rest and during treadmill hopping. For animals resting on the treadmill V(O)2 averaged 0.030+/-0.003 L min(-1). During hopping V(O)2 increased linearly with speed up to 2.5 m sec(-1). Above 2.5 m sec(-1) V(O)2 was independent of hopping speed and averaged 0.340+/-0.004 L min(-1). At rest, rectal temperatures and pulmonary blood temperatures averaged 36 degrees C. During treadmill hopping, rectal temperatures and pulmonary blood temperatures increased similarly, to 39 degrees C. The Pv(CO)2 increased and pHv decreased in proportion to the increased V(O)2. The Pa(CO)2 and pHa were not significantly changed from values for animals resting on the treadmill. Cardiac output (Vb) averaged 0.97+/-0.04 L min(-1) when the wallabies were at rest on the treadmill and increased linearly with treadmill speeds up to 2.5 m sec(-1). Above 2.5 m sec(-1) Vb was independent of hopping speed and averaged 2.9+/-0.04 L min(-1). When data for all speeds were combined, Vb increased linearly with V(O)2. Thus, in spite of their unique mode of locomotion wallabies have maintained relationships between pulmonary ventilation and V(O)2 and between Vb and V(O)2 that are similar to those reported for eutherian mammals.     

Oestrogen metabolism in the endometrium, corpus luteum and ovarian residual tissue of the rabbit

Reproductive tissues (uterine endometrium, corpus luteum and ovarian residual tissues) from pregnant and pseudopregnant rabbits were incubated with equimolar concentrations of [3H]oestrone and [3H]oestrone sulphate (0.375 pmol) to monitor the changes in oestrogen metabolism during the early stages of pregnancy (days 0, and 3-8 post coitum) and to investigate the embyonic effect upon maternal oestrogen metabolism. Oestradiol-17 beta was the major metabolite formed from oestrone and sulphoconjugation occurred in all tissues studied. Oestrone sulphate was converted primarily to oestradiol-17 beta-3-monosulphate. Endometrial 17 beta-oxidoreductase significantly decreased and sulphotransferase increased in activity during the preimplantation period, but no differences were noted between gravid and non-gravid horns in unilaterally pregnant animals, nor between pregnant or pseudopregnant animals. Significant decreases occurred in 17 beta-oxidoreductase and sulphotransferase activity in luteal tissue, but these were more than offset by increases in tissue weight. No differences in the activities in luteal tissues were detected between pregnant or pseudopregnant animals, nor between ovarian tissue adjacent to gravid or non-gravid uterine horns. The results show that significant changes occur in oestrogen metabolism in the rabbit endometrium and corpus luteum within 8 days after ovulation, and that these changes result from maternal factors expressed systemically rather than by the effects of the developing conceptus expressed locally.     

Oxytocin receptors in the ovine corpus luteum

There is inconclusive evidence that oxytocin acts directly on the corpus luteum and affects steroidogenesis. Since any such action would probably be mediated by oxytocin receptors, these should be present in luteal tissue. In this study, homogenates of corpora lutea from both pregnant and non-pregnant ewes were examined for oxytocin receptors by radioreceptor assay. Specific oxytocin binding was not observed in luteal tissue during the oestrous cycle. However specific binding was found in the corpora lutea of pregnant ewes; appearing at a fetal head length of approximately 0.65 cm (about 30 days of pregnancy) and persisting to a head size of 11 cm, the largest size examined in this study. The affinity (Kd) of the receptor was calculated as 2.9 +/- 0.3 nmol/l (S.E.M.; n = 9), a value similar to that obtained for the uterus. The receptor number ranged from a low of 8.7 +/- 3.2 fmol/mg protein (n = 6) at a head size of less than 0.65 cm, to a maximum of 40.1 +/- 6.5 fmol/mg protein (n = 25) at a head size of 2.5-3.75 cm. These values were lower than our estimate of 588 +/- 39 fmol/mg protein (n = 5) for the uterus. It is concluded that a direct action of oxytocin on the corpus luteum is possible but only after the first month of pregnancy and not in the corpus luteum of the oestrous cycle.     

Morphometric estimate of gas-exchange tissue in the new-born tammar wallaby,Macropus eugenii

The lung of the new-born marsupial is at the terminal air sac stage of development. The maturational status of the lung of new-born tammar wallaby was assessed using established morphometric techniques and the results were compared with data from a morphometric study of the lung of the rat. Volume densities of the parenchyma and non-parenchyma, conducting airways and blood vessels, the relative volumes of airspace and tissue, the thickness and the composition of the septa differed between the two species. In addition the volume of capillaries and the surface area of the effective gas-exchange tissue was greater in the new-born rat than in the new-born tammar pouch young. The lung of the new-born tammar appears to be at an earlier phase of the terminal air sac stage than that of the new-born rat. Lung development up to birth appears to be commensurate to the metabolic needs of the organism at birth.     

Production a corpus luteum angiogenic factor responsible for proliferation of capillaries and neovascularization of the corpus luteum.

Factors controlling the changes in the vascular pattern of the ovary that occur during the reproductive cycle have been investigated. By using the rabbit cornea, the abilities of ovarian corpus luteum and of follicles to induce neovascularization have been compared. While the corpus luteum is capable of inducing neovascularization, the follicles do not have this ability. It is therefore likely that the corpus luteum actively participates in its own neovascularization by secreting a factor that we have called "corpus leutem angiogenic factor" (CLAF).     

Convection requirement is established by total metabolic rate in the newborn tammar wallaby

Ventilation (VE) and metabolic rate, determined from both pulmonary and cutaneous gas exchange, were measured in 39 newborn tammar wallabies, Macropus eugenii, aged between 0 and 3 days. In 1-day-old animals both total metabolic rate (skin+lung exchange) and ventilation were approximately 50% of the values predicted for eutherian newborns of equivalent body mass. Hence, the convection requirement (VE/total metabolic rate) of the newborn tammar is close to predicted values for newborns and adult mammals in general. Metabolic rate in the newborn tammar is supported to some extent by cutaneous gas exchange, approximately 30% of the total in the 1-day-old animal. This ratio diminishes with increasing age as the lung takes on an increasingly more important role for respiratory exchange. The early establishment of the convection requirement in the newborn tammar, despite significant cutaneous gas exchange, provides supporting evidence that metabolic rate per se is important in establishing the level of ventilation.     

Angiogenesis in the human corpus luteum: changes in expression of angiopoietins in the corpus luteum throughout the menstrual cycle and in early pregnancy

CONTEXT: Blood vessel stabilization is regulated by angiopoietins and important for angiogenesis in the corpus luteum. OBJECTIVE: To study angiogenesis and blood vessel stabilization in the human corpus luteum, changes in expression of angiopoietin (Ang)-1, Ang-2, and their specific receptor, Tie-2, together with the number of blood vessels and pericytes were examined in the corpus luteum throughout the menstrual cycle and in early pregnancy. DESIGN: The number of blood vessels and pericytes was determined by immunohistochemistry for CD34 and alpha-smooth muscle actin, respectively. Ang and Tie-2 expression were examined by immunohistochemistry or RT-PCR. RESULTS: The number of blood vessels increased during the early luteal phase, whereas the number of pericytes was small in the early luteal phase and increased in the midluteal phase, suggesting that angiogenesis is undergoing during the early luteal phase and blood vessels are stabilized in the midluteal phase. Blood vessels and pericytes decreased in number during the late luteal phase. The increased number of both blood vessels and pericytes seen in the corpus luteum of early pregnancy suggests that angiogenesis is undergoing accompanied by blood vessel stabilization. Ang-2 expression with low Ang-1 expression was found during the early luteal phase. Thereafter, increasing Ang-1 expression during the midluteal phase, declining Ang-1 expression with continued Ang-2 expression during the late luteal phase, and relatively high Ang-1 expression in early pregnancy were observed. CONCLUSIONS: The change in Ang expression is closely associated with angiogenesis, blood vessel stabilization, and blood vessel regression during the divergent phases of luteal formation, luteal regression, and luteal rescue by pregnancy.