Immunoreactive forms of ACTH released by the adenohypophysis of the rat during the perinatal period in vivo and in vitro studies

Immunoreactive ACTH levels were determined in whole plasma, eluate fractions of plasma and perfusates of anterior lobe extracts subjected to chromatography on Sephadex G-50 fine. In the fetal plasma, ACTH levels were higher on day 19 than on days 17, 18, 20 and 21. After birth, ACTH concentrations dropped to reach the lowest values in one week old newborns; thereafter they increased until weaning on day 21. They were then similar to those of non pregnant adult females. Three peaks of immunoreactive ACTH were present in all the chromatograms; the first one eluted near the void volume ("big" ACTH, PM approximately 44,000), the third one coeluted with human ACTH (1-39) ("little" ACTH, PM approximately 4,500) and the second one eluted midway between the 2 previous peaks ("intermediate" ACTH, PM approximately 13,000). During the last days of pregnancy, the proportion of the "little" form of ACTH in the fetal plasma showed a gradual increase whereas that of the "big" one decreased. On days 17, 19 and 21 of gestation the anterior lobes of fetal pituitary glands released in vitro these 3 forms of immunoreactive ACTH in the same proportions as those observed in the anterior lobes. In contrast, the proportions of the circulating forms of ACTH were quite different; the "little" one gradually increased and the "big" one decreased as gestation progressed. In vitro controlled tryptic digestion of the isolated "big" form led to the appearance of "intermediate" and "little" forms suggesting some transformations in the circulation of the ACTH forms released by the fetal hypophysis in vivo.     

Cyclic nucleotides and haemoglobin concentration in rabbit bone marrow cell suspensions stimulated by purified erythropoietin

This study was conducted to determine the possible correlations between cyclic nucleotides cyclic adenosine monophosphate (cAMP) and cyclic guanine monophosphate (cGMP), and haemoglobin (Hb) concentration in nucleated cell suspensions of rabbit bone marrow incubated with erythropoietin (Ep). The levels of cAMP and cGMP were measured following the addition of different Ep concentrations to the suspensions. The Hb concentration was also measured in suspensions treated with Ep, dibutyryl cAMP (db-cAMP) or dibutyryl cGMP (db-cGMP), respectively. The following results were obtained: (1) upon the addition of 1 IU ml-1 Ep, an increase of cAMP levels was related to an increase in Hb concentration; while a decrease of Hb concentration was related to an increase of cGMP levels obtained when 0.1 IU ml-1 Ep was present in the incubation mixture. (2) A mimetic effect on Hb concentration was obtained upon the addition of db-cAMP or db-cGMP to the suspensions. (3) A quantitative correlation was found between the cAMP/cGMP ratio and Hb levels in cellular suspensions. This rapport was reviewed with respect to the controls as a decrease in Hb concentration when the ratio is less than one and an increase in Hb concentration when the ratio is greater than one.     

Changes in blood flow distribution during the perinatal period in fetal sheep and lambs.

We have measured total blood flows and blood flows per 100 g tissue to major tissues at 120 and 140 days gestation in fetal sheep and at 3 and 21 days of age in lambs (gestation period = 144 +/- 2 days). Between 120 and 140 days gestation, flow per 100 g tissue increased by 74, 150, and 317% in the renal, intestinal, and hepatic arterial beds, but no further significant change in flow was observed at 3 or 21 days postpartum. Blood flows per 100 g to cerebral hemispheres and cerebellar tissues also increased dramatically during late gestation (142 and 121%, respectively), but declined sharply by 3 days postpartum (73 and 75%, respectively). Brain blood flows at 21 days postpartum remained substantially below late gestational levels. Adrenal blood flows per 100 g more than doubled during late gestation, fell by more than half at birth, and only partially recovered by 21 days of age. Blood flows to carcass tissues did not change in late gestation, fell at birth, then partially recovered. Pre- and post-natal increases in brain blood flows were almost entirely attributable to increased perfusion rather than tissue growth, whereas large perinatal increases in flow to the diaphragm paralleled tissue growth. Tissue growth and increased perfusion per 100 g contributed almost equally to increased blood flows to kidneys postnatally, and to adrenal glands and the gastrointestinal tract prenatally.     

Dynamics of alpha-fetoprotein concentration in mice of different genotypes during the neonatal period]

A comparative assay of the alpha-fetoprotein (alpha-FP) level in mice of various genotypes (CBA, C3H, C57BL/Se/Sn, BALB/c, CC57W, AKR and nude--nu/nu) was conducted in the course of 3 weeks of postnatal period. The concentration of alpha-FP reached the following levels: the first day 2(-10)-2(-9); the 5th day 2(-8); the 8th day 2(-7); the 15th day 2(-4); on the 22nd day the level was zero. Nude mice which showed the alpha-FP concentration of 2(-2) on the 15th day were an exception. A conclusion was drawn that the alpha-FP synthesis was based not on the athymia of nude mice per se, but upon other unknown factors.     

Immunoreactive and biologically active somatostatin in human and sheep milk

The presence of immunoreactive and biologically active somatostatin in sheep and human milk has been demonstrated. Milk somatostatin exhibits similar chromatographic behavior to that of synthetic somatostatin-14 on both reversed-phase C18 and cation-exchange high-performance liquid chromatography columns. Milk, in contrast to plasma, contains only somatostatin-14-like material. Milk somatostatin was capable of inhibiting the basal and the prostaglandin-induced release of growth hormone from anterior pituitary cell cultures in a pattern similar to synthetic somatostatin-14. The concentrations of the peptide, as determined by radioimmunoassay, were found to be 113 pg/ml in human milk and 150 +/- 4.8 pg/ml (mean +/- range) in sheep milk. These values are severalfold higher than the corresponding concentration of the peptide in the plasma of these species. These findings are analogous to our previous observations concerning two other hypothalamic hormones, luliberin and thyroliberin [Baram, T., Koch, Y., Hazum, E. and Fridkin, M. (1977) Science (Wash. DC) 198, 300-302]. The high concentration of somatostatin and other neuropeptides in milk implies either an active concentrating mechanism in the mammary gland or an additional extrahypothalamic source for the synthesis and release of these peptides.     

A quantitative histological study of adrenal development during late gestation and the perinatal period in intact and hypophysectomized fetal sheep

Growth of the sheep adrenal was studied during late gestation and the perinatal period. Adrenals were recovered from 28 fetuses taken at D100 (D0: day of mating), D120, D132 and D144, and in 4 newborn animals 3 days after birth (equivalent to D151). Animals were of the Ile de France breed. Cortex (without capsule) and medulla volumes increased respectively by 7.2 and 2.4 between D100 and D151. The 2 parts had the same volume between D100 and D120; thereafter the cortex became predominant, representing 74% of the whole gland at D151. Hypertrophy and hyperplasia were shown after D132 in the cortex (zona fasciculata); they were observed later after D144 in the medulla (central zone). Fifteen fetuses hypophysectomized at D100 or D120 were recovered at D120 or D144. The lack of pituitary inhibited cortical growth (zona fasciculata) and suppressed hypertrophy and hyperplasia. The medulla continued to grow after hypophysectomy but to a lesser extent than in controls.     

Control of glucose metabolism in the perinatal period

The central importance of glucose as a fuel for energy metabolism and growth of the fetus is clear as is the role of insulin in coordinating its utilisation by many fetal tissues. What is less clear is the qunatitative nature of the interaction between the fetus and placenta in organising glucose metabolism. Increasingly there is evidence that the fetus coordinates some of the supply of glucose to the placenta and that this is particularly important when uterine blood flow is reduced. It is unclear how this is regulated, but substrate cycles of glucose and lactate appear to make a significant contribution to carbohydrate metabolism in fetus and placenta. Another area as yet unresolved in the control of fetal glucose metabolism is the coordination of the changes that occur around the time of birth. Notable of these is the activation of glycogen mobilisation and of glucose synthesis and changes in the setting of glucose regulatory mechanism. These are briefly reviewed.     

Rat liver glycogen metabolism in the perinatal period

The correlation between blood glucose levels, the concentration of glycogen, the activities of glycogen sythase and phosphorylase and their respective kinases and phosphatases was examined in liver of rat fetuses between day 18 of gestation and one day after birth. Between day 18 and 21 there is a rapid increase in the concentration of glycogen and in the activity of synthase a and a much slower increase in the activity of phosphorylase a. The activity of the respective kinases increased rapidly during this period and reached maximun on day 21. The activity of synthase phosphatase and phosphorylase phosphatase increased after day 18, to reach a maximum on day 19 and 20, respectively, but decreased again towards day 21. The possibility that the changes in glycogen concentration and enzyme activities were related to an effect of glucose of AMP on the respective phosphatases was considered. It was found that the Km of phosphatase for glucose in the prenatal period was 5–7 mM, as in the adult. Since the level of blood glucose during this period was constant (2.8 mM), an effect of glucose on phosphatase activity seems unlikely. AMP concentration increased between day 18 and 21 from 6–15 nmol/g. In view of the low level of phosphorylase a activity during this period, the increase in AMP concentration is not considered to be important in the regulation of glycogen breakdown at this time.Immediately after birth blood glucose levels dropped to 5 mg/dl. This was accompanied by a rapid decrease in glycogen concentration and in the activity of glycogen synthase and a rise in phosphorylase activity. Blood glucose levels returned to the initial level within 1 h after birth, whereas the changes in glycogen concentration and enzyme activities continued for at least 3 h after birth. On day 22 all parameters examined had reached the level found in adult rat liver.It is suggested that the rapid changes observed immediately after birth are due to an effect of hypoglycemia mediated by hormones and cannot be ascribed to direct effects of metabolites on the enzyme systems involved.     

The renin-angiotensin system in the perinatal period in rats

Plasma concentrations of renin, angiotensinogen, kininogen, total protein, and renal renin concentration were measured in rats before spontaneous birth, immediately after vaginal delivery, during the subsequent 48 h, as well as at the ages of 10, 20 and 80 days. Preterm rats had a plasma renin concentration about 15 times higher than adults, which increased further in 1 h-old vaginally-delivered rats. Thereafter renin fell to very low levels within 2 h, rose again during the first day and remained at 4 times the adults level until day 10. Renal renin content and concentration increased over the whole observation period, except for a slight fall of renin concentration in the first 3 h after birth. In pre- and full-terms rats, angiotensinogen concentration was only 20% that of adults, reaching even lower values immediately after delivery, due to excessive consumption by renin. Thereafter, angiotensinogen increased more than 10 fold within 48 h. Kininogen concentration in plasma was higher than in adults and stable up to the 10th postnatal day. We conclude that vaginal delivery is a strong stimulus for renin release, the resulting high concentration of renin being responsible both for the increased turnover of angiotensinogen and the subsequent inhibition of renin release. The cause and biological significance of the dramatic increase of angiotensinogen during the first 48 h of life remains obscure.     

Plasticity of haemoglobin concentration and thermoregulation in a mountain lizard

The plastic capability of species to cope with the new conditions created by climate change is poorly understood. This is particularly relevant for organisms restricted to high elevations because they are adapted to cold temperatures and low oxygen availability. Therefore, evaluating trait plasticity of mountain specialists is fundamental to understand their vulnerability to environmental change. We transplanted mountain lizards, Iberolacerta cyreni, 800 m downhill to evaluate the plastic response in body condition, thermoregulation traits, haemoglobin level, and haemoparasite load. Initial measurements of body mass, total haemoglobin concentration ([Hb]), hematic parasite intensities, dorsal luminance, and thermoregulatory behaviour were resampled after two and four weeks of acclimation. We also tested whether an anti-parasitic drug reduced haemoparasite intensity. After only two weeks of acclimation to a lower elevation, lizards decreased 42% in [Hb], had 17% less parasite intensities, increased body condition by 25%, and raised by ~3% their mean preferred temperatures and their voluntary thermal maximum. The anti-parasitic treatment had no significant effect on the intensity of hematic parasites, but our results suggest that negative effects of haemoparasites on [Hb] are relaxed at lower elevation. The rapid plastic changes observed in thermal preferences, body condition, [Hb], and parasite intensity of I. cyreni demonstrate a potential plastic response of a mountain specialist. This may be adaptive under the climatic extremes typical of mountain habitats. However, there is uncertainty in whether the observed plasticity can also help overcome long term environmental changes.     

The control of cardiovascular shunts in the fetal and perinatal period

The fetal circulation has two major vascular shunts, the ductus arteriosus and the ductus venosus. The ductus arteriosus connects the pulmonary artery with the descending portion of the aortic arch, hence shunting most of the right ventricular output away from the unexpanded lungs. The ductus venosus connects instead the portal sinus with the inferior vena cava and allows well-oxygenated umbilical vein blood to bypass the liver and reach the central circulation rapidly. Both blood vessels cease their function after birth and undergo permanent closure. It is now well established that prenatal patency of the ductus arteriosus is an active state sustained by a prostaglandin. A similar mechanism has been recently recognized in the fetal ductus venosus. Evidence is presented indicating that prostaglandin E2 and prostaglandin I2 are natural relaxants, respectively, for the ductus arteriosus and the ductus venosus. In addition, both vascular shunts share the dependence on an endogenous cytochrome P-450 mechanism to develop their contractile tone. This mechanism may be important in the normal process of shunt closure at birth. While broadening the knowledge of fetal cardiovascular homeostasis, advances in this field have important implications for the prevention and management of certain pathological conditions affecting the newborn.