The effects of stress on plasma ACTH and corticosterone in young and aging pregnant rats and their fetuses

Compared to younger rats, old rats exhibit prolonged elevations of plasma ACTH and corticosterone (CORT) in response to stress. In addition, CORT crosses the placenta. To investigate whether fetuses of older rats may be exposed to higher concentrations of CORT during development than fetuses of young rats, we compared the effects of stress on hypothalamic-pituitary-adrenal (HPA) axis function in young and aging pregnant rats and their 19-day-old fetuses. The plasma of the mothers and fetuses was assayed for ACTH and CORT by radioimmunoassay. Both young and aging pregnant rats showed a significant increase in plasma ACTH and CORT immediately after exposure to stress. However, aging rats had more prolonged elevations of ACTH and CORT than young rats. This suggests that, like old male rats, aging pregnant rats have an alteration in feedback inhibition of the HPA axis. Prolonged elevation of CORT was also seen in fetuses of aging mothers. These results have important implications concerning the effects of stress during pregnancy at different maternal ages, and for the potential deleterious consequences of prolonged prenatal elevation in stress hormones on the offspring of aging females.     

Acute effects of caffeine injection on neutral amino acids and brain monoamine levels in rats

The injection of caffeine (100 mg/kg, i.p.) into male rats acutely increased brain levels of trytophan, serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA). Blood levels of glucose, nonesterified fatty acids (NEFA) and insulin also increased, while those of the aromatic and branched-chain amino acids fell. Serum tryptophan levels either did not fall, or increased. Consequently, the serum ratio of trypthopahn to the sum of other large neutral amino acids (LNAA) increased. Less consistently noted were increases in serum free tryptophan levels. Brain tyrosine levels were not appreciably altered by caffeine, nor was the serum tyrosine ratio. In dose-response studies, 25 mg/kg of caffeine was the minimal effective dose needed to raise brain tryptophan, but only the 100 mg/kg dose elevated all three indoles in brain. In no experiments did caffeine, at any time or dose, alter brain levels of dopamine or norepinephrine. Caffeine thus probably raises brain tryptophan levels by causing insulin secretion, and thereby changing plasma amino acid levels to favor increased tryptophan uptake into brain. The rises in brain 5-HT and 5-HIAA may follow from the increase in brain tryptophan, although further data are required clearly to establish such a mechanism.     

Contractile effects of vanadate on aorta rings from virgin and pregnant rats

The present study was undertaken to characterize the contractile effects of vanadate on thoracic aorta rings from virgin and term-pregnant rats. Vanadate caused concentration-dependent contraction in rat aortic rings with an EC₅₀ (concentration producing 50% maximum response) of 0.10 mM. Contractions in response to vanadate were equivalent to the ones measured with 1 M phenylephrine. The effects of vanadate were not affected by indomethacin (up to 10 M), an inhibitor of prostanoid cyclooxygenase, but were blocked in a concentration-dependent manner by staurosporine (0.1–1.0 M), an inhibitor of protein kinase C. Vanadate exhibited a significant decrease of contractile responses in aorta of pregnant as compared to virgin rats. When aortic rings were bathed in presence of different concentrations of vanadate, the concentration-response curve to phenylephrine was shifted to the left, but maximum response was not affected. The potentiation of the contractions to phenylephrine by vanadate was significantly more prominent in aorta of virgin than of pregnant rats. These results suggest that the contractile effect of vanadate on rat aorta is independent of endogenous prostanoids and may be mediated by protein kinase C-dependent pathway. These results also show that the contractile response to vanadate on the rat aorta is impaired during pregnancy.     

Effect of starvation on nutrition and insulin secretion in pregnant rats and their fetuses

Pregnancy is thought to create a metabolic condition of accelerated starvation. To clarify this idea, the effect of fasting on pregnant rats (day 21 of gestation) and their fetuses was examined. Although pregnancy significantly increased plasma insulin, plasma ketone body concentrations in fed pregnant rats were higher than those of age-matched fed virgin rats. After 48 hr fasting (i.e., fasting during days 19-21 of gestation), plasma insulin was markedly decreased in virgin rats compared with term pregnant rats, while ketone bodies were significantly higher in pregnant rats than in virgin rats. Body weight was lower in fetuses from fasted mothers than those from fed mothers. Starvation also markedly diminished the insulin response to glucose in isolated, perfused pancreases in both virgin and pregnant rats. The amount of insulin released during glucose stimulation was greater in pregnancy, and the inhibitory effect of 48 hr fasting on insulin release was greater in virgin rats than in pregnant rats. It is possible, therefore, that in term pregnant rats a decrease in insulin release caused by fasting may cause more profound catabolism than in nongravid rats.     

Effect of L-propargylglycine on metabolism of sulfur-containing amino acids in pregnant rats and their fetuses

Cystathionine accumulated in several tissues of dams and fetuses by a single intraperitoneal administration of L-proparglyglycine to pregnant rats. Cystathionine in the liver of dams reached its maximal level at about 15 hrs after L-proparglyglycine injection (10 mg/300g), while that in the kidney and brain of dams, and in the liver, kidney, and brain of fetuses reached a maximum at about 21 hrs. The content of cystine in the liver of fetuses decreased gradually in proportion to the amount of L-proparglyglycine administered. Cystathionine gamma-lyase activity in the liver of dams and fetuses decreased to about 2-4% of that of control rats at 15 hrs after L-proparglyglycine injection, and that in the kidney and pancreas of dams to about 10-20% of that of control rats. On the other hand, cystathionine beta-synthase activity did not show significant changes from that of control rats.     

Chronic stress and its effects on adrenal cortex apoptosis in pregnant rats

The model of chronic intermittent stress by immobilization during pregnancy may produce alterations in the mechanisms that maintain adrenal gland homeostasis. In earlier investigations using this model, significant variations in plasma prolactin and corticosterone levels, and adrenal gland weights were observed. We hypothesized that chronic stress causes changes in apoptosis in the adrenal glands of pregnant rats. We identified and quantified apoptotic cells in the adrenal cortex and examined their ultrastructural characteristics using transmission electron microscopy. Adrenal glands of pregnant rats at gestation days 12, 17 and 21 were studied for control and experimental (stressed) rats. Immunolabelling techniques, stereological analysis and image quantification of adrenal gland sections were combined to determine differences in apoptosis in the different cell populations of the adrenal cortex. The apoptotic index of the experimental rats showed a significant reduction at gestation day 17, while at days 12 and 21 there were no differences from controls. Moreover, the apoptotic index of the reticular zones in control and experimental animals showed a significant increase compared to the glomerular and fascicular zones at the three gestation times studied. Chronic stress by immobilization reduced the caspase-dependent apoptotic index at gestation day 17, which may be related to variations in plasma concentrations of estrogens and prolactin.     

Effect of tryptophan on livers of pregnant and lactating rats and their fetuses and pups

The effect of the administration of L-tryptophan on hepatic polyribosomes and protein synthesis in pregnant rats and their fetuses and in lactating rats and their pups was investigated. Pregnant rats tube-fed tryptophan 1 hr before killing revealed increased hepatic protein synthesis but essentially unmodified polyribosomal aggregation of maternal livers while no changes were observed in fetal livers in comparison to controls (water-treated). Lactating rats tube-fed tryptophan 1 hr before killing revealed increased polyribosomal aggregation and protein synthesis of the livers in comparison to controls. Pups of these mothers that received tryptophan intraperitoneally 1 hr before killing did not reveal a significant change in the hepatic polyribosomes or protein synthesis.     

Effects of ethanol on brain monoamine content of spontaneously hypertensive rats (SHR)

Spontaneously hypertensive rats (SHR) were administered either 2.4 g/kg ethanol or an isocaloric glucose daily for 4 weeks and the levels of norepinephrine (NE), epinephrine (EP), dopamine (DA), serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) in different brain regions were determined. Results indicated a 3-fold increase in NE level in brain stem and hypothalamus and more than 2-fold increase in DA in corpus striatum in alcohol-treated rats as compared to controls. There was a significant increase in the level of DA in the corpus striatum but the levels in cerebral cortex, brain stem and hippocampus were decreased instead. Decreases in 5-HT levels were found in hypothalamus, brain stem, cortex and cerebellum of alcohol-treated brain as compared to untreated controls. These results indicate alterations of the biogenic amine contents in different regions of the SHR brain after chronic ethanol ingestion. Since stimulated release of biogenic amines in the SHR brain has been implicated in the regulation of blood pressure, changes due to ethanol ingestion may be a risk factor in hypertensive patients.     

Amelioration of some metabolic effects produced by hyperthyroidism in late pregnant rats and their fetuses. Effects on lipids and proteins.

We have studied the effect of 40-45 days administration of 1 mg/kg thyroxine on protein and lipid metabolism in liver, heart, lungs, kidneys and adrenal glands of virgin and 21-day pregnant rats and their fetuses and placentae. The chronic administration of thyroid hormone produced significant increases in serum T3 and T4 in both groups as well as in organ weights and protein concentrations in virgin rats, but much smaller modifications in pregnant ones. Hyperthyroidism decreased the weight of fetal livers and increased that of placentae; protein content was increased in all fetal organs. Hyperthyroidism induced increases in phospholipid concentrations in all the organs and in total lipids only in liver and heart of adult rats, which were not counteracted by pregnancy. Pregnant rats had increases in total lipids in liver and kidneys and in adrenal phospholipids. In hyperthyroid fetuses there was an increase in hepatic total lipids and no changes in phospholipids. Hepatic lipogenesis (measured by in vivo incorporation of 3H2O into lipids) was increased by hyperthyroidism in virgin and pregnant rats, but the increase was significantly smaller in the pregnant hyperthyroid rats compared with the virgin ones. Fetal lipogenesis in liver and lung was not changed. In addition, an increase was observed in lipogenic enzyme (fatty acid synthetase and glucose-6-phosphate dehydrogenase) activities in hyperthyroid virgin rats which was prevented by pregnancy. In fetuses only pulmonary glucose-6-phosphate dehydrogenase was increased when expressed in terms of tissue weight. Our results indicate that the metabolic effect of hyperthyroidism is attenuated in pregnant rats and their fetuses, when compared with adult virgin rats, in most of the parameters studied.(ABSTRACT TRUNCATED AT 250 WORDS)     

Acute and short term effects of ethanol on membrane enzymes in rat brain

Changes in the biophysical and biochemical character of membranes brought about by ethanol have been emphasized in the underlying mechanism of alcohol toxicity. Membrane enzymes such as Na⁺, K⁺ activated ATPase, 5-nucleotidase, and -glutamyl transpeptidase were studied in cerebral cortex, cerebellum, and brain stem of rats subjected to acute and short term ethanol toxicity. Acute ethanol toxicity was induced by intraperitoneal injection of 1 ml of 7M ethanol per 100 g body weight of rat and the animals were sacrificed half an hour after the administration. Short term ethanol toxicity was induced by intraperitoneal injections of 0.5 ml (7 M ethanol) per 100 g weight of the rat for 7 days and the animals were sacrificed half an hour after the last injection. In acute ethanol toxicity the activity of Na⁺, K⁺-activated ATPase was found to decrease significantly in cerebral cortex and brain stem, while in short term alcohol toxicity, the activity was found to increase in cerebral cortex and cerebellum. The activity of -glutamyl transpeptidase was found to increase in all the three regions in acute and short term ethanol toxicity. No change in the activity of 5-nucleotidase was observed in any of the regions either in acute or in chronic ethanol toxicity. While a significant increase in the activity of adenosine deaminase was found in cerebral cortex, cerebellum, and brain stem in acute ethanol toxicity, the same was found to decrease significantly in cerebral cortex and a persistent increase in brain stem in short term ethanol toxicity. The above changes in the activities of the enzyme were discussed with reference to the well known changes in the membrane structure and consequent alteration in brain function.This work forms part of a Ph.D. thesis.     

Changes of glucocorticoid level in plasma and adrenal glands of pregnant rats and their offspring after antenatal stress of different etiology

We investigated the glucocorticoid level in plasma and adrenal glands of pregnant rats in the period of early organogenesis. Tests were performed 24 hours after stress of different etiology (acute hypobaric hypoxia, intermittent normobaric hypoxia, and immobilization) and then repeated in the adult offspring. There was a significant decrease in the glucocorticoid level in pregnant rats 24 hours after hypoxic stress. Various changes of the basal glucocorticoid level were found in the offspring after antenatal stress. Changes were mostly found in female offspring.     

Effects of neurotrophic stress on maternal metabolism and binding of plasma cortisol in late pregnant guinea-pigs and their fetuses

The maternal metabolic clearance rate (MCR) and the binding of cortisol in the plasma of fetus and mother were estimated 8 days before term in conscious pregnant guinea-pigs, control or subjected to a neurotrophic stress (they were immobilized for 3 h in a dark room in front of an intermittent luminous flash). The maternal MCR of cortisol dropped during pregnancy related to an increase in corticosteroid binding globulin (CBG) and the stress mimicked these changes with a decrease in MCR and a rise in CBG. The increase in cortisol concentrations occurring in the fetal plasma after maternal stress could be related to the increase in free cortisol in the stressed mother. Nevertheless, although the fetal CBG did not rise after maternal stress, the increase in free cortisol in the fetal plasma remained moderate, due to the numerous free sites of CBG and albumin for cortisol.     

Mechanical stimulation of mammary gland development in virgin and pregnant rats

The mammary glands of pregnant Sprague-Dawley rats prevented from self-licking by collars around the neck (n = 9) were only about 50% as developed as those of pregnant rats without collars (n = 9). However, when the ventral body surface of pregnant collared rats was stimulated mechanically (n = 9), mammary glands increased in size, underwent lobuloalveolar growth, and secreted milk in a manner similar to normal pregnant rats.When virgin rats were stimulated mechanically for 22 days (n = 9), their mammary glands had significantly greater secretory tissue and lobuloalveolar development than did the glands of nonstimulated virgin rats (n = 9).Stimulated rats were tied to the stimulating device under ether anesthesia. Because ether and restraint have been shown under some conditions to be Stressors that increase development of the mammary glands, two additional pregnant collared control groups were added. One was restrained on the device without stimulation (n = 9). The other was lightly anesthetized with ether (n = 10). Mammary underdevelopment occurred in both groups. This suggested that stress of ether and restraint were unlikely causes of the mammary development in the stimulated group.     

Strontium-85 in the fetuses of pregnant rats and mice

Pregnant SPF Wistar rats and ICR/Swiss albino mice were injected in the tail vein with 85SrCl2 with 0.05 mM inactive carrier (SrCl2) given in volumes of 0.1 ml. The activity in the injected volume was about 14 MBq per kg of rat and 13 MBq per kg of mouse. The animals were injected at 2 or 13 days of gestation. The activity retained by the fetuses was quantitatively determined at three stages of the fetal intrauterine development: in rats at 14, 16 and 21 days of gestation, in mice at 14, 16 and 20 days of gestation. The activity of fetuses and/or placentas with fetal membranes was measured using a TESLA automatic gamma counter. Results indicate that fetuses of mice retained a significantly (P less than 0.01) greater percent of strontium activity than fetuses of rats. The highest specific activities (the percentage of total activity retained per gram of fetal tissue) were found in the late pregnancy period (at 21 days of gestation in rats and 20 days of gestation in mice) in animals that were injected with the radionuclide at 13 days of gestation.     

Lipogenesis in interscapular brown adipose tissue of virgin, pregnant and lactating rats. The effects of intragastric feeding.

Lipogenesis in brown adipose tissue of virgin rats increased 8--10-fold after intragastric feeding with glucose or medium-chain triacylglycerol, and this increase was prevented by short-term insulin deficiency. Brown adipose tissue increased in weight during pregnancy, regressed during lactation and hypertrophied again on weaning; the rate of lipogenesis paralleled these changes. Glucose did not increase brown-adipose-tissue lipogenesis at mid-lactation.     

Effect of propranolol on plasma renin activity, renal cortex renin, and adrenal and brain isorenins in rats.

Propranolol administration to rats was studied for its effects on plasma renin activity, renal renin content, and adrenal and brain isorenins. Propranolol was given intraperitoneally at 6 and 30 mg/kg/day for a 15-day period. Pulse rate was significantly decreased. There were no effects on the isorenin content of adrenal or brain tissue or on renal renin content. Rats responded in two completely different ways with respect to plasma renin activity. Two-fifths had a total suppression of plasma renin activity; the rest had concentrations similar to those in controls. These observations are consistent with those seen during chronic administration of propranolol to hypertensive patients and suggest that its antihypertensive effect may in some patients be through the suppression of renin release. Its mechanism of action in most patients remains at present unclear.     

Enzymatic and morphological response of the thymus to drugs in normal and zinc-deficient pregnant rats and their fetuses

In the thymus of normally fed pregnant rats the plasma membrane enzymes dipeptidyl peptidase IV (DPP IV) and alkaline phosphatase (alP) were found in cortical and medullary lymphocytes (thymocytes). Plasma membrane aminopeptidase A (APA) and adenosine monophosphate hydrolysing phosphatase (AMPP) were present in cortical reticular cells. In medullary reticular cells, aminopeptidase M (APM), gamma-glutamyl transferase (GGT), adenosine triphosphate (ATPP) and thiamine pyrophosphate (TPPP) cleaving phosphatases were detected. Medullary reticular cells did not contain APA. Lysosomal DPP I and II, acid phosphatase, acid beta-D-galactosidase, beta-D-N-acetyl-glucosaminidase, beta-D-glucuronidase and non-specific esterases occurred especially in macrophages at the corticomedullary junction. The 21-day-old fetal thymus showed a similar reaction pattern as the maternal organ except for APA which was absent before birth. After treatment of the pregnant rats with valproic acid (VPA), salicylic acid (SA), streptozotocin (ST) and retinoic acid (RA) APA showed an increase in activity in the thymic cortex. In addition, ST and RA induced AMPP, ATPP and TPPP activity in cortical reticular cells up to the same pattern as in medullary reticular cells. After ethanol (ET) administration severe damages occurred. The thymic cortex was free of DPP IV-positive lymphocytes; the medullary reticular cells showed reduced or no GGT and occasionally an increased APM activity. Dexamethasone (DEXA) given to normal or zinc-deficient rats produced the most severe lesions; thymocytes with DPP IV activity were completely absent in the cortex and medulla. In Zn-deficient pregnant rats similar alterations were observed as after ET.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Enzymatic and morphological response of the thymus to drugs in normal and zinc-deficient pregnant rats and their fetuses

Summary In the thymus of normally fed pregnant rats the plasma membrane enzymes dipeptidyl peptidase IV (DPP IV) and alkaline phosphatase (alP) were found in cortical and medullary lymphocytes (thymocytes). Plasma membrane aminopeptidase A (APA) and adenosine monophosphate hydrolysing phosphatase (AMPP) were present in cortical reticular cells. In medullary reticular cells, aminopeptidase M (APM), -glutamyl transferase (GGT), adenosine triphosphate (ATPP) and thiamine pyrophosphate (TPPP) cleaving phosphatases were detected. Medullary reticular cells did not contain APA. Lysosomal DPP I and II, acid phosphatase, acid -d-galactosidase, -d-N-acetylglucosaminidase, -d-glucuronidase and non-specific esterases occurred especially in macrophages at the corticomedullary junction. The 21-day-old fetal thymus showed a similar reaction pattern as the maternal organ except for APA which was absent before birth.—After treatment of the pregnant rats with valproic acid (VPA), salicylic acid (SA), streptozotocin (ST) and retinoic acid (RA) APA showed an increase in activity in the thymic cortex. In addition, ST and RA induced AMPP, ATPP and TPPP activity in cortical reticular cells up to the same pattern as in medullary reticular cells. After ethanol (ET) administration severe damages occurred. The thymic cortex was free of DPP IV-positive lymphocytes; the medullary reticular cells showed reduced or no GGT and occasionally an increased APM activity. Dexamethasone (DEXA) given to normal or zinc-deficient rats produced the most severe lesions; thymocytes with DPP IV activity were completely absent in the cortex and medulla. In Zn-deficient pregnant rats similar alterations were observed as after ET. When the drugs were applied to Zn-deficient pregnant rats, the alterations resembled those observed after drug treatment alone. In all cases of severe thymus degeneration, i.e. ET and DEXA treatment and Zn-deficiency, the number of macrophages and activities of lysosomal hydrolases in macrophages and reticular cells were increased; the lysosomal hydrolases were often homogeneously distributed over the cortex. Cell contacts between reticular cells and lymphocytes were reduced. Vacuoles occurred within the reticular cells.—The fetal thymus was reduced in size and the number of macrophages and the activities of their lysosomal enzymes were increased after Zn-deficiency, DEXA treatment and Zn-deficiency combined with ET administration.Supported by the Deutsche Forschungsgemeinschaft (Sfb 174)