Virgin Control Rats Research Articles

Serum immunoreactive erythropoietin and red blood cell mass during pregnancy in conscious rats

Serum erythropoietin concentration increases during human pregnancy and presumably accounts for expansion of red blood cell mass. The mechanism(s) underlying gestational changes of serum erythropoietin are unknown. Moreover, if erythropoietin synthesis increases, then the organ(s) questions about erythropoietin in pregnancy, we first set out to establish an animal model. Chronically instrumented, conscious unrestrained rats were studied. 51Cr-labeled red blood cells and radioimmunoassay were used to assess red blood cell mass and serum erythropoietin, respectively. Except for a lower hematocrit (P < 0.05 vs. virgin rats) and a slightly higher plasma volume (P value not significant) for gravid rats on gestational day 6, all other variables measured in early pregnancy rats were comparable to those measured in virgin control animals. Significant increases in total blood volume, plasma volume, and red blood cell mass were observed by gestational day 13 (midpregnancy) when compared with virgin control rats. These changes were even more pronounced on gestational day 20. Serum immunoreactive erythropoietin was also significantly increased at both of these stages of pregnancy. We conclude that the gravid rat is a reliable animal model of human gestation in which to further investigate erythropoietin in pregnancy.

Role of renal nerves in renal responses to acute volume expansion during pregnancy in rats.

To examine the role of the renal nerves in renal responses to acute volume expansion (VE) at Days 17-19 of pregnancy in rats, the diuretic and natriuretic responses to acute VE were measured from intact and denervated kidneys. One group of pregnant rats (Pregnant 1) was treated with the same amount of VE (1 ml/min for 15 min) as age -and sex-matched virgin control rats, and a second group of pregnant rats (Pregnant 2) was treated with a VE corrected for the higher body weight (presumably expanded blood volume) normally observed in late pregnancy (1.38 ml/min for 15 min). Urine flow and sodium excretion were measured before and after VE from innervated and denervated kidneys in anesthetized (Inactin) rats. Mean arterial pressure was not significantly different among the groups. During VE, the increments in urinary flow (UV) rate and sodium excretion (UNaV) from the innervated kidneys of Pregnant 1 rats were significantly smaller (26.5% for UV and 17.0% for UNaV) than those from the innervated kidneys of virgin rats. Although the UV and UNaV were greater in the Pregnant 2 group than in the Pregnant 1 group, these differences were not statistically significant. However, the values were still significantly smaller than those observed in the control group (39.1% for UV and 52.8% for UNaV). Urine flow and sodium excretion from the denervated kidneys of pregnant rats (both groups) were not significantly different from those of denervated kidneys of control rats. These results demonstrate that the reduced diuresis and natriuresis observed during acute volume expansion in pregnant rats may be due to the contribution of tonic renal nerve activity during the third week of pregnancy.

Changes in rates of glucose utilization and regulation of glucose disposal by fast-twitch skeletal muscles in late pregnancy

Glucose utilization indices (GUI) were measured in vivo in conjunction with active pyruvate dehydrogenase complex (PDH(a) and glycogen synthase (GS) activities in fast-twitch skeletal muscles [extensor digitorum longus (EDL), tibialis anterior and gastrocnemius] of late-pregnant rats and age-matched virgin control rats in the fed state, after 24 h starvation and at 2 h after re-feeding with standard laboratory chow ad libitum after 24 h starvation. As demonstrated previously [Holness and Sugden (1990) Biochem. J 277, 429-433], GUI values of fast-twitch skeletal muscles of virgin rats were low in the fed ad libitum and the 24 h-starved states, but dramatically increased after subsequent chow re-feeding. GUI values of fast-twitch skeletal muscles of late-pregnant rats were also low in the fed and starved states and were increased by re-feeding, but the increase in GUI values elicited by re-feeding was greatly attenuated. PDHa activities in EDL, tibialis anterior and gastrocnemius in the fed state were unaffected by late pregnancy, and skeletal-muscle PDHa activities were decreased after 24 h of starvation in both groups. Whereas re-feeding of virgin rats with standard diet for 2 h restored PDHa activities in fast-twitch skeletal muscles to values for rats continuously fed ad libitum, PDHa activities in fast-twitch skeletal muscles of late-pregnant rats, although increased in response to re-feeding, remained considerably less than the corresponding fed ad libitum values after 2 h of re-feeding. In contrast, neither skeletal-muscle GS re-activation nor rates of skeletal-muscle glycogen deposition after re-feeding were markedly affected by late pregnancy. The results are discussed in relation to the specific targeting of individual pathways of glucose disposal in fast-twitch skeletal muscles during re-feeding in late pregnancy.

In vivo glucose utilization by individual tissues in virgin and pregnant offspring of severely diabetic rats.

Adult offspring of diabetic rats or SDF rats are characterized by insulin resistance in the liver and extrahepatic tissues; this insulin resistance does not worsen during pregnancy. In this study, we determined the glucose metabolic index in tissues of anesthetized virgin and pregnant control and SDF rats in basal conditions and during a euglycemic hyperinsulinemic clamp. Tissues comprised insulin-sensitive tissues (five skeletal muscles, diaphragm, and periovarian white adipose tissue) and control tissues (duodenum and cerebrum). In addition, this study measured the GMI of placenta and fetuses. In basal conditions, SDF rats showed a slight decrease (9-29%) in the GMI of skeletal muscles compared with control rats; it was not altered by pregnancy in any of the tissues. During physiological hyperinsulinemia, virgin SDF rats exhibited a 25-70% decrease in the GMI of skeletal muscles compared with control rats; this decrease was not observed in diaphragm, or in adipose tissue in which the GMI was found to be increased. During pregnancy, SDF rats did not show an additional drop in the GMI of skeletal muscles, whereas the GMI of both skeletal muscles and adipose tissue was clearly diminished (25-60%) in control rats. The GMI of skeletal muscles was therefore comparable in pregnant control rats and SDF rats. The placental, but not fetal, GMI was increased by 24% during hyperinsulinemia in control rats; the placental and fetal GMIs, in basal and hyperinsulinemic conditions, were similar in control rats and SDF rats. In conclusion, skeletal muscles, but not white adipose tissue, are involved in the peripheral insulin resistance of the SDF rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Adriamycin nephropathy: a model to study effects of pregnancy on renal disease in rats.

The influence of pregnancy on the evolution of primary renal disease is still a matter of controversy. Hypertension and derangement of renal function may occur. The pathophysiology of these complications is poorly understood. In the present study, we assessed the influence of pregnancy on the evolution of adriamycin (Adr) nephropathy. Four groups of animals were studied: 1) control virgin rats (C), 2) normal pregnant rats (NP), 3) virgin rats with nephropathy (Adr), and 4) pregnant rats with nephropathy (Adr-P). Inulin clearance measured at the end of pregnancy in awake rats was similar in NP (1.68 +/- 0.20 ml/min) and C (1.39 +/- 0.03 ml/min). In Adr-P rats, it tended to decrease (1.22 +/- 0.7 vs. 1.93 +/- 0.44 ml/min in Adr rats). Mean arterial pressure was increased in Adr-P rats (137 +/- 2.5 vs. 95 +/- 3.2 mmHg in NP; P < 0.001). Urinary protein excretion was 216 +/- 61 mg/day in Adr-P compared with 28.7 +/- 18 mg/day in Adr (P < 0.001). A significant increase in the glomerular thromboxane B2-to-prostaglandin E2 ratio was found in Adr-P rats (1.15 +/- 0.26 vs. 0.52 +/- 0.12 in Adr rats; P < 0.03). In NP rats, no change was observed. Kidneys and placentas were normal on light and electron microscopy. Thus pregnant rats with adriamycin nephropathy developed a clinical picture with several features of preeclampsia. Changes in glomerular prostanoid synthesis might play a role in the development of this complication.

Circulating metabolite utilization by periuterine adipose tissue in situ in the pregnant rat

To study the use of glucose for lipid synthesis by the periuterine adipose tissue in situ, 14C-glucose was infused through the left uterine artery of anesthetized, fed pregnant and virgin control rats. A greater amount of 14C-lipid always appeared in the adipose tissue from the left uterine horn than in the tissue from the right uterine horn, indicating direct utilization of the infused 14C-glucose by the tissue. Glucose utilization for both glyceride glycerol and fatty acid synthesis increased from day 0 (virgin rats) to day 20 of gestation and then decreased dramatically on day 21. In virgin and 12-day pregnant rats, glucose was incorporated into either lipidic moiety at similar rates, whereas in late pregnant rats glucose utilization for glyceride glycerol synthesis was four to five times greater than for fatty acids. The utilization of circulating fatty acids and the lipoprotein triglyceride-derived fatty acids was studied by infusing 14C-palmitate or 14C-triolein-labeled very-low-density lipoprotein (VLDL) through the left uterine artery in both virgin and 20-day pregant rats. Incorporation of fatty acids from either one of these plasma sources was significantly higher in the pregnant than in virgin rats. This high amount of fatty acid acquisition did not account for the very active glyceride glycerol synthesis observed in pregnant rats and can only be explained by the intracellular reesterification of some lipolitic fatty acids. The results suggest a highly accelerated triacylglycerol/fatty acid substrate cycle in adipose tissue during late pregnancy, which would allow active esterification (contributing to fat accumulation) and responsive lipolysis (permitting rapid fat mobilization) by the mother.

Amino acid metabolism and protein synthesis in lactating rats fed on a liquid diet

1. Amino acid metabolism was studied in control virgin rats, lactating rats and virgin rats protein-pair-fed with the lactating rats (high-protein virgin rats). 2. Urinary excretion of nitrogen and urea was higher in lactating than in control virgin rats, and in high-protein virgin rats it was higher than in lactating rats. 3. The activities of urea-cycle enzymes (units/g) were higher in high-protein virgin than in lactating rats, except for arginase. In lactating rats the activities of carbamoyl-phosphate synthase, ornithine carbamoyltransferase and argininosuccinate synthase were lower than in control virgin rats. When the liver size is considered, the activities in lactating rats were similar to those in high-protein virgin rats, except for arginase. 4. N-Acetylglutamate content was higher in high-protein virgin rats than in the other two groups. 5. The rate of urea synthesis from precursors by isolated hepatocytes was higher in high-protein virgin rats than in the other two groups. 6. The flooding-dose method (L-[4-3H]phenylalanine) for measuring protein synthesis was used. The absolute synthesis rates of mammary gland, liver and small-intestinal mucosa were higher in lactating rats than in the other two groups, and in high-protein virgin rats than in control virgin rats 7. These results show that the increased needs for amino acids during lactation are met by hyperphagia and by a nitrogen-sparing mechanism.

On mast cell‐mediated mitogenesis in normal and hyperplastic mesenterial windows in female rats

The outcome of the mast cell-mediated mitogenesis in hyperplastic membranous mesenterial windows of lactating rats as well as in normal mesenterial windows of age-matched and young virgin female rats was studied quantitatively in vivo and in organ culture. Besides elucidating the effect of age and tissue hyperplasia on mitogenic responsiveness, this approach should provide some insight into the pathogenic mechanics of the previously reported supranormal mast cell-mediated mitogenic reaction that emerges in similarly hyperplastic mesenterial windows of diabetic rats. Mast cell secretion was elicited by Compound 48/80 and the histamine release, which was quantified fluorometrically, was unaffected by lactation. The young female rats showed a statistically significant mast cell-dependent mitogenesis taken as the mitotic index and the fraction of the predominating fibroblasts and mesothelial cells in the (S+G2) cell cycle phases after Feulgen-DNA absorption analysis of the cells in situ. Although there was an age-dependent decrease in mitogenesis, the older lactating and non-lactating virgin control rats also showed mast cell-mediated mitogenesis measured as the specific DNA activity. The hyperplastic mesenterial tissue of the lactating animals showed a virtually normal mitogenic reactivity following local mast cell secretion, but at a lower level than in the age-matched controls. This finding suggests that the supranormal mast cell-mediated mitogenesis previously found in the hyperplastic mesenterial windows of diabetic animals is causally related to the diabetic condition rather than to the hyperplastic state of the test tissue.

Elevated calcium, phosphorus, and magnesium retention in pregnant rats prior to the onset of fetal skeletal mineralization.

Intestinal absorption and renal excretion rates of Ca, P, and Mg were compared in pregnant and control virgin rats fed a purified diet containing 0.55% Ca, 0.54% P, and 0.067% Mg. Four consecutive balance intervals of 5 days each were examined, beginning on day 1 of gestation. During days 6-10 of gestation, pregnant rats had elevated fractional intestinal absorptions of Ca (29.6 +/- 1.7 versus 20.6 +/- 1.5%), P (65.4 +/- 2.0 versus 59.9 +/- 0.9%), and Mg (54.3 +/- 1.5 versus 42.4 +/- 2.6%). Since urinary excretions of these elements did not change, the pregnant rats retained more Ca, P, and Mg than virgin rats. Fractional intestinal absorptions of these elements during pregnancy were similar to control values during days 1-5 and 11-15 of gestation and then rose for each element during days 16-20 of gestation. Presumably because of an increase in glomerular filtration rate, urinary excretions of Ca and Mg were elevated by 40 and 26% during days 16-20 of gestation. In contrast, urinary P excretion was decreased by 30% during days 11-20 of gestation. Analyses of uterine mineral contents indicated the increased maternal Ca and Mg retentions during pregnancy were balanced by the transfers of these elements to the fetuses. In contrast, pregnancy was associated with a net maternal retention of P. These data are consistent with previous observations of increased maternal skeletal mineralization during early pregnancy before the onset of fetal osteogenesis and subsequent enhanced maternal Ca intestinal absorption concurrent with fetal skeletal mineralization.

Prolactin enhances cell-to-cell communication among beta-cells in pancreatic islets.

To determine the role of prolactin in increasing junctional communication among islet beta-cells, we studied dye coupling in pancreatic islets exposed to elevated levels of prolactin in vivo and in vitro. Islets were isolated from rats immediately after lactation or from rats bearing mammosomatotropic tumors (MtTW15), conditions involving high levels of prolactin (either 5-fold or 1000-fold control levels, respectively). When beta-cells were microinjected with the gap junction permeant dye Lucifer yellow CH, the mean number of dye-coupled cells per injection was approximately 10-fold greater than in islets from virgin control rats. As a more direct test of the effects of prolactin on beta-cell coupling, islets isolated from virgin rats were treated for 90 min with 500 ng/ml rat prolactin in the presence of low glucose (2.8 mM) and were microinjected with dye. The mean number of dye-coupled cells per injection increased by 6.7-fold over controls with low glucose, demonstrating a direct effect of prolactin on beta-cell coupling. In vitro treatment with high glucose (16.7 mM) resulted in a 2.7-fold increase in dye-coupled cells per injection. We discuss the possible relationship between the effects of glucose and of prolactin on coupling.

Impaired basal and noradrenaline-induced iodothyronine 5′-deiodinase activity in brown adipose tissue from pregnant and lactating rats

Basal iodothyronine 5'-deiodinase activity is lowered in brown fat from 20-day pregnant, 5 and 15-day lactating rats when compared with virgin controls. Acute noradrenaline treatment caused a seven fold increase in 5'-deiodinase activity in brown fat from virgin control rats. Late pregnant and lactating rats showed a reduction in noradrenaline-induced 5'-deiodinase activity in brown adipose tissue and the maximum impairment was observed in 15-day lactating rats. Lowered 5'-deiodinase activity in brown fat during late pregnancy and lactation correlates with the known reduction in the thermogenic activity of the tissue during these situations and agrees with the proposal that the rate of 3,5,3'-triiodothyronine generated in situ because of thyroxine 5'-deiodination could be an essential event related to thermogenesis in brown fat. Even though the relationship between 3,5,3'-triiodothyronine generation in the tissue and the specific thermogenic mechanisms of brown fat is unknown, present results indicate a close link between the thermogenic and 5'-deiodinase activities in physiological situations when brown adipose tissue needs to adapt to a low activity, such as that of the breeding cycle.

Ethanol and acetaldehyde concentrations in the rat foeto-maternal system after an acute ethanol administration given to the mother.

The metabolization curves for both ethanol and acetaldehyde after an acute intragastric or intravenous administration to the mother, have been studied. Metabolization of ethanol followed a very similar pattern in both the pregnant and their control virgin rats, whereas the levels of acetaldehyde derived from the metabolism of the administered ethanol were significantly higher in the pregnant animals, this fact implying that, in late gestation, there is a decrease in the mother's capacity for acetaldehyde metabolism. At the foetal side of the placenta, 150 min after the administration, the concentration of ethanol was similar to that found in the mother's circulation, thus proving a fluid transit of this metabolite through the placenta. The concentration of acetaldehyde in the foetus was relatively high, after the intragastric administration of the ethanol dose; we conclude that at certain ethanol concentrations, acetaldehyde can cross the rat placenta.

Brown adipose tissue metabolism in lactating rats: the effect of litter size.

Brown adipose tissue (BAT) thermogenic activity has been measured in lactating rats on day 11 post-partum. In order to assess the response of this tissue to different rates of milk production, litter sizes were adjusted to give 2, 4, 8 or 12 pups/dam. In all lactating animals, BAT mass and protein content were markedly reduced, but thermogenic activity was only fully suppressed in dams nursing large litters. In those with 4 pups, guanosine diphosphate-binding activity (expressed as pmol GDP bound/mg mitochondrial protein) was lower than that of virgin control rats, but remained well above values obtained from control animals acclimated at thermoneutrality. Other estimates of BAT thermogenesis, cytochrome oxidase activity and noradrenaline-stimulated increase in oxygen consumption supported the inverse relationship between litter size and BAT function. Possible control mechanisms for the observed changes in BAT activity in lactating animals are discussed.

Pathophysiological differences between paired and communal breeding of male and female Sprague-Dawley rats.

Sexually mature, male and female Sprague-Dawley rats were housed in large communal breeding cages or in smaller paired breeding cages. Virgin control rats of the same age were housed similarly but segregated by sex. Breeders became obese, developed a fatty liver, and showed elevated levels of triglycerides, free fatty acids, and cholesterol. Breeders had high blood pressure, enlarged hearts, hyperglycemia, and islet beta cell degranulation. Serum enzymes, creatine phosphokinase, serum glutamic oxalo-pyruvic transaminase, serum glutamic pyruvic transaminase, lactate dehydrogenase, and blood urea nitrogen levels were elevated in breeder rats. The adrenal glands of male breeders appeared hyperactive; the adrenal glands of female breeders were thrombosed and appeared to be hypoactive. Male breeder rats developed microscopic aortic lesions only; female breeders developed advanced calcific aortic sclerosis. Male breeders kept in active stud service manifested the most abnormal metabolic and pathophysiological changes. Female breeders developed similar pathophysiological changes after four pregnancies, irrespective of their paired or communal breeding environment. Virgin rats were normal regardless of housing conditions. Our findings suggest that repeated breeding in male and female rats causes resetting of the hypothalamic-pituitary-adrenal-gonadal axis. This may lead to disturbed hormonal and metabolic changes which culminate with the development of accelerated cardiovascular degenerative changes.

Relationship between previous reproductive history and chemically induced mammary cancer in rats.

AbstractThe relationship between a previous history of multiple pregnancies and/or lactations and 7,12‐dimethylbenz(a)anthracene‐induced mammary carcinogenesis was studied in groups of Sprague‐Dawley rats. All rats received the carcinogen at approximately 6 1/2 months of age. Virgin control rats which did not receive the carcinogen did not develop mammary cancer within 13 months although carcinogen‐fed virgin rats exhibited a mammary cancer incidence of 38.5%. A single pregnancy prior to feeding the carcinogen resulted in a decrease in the incidence and an increase in the mean time of appearance of the first palpable tumor. A single pregnancy followed by a subsequent nursing period reduced the percentage incidence and increased the mean time of appearance of the first palpable tumor when compared to the carcinogen‐fed virgin rats. A further increase in the number of pregnancies and/or lactations did not greatly influence the incidence of mammary cancer nor the number of cancers per rat. The mean time of tumor appearance was, however, increased. These data indicated that the relationship between party and carcinogen‐induced mammary cancer in the rat is similar to that existing in the human female but opposite to that found for the mouse.

Influence of Lactation on the Retention of a Single Dose of Strontium-90 in Rats

IN previous investigations1–3, it was found that the retention of radiostrontium after the lactation period (21 days post partum) was 45 per cent lower than that of virgin control rats when the isotope was injected into the pregnant rats 3 days before delivery. It was not clear, however, how much of this loss was due to a decrease in uptake of strontium-90 into bone or to other processes restricted to the time immediately after uptake or to a process still ruling after this initial period.