bluuebunny/crossref_metadata_2025 huggingface dataset:
Pituitary and testicular activity in chickens after embryonic testosterone treatment
Authors: Taylor R. L. , Glick B.
Published: December 2017

This study was conducted to explain the lower circulating testosterone levels in chickens exposed to testosterone propionate (TP) on the 3rd day of incubation. Control and TP males were injected with either luteinizing hormone (LH) or gonadotropin-releasing hormone (GnRH). Testosterone (T), dihydrotestosterone (DHT), and LH were measured by radioimmunoassay. Levels of circulating T in 20-wk-old TP males were significantly lower than controls, but the response to GnRH was similar in both groups. LH levels were also similar. Testosterone responses to LH in 20-wk-old TP males were lower than controls, possibly due to their lower initial T concentrations. Pituitary extracts from TP and control birds showed similar abilities to induce testicular 32P uptake in day-old cockerels. The data demonstrate that the anterior pituitary gland and testes of TP birds are capable of responding to GnRH and LH, respectively. The TP males are defective in that they fail to produce testosterone, which may reflect a reduced concentration of LH receptors in their testes.
Significance of hyperinsulinemia in ventromedial hypothalamus-lesioned rats
Authors: Hansen F. M. , Nilsson P. , Hustvedt B. E. , Nilsson-Ehle P. , Lovo A.
Published: December 2017

The significance of the hyperinsulinemia on the altered metabolism in ventromedial hypothalamus (VMH)-lesioned rats was tested. VMH lesions induced increases in fatty acid synthesis, lipoprotein lipase (LPL) activity, and plasma urea levels, and a decrease in plasma triglyceride concentrations. Similarly, in normal rats treated with pharmacological doses of insulin (14 U X rat-1 X day-1), fatty acid synthesis and LPL activity in fat tissue and the plasma urea were considerably elevated and plasma triglyceride was lowered compared with untreated controls. When endogenous insulin production was abolished by streptozotocin treatment, the four metabolic variables in the VMH-lesioned rats did not differ from those in diabetic controls. Substitution with 2 U of insulin X rat-1 X day-1, however, restored the differences in metabolism between VMH-lesioned diabetic and control diabetic rats. Substitution with 3 or 4 U insulin X rat-1 X day-1 showed the same differences. In VMH-lesioned rats the insulin level increased significantly from the 10th to the 70th day postoperatively; however, the rate of fatty acid synthesis, LPL activity, and plasma urea levels decreased, whereas plasma triglyceride concentrations increased. The results strongly suggest that the metabolic changes occurring after VMH lesions are only in part explained by the hyperinsulinemia associated with the VMH syndrome and indicate that other hormonal and/or nervous factors are involved.
Studies of origins of abnormal sympathetic function in obese Zucker rats
Authors: Levin B. E. , Triscari J. , Sullivan A. C.
Published: December 2017

To explain previously described abnormalities of sympathetic function in the genetically obese Zucker rat, the kinetics of norepinephrine (NE) metabolism were examined. Lower stress-induced levels of plasma NE in the obese rat were shown to be due to decreased release of NE because uptake of [3H]NE tracer from plasma was 45% slower in the obese compared with the lean rat. NE levels were 15, 20, and 34% lower in heart, pancreas, and interscapular brown adipose tissue (IBAT), and NE turnover rates (determined by two independent methods) were 39-48 and 43-69% lower in the pancreas and IBAT of obese compared with lean rats. In vivo synthesis of [3H]NE from [3H]dopamine by the enzyme dopamine beta-hydroxylase was 60% lower in IBAT but was equal in hearts of obese compared with lean rats. These results suggest an organ-specific decrease of NE synthesis in the obese Zucker rat, possibly due to decreased dopamine beta-hydroxylase activity in IBAT.
Uterine and adipose lipoprotein lipase activity in hormone-treated and pregnant rats
Authors: Gray J. M. , Greenwood M. R.
Published: December 2017

We report for the first time the presence of lipoprotein lipase (LPL) activity in the uteri of adult rats. This uterine LPL activity was significantly elevated by administration of the estrogen, estradiol benzoate (EB), but not by treatment with the antiestrogen, nafoxidine. Both EB and nafoxidine, however, greatly reduce both parametrial and retroperitoneal adipose tissue LPL activities. Uterine LPL activity also changed as a function of the duration of pregnancy, increasing severalfold between days 14 and 18 and then decreasing by about 20% on day 21. These data suggest the existence of a hormone-dependent system of providing lipids for storage and/or utilization by metabolic and reproductive tissues as needed.
Regulation of protein synthesis in lung by amino acids and insulin
Authors: Besterman J. M. , Watkins C. A. , Rannels D. E.
Published: December 2017

Acute effects of amino acid availability and insulin on protein synthesis were investigated in rat lungs perfused in situ with buffer containing either 4.5% fraction V bovine serum albumin (FrV BSA), 4.5% essentially fatty acid-free (FAF) BSA, or 4.5% dextran to maintain colloid osmotic pressure. In the presence of FrV BSA, protein synthesis was unaffected by perfusion for 1 or 3 h with buffer containing no added amino acids (0X), as compared with amino acids at concentrations one (1X) or five (5X) times those in rat plasma. Regardless of the amino acid concentration, addition of insulin was without effect. Likewise, in lungs perfused for 1 h with either FAF BSA or dextran, protein synthesis was insensitive to amino acid availability or to insulin. After 3 h, however, protein synthesis decreased 34 and 37%, respectively, when these lungs were perfused in the absence of both amino acids and insulin. In both cases, the inhibition was prevented by addition of insulin to the perfusate; addition of the hormone to perfusate containing 1X amino acids or elevating perfusate amino acids to 5X did not affect protein synthesis. The deficit in protein synthesis observed in the absence of both amino acids and insulin was not accompanied by ATP depletion or by lower intracellular concentrations of amino acids. Similarly, the effect of insulin was not associated with a general elevation in intracellular amino acid concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)
Effects of housing and chronic cannulation on plasma ACTH and corticosterone in the rat
Authors: Fagin K. D. , Shinsako J. , Dallman M. F.
Published: December 2017

Effects of housing and cannulation on plasma ACTH and corticosterone levels were evaluated to validate a method for obtaining repeated blood samples for ACTH from conscious rats. Resting morning, evening, or noise-stimulated (4 min of metal rattling) levels of plasma ACTH or corticosterone obtained by decapitation were similar in rats housed alone or in pairs. Individually housed, cannulated rats had slightly elevated morning levels of plasma ACTH and corticosterone, but the evening and noise-stimulated increases in hormone levels were similar to those in unoperated, decapitated rats. A hemorrhage stimulus administered on separate days to cannulated rats evoked highly replicable elevations in plasma ACTH and corticosterone. These results demonstrate that 1) ACTH and corticosterone secretion are similar in rats housed alone versus in pairs, and 2) long-term cannulation permits multiple intra-animal comparisons of basal and acutely stimulated plasma ACTH and corticosterone levels that are useful reflections of those measured in decapitated rats.
Diabetogenic activity of native and biosynthetic human growth hormone in obese (ob/ob) mouse
Authors: Kostyo J. L. , Gennick S. E. , Sauder S. E.
Published: December 2017

It has been repeatedly suggested that diabetogenic activity is not an intrinsic property of native pituitary growth hormone (GH) and that the diabetogenic effects produced by GH preparations are due to low-molecular-weight contaminants or degradation products of the hormone. This possibility was evaluated in this study by assessing the ability of purified native human GH (hGH) and biosynthetic methionyl-hGH to exacerbate fasting hyperglycemia and glucose intolerance in the obese (ob/ob) mouse. Native hGH that had been purified by DEAE-cellulose chromatography (A-type; 1.8 IU/mg) produced fasting hyperglycemia and glucose intolerance in the ob/ob mouse when injected subcutaneously at doses of 50 micrograms/day or greater for 3 days. It had no effect when a single subcutaneous dose of 200 micrograms was administered 24 h previously. To eliminate possible contamination with smaller peptides, the hGH was gel-filtered on a column of Sephacryl S-200 in 6 M guanidine-HCl. When injected subcutaneously into ob/ob mice at a dose of 50 micrograms/day or greater for 3 days, the guanidine-treated hGH produced glucose intolerance. Also biosynthetic methionyl-hGH produced marked fasting hyperglycemia and glucose intolerance when injected subcutaneously at doses of 50 or 100 micrograms/day for 3 days. These results support the conclusion that hGH itself is indeed diabetogenic but that chronic exposure of the organism to the hormone is required for its effects on glucose metabolism to become clearly manifest.
Prolactin profiles during estrous cycle and pregnancy in hamster as measured by homologous RIA
Authors: Talamantes F. , Marr G. , DiPinto M. N. , Stetson M. H.
Published: December 2017

The development of a homologous radioimmunoassay (RIA) for the measurement of serum hamster prolactin (PRL) has facilitated our studies on monitoring the secretion patterns of this hormone during different states of reproduction. Four salient findings emerge from the present studies. First, rapid cardiac puncture without anesthesia does not affect serum PRL levels during selected times in the estrous cycle or during pregnancy. Second, serum PRL levels during the estrous cycle describe a daily rhythm of PRL release with maximum serum concentrations each day occurring in the afternoon and the highest of these occurring on proestrus (day 4 of cycle). Third, PRL release during pregnancy is characterized by a single surge each day; although the time of maximum PRL release varies, levels increase during the hours of darkness and decline to basal during the light. Fourth, the amount of PRL released during pregnancy diminishes with time; on day 15 the total amount of PRL in the serum is less than 10% of that measured on day 5.
Mapping episodic fluctuations in plasma LH in orchidectomized rats
Authors: Ellis G. B. , Desjardins C.
Published: December 2017

Reports of the frequency of pulsatile LH release in orchidectomized rats are surprisingly variable. Estimates of the period between LH pulses vary from 18.5 to 250 min in nine reports from six laboratories, published in 1981-1983. In these studies, blood samples were drawn at intervals ranging from 2.5 to 60 min. We examined the relationship between estimated LH pulse frequency and blood-sampling interval. Six castrated male rats were cannulated, and blood samples were drawn at 2.5-min intervals through 4 h. Plasma LH levels were determined by radioimmunoassay. Using the data obtained at 2.5-min intervals, we simulated blood-sampling intervals of 5, 7.5, 10, 12.5, and 15 min by sequentially deleting data points. The original and simulated data sets were analyzed by both the PULSAR and Cycle Detector computer programs. The results show that as the sampling interval increased from 2.5 to 15 min, the apparent period of pulsatile LH release rose steadily from about 20 min to about 100 min. We found a high, positive correlation between blood-sampling interval and the apparent period of LH pulses. Estimates of LH pulse frequency in castrated male rats vary directly with the frequency at which blood samples are taken. Sampling intervals greater than 5 min in orchidectomized rats yield an LH pulse period that is most likely exaggerated.
Effects of catecholamines and ammonia on plasma and brain amino acids in dogs
Authors: Strombeck D. R. , Harrold D. , Rogers Q. R.
Published: December 2017

Investigations were made on the effects of catecholamine (Cat) infusions with and without ammonia (NH3) on plasma and brain amino acids (AA) and brain neurotransmitters in dogs. Groups of four dogs were infused for 5 h with epinephrine (E), epinephrine + norepinephrine (E + NE), epinephrine + norepinephrine with NH3 during h 4 and 5 (E + NE + NH3), epinephrine + norepinephrine + tryptophan with NH3 during h 4 and 5 (T + E + NE + NH3), or saline (C). Cat decreased (P less than 0.05) plasma Gly, Thr, Lys, Pro, Val, Ser, Arg, Leu, Trp, Phe, Asn, Tyr, Met, Ile, Cit, and Asp. The decreases at h 3 for all were to a mean of 45% of 0 h and were associated with no changes in plasma insulin or glucagon. Cat increased plasma Tau and Orn. Of the most abundant brain AA (82% of total), E + NE + NH3 had no effect (GABA, Asp, Gly, Ala, p-ethanolamine) or increased (Glu, Gln, Tau) brain levels. These AA were unchanged by Cat alone. Of the remaining brain AA, most were decreased by Cat (7 of 16, P less than 0.05) and E + NE + NH3 increased brain Trp but had no effect on brain serotonin, 5-hydroxyindoleacetic acid, or NE. Cat changed plasma AA in a way similar to changes produced by NH3 infusion and seen with hepatic insufficiency due to portacaval shunts and nitrosamine-induced pathology. Cat reduced brain AA levels, and this was partially restored by NH3.
Effect of epinephrine and norepinephrine on zinc thionein levels and induction in rat liver
Authors: Brady F. O. , Helvig B.
Published: December 2017

Hepatic zinc metallothionein (MT) levels are increased in response to a variety of stresses. Glucocorticoid induction of zinc thionein is insufficient in accounting for the levels attained. The potential involvement of catecholamines in the modulation of rat hepatic zinc metabolism and zinc thionein levels has been systematically studied. Eleven hours after multiple injections (6) of epinephrine, norepinephrine, or isoproterenol, zinc thionein levels of 4.01 +/- 0.74, 6.83 +/- 0.67, and 11.75 +/- 0.96 micrograms Zn in MT/g liver, respectively, were attained (untreated, 1.04 +/- 0.14). The levels of hepatic zinc thionein thus reached the range of stress response-induced levels (4–10 micrograms Zn in MT/g liver), attained 11 h after the onset of the stress. Multiple injections of isoproterenol and norepinephrine induced the formation of isoforms MT-I and MT-II in roughly equal amounts. The alpha-adrenoceptor blocker phentolamine blocked the 11-h increase in norepinephrine-stimulated (6) zinc thionein levels by 88%. The beta-adrenoceptor blocker propranolol blocked the 11-h increase in isoproterenol-stimulated (6) zinc thionein levels by 55%. This inhibition could be increased to 72% by previous administration of both phentolamine and propranolol. Catecholamines stimulated increases in both the zinc and the protein of MT, the latter as assessed by [35S]cysteine incorporation. Both of these increases were blocked by cycloheximide, confirming the requirement for de novo protein synthesis in this induction response.
Extrapancreatic effect of somatostatin infusion to increase glucose clearance
Authors: Bergman R. N. , Ader M. , Finegood D. T. , Pacini G.
Published: December 2017

Constant intraportal insulin, coupled with variable intraportal glucagon, was used in the attempt to reestablish basal metabolic conditions in dogs during somatostatin (SRIF) infusion (0.8 micrograms X min-1 X kg-1). SRIF alone lowered glucose (G), insulin (I), and glucagon (GN) (G: 90 +/- 5 to 69 +/- 1 mg/dl; I: 18 +/- 4 to 4 +/- 1 microU/ml; GN: 257 +/- 52 to 168 +/- 40 pg/ml; P less than 0.05 or better). Hormone replacement. Hypoglycemia persisted (G at steady state, SS, 60–150 min): 12 +/- 3 mg/dl below basal; P = 0.006) despite intraportal insulin replacement (200 microU X min-1 X kg-1; insulin at basal: 14 +/- 1; at SS: 14 +/- 2 microU/ml; P greater than 0.9) and glucagon overreplacement (basal: 341 +/- 42; SS: 486 +/- 80 pg/ml; P less than 0.05). Glucose clearance was increased 65% above basal (P less than 0.0001). Insulin underreplacement. With a lower intraportal insulin infusion rate (50 microU X min-1 X kg-1), insulin fell from basal (10 +/- 2 microU/ml) to 4 +/- 1 microU/ml during steady state (P = 0.03). Glucose and glucose clearance were normalized to basal values (G: 85 +/- 3 mg/dl, P = 0.3; clearance: 5.7 +/- 0.5 ml X min-1 X kg-1; P = 0.2) with full glucagon replacement (basal: 281 +/- 120; SS: 264 +/- 80 pg/ml; P greater than 0.9). Thus, during constant SRIF infusion, normoglycemia was reattained when insulin was underreplaced via the portal vein. The failure to reattain euglycemia with normoinsulinemia was due to a SRIF-induced increase in extrahepatic glucose clearance. Insulin replacement and growth hormone (GH) infusion. GH (15 ng X min-1 X kg-1) partially reversed the hypoglycemia during SRIF, with full insulin replacement. The SRIF-induced increase in glucose clearance may be partially mediated by a decrease in GH.
Trophic response of hamster brown adipose tissue: roles of norepinephrine and pineal gland
Authors: Triandafillou J. , Hellenbrand W. , Himms-Hagen J.
Published: December 2017

The objective was to find out whether the reduced amount of brown adipose tissue in myopathic hamsters [Am. J. Physiol. 239 (Cell Physiol. 8): C18–C22, 1980] was secondary to a refractoriness to the trophic influence of norepinephrine. However, no evidence for any trophic influence of norepinephrine on brown adipose tissue of either normal or myopathic hamsters could be detected under experimental conditions that have demonstrated such an influence in rats. A mediator other than norepinephrine, melatonin, secreted by the pineal gland, is known to mediate the control of brown adipose tissue growth induced in hamsters by short photoperiod. Further studies of intact or pinealectomized hamsters showed that the pineal gland was not required for either cold- or diet-induced growth of brown adipose tissue. It is concluded that the defect in control of brown adipose tissue size in the hamster with muscular dystrophy is not due either to abnormal control by norepinephrine or to the pineal gland since neither of these appears to participate in the normal regulation of brown adipose tissue size in relation to environmental temperature or to diet.
cAMP response of vascular smooth muscle cells to bovine parathyroid hormone
Authors: Stanton R. C. , Plant S. B. , McCarron D. A.
Published: December 2017

Parathyroid hormone (PTH) is a vasodilator of vascular smooth muscle tissue. It has been shown to produce this vasodilation in normotensive and hypertensive laboratory rats. The effect is log dose dependent, maximal at 1 min and persists for 3–5 min. The cellular mechanisms involved in PTH-mediated vasodilation are unknown. In this study, we sought to determine the cellular changes of cAMP after administration of bovine (b)PTH (1–34). cAMP content of vascular smooth muscle cells was measured at 30 s, 1, 3, and 5 min after incubation with synthetic bPTH (1–34). Tissue cAMP content was decreased by 55% at 1 min (4.1 +/- 0.5 pmol/mg protein at time 0 vs. 1.9 +/- 0.2 pmol/mg protein at 1 min, P less than 0.001). After 5 min, cAMP levels returned to base-line values and increased over the next 5–10 min to levels above base line (P less than 0.01). In conclusion, our data suggest that the initial response of vascular smooth muscle cells to short-term incubation with bPTH (1–34) is an acute decrease in cAMP content.
Regulation of bone mineral loss during lactation
Authors: Brommage R. , DeLuca H. F.
Published: December 2017

The effects of varying dietary calcium and phosphorus content, vitamin D deficiency, oophorectomy, adrenalectomy, and simultaneous pregnancy on bone mineral loss during lactation were examined in rats. Unless otherwise stated, the diet contained 0.47% calcium and 0.3% phosphorus and the rats were given 26 nmol of vitamin D3. Femur ash weights were determined after 21 days of lactation and on age-matched nonlactating rats. Decreasing dietary calcium to 0.02% caused an increased loss of bone mineral, whereas increasing dietary calcium to 1.4% increased plasma calcium levels to 12 mg/100 ml but did not diminish the bone mineral loss observed during lactation. Varying dietary phosphorus did not have a major effect on bone mineral loss during lactation. In vitamin D-deficient rats, bone mineral loss during lactation was independent of dietary calcium levels and slightly greater than the loss observed in vitamin D-replete rats fed the normal calcium diet. Oophorectomy and adrenalectomy did not produce changes in femur ash weights of nonlactating rats or reduce bone mineral loss during lactation. Rats mated during their postpartum estrus and thus simultaneously pregnant and lactating, lost the same amount of bone mineral as caused by lactation alone.
cAMP metabolism and lipolysis in brown adipocytes of hamsters consuming a cafeteria diet
Authors: Schimmel R. J. , McCarthy L. , McMahon K. K.
Published: December 2017

Feeding animals cafeteria diets causes increased sympathetic activity to brown adipose tissue and this is believed to be responsible for the concomitant activation of thermogenesis. Because chronic catecholamine stimulation in other systems leads to a desensitization of beta-adrenergic receptors, we examined lipolysis and cAMP production in brown adipocytes of hamsters eating cafeteria diets for evidence of diminished beta-adrenergic responses. Basal cAMP levels were similar in chow- and cafeteria-fed hamsters. However, adipocytes from overfed animals formed less cAMP in response to isoproterenol than those of control animals. Isoproterenol-stimulated adenylate cyclase activity was similarly decreased in membrane preparations from cafeteria-fed hamsters. However, when the diterpene forskolin was used, equal amounts of cAMP were formed in cells and membrane preparations from control and overfed animals. In contrast to the reduced responses of the cAMP system to isoproterenol stimulation observed in overfed hamsters, isoproterenol-stimulated lipolysis was greater in cells from overfed animals than in cells from control animals. These results are consistent with a desensitization of the adenylate cyclase system in brown adipocytes occurring during chronic hyperphagia. Because eating cafeteria diets has been reported to increase sympathetic activity to brown fat depots, the apparent desensitization of brown adipocytes observed in this study may result from a persistent stimulation of the brown fat with norepinephrine in vivo. Our data also suggest the existence of mechanisms that preserve lipolysis in the face of low cAMP levels.
Ischemic conduction failure and energy metabolism in experimental diabetic neuropathy
Authors: Low P. A. , Ward K. , Schmelzer J. D. , Brimijoin S.
Published: December 2017

We examined the effect of ischemia on nerve conduction in experimental diabetic neuropathy (EDN) and related electrophysiological changes to nerve adenosine triphosphate (ATP), creatine phosphate (CP), and lactate under anoxic conditions. Rats rendered diabetic with streptozotocin had a resistance to ischemic conduction block (RICB). Caudal nerve action potential (NAP) was well maintained for 10 min in controls and for 15 min in EDN, after which time NAP declined in both groups but more rapidly in normal rats. Time to 50% reduction in nerve ATP and CP was 10 and 3 min, respectively, in controls and delayed to 20 and 8 min in EDN. Rate of utilization of high-energy phosphate (approximately P) was linear for 5 min in controls to be followed by a progressive decline. In EDN rate of utilization of approximately P was linear to 15 min to be followed by a more gradual decline than in normal nerves. These findings suggest that the maintenance of nerve transmission in anoxic-ischemic states depends on anaerobic metabolism and that RICB in EDN is due in part to the ability of diabetic nerves to maintain a higher level of anaerobic glycolysis and for a longer time than normal nerves.
Indomethacin and salicylate modulate effect of insulin on glucose kinetics in dogs
Authors: Miller J. D. , Ganguli S. , Sperling M. A.
Published: December 2017

We studied insulin's effects on glucose production (Ra) and utilization (Rd) in trained, conscious dogs before and during treatment with indomethacin (Indo) and salicylate (S). Ra and Rd (mg X kg-1 X min-1) were calculated by isotope dilution using [3-3H]glucose. Animals were treated with either oral Indo or acetylsalicylic acid for 1 day before the respective studies. On the study day, experimental animals were given a continuous infusion of either saline (control), Indo (5 mg/kg bolus followed by 0.05 mg X kg-1 X min-1), or sodium salicylate (0.45 mg X kg-1 X min-1) for 330 min on separate days; each animal participated in all three protocols. After establishing steady-state specific activity, control (C) and experimental animals (n = 6/group) received insulin, 0.275 mU X kg-1 X min-1 for 150 min, raising serum insulin levels two- to threefold above basal. During insulin infusion in C, plasma glucose (G) fell from 99 +/- 2 to 82 +/- 6 ml/dl (P less than 0.01), associated with a transient fall in Ra from 2.5 +/- 0.3 to 1.9 +/- 0.2 (P less than 0.01) at 30 min, returning to base line at 45 min; Rd did not change. In the Indo and S groups, G also fell by a similar extent. In contrast to C, however, the fall in G was associated with a rise in Rd, commencing at 30 min in the Indo group (P less than 0.05) and at 45 min in the S group (P less than 0.01); Ra did not fall and actually rose above basal (P less than 0.05), although it did not match the rise in Rd.(ABSTRACT TRUNCATED AT 250 WORDS)
Long-term regulation of hexose uptake by isoproterenol in cultured 3T3 adipocytes
Authors: van Putten J. P. , Krans H. M.
Published: December 2017

Catecholamines are known to have short-term regulatory effects on fat cell hexose uptake. We examined the long-term effects of catecholamines on the insulin-sensitive 2-deoxyglucose (dGlc) uptake in cultured 3T3-L1 adipocytes. Prolonged exposure (48 h) to isoproterenol (beta-adrenergic agonist) stimulated the basal dGlc uptake up to 90%. The effect was specific, time, concentration, and protein synthesis dependent and reversible. The effect of insulin was unaltered and superimposed on the increase in basal dGlc uptake. The long-term effect of isoproterenol was mimicked by epinephrine, dibutyryl cAMP (DBcAMP), and 1-methyl-3-isobutylxanthine (IBMX). By contrast, short-term exposure to isoproterenol (and epinephrine) induced a protein synthesis-independent increase in basal dGlc uptake (30%) not accompanied by an increase in insulin responsiveness. Moreover, on short-term basis, DBcAMP and IBMX suppressed both the basal and insulin-stimulated uptake up to 50%. Determination of the intracellular nonphosphorylated dGlc during the uptake and of the hexokinase activity revealed that the long-term effect of isoproterenol was most likely due to alterations low in dGlc transport. In conclusion, long-term regulators of hexose uptake are in cultured 3T3-L1 adipocytes, isoproterenol, and other cAMP stimulators. The long-term effect is independent from the short-term regulatory effect of the agents and from the effect of insulin.
Decreases in ovine fetal cartilage sulfate uptake and serum sulfate during gestation
Authors: Morriss F. H. , Marshall R. N. , Crandell S. S. , Fitzgerald B. J. , Riddle L.
Published: December 2017

In vitro assays for [35S]sulfate uptake by ovine fetal costal cartilage were used to assess gestational changes in cartilage metabolism. Addition of 20% normal human serum to the incubation medium increased fetal cartilage [35S]sulfate incorporation into glycosaminoglycans. Both basal and human serum-stimulated uptakes of [35S]sulfate by fetal sheep cartilage decreased from midgestation to full term. The incremental response in [35S]sulfate uptake that was stimulated by human serum decreased as gestation proceeded to full-term. Fetal serum sulfate concentration decreased logarithmically during gestation, raising the possibility that cartilage sulfate uptake might become substrate limited as full term is approached. Perfusion of seven late gestation sheep fetuses for 7 days with Na2SO4 to achieve serum sulfate concentrations similar to those observed earlier in gestation resulted in a 33% increase in mean cartilage [35S]sulfate uptake compared with that of control twin fetuses, but uptake was not increased to values that occurred spontaneously earlier in gestation. These results suggest that the decreasing rate of [35S]sulfate uptake by fetal cartilage during the last half of gestation is associated only minimally with decreasing serum sulfate levels and is most consistent with intrinsic change in resting chondrocyte metabolism during gestation.
Somatomedin C augments FSH-induced differentiation of cultured rat granulosa cells
Authors: Davoren J. B. , Hsueh J. W. , Li C. H.
Published: December 2017

Growth hormone (GH) deficiency in rats is associated with decreased ovarian steroidal responsiveness to gonadotropins, possibly through a reduction in the production of the GH-dependent Somatomedin C/insulinlike growth factor I (SM C/IGF I). We have investigated the direct effects of synthetic SM C/IGF I on gonadotropin-stimulated ovarian steroidogenesis in vitro. Granulosa cells were cultured in a serum-free medium for 48 h in the presence of follicle-stimulating hormone (FSH), with or without SM C/IGF I. FSH dose-dependently increased both estrogen and progestin production. Concomitant treatment with SM C/IGF I led to a dose-dependent augmentation of progestin secretion over the full range of FSH doses tested, by a maximum of 2.3- to 2.6-fold. FSH-stimulated estrogen was enhanced by up to 2.4-fold but only at low doses of FSH. SM C/IGF I-enhanced progestin production was associated with increased pregnenolone production and 3 beta-hydroxysteroid dehydrogenase activity, whereas augmented estrogen production appeared to be due to enhanced aromatase activity. The actions of SM C/IGF I, at physiologically relevant concentrations were correlated with increased extracellular cAMP accumulation and cellular protein content but were independent of any change in cell number or viability. In contrast to SM C/IGF I, the closely related peptide multiplication-stimulating activity decreased estrogen production while increasing progestin metabolite accumulation. The present results indicate that the GH-dependent peptide SM C/IGF I may play a role in ovarian development by enhancing gonadotropin-stimulated granulosa cell steroidogenesis.
In vitro recovery of insulin binding after downregulation in cultured normal human monocytes
Authors: Whitson R. H. , Kaplan S. A.
Published: December 2017

When normal human peripheral blood monocytes were treated with insulin in vitro, surface insulin receptors disappeared rapidly, but total insulin receptors (surface and internalized receptors), measured in detergent-solubilized extracts of total cellular membranes, decreased slowly. Surface receptors decreased to 51 +/- 4, 36 +/- 12, and 34 +/- 12%, of control levels after 2, 6, and 18 h of insulin pretreatment, respectively. Total receptors decreased to 86 +/- 12, 69 +/- 17, and 34 +/- 12% of control levels in the same periods. Chloroquine, a lysosomotropic agent, inhibited the removal of surface receptors, indicating that lysosomal proteases play a role in this process. Unlike monocytes, IM-9 lymphocytes lost surface receptors and total receptors at the same rate when incubated with insulin. Monocytes treated with insulin for 18 h, washed free of unbound insulin and recultured for 48 h regained 94 +/- 7% of control insulin binding, indicating that cultured monocytes are competent to regenerate their insulin receptors. Monocytes treated with insulin for 6 h also required 48 h to recover their insulin binding, despite the fact that substantial numbers of insulin receptors remained intact within these cells. Two-hour pretreated monocytes recovered somewhat faster, attaining control levels of receptors after 24 h of reculture. This suggests that internalized insulin receptors pass from a recyclable pool to a nonrecyclable one.
Premature weaning-induced changes of cholesterol metabolism in guinea pigs
Authors: Subbiah M. T. , Yunker R. L. , Menkhaus A. , Poe B.
Published: December 2017

Premature weaning in animals causes higher serum cholesterol levels and a relative inability to handle cholesterol challenge in adult life. The mechanisms responsible for these delayed effects of premature weaning are not clear. The aims of this investigation were to 1) determine the effect of premature weaning on the activity of hepatic cholesterol 7 alpha-hydroxylase and plasma cholesterol changes and 2) determine whether addition of cholesterol to the diet will restore premature weaning-induced changes. Premature weaning did not prevent the phenomenon of neonatal hypercholesterolemia noted in day 5 of normally weaned guinea pigs. At day 10, prematurely weaned animals had lower serum cholesterol when compared with normally weaned animals. Premature weaning caused (without or with cholesterol in diet) a significant decrease in the hepatic activity of cholesterol 7 alpha-hydroxylase, and this difference persisted (after 1 mo) when fed the stock diet. These studies demonstrate 1) premature weaning on day 2 does not prevent neonatal hypercholesterolemia on day 5, 2) premature weaning causes a decrease in hepatic cholesterol 7 alpha-hydroxylase activity that persists into adult life, and 3) dietary cholesterol intake in early life is unable to negate premature weaning-induced changes.
Effects of in vivo gonadal hormone environment on in vitro hGRF-40-stimulated GH release
Authors: Evans W. S. , Krieg R. J. , Limber E. R. , Kaiser D. L. , Thorner M. O.
Published: December 2017

The effects of gender and the gonadal hormone environment on basal and stimulated growth hormone (GH) release by dispersed and continuously perifused rat anterior pituitary cells were examined. Cells from intact male and diestrus day 2 female rats and from castrate male rats either untreated or treated with testosterone (T) or 17 beta-estradiol (E2) were used. Basal GH release (ng/min per 10(7) cells; mean +/- SE) by cells from diestrus day 2 female rats was less than by cells from castrate rats treated with T (4.3 +/- 0.6 vs. 11.4 +/- 2.7, respectively; P less than 0.025). No other differences in basal release were detected. Concentration-response relationships were documented between human GH-releasing factor 40 (hGRF-40; 0.03-100 nM given as 2.5-min pulses every 27.5 min) and GH release. Mean (+/- SE) overall GH release (ng/min per 10(7) cells) above base line was greater by cells from intact male rats (496 +/- 92) than by cells from castrate (203 +/- 37.3; P less than 0.0001), castrate and T-treated (348 +/- 52.8; P = 0.008), or castrate and E2-treated (58.1 +/- 6.8; P less than 0.001) male rats or by diestrus day 2 rats (68.6 +/- 9.5; P = 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)
Calcium channel agonist BAY k8644 enhances anterior pituitary secretion in rat and monkey
Authors: Cronin M. J. , Anderson J. M. , Rogol A. D. , Koritnik D. R. , Thorner M. O. , Evans W. S.
Published: December 2017

A calcium channel agonist BAY k8644 was applied to anterior pituitary (AP) cells in vitro. BAY k8644 (0.1-10 microM) stimulated prolactin and growth hormone (GH) release from monolayer AP cultures; the calcium channel antagonist D-600 (1-10 microM) completely blocked this effect. By utilizing a perifusion system, we observed an immediate and sustained amplification of prolactin (2.9-fold), growth hormone (2.3-fold), and luteinizing hormone (LH, 1.6-fold) release during the 1-h application of BAY k8644 (3 microM). A hypophysiotrophic peptide pulse 4 h after the BAY k8644 was removed confirmed that the cells remained responsive to their natural secretagogues. In another perifusion study 10-3,000 nM BAY k8644 produced a graded increase in prolactin release that was maintained over the 30-min exposure period. Finally, individual primate mammotrophs and somatotrophs showed a marked enlargement of hemolytic plaque area, an index of hormone release, 1 h after BAY k8644 (1 microM). We conclude that this synthetic dihydropyridine enhances the rate of prolactin, GH, and LH release from AP cells of two species. Because this is the first synthetic calcium channel agonist, structure-function studies characterizing calcium channel activation and exocytosis are now feasible.
Influence of training on skeletal muscle enzymatic adaptations in normal and diabetic rats
Authors: Noble E. G. , Ianuzzo C. D.
Published: December 2017

Muscle homogenates representing slow-twitch oxidative, fast-twitch oxidative-glycolytic, fast-twitch glycolytic, and mixed fiber types were prepared from normal, diabetic, and insulin-treated diabetic rats. Diabetes was induced by injection of 80 mg . kg-1 of streptozotocin. The activities of citrate synthase, succinate dehydrogenase, and 3-hydroxyacyl-CoA dehydrogenase were employed as markers of oxidative potential, whereas phosphorylase, hexokinase, and phosphofructokinase activities were used as an indication of glycolytic capacity. Diabetes was associated with a general decrement in the activity of oxidative marker enzymes for all fiber types except the fast-twitch glycolytic fiber. In contrast, the fast-twitch glycolytic fibers demonstrated the greatest decline in glycolytic enzymatic activity. Insulin-treated animals, either trained or untrained, exhibited enzyme activities similar to their normal counterparts. Exercise training of diabetic rats mimicked the effect of insulin treatment and caused a near normalization of the activity of the marker enzymes. These findings suggest that the enzymatic potential of all skeletal muscle fiber types of diabetic rats may be normalized by exercise training even in the absence of significant amounts of insulin.
Adenohypophyseal response to hypophysiotropic hormones in male obese Zucker rats
Authors: Heiman M. L. , Porter J. R. , Nekola M. V. , Murphy W. A. , Hartman A. D. , Lance V. A. , Coy D. H.
Published: December 2017

Description of the recessive, homozygote obese Zucker rat (fafa) includes disorders of growth and reproduction. The aim of this study was to compare responsiveness of adenohypophyseal cells, obtained from male fafa rats and from their lean siblings, to growth hormone-releasing factor (GRF) and to luteinizing hormone-releasing hormone (LHRH). Pituitary cells were cultured for 4 days and were then challenged with either GRF-29 (the NH2-terminal 29 amino acid GRF peptide that expresses full biological activity of its parent 44 amino acid molecule) or [D-Trp6]LHRH (LHRH-A, an LHRH agonist). Medium was assayed for growth hormone (GH), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) by radioimmunoassay. Dose-response curves were compared using the computer program ALLFIT. The median effective GRF-29 concentration (EC50) computed for hypophyseal cells cultured from lean animals (0.30 +/- 0.01 fM; means +/- SE of 4 experiments) was less (P less than 0.01) than that calculated for cells obtained from fafa rats (15.8 +/- 6.7 fM). In contrast, cells derived from lean littermates required a larger (EC50) concentration of LHRH-A than did gonadotrophs cultured from obese rats [58.2 +/- 1.2 vs. 10.7 +/- 1.2 pM (P less than 0.01) and 59.4 +/- 10.4 vs. 15.7 +/- 7.6 pM (P less than 0.05)] to secrete LH and FSH, respectively. Our data describe an attenuated pituitary response to GRF-29 and an enhanced response to LHRH-A in the fafa.
Differential effects of forskolin on LH and GH release
Authors: Evans W. S. , Brannagan T. H. , Limber E. R. , Cronin M. J. , Rogol A. D. , Hewlett E. L. , Thorner M. O.
Published: December 2017

The effects of forskolin, an agent which increases intracellular levels of cAMP, on basal luteinizing hormone (LH) and growth hormone (GH) release and on gonadotropin-releasing hormone (GnRH)-stimulated LH release were documented. Continuously perifused dispersed anterior pituitary cells from female rats at random stages of the estrous cycle were used. Secretory rates of both LH and GH increased in a concentration-dependent manner in response to a 1-h challenge with 0.03, 0.1, 0.3, 1, or 3 microM forskolin. In response to 0.3 microM forskolin, maximum GH release was achieved within 15-20 min, after which secretion decreased. In contrast, LH release increased gradually, became maximal at 1.5-2 h, and remained constant until the forskolin was withdrawn. Cells exposed to 10 nM GnRH for 4 h exhibited a biphasic release of LH with the interphase nadir occurring at 30 min. The second phase of LH release was enhanced by simultaneous addition of forskolin with the GnRH. Whereas second phase release did not increase further, exposure of the cells to forskolin for 60 or 120 min before GnRH resulted in increased first-phase LH release. We suggest that, whereas our data are consistent with a role for cAMP in mediating the acute release of GH, cAMP may be involved in the process through which nonimmediately releasable LH becomes available for release.
Effect of pentobarbital on fructose 2,6-bisphosphate metabolism in isolated rat hepatocytes
Authors: Nyfeler F. , el-Maghrabi M. R. , Pilkis S. J.
Published: December 2017

Addition of the commonly used anesthetic pentobarbital to hepatocytes from fed rats resulted in a dose-dependent decrease in the level of fructose 2,6-bisphosphate. At a concentration of pentobarbital (0.4 mM) that lowered fructose 2,6-bisphosphate by 60%, there was no significant change in the level of fructose 6-phosphate, ATP, or L-glycerol 3-phosphate. Higher concentrations of pentobarbital (2 mM) enhanced both glycolysis and glycogenolysis and fructose 2,6-bisphosphate levels were reduced to less than 10% of the control. Concomitant with these changes there was a decrease in ATP, glucose 6-phosphate, and fructose 6-phosphate and a two- and fivefold increase in ADP and AMP, respectively. In hepatocytes from starved rats pentobarbital also lowered ATP levels and inhibited gluconeogenesis but had no effect on either lactate production or the already low level of sugar diphosphate. However, in the fasted case pentobarbital completely prevented the 10-fold elevation of fructose 2,6-bisphosphate brought about by 30 mM glucose. The anesthetic had no effect on cAMP-dependent protein kinase activity or on pyruvate kinase activity in hepatocytes from fed or starved rats but caused reciprocal changes in the activities of the bifunctional enzyme 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase. Kinase activity was decreased and bisphosphatase activity was increased. These results suggest that the effects of pentobarbital on gluconeogenesis and glycolysis are due to inhibition of energy metabolism with elevated AMP levels causing activation of 6-phosphofructo-1-kinase and inhibition of fructose 1,6-bisphosphatase.(ABSTRACT TRUNCATED AT 250 WORDS)
cAMP augmentation of secretagogue-induced luteinizing hormone secretion
Authors: Turgeon J. L. , Waring D. W.
Published: December 2017

Whether adenosine 3',5'-cyclic monophosphate (cAMP) acts as a mediator for luteinizing hormone-releasing hormone (LHRH) in either its immediate LH release action or in its self-priming action was investigated. Pituitary pieces from either proestrous or estrous rats were superfused in vitro in the presence of dibutyryl cAMP [(Bu)2cAMP], 8-bromo-cAMP (8BrcAMP), forskolin, or control for 2-3 h. For proestrous but not estrous pituitary pieces, a slight increase in base-line LH secretion rate occurred at approximately 70 min of exposure to elevated cAMP; in the same system LHRH caused an increase in LH secretory rate within 2 min in either proestrous or estrous tissue. In contrast to its ineffectiveness as a secretagogue, cAMP elevation resulted in a severalfold augmentation of both LHRH- and elevated K+-induced LH secretion from proestrous but not estrous pituitary pieces; for these experiments, superfusion with a cAMP analogue or forskolin for varying times preceded a 10-min pulse of either 8 nM LHRH or 47 mM K+. Augmentation was evident after 30 min of cAMP elevation; longer exposures were coincident with greater potentiation. Cycloheximide prevented (Bu)2cAMP augmentation of LHRH-induced secretion. These data show that cAMP does not mediate the immediate LH release action of LHRH, but cAMP does augment LHRH- or K+-induced LH secretion with characteristics in common with the self-priming action of LHRH.
Beta-adrenergic modulation of insulin binding in skeletal muscle
Authors: Webster B. , Vigna S. R. , Paquette T. , Koerker D. J.
Published: December 2017

Both a high physiological concentration (13.1 nM) of epinephrine (E) and acute exercise (AEx) have previously been shown to increase 125I-insulin binding in skeletal muscle. To investigate the site and mechanism of the effect of epinephrine on binding and the possible link between epinephrine- and AEx-enhanced insulin binding, we measured insulin binding in three different preparations: 1) crude membranes derived from whole soleus muscle incubated in vitro with 13.1 nM E, 2) crude membranes with E present in the binding assay, and 3) purified plasma membranes with E present. Epinephrine enhanced binding in all three preparations by 169, 144, and 164%, respectively, at low concentrations of insulin but had little effect at high concentrations. Epinephrine, therefore appears to have its effect at the plasma membrane. Propranolol (10 microM), a beta-adrenergic antagonist, blocked E-enhanced insulin binding and when added to crude membranes made from soleus and extensor digitorum longus muscle of AEx rats reversed the increase in binding seen with exercise. This indicates that E-enhanced insulin binding is mediated by beta-adrenergic receptors and that AEx enhances insulin binding via beta-adrenergic receptors. Sodium orthovanadate (3 mM), a phosphotyrosyl-protein phosphatase inhibitor, also inhibited the increase in insulin binding due to E, implying that E may increase insulin binding by activating a phosphotyrosyl-protein phosphatase which decreases the phosphorylation of a plasma membrane protein, presumably the insulin receptor.
Functional changes in luteinizing hormone-secreting cells from pre- and postpartum ewes
Authors: Wise M. E. , Sawyer H. R. , Nett T. M.
Published: December 2017

Luteinizing hormone (LH)-containing cells from ovine pituitaries obtained during gestation and at various times after parturition were examined to determine whether the ability to store and secrete LH in vitro was correlated with morphological changes. Pituitaries collected on days 50 and 140 of gestation and on days 2, 13, 22, and 35 after parturition were enzymatically dissociated and the resulting cells cultured in media containing estradiol (12 pg/ml), cortisol (12 ng/ml), or no steroid. After 4, 7, or 10 days of culture, cells were washed and basal LH, gonadotropin-releasing hormone (GnRH)-stimulated release of LH, and cellular content of LH were determined. The content of LH (ng/10(6) cells) was lowest on day 140 of gestation (2.7 +/- 0.3) and day 2 postpartum (2.2 +/- 0.6) and then increased (P less than 0.05) on days 13 (36.6 +/- 8.3), 22 (59.9 +/- 14.4), and 35 (54.6 +/- 19.3) postpartum. The percentage of pituitary cells containing immunoreactive LH nearly doubled (P less than 0.05) between days 2 (5.6 +/- 0.2%) and 35 (10.6 +/- 1.1%) postpartum. Moreover, LH-containing cells were smaller, and the percent total cellular volume occupied by secretory granules was less on day 2 than on days 22 and 35 after parturition. Secretion of LH after 4, 7, or 10 days of culture reflected the cellular content of LH and was not influenced by the presence of steroids in the media. These data indicate that decreased synthesis of LH during gestation is associated with hypoplasia of the LH-secreting cells. These cells are reactivated during the postpartum period and their capacity to synthesize LH gradually returns to normal.