bluuebunny/crossref_metadata_2025 huggingface dataset:
Metabolic clearance rate of insulin-like growth factor-I in fed and starved sheep
Authors: Hodgkinson S. C. , Davis S. R. , Burleigh B. D. , Henderson H. V. , Gluckman P. D.
Published: December 2008

The metabolic clearance of insulin-like growth factor-I (IGF-I) has been examined in sheep using a radioiodinated hormone preparation (131I-labelled IGF-I). Following i.v. administration, 131I-labelled IGF-I was distributed in a volume equivalent to plasma (60 ml whole blood/kg liveweight) and demonstrated a triphasic pattern of clearance with apparent half-lives (t½) of 4·0 ± 0·4 (s.e.m.), 52·4 ± 3·4 and 792 ± 26·5 min (n = 10). No significant differences in the t½ of the three phases were identified in fed compared with starved animals (fed, n = 4, phase 1 = 3·1 ± 0·64, phase 2 = 46 ± 5·9 and phase 3 = 756 ± 27 min; starved, n = 6, phase 1 = 4·6 ± 0·58, phase 2 = 57 ± 3·2 and phase 3 = 816 ± 38·5 min). Similarly, no significant differences in the distribution volume (fed, n = 4, 44 ± 4 ml/kg liveweight; starved, n = 6, 39 ± 2 ml/kg liveweight) or metabolic clearance rate (fed, n = 4, 2·9 ± 0·15 ml/min; starved, n = 6, 3·2 ± 0·5 ml/min) of the IGF-I were found in fed compared with starved animals. Highperformance gel filtration chromatography of sequential plasma samples following injection of 131I-labelled IGF-I revealed three clear peaks of radioactivity which demonstrated markedly different patterns of clearance. These correspond to hormone complexed to binding proteins of 150 000 and 50 000 daltons and to 'free' hormone. While radioactivity eluting in the position of free IGF-I had reduced to negligible levels by 12 min, 131I-labelled IGF-I associated with the intermediate molecular weight binding protein was still detectable 6 h after injection. In contrast, an initial increase in radioactivity associated with the 150 000 dalton binding protein was revealed. This form was cleared from the circulation very slowly, by 8 h it was the only species remaining and it was still detectable 26 h after injection. No significant differences in this pattern of bound and free hormone clearance were found in fed compared with starved animals. While there was apparently no interchange of hormone between the binding proteins in vitro the nature of the decay in vivo indicated the possibility of exchange of IGF-I between the different binding proteins.

J. Endocr. (1987) 115, 233–240
Pituitary responsiveness to LHRH during pregnancy in the rat: effect of progesterone
Authors: Koiter T. R. , van der Schaaf-Verdonk G. C. J. , Schuiling G. A.
Published: December 2008

The LH and FSH responses to a standard infusion of LHRH were studied on days 8, 12, 15, 17, 19, 21 and 22 after conception as well as on day 23, i.e. after parturition. Groups of rats were also killed on days 8, 15, 19, 22 and 23 and on the day of pro-oestrus of the 4-day cycle for the assay of progesterone, 20α-dihydroprogesterone (DHP), oestradiol-17β, LH and FSH. Finally, the post-partum surges of LH and FSH were compared with those at pro-oestrus in 4-day cyclic rats.

The LH and FSH responses to LHRH were relatively low on days 8 and 12, twice as high on days 15, 17 and 19, had increased further on day 21 and reached maximal values on day 22. The gonadotrophin responses were low on day 23. The post-partum surges of LH and FSH were much higher than the pro-oestrous surges. Pituitary contents of LH and FSH were higher than on pro-oestrus of the 4-day cycle. On day 23, however, the pituitary contents had declined by 60–80%. No apparent relationship was found between plasma concentrations of LH and FSH and LHRH responsiveness during pregnancy. Concentrations of oestradiol-17β on day 22 were higher than on all other days of pregnancy, but lower than on pro-oestrus of the 4-day cycle. Concentrations of progesterone were high until day 19 and low on days 22 and 23; the concentration of DHP was low until day 19 and very high on days 22 and 23. It was concluded that the increase of the gonadotrophin response to LHRH half-way through pregnancy, in contrast to the increase at the end of pregnancy, cannot be explained by an increase in the concentration of oestradiol-17β, while the pituitary LH and FSH contents play, if any, only a permissive role.

The effect of s.c. implants of progesterone on various hormone parameters during the last week of pregnancy was also investigated. It was found that progesterone not only delayed parturition but also prevented the rise of the gonadotrophin response at the end of pregnancy. Concentrations of oestradiol-17β and DHP were also lower on day 23 than in untreated rats, while the pituitary gonadotrophin contents were higher. It is suggested that delay of luteolysis (in the present study by progesterone treatment) at the end of pregnancy prevents the occurrence of a series of interrelated events: a rise in oestradiol-17β production, a rise in the gonadotrophin responses to LHRH, parturition and the generation of the post-partum gonadotrophin surges.

J. Endocr. (1987) 115, 247–254
The role of calcium and calmodulin in mediating release of thyrotrophin-releasing hormone by cultured hypothalamic cells
Authors: Lewis M. D. , Foord S. M. , Scanlon M. F.
Published: December 2008

We have developed a fetal rat hypothalamic cell culture system for the study of factors controlling the acute release of TRH. Release of TRH by the cells has been characterized by reversed-phase high pressure liquid chromatography and about 86% of the total immunoreactivity in the medium co-eluted with synthetic TRH. Release of TRH by the cells in response to 56 mmol K+/l increased between days 5 and 9 of culture but reached a plateau thereafter. Cell contents of TRH did not change significantly between days 5 and 14 of culture.

Release of TRH from the cells was stimulated by K+ (56 mmol/l), veratridine (100 μmol/l) and ouabain (100 μmol/l) to 550, 480 and 335% of basal release respectively over a 1-h period. Release of TRH was dependent upon calcium in that it was absent when calcium-free medium was used and could be blocked by verapamil (20 μmol/l); however it could not be blocked by nifedipine (50 μmol/l). The calcium ionophore A23187 (1 μmol/l) stimulated TRH release to 340% of basal release. Tetrodotoxin (1 μmol/l) completely abolished the release in response to veratridine but had no effect on the release stimulated by K+ (56 mmol/l).

The calmodulin antagonists trifluoperazine and triflupromazine (50 μmol/l) inhibited veratridine-stimulated TRH release. This was at a site after calcium influx as they also inhibited A23187-stimulated TRH release. The highly specific calmodulin antagonist W7 (10 μmol/l) also inhibited both veratridine and A23187-stimulated TRH release whereas, at the same concentration, its inactive analogue W5 did not significantly inhibit TRH release in response to either stimulus.

These results confirm that fetal rat hypothalamic cell cultures release authentic TRH which can be stimulated by a number of depolarizing agents. Calcium is essential for depolarization-induced release which is also dependent on calmodulin. Fetal rat hypothalamic cell cultures are a valid model for the study of factors controlling the release of TRH.

J. Endocr. (1987) 115, 255–262
Immunoaffinity chromatography of bovine FSH using monoclonal antibodies
Authors: Miller K. F. , Goldsby R. A. , Bolt D. J.
Published: December 2008

Bovine FSH (bFSH) was used to immunize BALB/c mice. Spleen cells were fused to the SP 2/0 cell line to produce hybridomas that secreted monoclonal antibodies to bFSH. One of these antibodies (USDA-bFSH-MC28) was extensively characterized and found to be a gamma 1 with kappa light chains, having extremely low cross-reactivity with other bovine pituitary hormones and with ovine and porcine FSH. The dissociation constant as measured by Scatchard analysis was 4·3 nmol/l, and proved to be in a very useful range for affinity chromatography. In an essentially one-step immunoaffinity chromatography procedure, bFSH was easily isolated in a single chromatographic step from crude anterior pituitary homogenate with better yield and with the same purity as classical chromatographic techniques.

J. Endocr. (1987) 115, 283–288
Alpha1-adrenoceptor production of inositol phosphates mediates the inhibition of thyroxine release from the mouse thyroid
Authors: Muraki T. , Nakaki T. , Kato R.
Published: December 2008

To examine the mechanism of the α1-adrenoceptor-mediated inhibition of the release of thyroid hormones in the mouse thyroid, the effect of noradrenaline on the production of inositol phosphates was investigated using thyroids preloaded with [3H]inositol. Noradrenaline increased [3H]inositol accumulation into inositol phosphates in the mouse thyroid during a 30-min incubation whereas TSH (10 mU/ml) was without effect. The effect of noradrenaline was mimicked by phenylephrine, but not by clonidine or isoprenaline. Prazosin antagonized the effect of noradrenaline, while yohimbine had no effect. These results suggest that noradrenaline-induced production of inositol phosphates is mediated by α1-adrenoceptors. The inhibition of release of thyroid hormones elicited by α1-adrenergic agonists in the mouse thyroid is possibly mediated through the hydrolysis of membrane phosphoinositides followed by the generation of second messengers such as inositol phosphates and diacylglycerol.

J. Endocr. (1987) 115, 289–293
Studies of the rapid effects of parathyroid hormone and prostaglandins on 45Ca uptake into chick and rat bone in vivo
Authors: Dacke C. G. , Shaw A. J.
Published: December 2008

The rapid effects of parathyroid hormones and a variety of prostaglandins on net uptake of 45Ca into the skeleton have been investigated in chicks and, in a limited parallel study, in immature rats.

Intravenous injection of bovine (b) parathyroid hormone(1–34) bPTH(1–34)) or 16,16-dimethyl prostaglandin E2 (16,16-dimethyl PGE2) in a 45Ca-labelled vehicle, combined with subsequent microwave fixation of tissue isotope levels, resulted in rapid (3–15 min) net inhibition of 45Ca uptake into endochondral bone (femur) in chicks (12 days old) and rats (4 weeks old). Use of 125I-labelled albumin and [14C]mannitol indicated that these responses were not a reflection of gross changes in tissue vascular or extracellular space. In rats, bPTH(1–84) also caused significant net inhibition of 45Ca uptake into femur at 10 min. Both bPTH(1–34) and 16,16-dimethyl PGE2 produced generally smaller decreases in 45Ca uptake into chick dermal bone (calvarium) at 3–15 min. In rat calvarium, however, these agents stimulated net uptake of 45Ca at these times. When microwave fixation was omitted, inhibitory responses were reduced or disappeared, while the stimulatory response in rat calvarium was enhanced. Responses to natural prostaglandins (PGE1, PGE2, PGF2α and PGI2) in chicks at 3 min were similar but less marked than those to 16,16-dimethyl PGE2; 45Ca uptake into femur and, to a lesser extent in calvarium, being inhibited. In rats, PGE1, PGE2 and PGF2α showed a tendency to decrease 45Ca uptake into femur while PGE1 and PGE2 both increased 45Ca uptake into calvarium.

J. Endocr. (1987) 115, 369–377
Difference in luteal and placental P45017α: substrate preference and hormonal regulation in the rat
Authors: Eckstein B. , Khan I. , Gibori G.
Published: December 2008

The purpose of this study was to assess the substrate specificity of P45017α in both the corpus luteum and placenta of pregnant rats, and to analyse the site at which LH/human chorionic gonadotrophin (hCG) regulates the activities of this enzyme. To distinguish the substrate preference, placentas and corpora lutea were obtained from rats on day 15 of pregnancy. Tissues were homogenized and the 10 000 g supernatants incubated in the presence of equimolar concentrations of [14C]progesterone and [3H]17α-hydroxyprogesterone as substrate with either NADH or NADPH as cofactors for 2, 8, 16 and 24 min. The labelling pattern of both 17α-hydroxyprogesterone and testosterone indicated that the corpus luteum produced testosterone preferentially from progesterone, whereas the placenta principally used 17α-hydroxyprogesterone and synthesized six times as much testosterone from 17α-hydroxyprogesterone than from progesterone. Addition of either NADPH or NADH as cofactors had no effect on substrate preference. The products of the two enzymatic activities were identified by recrystallization to constant 14C/3H ratios. The ratio of 14C/3H in testosterone produced by the corpus luteum was 16-fold higher than in that produced by the placenta. To explore which of the two activities of P45017α is regulated by the gonadotrophin, rats were treated with either 1·5 IU hCG or vehicle between days 13 and 15 of pregnancy. Hydroxylase and lyase activities were determined on day 15 after incubation for 2,8,16 or 24 min in the presence of either NADH or NADPH. Administration of hCG significantly inhibited NADH-dependent 17α-hydroxylase in the placenta at each time-point studied. The inhibition reached 69% at 24 min. Human chorionic gonadotrophin did not affect the NADPH-dependent 17α-hydroxylase and had only a slight inhibitory effect on both NADH- and NADPH-dependent 17,20-lyase activities in the placenta. In contrast to its effect on the placenta, hCG stimulated both NADH- and NADPH-linked 17,20-lyase activities but had no measurable effect on 17α-hydroxylase activities in the corpus luteum.

In summary, the results of the present investigation have revealed a significant difference in the behaviour of 17α-hydroxylase/17,20-lyase activities in the placenta and corpus luteum. The substrate preference and the control of both enzyme activities by LH/hCG differs dramatically.

J. Endocr. (1987) 115, 387–393
Mechanism of replenishment of androgen receptors in cytosol of mouse submandibular gland
Authors: Kyakumoto S. , Kurokawa R. , Ota M.
Published: December 2008

Female mice were used to examine the process of depletion and replenishment of cytosolic androgen receptors in submandibular glands, and to investigate the effects of cycloheximide and actinomycin D on these processes. The dose-dependence of receptor depletion and replenishment in the cytosolic fraction, and that of receptor accumulation in the nuclear fraction were investigated. Almost 100% depletion was revealed 1 h after the injection of testosterone propionate at a dose of 500 or 50 μg testosterone/100 g body weight. With a 5 μg dose, depletion of cytosolic receptors was not complete and replenishment proceeded rapidly compared with that which occurred with the 50 or 500 μg dose. The nuclear receptor level increased 1 h after injection of testosterone, and the raised level was gradually reduced to the pretreatment level with all doses. However, the time required for this return to pretreatment level was dependent on the dose of testosterone. The change in the levels of cytosolic and nuclear androgen receptors following injection of testosterone was parallel to the level of circulating androgen. To determine the requirements for transcriptional and translational events in the replenishment process, cycloheximide and actinomycin D were given in vivo. The process of replenishment of cytosolic receptors was inhibited by the injection of cycloheximide. However, actinomycin D exerted no inhibitory effect on receptor replenishment. Neither cycloheximide nor actinomycin D had any effect on the nuclear receptor level until 6 h after the injection of testosterone. Cycloheximide or actinomycin D alone had no effect on the cytosolic or nuclear receptor level. These results suggest that receptor replenishment involves protein synthesis.

J. Endocr. (1987) 115, 411–418
Dopamine stimulates release of thyrotrophin-releasing hormone from perfused intact rat hypothalamus via hypothalamic D2-receptors
Authors: Lewis B. M. , Dieguez C. , Lewis M. D. , Scanlon M. F.
Published: December 2008

We have studied the effect of dopamine together with agonist and antagonist drugs of different specificities on the release of TRH from the perfused, intact hypothalamus of the adult rat in vitro. Dopamine produced a dose-related stimulatory effect on TRH release with maximal effect being achieved at 1 μmol/l (increase over basal, 118 ±16·5 (s.e.m.) fmol TRH; P <0·001 vs basal). This effect was mimicked by the specific D2-agonist drugs bromocriptine (0·1 μmol/l) and LY 171555 (0·1 μmol/l) (increase over basal values, 137·5±13·75 fmol and 158·6± 10·7 fmol respectively; P <0·001 vs basal), but not by the D1-agonist SKF 38393A. The stimulatory effect of dopamine (1 μmol/l) was blocked in a stereospecific manner by the active (d) but not by the inactive (l) isomers of the dopamine antagonist butaclamol. Similar blockade was achieved with the specific D2-antagonist domperidone (0·01 μmol/l) whereas the D1-antagonist SCH 23390 was only effective when used at a concentration 100 times greater. Lower concentrations (0·01 μmol/l) of this D1 -antagonist did not block the stimulatory effect of dopamine. High-performance liquid chromatography characterization of the material secreted within the hypothalamus showed one single peak of immunoreactive material which coeluted with synthetic TRH. These data suggest that dopamine exerts a stimulatory role in the control of hypothalamic TRH release by acting at specific D2-receptors.

J. Endocr. (1987) 115, 419–424
Relative occupation of type-I and type-II corticosteroid receptors in rat brain following stress and dexamethasone treatment: functional implications
Authors: Reul J. M. H. M. , van den Bosch F. R. , de Kloet E. R.
Published: December 2008

The rat brain contains two receptor systems for corticosterone: the type-I corticosterone-preferring receptor and the classical type-II glucocorticoid receptor. The two receptor populations can be distinguished in binding studies with the 'pure' synthetic glucocorticoid 11β,17β-dihydroxy-6-methyl-17α (1-propynyl)-androsta-1,4,6-trione-3-one (RU 28362). In-vitro autoradiography and quantitative image analysis showed that the type-I receptor was localized almost exclusively in the hippocampus, whereas the type-II receptor extended throughout the brain, with the highest levels in the nucleus paraventricularis, nucleus supraopticus and in the thalamic, amygdaloid, hippocampal and septal regions. Unoccupied type-I and type-II receptor sites, as measured in vitro by cytosol binding of 3H-labelled steroids, displayed a large difference in the rate of appearance after adrenalectomy. The availability of type-I receptors exhibited a marked increase, reaching maximal levels within 4–7 h, and then remained constant until 2 weeks after adrenalectomy. The availability of type-II receptors did not change considerably during the first 24 h after adrenalectomy, but displayed a large increase in capacity during the subsequent 2 weeks. After adrenocortical activation as a consequence of exposure to a novel environment, plasma concentrations of corticosterone increased to reach a peak of 811 nmol/l after 30 min and attained the basal concentration (43 nmol/l) after 240 min. During this time, occupation of type-I receptors increased from 77·8% at 0 min to 97% at 30–60 min and then declined to 84·8% after 240 min. Occupation of the type-II receptors was 28·1% at 0 min, 74·5% after 30 min and 32·8% after 240 min. Injection of dexamethasone (25 μg/100 g body wt) at 08.00 h resulted in suppression of basal plasma concentrations of corticosterone and prevented the circadian-driven rise in circulating corticosterone. Occupation of type-I receptors did not change considerably as a result of injection of dexamethasone, but occupation of type-II receptors was markedly increased till 16.00 h compared with that after injection of vehicle.

It was concluded that the type-I and type-II receptors are not only localized differently in the rat brain, but also exhibit a striking difference in occupation after manipulation of the pituitary-adrenocortical system. The data further support the concept of a type-I receptor-mediated tonic activating influence and a type-II receptor-mediated feedback action of corticosterone on brain function.

J. Endocr. (1987) 115, 459–467
INHIBIN BIOACTIVITY IN HUMAN TESTICULAR EXTRACTS
Authors: Simpson B.J.B. , Tsonis C.G. , Wu F.C.W.
Published: December 2008

Inhibin bioactivity was measured in human testicular extracts by a sensitive sheep pituitary cell bioassay. The relationship between testicular inhibin bioactivity, daily sperm production (DSP) and plasma concentrations of FSH, LH, testosterone and oestradiol were examined. The mean level of testicular inhibin bioactivity was 4.4 ±1.3 U/g (mean ± SD) with a significantly lower value in those who received radiotherapy (3.2 ± 1.4 U/g) than in the untreated group (4.8 ± 1.1 U/g). In contrast to the rat, human testicular inhibin bioactivity was not significantly correlated to FSH or DSP. These findings suggest that inhibin may have a complex role in normal and/or pathological testicular function.
LEYDIG CELL STEROID PRODUCTION IN THE PRESENCE OF LH IS FURTHER STIMULATED BY RAT TESTICULAR FLUID, FETAL CALF SERUM AND BOVINE FOLLICULAR FLUID, BUT NOT BY RAT AND BOVINE SERUM
Authors: Melsert R. , Rammerts F.F.G.
Published: December 2008

LH-dependent steroid production by isolated rat Leydig cells is fourfold stimulated by the addition of charcoal–stripped rat testicular fluid. Fetal calf serum and bovine follicular fluid have similar stimulatory effects. In contrast, rat and bovine serum suppress steroid production in the presence of LH in a dose–dependent manner. The results show that stimulatory and inhibitory factors can modulate Leydig cell steroidogenesis and these factors appear not to be organ or species specific.
Metabolic actions of cortisol in a macropodid marsupial Thylogale billardierii
Authors: Martin I. K. , McDonald I. R.
Published: December 2008

In undisturbed pademelon wallabies (Thylogale billardierii) with indwelling jugular venous catheters, an increase in the plasma cortisol concentration from 0.25±0.05 to 1.35±0.15 (s.e.m.) μmol/l in 2 h, during i.v. infusion of cortisol at 1.0 mg/kg per h, caused no significant change in the plasma glucose concentration from the control value of 4.26±0.25 mmol/l. The rates of appearance (Ra) and metabolic clearance (MCR) of glucose, measured by steady-state isotope dilution, also did not change significantly from the control values of 14.9±0.7 μmol/kg per min and 3.52±0.19 ml/kg per min respectively. Twice-daily i.m. injections of 7 mg cortisol/kg for 7 days caused increases in plasma concentrations of cortisol, from 0.26±0.02 to 0.66±0.04 μmol/l on day 7, and glucose, from 5.1±0.1 to 7.2±0.6 mmol/l by day 5. The concentration of glycogen in the liver of wallabies fasted for 24 h increased from the control level of 1.17±0.56 to 5.92±1.14 g/100 g on day 7 (P<0.01), but mean glucose Ra and MCR did not change significantly. Plasma concentrations of α-amino nitrogen rose from 2.73±0.13 to 3.22±0.12 mmol/l on day 1 and remained at this level. Plasma concentrations of urea rose from 8.59±0.62 to 9.70±0.32 mmol/l on day 1, but then declined below the control level. Food intake and urinary excretion of nitrogen did not change in undisturbed animals. However, fasting followed by liver biopsy was accompanied by urinary excretion of nitrogen in excess of food intake, persisting until day 2 of treatment.

The transient effect of cortisol on the plasma concentration of urea and lack of effect on urinary excretion of nitrogen could be explained by urea recycling, as indicated by a low urinary urea nitrogen: total nitrogen ratio and failure to excrete more than a mean of 26% of infused urea.

It is concluded that cortisol has no short-term effect on carbohydrate metabolism in this marsupial. In the long term it can increase hepatic carbohydrate reserves through utilization of tissue amino nitrogen, the resulting urea being conserved by recycling.

J. Endocr. (1988) 116, 71–79
Induced development of ovulatory follicles during the early stages of lactation by the administration of LH in the rat
Authors: Taya K. , Sasamoto S.
Published: December 2008

To determine whether failure of follicular maturation during the early stages of lactation in rats is due to inadequate LH stimulation, lactating rats nursing eight pups were injected twice daily for 1–3 days (days 2–5 of lactation) with various doses of ovine LH. Follicular maturation was determined by the ability of the follicles to ovulate in response to 10 IU human chorionic gonadotrophin (hCG), endogenous oestradiol-17β and inhibin production. Ovulation was not induced in control animals in response to 10 IU hCG given between days 2 and 5 of lactation. On the other hand, an injection of 10 IU hCG could induce ovulation in LH-treated animals, in which 25 and 50 μg LH per injection were given s.c. from days 2 to 5 of lactation. Concentrations of oestradiol-17β and inhibin activity in ovarian venous plasma increased progressively after the administration of LH, indicating that induced development of ovulatory follicles had occurred. Plasma concentrations of FSH declined in LH-treated animals compared with those in control animals. The decrease in plasma concentrations of FSH was not observed when lactating rats were ovariectomized before the first injection of LH, indicating that ovarian products, probably inhibin, from developing follicles may suppress the secretion of FSH from the pituitary gland. In both LH-treated and control animals, concentrations of prolactin and progesterone remained increased during the period of LH administration. The present results, therefore, suggest that the plasma levels of LH are an important determinant of follicular maturation during lactation in rats.

J. Endocr. (1988) 116, 115–122
Peripheral progesterone concentrations in the luteal-phase ewe: effects of a β-adrenergic receptor antagonist and two β2-adrenergic agonists
Authors: Wheeler A. G. , Lean J. , Walker M.
Published: December 2008

Peripheral blood samples were collected at 10-min intervals from three conscious sheep in which ovulation had been induced 6–10 days previously using exogenous hormones. Saline was infused into a jugular vein for about 1 h, followed by the experimental drug for 1–2 h and followed by saline again for a further 2 h. The experiments were repeated following induced luteolysis and ovulation. The infusion of a β-adrenergic antagonist (propranolol) into three conscious luteal-phase ewes decreased (P<0·05) the peripheral progesterone concentration in each animal. Infusions of β2-adrenergic agonists (ritodrine and salbutamol) increased (P<0·05) the progesterone concentration in four out of eight experiments. The β-adrenergic antagonist decreased the heart rate and the β2-adrenergic agonist increased it; the arterial blood pressure and respiratory rate were unaffected. The decrease in the prosgesterone concentration in response to the β-adrenergic antagonist suggests that the normal ovarian secretion of progesterone is partly the result of sympathetic stimulation, and that the sympathetic innervation of the ovary may have a physiological role in modulating progesterone secretion.

J. Endocr. (1988) 116, 137–142
Ovarian steroids modulate the action of calcitonin in women
Authors: Pansini F. , Bergamini C. M. , Bellinazzi A. , Andreoli M. , Perri G. , Bagni B. , Mollica G.
Published: December 2008

Slow i.v. infusion of salmon calcitonin into normal women led to a transient increase in plasma concentrations of cyclic AMP and to a decrease in plasma calcium, which was maintained for at least 2 h. These responses were significantly reduced in the same patients after hysterectomy and ovariectomy. In contrast, maintenance of the ovaries at the time of surgery, or substitutive hormonal therapy, seemed to restore the sensitivity to exogenous calcitonin, suggesting a permissive role of ovarian steroids in the effect of calcitonin. The most likely explanation for this phenomenon is that these hormones influence either the number of calcitonin receptors or the coupling of the receptors to the functional unit of bone adenylate cyclase.

J. Endocr. (1988) 116,155–159
The somatotrophic axis in young steers: influence of nutritional status and oestradiol-17β on hepatic high-and low-affinity somatotrophic binding sites
Authors: Breier B. H. , Gluckman P. D. , Bass J. J.
Published: December 2008

The binding of bovine GH (bGH) to hepatic membranes obtained from steers on either high (3% dry matter of body weight per day) or low (1% dry matter of body weight per day) planes of nutrition with or without an oestradiol-17β implant was studied (n = 5 per group). Binding studies were performed on both crude membrane homogenates and on 100 000 g microsomal membrane fractions; identical results were obtained using both preparations. In all four groups of animals, linear Scatchard plots were obtained, but following pretreatment of the membranes with MgCl2 to remove endogenously bound hormone, curvilinear plots were obtained in the groups on the high plane of nutrition. Analysis of these curves suggested the presence of a high- and low-affinity binding site, the high-affinity site being fully occupied in the absence of MgCl2 pretreatment.

The specific binding of bGH in MgCl2-pretreated crude membranes was greater (P < 0·01) in well-fed steers (14·8 ± 1·6%) than in poorly fed steers (9·8 ± 0·9%). Scatchard analysis showed this to be due to the presence of a high-affinity site (dissociation constant (Kd) = 11·6 ± 3·3 pmol/l) in the well-fed animals only. In addition, there was an increase (P < 0·01) in the affinity, but not in the capacity, of the low-affinity site (Kd = 106·4 ± 22·8 pmol/l in well-fed steers and 197·0 ± 23·8 pmol/l in poorly fed steers).

Oestradiol treatment was associated with an increase (P < 0·01) in specific binding at both planes of nutrition, but binding was higher (P < 0·01) in well-fed (24·8 ± 2·9%) than in poorly fed (15·6 ± 3·7%) steers. Scatchard analysis after MgCl2 pretreatment again showed a curvilinear plot at the high and a linear plot at the low nutritional plane. The effect of oestradiol was to increase (P < 0·001) the capacity of the high-affinity site from 1·87 ± 0·61 pmol/100 mg in the control well-fed group to 6·56 ± 1 ·2 pmol/100 mg. The capacity of the low-affinity site was increased (P < 0·01) from 20·1 ± 2·6 to 30·1 ± 3·2 pmol/100 mg in the well-fed group, with a similar change in the poorly fed group. Oestradiol had no effect on the apparent affinity of either binding site.

These studies demonstrate a heterogeneity of somatotrophic binding sites of hepatic membranes in steers. The presence of a high-affinity site is determined by nutritional status, whereas oestradiol primarily affects receptor capacity. Thus nutrition and oestradiol have independent and qualitatively different effects on somatotrophic binding. As the rate of weight gain correlated (P < 0·01) with the capacity of the high-affinity site, it is suggested that somatotrophic receptor modulation is a primary factor in the regulation of somatic growth in the ruminant.

J. Endocr. (1988) 116, 169–177
The role of protein kinase C in LHRH-induced LH and FSH release and LHRH self-priming in rat anterior pituitary glands in vitro
Authors: Johnson M. S. , Mitchell R. , Fink G.
Published: December 2008

We have investigated the role of protein kinase C (PKC) in LHRH-induced LH and FSH secretion and LHRH priming. Hemipituitary glands from pro-oestrous rats were incubated with agents known to affect PKC and with or without LHRH, during which time the secretion of gonadotrophins was measured. Phorbol esters and phospholipase C, activators of PKC, released LH and FSH in a concentration-dependent manner and potentiated the LHRH-induced secretion of gonadotrophins in parallel with their ability to release these hormones alone. Inhibitors of PKC had either no effect on LH release (1-(5-isoquinolinesulphonyl)-2-methylpiperazine hydrochloride) or they augmented LHRH-induced gonadotrophin release (polymyxin B and 8-(N,N-diethylamino) octyl-3,4,5-trimethoxybenzoate). Neither the activators nor the inhibitors of PKC, when present with LHRH, caused any change in LHRH priming, even though the activators alone produced a release of gonadotrophins that showed a temporal pattern similar to that produced by LHRH priming. The profiles of effects on LH and FSH secretion were always qualitatively similar. These results show that PKC may be involved in general regulation of gonadotrophin release but that it is not important in acute responses to LHRH nor in LHRH self-priming.

J. Endocr. (1988) 116, 231–239
Inhibin bioactivity and pituitary cell mitogenic activity from cultured chicken ovarian granulosa and thecal/stromal cells
Authors: Tsonis C. G. , Sharp P. J. , McNeilly A. S.
Published: December 2008

A sensitive bioassay for inhibin based on the suppression of FSH release from cultured sheep anterior pituitary cells was used to determine whether inhibin is present in the preovulatory follicle in the domestic hen. Granulosa and thecal/stromal layers were separated from the five largest (F1–F5) yellow yolky follicles in the ovary and incubated in culture medium for 18 h. Inhibin was found predominantly in the media in which granulosa layers had been incubated. There was a progressive increase in the amount of inhibin produced per mg granulosa layer protein during the 5–6 days before ovulation.

The ovary was observed to contain a growth factor which stimulated the proliferation of ovine pituitary cells. Thecal/stromal layer-conditioned medium (ThCM) but not granulosa layer-conditioned medium had a dose- and time-dependent mitogenic effect on cultured sheep pituitary cells. The maximal mitogenic effect achieved for ThCM was four to fivefold greater than control media and was significantly higher than the maximal mitogenic effects of epidermal growth factor (250 ng/ml; 1·5 × control) and transforming growth factor-β (500 ng/ml; 1·2 × control).

It is concluded that inhibin is produced by the granulosa layers in the large yellow yolky preovulatory ovarian follices of the domestic hen. The thecal/stromal layers in these follicles produce a potent mitogenic factor, not produced by the granulosa layers, which stimulates the division of ovine anterior pituitary cells in vitro.

J. Endocr. (1988) 116, 293–299
Effect of isolation stress on concentrations of arginine vasopressin, α-melanocyte-stimulating hormone and ACTH in the pituitary venous effluent of the normal horse
Authors: Alexander S. L. , Irvine C. H. G. , Livesey J. H. , Donald R. A.
Published: December 2008

A non-surgical, non-stressful technique was used for collection of pituitary venous blood from five conscious horses every minute for two 10-min periods before and during isolation from the herd, which caused a predictable, yet humane and physiological, emotional stress. Pituitary blood was also sampled every 5 min for two approximately 90-min periods before and after isolation, while jugular blood was sampled every 15 min throughout the experiment.

During isolation, all horses became agitated, hyperventilating and sweating. Packed red cell volume increased, as did pituitary venous concentrations of adrenaline (mean ± s.e.m. concentration before isolation, 621·5±112·3 pmol/l; peak during isolation, 2665·4 ± 869·8 pmol/l; P <0·05) and noradrenaline (before, 871·8 ± 111·8 pmol/l; peak, 2726·1 ± 547·4 pmol/l; P<0·02). Concentrations of arginine vasopressin (AVP) were higher in pituitary venous but not in jugular blood during isolation than during the preceding 10-min period (P <0·05). Although AVP secretion increased in all horses, in three of the five it rose dramatically in the first minute of isolation to 25·7 (horse 1), 13·6 (horse 4) and 145·1 (horse 5) times the level in the last sample collected before isolation. Mean pituitary venous concentrations of ACTH and α-MSH increased during isolation in the three horses which had large increases in AVP secretion, but, overall, stress did not significantly affect ACTH or α-MSH secretion. Similarly, mean jugular cortisol levels were not significantly altered by isolation. However, the magnitudes of ACTH, AVP and α-MSH responses to isolation were negatively correlated with the jugular cortisol level before isolation. The changes in pituitary venous concentrations of ACTH and AVP were synchronous under resting conditions, whether samples were collected at intervals of 1 (P <0·01) or 5 (P <0·005) min; however, this synchrony was lost during isolation. The changes in pituitary venous concentrations of ACTH and α-MSH were synchronous both at rest (P <0·025 for 1-min sampling, P <0·01 for 5-min sampling) and during isolation (P<0·01).

We conclude that isolation stress increases AVP secretion and may alter the temporal relationship between pituitary venous concentrations of AVP and ACTH. Furthermore, the magnitude of the responses of AVP, ACTH and α-MSH to isolation is significantly affected by the prevailing cortisol level.

J. Endocr. (1988) 116, 325–334
Role of calcium in the secretomotor response of the thyroid: effects of calcium ionophore A23187 on radioiodine turnover, membrane potential and fluid transport in cultured porcine thyroid cells
Authors: Manley S. W. , Rose D. S. , Huxham G. J. , Bourke J. R.
Published: December 2008

The calcium ionophore A23187 (0·1–1 μmol/l) inhibited membrane electrical polarization, uptake of 125I, fluid transport and TSH-stimulated release of radioiodine from the organic pool in follicular cultures of porcine thyroid cells. At higher concentrations (1–30 μmol/l), A23187 promoted release of radioiodine from the organic pool. Stimulation of release of radioiodine from the organic pool by veratridine (a sodium channel agonist, 0·4–1 mmol/l) and A23187 was dependent on the calcium concentration of the medium, while TSH action was independent. Incubation in medium of very low calcium concentration (0·0177 mmol/l) resulted in enhanced release from the organic pool, which was inhibited by TSH (256 μU/ml), A23187 (25 μmol/l) or veratridine (0·5 mmol/l). These data therefore do not support the hypothesis that calcium acts as a mediator of the secretomotor action of TSH, but suggest the possibility of a TSH-induced increase in intracellular calcium as a regulatory negative-feedback mechanism.

J. Endocr. (1988) 116, 373–380
RNA synthesis in the Leydig cells of the mature rat: effect of oestradiol-17β
Authors: Ronco A. M. , Pino A. M. , Valladares L. E.
Published: December 2008

In view of the evidence that there may be an effect of high concentrations of oestradiol on testicular steroidogenic function, we have investigated the effect of this steroid on [3H]uridine incorporation into RNA by testicular cells. Our results have shown that oestradiol in vitro induced a marked dose-dependent inhibition of RNA synthesis by purified Leydig cells. The concentrations of oestradiol tested varied from 2 to 40 μmol/l; these concentrations also impaired net testosterone synthesis in vitro after human chorionic gonadotrophin (hCG) stimulation. Under the effect of oestradiol, the kinetics of [3H]nucleoside incorporation into RNA were impaired early and the inhibition of RNA synthesis was specific for oestrogenic compounds. It was concluded that, in Leydig cells, oestradiol, in addition to its known inhibitory action on the response of testosterone to hCG, triggers a more extensive response that also includes RNA synthesis in vitro.

J. Endocr. (1988) 116, 387–392
Plasma levels of neuropeptide tyrosine Y (NPY) are increased in human sepsis but are unchanged during canine endotoxin shock despite raised catecholamine concentrations
Authors: Watson J. D. , Sury M. R. J. , Corder R. , Carson R. , Bouloux P. M. , Lowry P. J. , Besser G. M. , Hinds C. J.
Published: December 2008

Alterations in levels of adrenal vein and circulating immunoreactive neuropeptide tyrosine Y (NPY), as well as systemic catecholamine concentrations, have been investigated in the dog after endotoxin administration. Eighteen dogs were anaesthetized with α-chloralose and allowed to breathe spontaneously. Severe shock was produced by the administration of a large bolus of Escherichia coli endotoxin followed by a continuous infusion. In two further animals the left lumbar adrenal vein was cannulated and an intermittent choke allowed retrograde sampling of the adrenal effluent. The production of hypodynamic shock was associated with marked increases in systemic catecholamine concentrations but there were no consistent changes in adrenal vein or systemic immunoreactive NPY levels. In contrast, circulating concentrations of immunoreactive NPY were markedly raised above the normal range in five patients with septic shock.

J. Endocr. (1988) 116, 421–426
Relationship between external and cytoplasmic calcium concentrations, parathyroid hormone release and weight of parathyroid glands in human hyperparathyroidism
Authors: Wallfelt C. , Gylfe E. , Larsson R. , Ljunghall S. , Rastad J. , Åkerström G.
Published: December 2008

Parathyroid hormone (PTH) release and cytoplasmic calcium concentrations were investigated at ambient calcium concentrations of 0·5–3·0 mmol/l in dispersed parathyroid cells from 44 hypercalcaemic patients with primary or uraemic hyperparathyroidism (HPT). In comparison with parathyroid cells from adult cattle, release of PTH by human preparations was reduced and values of the ambient calcium concentration causing half-maximal inhibition of PTH release (median effective dose, ED50) were significantly increased. Half-maximal inhibition of PTH release was obtained with concentrations of cytoplasmic calcium almost identical to the concentrations of ionized calcium in the plasma of the individual patients. Cytoplasmic concentrations of calcium in the parathyroid cells were inversely related to release of PTH. Concentrations of cytoplasmic calcium were significantly lower in human than in bovine cells and the ED50 for ambient calcium increase on cytoplasmic calcium was raised to the same extent as the ED50 for ambient calcium inhibition of PTH release in human compared with bovine cells. The magnitude of the increased ED50 for ambient calcium inhibition of PTH release and increase of cytoplasmic calcium concentration was similar in adenomas and sporadic as well as hereditary primary hyperplasias, but the secretion was the least aberrant in uraemic hyperplasias, although they had by far the largest glandular mass. Serum concentrations of total calcium before surgery correlated with the ED50 for ambient calcium effects of PTH release and cytoplasmic calcium, but not with glandular weight. These findings demonstrate a universally abnormal regulation of cytoplasmic calcium in HPT and its importance for PTH release, and that disturbance of cytoplasmic calcium rather than the increased glandular mass contributes to the hypercalcaemia in adenomatous and hyperplastic HPT.

J. Endocr. (1988) 116, 457–464
GALANIN ABOLISHES THE INHIBITORY EFFECT OF CHOLINERGIC BLOCKADE ON GROWTH HORMONE-RELEASING HORMONE-INDUCED SECRETION OF GROWTH HORMONE IN MAN
Authors: Chatterjee V.K.K. , Ball J.A. , Proby C. , Burrin J.M. , Bloom S.R.
Published: December 2008

In five healthy normal male volunteers, pretreatment with the cholinergic muscarinic antagonist pirenzepine (30 mg i.v.) almost abolished the growth hormone (GH) response to a maximal dose (120 μg i.v.) of growth hormone-releasing hormone (GHRH) (GH response at 40 min 5.6 ± 1.3 mU/l with GHRH and pirenzepine vs 40.8 ± 5.3 mU/l with GHRH alone, P <0.02). Concomitant i.v. infusion of galanin (40 pmol/kg/min) with pirenzepine not only restored but significantly potentiated the GH response to GHRH (GH at 40 min 72.2 ± 10.5 mU/l, P <0.001 vs GHRH and pirenzepine, P <0.02 vs GHRH alone). Previous studies have proposed that cholinergic pathways control GH release via samatostatin and this study suggests that galanin may act by modulating hypothalamic somatostatinergic tone either directly or, possibly, by facilitating cholinergic neurotransmission.
THE SHEEP CORPUS LUTEUM SECRETES INHIBIN
Authors: Tsonis C. G. , Baird D. T. , Campbell B. K. , Leask R. , Scaramuzzi R. J.
Published: December 2008

An experiment was performed in 20 Merino ewes in which ovarian venous blood was collected by venepuncture at surgery and at two stages of the oestrous cycle. The ovarian venous concentrations of inhibin, oestradiol-17β and progesterone were determined. The results demonstrate that during the luteal phase of the oestrous cycle the ovarian venous blood draining an ovary containing luteal tissue contains significantly more inhibin bioactivity than ovarian venous blood from an ovary not containing luteal tissue. During the follicular phase the concentration of inhibin bioactivity in ovarian venous blood was reduced compared with the luteal phase. From this data we conclude that the sheep corpus luteum secretes inhibin bioactivity into the ovarian venous blood.
A COMPARATIVE STUDY OF ENDOMETRIAL ACID PHOSPHATASE ACTIVITY IN THE MID-LUTEAL PHASE OF THE MENSTRUAL CYCLE AND IN THE PERI-IMPLANTATION STAGE IN THE RHESUS MONKEY (MACACA MULATTA)
Authors: Ghosh D. , Sengupta J.
Published: December 2008

Acid phosphatase activity in both total and subcellular fractions of endometrium of rhesus monkey on day 5 of gestation was significantly (P <0·01) lower than levels found in tissues collected from normally menstruating monkeys on day 5 after ovulation. This endometrial response to preimplantation embryo may be related to the menstrual withdrawal in a fertile cycle.
Characterization of the regenerated Leydig cell population of the rat after destruction by ethylene-1,2-dimethanesulphonate
Authors: Edwards G. , Robertson W. R. , Morris I. D.
Published: December 2008

The Leydig cells repopulating the adult rat testis after destruction by a single injection of the cytotoxic ethylene-1,2-dimethanesulphonate (EDS) were investigated. After 14 days, serum concentrations of LH and FSH were significantly raised and concentrations of testosterone in the serum and testis reduced. At 21 days, hormone concentrations had returned to within the normal range. Binding of 125I-labelled human chorionic gonadotrophin (hCG) to testis homogenate, however, was still less than 10% of normal. After 21 or 28 days the 125I-labelled hCG binding profiles of isolated Leydig cells from EDS-treated rats, separated on a Percoll gradient, showed a single peak similar to that of immature (25 days old) rats. After 49 days, 125I-labelled hCG binding resolved into two peaks more like that of normal adult rats. Using a quantitative cytochemical method, 3β-hydroxysteroid dehydrogenase activity in individual Leydig cells of unfixed testis sections was determined. Activity was increased by 70% (P < 0·05) in repopulating Leydig cells 21 days after EDS treatment compared with cells from vehicle-treated rats. In addition, Leydig cells were still capable of further 'in-vivo' stimulation by pharmacological doses of hCG. These data indicate that Leydig cells repopulating the testis are homogenous. Fewer cells from the newly formed population are capable of maintaining normal serum concentrations of testosterone and must thus be individually more active in secreting testosterone. In these respects, the Leydig cells repopulating the adult rat testis after EDS treatment more closely resemble those of the fetal rat testis.

J. Endocr. (1988) 117, 11–18
Stimulatory effect of insulin-like growth factor-I on [3H]thymidine incorporation, DNA content and insulin biosynthesis and secretion of isolated pancreatic rat islets
Authors: Sieradzki J. , Fleck H. , Chatterjee A. K. , Schatz H.
Published: December 2008

The influence of insulin-like growth factor-I (IGF-I) on both the growth and function of isolated pancreatic rat islets was studied. As a measure of islet cell replication, [3H]thymidine incorporation and DNA content was estimated at the same time as parameters for (pro)insulin biosynthesis and secretion as a measure of the functional capacity of B cells. Incorporation of [3H]thymidine was measured after tissue solubilization; DNA was determinated fluorometrically. (Pro-)insulin biosynthesis was determinated by incorporation of [3H]leucine after immunoprecipitation and binding to protein A–Sepharose. There was a significant increase in both [3H]thymidine incorporation and DNA content after the culture of islets with IGF-I. IGF-I resulted also in an increase in insulin biosynthesis and secretion from isolated pancreatic islets. These results demonstrate a role of IGF-I in regulating the growth and function of pancreatic islets.

J. Endocr. (1988) 117, 59–62