bluuebunny/crossref_metadata_2025 huggingface dataset:
Changes in serum concentrations of immunoreactive inhibin during the oestrous cycle of the rat
Authors: Hasegawa Y. , Miyamoto K. , Igarashi M.
Published: December 2008

Serum and ovarian concentrations of inhibin during the oestrous cycle of rats were determined by a radioimmunoassay (RIA) based on a porcine inhibin RIA. Serum concentrations of LH, FSH, oestradiol-17β, progesterone and testosterone were also determined during the cycle. Serum concentrations of inhibin were high (8·47 ± 0·58 μg/l) during the morning of pro-oestrus and then dropped sharply to the lowest level (3·21 ± 0·38 μg/l) at 24.00 h on the day of pro-oestrus after the preovulatory LH and FSH surge. Inhibin concentrations then recovered rapidly, with a peak (7·94 ± 0·97 μg/l) at 15.00 h on the day of oestrus. Inhibin concentrations in serum dropped again to their second lowest value (4·00 ± 0·12 μg/l) at 24.00 h on the day of oestrus. The largest FSH surge observed from midnight of pro-oestrus to the morning of oestrus (the so-called second FSH surge) was shown to be inversely related to the changes in serum concentrations of inhibin, suggesting a close relationship between the serum FSH and inhibin concentrations in cyclic rats.

Journal of Endocrinology (1989) 121, 91–100
Effects of cholesterol and lipoproteins on aldosterone secretion by bovine zona glomerulosa cells
Authors: Simpson H. D. , Shepherd R. , Shepherd J. , Fraser R. , Lever A. F. , Kenyon C. J.
Published: December 2008

Freshly isolated bovine adrenocortical cells were pretreated with various concentrations of cholesterol and of high- (HDL) and low-density lipoprotein (LDL) fractions of known cholesterol content and then incubated in medium alone with and without angiotensin II. Preincubation with cholesterol (323 μmol/l) caused basal aldosterone synthesis to increase from 0·89 ± 0·08 to 2·77 ± 0·22 pmol/106 cells per hour (±s.e.m.) but did not significantly affect angiotensin-stimulated synthesis. Human HDL containing cholesterol at a final concentration of 129–647 μmol/l increased both basal and angiotensin-stimulated aldosterone synthesis. In HDL-treated cells, both the threshold response and responses to increasing concentrations of angiotensin were raised. Human LDL had no effect on basal or stimulated aldosterone synthesis nor did LDL alter the effects of HDL when cells were incubated with HDL and LDL in combination. Qualitatively similar results were obtained with bovine lipoproteins.

These studies show that, in short-term incubations of fresh tissue, the supply of cholesterol may be a limiting factor in aldosterone synthesis and that HDL rather than LDL is the preferred source. These observations are discussed in relation first to the mechanisms by which cholesterol/HDL might augment steroid responses and secondly to other studies with cultured cells which have demonstrated a role for LDL.

Journal of Endocrinology (1989) 121,125–131
LH in polycystic ovary syndrome: reproducibility and pulsatile secretion
Authors: Murdoch A. P. , Diggle P. J. , White M. C. , Kendall-Taylor P. , Dunlop W.
Published: December 2008

Serum concentrations of LH are increased in polycystic ovary syndrome (PCOS). We have investigated two aspects of LH secretion which have not previously been reported: its reproducibility within individuals and the pattern of superimposed pulses of LH secretion. In nine patients with PCOS the mean concentration of LH was calculated from 24 blood samples taken at 15-min intervals for 6 h on two or three occasions over 1 year. Results showed differences in mean LH between subjects but reproducible concentrations within subjects over that period. It has been shown that LH is secreted in a complicated pattern of superimposed pulses which can be characterized by using the statistical methods of time-series analysis. To evaluate these pulse patterns of LH we studied nine patients with PCOS and compared the results with those of 12 normal women in the early follicular phase of the ovarian cycle. Blood samples were taken at either 5-min intervals for 6 h or 1-min intervals for 1 h. Pulses were detected in both groups at frequencies of about 1 h and 2 to 3 min. There was no significant difference in the pulse frequencies between the patients and controls but the amplitude of both groups of pulses was increased in the PCOS patients.

Journal of Endocrinology (1989) 121, 185–191
Macrophage activation enhances the human chorionic gonadotrophin-induced disruption of spermatogenesis in the rat
Authors: Kerr J. B. , Sharpe R. M.
Published: December 2008

The objective of this study was to examine the role of intertubular macrophages in modifying the response of the rat testis to stimulation by human chorionic gonadotrophin (hCG). The phagocytic activity of macrophages was stimulated by a unilateral intratesticular injection of polystyrene latex beads. Latex beads were engulfed by the resident macrophages and retained within their cytoplasm. Contralateral testes received injection of vehicle alone. A group of control rats was killed 3 days later; other groups received 100 IU hCG s.c. and the morphological and functional responses of the testes were examined 12, 24 and 48 h later. Spermatogenesis was unaffected in control rats, whereas in the testes of all hCG-treated rats leukocytes infiltrated into the intertubular tissue and the seminiferous tubules exhibited focal disruptions of spermatogenesis which were more severe in testes containing activated macrophages. Spermatogenic disruption was dependent upon the stage of the spermatogenic cycle, with the maximum tubule degeneration occurring at or near stages III and IX–XI. However these changes were not a consequence of androgen deprivation, since no consistent correlation was demonstrated between alterations in testosterone levels in testicular interstitial fluid and the accompanying damage to germ cells. It is concluded that hCG alone or in combination with activated macrophages induces an inflammatory-type response of the intertubular tissue and localized degeneration of the seminiferous epithelium. The antispermatogenic effects of hCG may have important implications for in-vivo investigations of Leydig cell function in laboratory animals and for the efficacy of hCG administration used in the clinical treatment of male hypogonadism.

Journal of Endocrinology (1989) 121, 285–292
Effect of the glycogenolytic gluconeogenetic hormones, glucagon, vasopressin and angiotensin II, on biliary excretion of iodothyronines in rats is possibly related to the inhibition of 5′-monodeiodination in the liver
Authors: Langer P. , Földes O. , Gschwendtová K.
Published: December 2008

Polyethylene tubes were inserted into the bile duct and femoral vein of rats under pentobarbital anaesthesia and bile was collected for three 2-h periods. After the first (control) period the animals were infused intravenously at a rate of 1·2 ml/h with the following compounds: (1) 0·9% (w/v) NaCl (control group), (2) glucagon (1200 ng/h), (3) vasopressin (1200 ng/h) or (4) angiotensin II (600 ng/h). The concentrations of thyroxine (T4), tri-iodothyronine (T3) and reverse tri-iodothyronine (rT3) in the bile were estimated by radioimmunoassay. No significant differences between groups were found in the biliary excretion of T4 and T3, while the excretion of rT3 after the infusion of all the hormones used was significantly (P < 0·001 at 2 to 4 h of the infusion) increased, no such increase being found in the controls. It may be concluded therefore that the administration of the above hormones resulted in some changes in iodothyronine metabolism in the liver. These may be explained by an inhibition of iodothyronine 5′-monodeiodination related to the glycogenolytic and gluconeogenetic effects of these hormones.

Journal of Endocrinology (1989) 121, 299–302
Effect of a single injection of human chorionic gonadotrophin on testosterone levels in testicular interstitial fluid, and in testicular and peripheral venous blood in adult rats
Authors: Maddocks S. , Setchell B. P.
Published: December 2008

We have used a push–pull cannula to collect interstitial fluid from the testes of anaesthetized rats at various times after a single injection of human chorionic gonadotrophin (hCG; 50 IU), and compared the levels of testosterone in this fluid with the levels in testicular and peripheral venous blood collected at the same times. Following hCG injection, significant increases in testosterone concentrations were observed in all fluids with notable peaks occurring in interstitial fluid at 2, 8 and 24 h, in testicular venous blood at 2, 8 and 30 h, and in peripheral venous blood at 2, 8, 24 and 72 h. The results demonstrate for the first time that changes in testosterone concentrations in interstitial fluid can be different from those in testicular venous blood. In addition, when testosterone levels in interstitial fluid were compared with levels in testicular venous blood at each time-point, the results suggested that the partitioning of testosterone between these two compartments can be regulated. Furthermore, the changes in both interstitial fluid and testicular venous blood levels of testosterone do not always parallel those in peripheral venous blood, suggesting that changes in testicular blood flow and peripheral clearance rates of testosterone may also be important in the control of circulating testosterone concentrations.

Journal of Endocrinology (1989) 121, 311–316
Possible role of histones in the organization of rat liver thyroid hormone receptors in chromatin
Authors: Sakurai A. , Ichikawa K. , Hashizume K. , Miyamoto T. , Yamauchi K. , Ohtsuka H. , Nishii Y. , Yamada T.
Published: December 2008

The effects of histone subfractions on rat liver thyroid hormone receptor–DNA interaction were examined using an in-vitro DNA-cellulose binding assay. H1 histones bound to DNA showed reversible and potent inhibition of receptor–DNA binding without affecting receptor–hormone binding. Poly-lysine, bovine serum albumin, ovalbumin and cytochrome c did not alter receptor–DNA binding. H1 histone subfractions (calf thymus lysine-rich histone (CTL)-1, CTL-2 and CTL-3) showed potent inhibition of receptor–DNA binding indistinguishable from each other. The quantity of H1 histone subfractions bound to DNA was the same. Although each subfraction has different functional properties, inhibition of receptor–DNA binding was a common feature of all the H1 histone subfractions, which is important for the non-random distribution of the receptor in chromatin.

Binding of the receptor to core histones was investigated; it was found to bind to core histones more potently than to other proteins (H1 histone, ovalbumin and cytochrome c). Among core histone subfractions, H4 histone bound to the receptor most potently and is the candidate to be one of the acceptor sites of the receptor in chromatin.

Journal of Endocrinology (1989) 121, 337–341
Inhibition of arginine vasopressin-stimulated Na+K+ATPase activity by difluoromethyl ornithine in the rat renal medulla
Authors: Charlton J. A. , Baylis P. H.
Published: December 2008

Arginine vasopressin (AVP) stimulates Na+K+ ATPase and ornithine decarboxylase (ODC) activity in the rat medullary thick ascending limb. The effect of difluoromethyl ornithine (DFMO), a specific inhibitor of ODC activity, on AVP-stimulated Na+K+ATPase activity was evaluated using a cytochemical bioassay. Peaks in Na+K+ATPase activity in cultured rat renal segments which occurred after tissue had been exposed to 1 fmol AVP/l were completely inhibited by the addition of 20 mmol DFMO/l to the culture medium containing AVP. The addition of 20 mmol DFMO/l to the culture medium containing AVP in the concentration range 0·001–10 fmol/l inhibited completely the stimulation of Na+K+ATPase activity by AVP. The response of Na+K+ATPase to increasing doses of ATP (10–40 g polypeptide/l) was not influenced by the addition of 20 mmol DFMO/l to the culture medium containing AVP, suggesting that the prevention of AVP-stimulated Na+K+ATPase activity by DFMO was not due to a direct effect on the enzyme.

Journal of Endocrinology (1989) 121, 435–439
Vitamin D-enhanced thyrotrophin release from rat pituitary cells: effects of Ca2+, dihydropyridines and ionomycin
Authors: d'Emden M. C. , Wark J. D.
Published: December 2008

Vitamin D may regulate pituitary function, as there are selective effects of 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) on gene expression in clonal pituitary tumour cells, and on TRH-induced TSH release in normal rat pituitary cells in vitro. The role of Ca2+ in 1,25-(OH)2D3-enhanced TSH release from primary rat pituitary cell cultures was investigated. Pretreatment with 10 nmol 1,25-(OH)2D3/l for 24 h augmented KCl (3–60 mmol/l)-induced TSH release over 1 h at all KCl concentrations greater than 7·5 mmol/l (P< 0·001), with a 76% enhancement of TSH release induced by 30 mmol KCl/l (P<0·001). The Ca2+ channel antagonist nifedipine (10 nmol/l–10 μmol/l) caused a concentration-dependent inhibition of KCl (60 mmol/l)-induced TSH secretion. Pretreatment with 1,25-(OH)2D3 enhanced KCl-induced release at all concentrations of nifedipine (P<0·001). The Ca2+ selective divalent cation ionophore ionomycin (1 nmol/l–1 μmol/l), and the Ca2+ channel agonist BAY K 8644 (10 nmol/l–1 μmol/l) increased prolactin secretion but did not increase TSH release, and 1,25-(OH)2D3 had no effect. At an extracellular Ca2+ concentration of less than 500 nmol/l, TRH-induced TSH release was observed only after treatment with 1,25-(OH)2D3 (P<0·01). As the extracellular Ca2+ concentration was increased, greater increments of TRH-induced TSH release were observed following pretreatment with 1,25-(OH)2D3 (P<0·01). However, the effect of 1,25-(OH)2D3 in the thyrotroph was independent of the pretreatment extracellular Ca2+ concentration. We have shown that 1,25-(OH)2D3 acts selectively on the thyrotroph to enhance in-vitro responsiveness to TRH and KCl. These data suggest that the action of 1,25-(OH)2D3 in the thyrotroph is to enhance intracellular signal transduction. They further support a permissive or regulatory role of vitamin D in the normal pituitary gland.

Journal of Endocrinology (1989) 121, 441–450
A peptidase activity from primate liver that inactivates oxytocin in vitro: purification and partial characterization
Authors: Sri Krishna G. , Kanagasabapathy A. S.
Published: December 2008

An aminopeptidase from monkey (Macaca radiata) liver, inactivating oxytocin in vitro and located predominantly in the lysosomal and microsomal fractions, was purified by chromatography on Bio-Gel HTP, DEAE-Sephacel and nickel ion chelate gel and gel filtration on Sephacryl S300. Absence of binding to nickel ion chelate gel indicated the absence of exposed histidine and thiol residues on the enzyme. The enzyme appeared to be a high molecular weight (Mr 106 000) monomeric protein. It was sensitive to inhibition by metal chelators and was found to be a zinc metalloprotein by atomic absorption spectrophotometry. Divalent metal ions Ni2+ and Co2+, and sulphydryl activators glutathione and 2-mercaptoethanol had activating effects, while 4-chloro mercuribenzoate, amino acids with large hydrophobic side chains and l-cystine, β-lactam antibiotic cloxacillin and peptidase inhibitor amastatin had inhibitory effects on the enzyme activity. The enzyme was most active against S-benzyl l-cysteine 4-nitroanilide substrate. The properties of the enzyme were distinct from those of the well-characterized alanine and leucine aminopeptidases (EC 3.4.11.2 and EC 3.4.11.1 respectively) of liver, and of primate placental cystine aminopeptidases (EC 3.4.11.3).

Journal of Endocrinology (1989) 121, 537–544
EFFECT OF THE TFM MUTATION ON HANDEDNESS IN MICE
Authors: Nosten M. , Roubertoux P. , Degrelle H. , Leboyer M.
Published: December 2008

The hyposthesis has been proposed that testosterone is involved in the determination of handedness in man: a high sensitivity to testosterone being associated with left handedness. Handedness in mice is tested according to Collins' paradigm: most mice present either a right or a left paw preference but others are ambilateral. The hypothesis that there is an association between a low neonatal imprinting by testosterone and a strong handedness (right or left) is tested here using Tfm male mice which are testosterone insensitive. Our results confirmed the hypothesis, since Tfm males were as well lateralized as their female siblings and significantly more strongly lateralized than their male siblings not carrying the mutation.
Effects of recombinant human interleukin-1α, -1β, 2 and 6 on ACTH synthesis and release in the mouse pituitary tumour cell line AtT-20
Authors: Fukata J. , Usui T. , Naitoh Y. , Nakai Y. , Imura H.
Published: December 2008

The effects of recombinant human interleukin (rhIL)-1α, -1β, 2 and 6 on the release of ACTH from the ACTH-producing tumour cell line AtT-20 of the mouse were studied during relatively long periods of incubation. Levels of ACTH in the media, measured by radioimmunoassay, were increased by the addition of rhIL-1α or -1β after latent periods of more than 4 h. RhIL-1α and -1β were almost equally potent in this experiment and the minimum, half-maximum and maximum effective concentrations of both rhIL-1α and -1β were about 0·1 pmol/l, 1–3 pmol/l and 10–100 pmol/l respectively. During incubation with rhIL-1β, immunoreactive ACTH levels and mRNA levels of the ACTH precursor pro-opiomelanocortin in cells also increased without apparent changes in the growth rate of the cells. Although the AtT-20 cells used in this study were quite insensitive to human/rat corticotrophin-releasing hormone (CRH), the cells showed a significant response to CRH after incubation with rhIL-1β. RhIL-6 showed similar effects to those of rhIL-1β on ACTH synthesis and release; increasing ACTH in cells and media after a certain latent period. On the other hand, rhIL-2 did not change ACTH levels in the AtT-20 cells in this study. These observations indicate that rhIL-1α, -1β and rhIL-6 have direct effects on ACTH-producing cells to stimulate the release and synthesis of ACTH after a latent period.

Journal of Endocrinology (1989) 122, 33–39
Growth hormone (GH) release after administration of GH-releasing hormone in relation to endogenous 24-h GH secretion in short children
Authors: Gelander L. , Albertsson-Wikland K.
Published: December 2008

Endogenous GH secretion was measured every 20 min for 24 h in 36 short children. This was immediately followed by an i.v. injection of GH-releasing hormone (GHRH)(1–29)-NH2 (1 μg/kg), and GH was estimated every 15 min for the following 2 h. The aim was to determine whether endogenous pulsatile GH secretion had any relation to, or influence on, the GH release induced by GHRH. A high variability was found both in the 24-h GH secretion expressed as area under the curve above the baseline (0–1588 mU/l × 24 h) and the maximal GH response to GHRH (5–296 mU/l), as well as after an arginine–insulin tolerance test (4–59 mU/l). We found a positive correlation (correlation coefficient of Spearman (rs) = 0·49; P < 0·01) between the GH response to GHRH and the spontaneous GH secretion over a 24-h period, in spite of a negative correlation (rs = −0·80; P < 0·01) with the GH secretion during the preceeding 3 h. We conclude that the GH response to a GHRH test correlates with endogenous GH secretion in short children, and may be helpful in estimating the ability to release GH. It is important, however, to be aware of the influence of the spontaneous GH secretion during the 3 h immediately preceeding administration of GHRH.

Journal of Endocrinology (1989) 122, 61–68
Insulin-like growth factor (IGF)-binding protein release by human fetal fibroblasts: dependency on cell density and IGF peptides
Authors: Hill D. J. , Camacho-Hubner C. , Rashid P. , Strain A. J. , Clemmons D. R.
Published: December 2008

Insulin-like growth factors (IGFs) are bound to specific binding proteins in extracellular fluids in vivo and when released by cells in vitro. One class of binding protein (IGF-BP), a peptide of 26 kDa purified from amniotic fluid, has been shown to modulate IGF bioactivity on isolated human fibroblasts. We have determined the factors that control release of IGF-BP from monolayers of human fetal fibroblasts using a radioimmunoassay, and have compared this with the effects of these factors on the release of IGF-I and -II. Separation of cell-conditioned cultured medium on SDS-PAGE, and subsequent immunoblotting with antibody against IGF-BP showed that fibroblasts released a single species of immunoreactive protein of an estimated molecular weight of 30 kDa. This was not the predominant binding protein released by cells since major bands of approximately 42 kDa and 39 kDa were visualized following separation by SDS-PAGE and ligand blotting with 125I-labelled IGF-I. The 30 kDa IGF-BP was released in parallel with radioimmunoassayable IGF-I and -II over 48 h. However, a significant inverse correlation was found between the release of IGF-BP, IGF-I, IGF-II and cell density. The exposure of fibroblasts to 1·3 nmol/l or greater of IGF-I or -II caused a significant release of IGF-BP. Maximum release was seen in sparse cultures with little or no release from confluent cultures. IGF-I and -II were approximately equipotent with a fourfold increase in IGF-BP release at 19·7 nmol/l. Insulin caused a release of IGF-BP and IGF-I and -II from fibroblasts at supraphysiological concentrations (16·7 nmol/l) which again was maximal on sparse cell cultures. Increases in IGF-BP, IGF-I and -II release were also found in the presence of human placental lactogen (23·3 nmol/l), but human GH, epidermal growth factor, fibroblast growth factor and platelet-derived growth factor were without effect. The results show that human fetal fibroblasts released an IGF-BP immunologically similar to that seen in amniotic fluid together with IGF-I and -II, that IGF-BP release was enhanced by exogenous addition of IGF peptides, and that the release of all three peptides was a property of sparsely plated, rapidly growing cells. These findings strengthen the hypothesis that the cellular expression of IGF-binding proteins may represent an important level of control in IGF physiology.

Journal of Endocrinology (1989) 122, 87–98
Peripheral metabolism of [35S]parathyroid hormone in vivo: influence of alterations in calcium availability and parathyroid status
Authors: Bringhurst F. R. , Stern A. M. , Yotts M. , Mizrahi N. , Segre G. V. , Potts J. T.
Published: December 2008

Parathyroid hormone (PTH) is rapidly metabolized, mainly by liver and kidney, to smaller fragments that are believed to be biologically inactive. The significance of this peripheral metabolism in the overall actions of PTH is unclear. Generation of circulating biologically active amino-terminal PTH fragments during metabolism in vivo has been suggested by certain observations in vitro, and what are believed to be amino-terminal fragments may be detectable in blood under pathological circumstances in vivo (such as renal failure and coexistent hyperparathyroidism) when highly sensitive assays are employed. We recently reported, however, that administration to normal rats of [35S]bovine PTH ([35S]bPTH) directly labelled at amino-terminal methionines, followed by high-resolution chromatographic analysis of extracted [35S]peptides, does not result in appearance of radioactive amino-terminal fragments in blood, even when the tracer is continuously infused to near-physiological plasma concentrations. We have now employed these techniques to address a second question regarding hormonal metabolism: is hormonal metabolism modified during metabolic perturbations such as changing calcium availability or altered levels of calciotrophic hormones? Metabolism of [35S-Met]bPTH (900 Ci/mmol), either alone or together with [3H-Pro]bPTH, however, did riot lead to alterations in the rate of hormonal clearance nor to detectable circulating amino-terminal fragments, either in calcium-deprived or thyroparathyroidectomized rats or when animals were first rendered intoxicated with vitamin D or maintained on a high calcium intake. Likewise, tissue localization and specific cleavage patterns of intact hormone in liver or kidney were all unaltered by these various manoeuvres. We conclude that regulation of peripheral metabolism of PTH does not exert important physiological control over the concentration of circulating biologically active PTH and that active amino-terminal fragments of the hormone are not released into blood even under circumstances of calcium deprivation or hypocalcaemia.

Journal of Endocrinology (1989) 122, 237–245
Effect of sodium intake on phosphoinositides and inositol trisphosphate response to angiotensin II, K+ and ACTH in rat glomerulosa cells
Authors: Underwood R. H. , Menachery A. I. , Williams G. H.
Published: December 2008

To assess the impact of sodium intake on the adrenal phosphoinositide system, rats were maintained on a low or normal salt diet for 5 days, and glomerulosa cell preparations (2×105 cells) were stimulated by angiotensin II (AII; 10 nmol/l), potassium (K+; 8·7 mmol/l) or ACTH (0·1 nmol/l) for 0, 2, 4, 6, 12, 15 and 60 s. Levels of phosphatidylinositol (PtdIns), phosphatidylinositol 4-phosphate (PtdIns 4-P), phosphatidylinositol 4,5-bisphosphate (PtdIns 4,5-P2) and inositol 1,4,5-trisphosphate (Ins 1,4,5-P3)+inositol3) + inositol 1,3,4-trisphosphate (Ins 1,3,4-P3) were assayed by a microspectrophotometric procedure. Non-stimulated levels of PtdIns, PtdIns 4-P, PtdIns 4,5-P2 and Ins 1,4,5-P3 (+Ins 1,3,4-P3) (means ± s.e.m.; n = 36) in cells from rats on the low Na+ intake were 580 ± 6·5, 187 ± 2·6, 82 ± 3 and 95 ± 1·2 pmoler incubate respectively, indistinguishable from those observed in rats on a normal Na+ intake, except for the significantly (P<0·025) greater Ptdlns 4,5-P2 level. In response to AII stimulation, all four compounds showed an earlier and greater peak response when cells were obtained from animals on a low rather than a high sodium intake. All values had returned to control levels by 12–15 s, regardless of the level of sodium intake. In contrast, with K+ stimulation there were no differences in the peak response of cells from rats on the two dietary intakes, but there was a shift of the peak to a longer time-interval (6 versus 8 s) in animals maintained on a low sodium intake. In addition, PtdIns 4,5-P2 did not return to control levels in cells obtained from animals on a low sodium intake. However, the most striking differences were observed in response to ACTH. In animals maintained on a normal sodium intake, there were no changes in any of the four compounds. In contrast, there was a sharp increase in all four substances in response to ACTH when the cells were obtained from animals on a low sodium intake, with peak levels occurring at 8 s, similar to that observed with potassium. Thus, changes in the response pattern of the phosphoinositides may mediate the altered adrenal responsiveness to AII and ACTH with sodium restriction.

Journal of Endocrinology (1989) 122, 371–377
Reduced responses of prolactin and catecholamine to stress in the lactating rat
Authors: Higuchi T. , Negoro H. , Arita J.
Published: December 2008

Prolactin, GH, TSH, adrenaline and noradrenaline responses to the stress of immobilization were compared between lactating and non-lactating dioestrous rats. The concentrations of GH in plasma were reduced to a similar degree by the immobilization of lactating and non-lactating rats, and TSH levels were unchanged in both groups. The increases in plasma concentrations of adrenaline and noradrenaline induced by stress were significantly smaller in lactating than in non-lactating rats. Immobilization caused a marked increase in prolactin levels in the plasma of non-lactating rats but no increase in lactating rats. These changes may help to save energy and maintain milk production during the period of lactation.

Journal of Endocrinology (1989) 122, 495–498
Seasonal and steroid-dependent effects on the modulation of LH secretion in the ewe by intracerebroventricularly administered β-endorphin or naloxone
Authors: Horton R. J. E. , Francis H. , Clarke I. J.
Published: December 2008

The natural opioid ligand, β-endorphin, and the opioid antagonist, naloxone, were administered intracerebroventricularly (i.c.v.) to evaluate effects on LH secretion in ovariectomized ewes and in ovariectomized ewes treated with oestradiol-17β plus progesterone either during the breeding season or the anoestrous season. Ovary-intact ewes were also studied during the follicular phase of the oestrous cycle. Jugular blood samples were taken at 10-min intervals for 8 h and either saline (20–50 μl), 100 μg naloxone or 10 μg β-endorphin were injected i.c.v. after 4 h. In addition, luteal phase ewes were injected i.c.v. with 25 μg β-endorphin(1–27), a purported endogenous opioid antagonist.

In ovariectomized ewes, irrespective of season, saline and naloxone did not affect LH secretion, but β-endorphin decreased the plasma LH concentrations, by reducing LH pulse frequency. The effect of β-endorphin was blocked by administering naloxone 30 min beforehand. Treating ovariectomized ewes with oestradiol-17β plus progesterone during the breeding season reduced plasma LH concentrations from 6–8 μg/l to less than 1 μg/l. In these ewes, saline did not alter LH secretion, but naloxone increased LH pulse frequency and the plasma concentrations of LH within 15–20 min. During anoestrus, the combination of oestradiol-17β plus progesterone to ovariectomized ewes reduced the plasma LH concentrations from 3–5 μg/l to undetectable levels, and neither saline nor naloxone affected LH secretion.

During the follicular phase of the oestrous cycle, naloxone enhanced LH pulse frequency, which resulted in increased plasma LH concentrations; saline had no effect. In these sheep, β-endorphin decreased LH pulse frequency and the mean concentrations of LH, and this effect was prevented by the previous administration of naloxone. The i.c.v. administration of β-endorphin(1–27) to luteal phase ewes did not affect LH secretion.

These data demonstrate the ability of a naturally occurring opioid peptide to inhibit LH secretion in ewes during the breeding and non-breeding seasons, irrespective of the gonadal steroid background. In contrast, whilst the gonadal steroids suppress LH secretion in ovariectomized ewes during both seasons, they only appear to activate endogenous opioid peptide (EOP)-mediated inhibition of LH secretion during the breeding season. Furthermore, these data support the notion that LH secretion in ovariectomized ewes is not normally under the control of EOP, so that naloxone has no effect.

Journal of Endocrinology (1989) 122, 509–517
Growth hormone and growth in diabetic rats: effects of insulin and insulin-like growth factor-I infusions
Authors: Carlsson L. M. S. , Clark R. G. , Skottner A. , Robinson I. C. A. F.
Published: December 2008

The effects of streptozotocin-induced diabetes on weight gain, bone growth and GH secretion have been studied in conscious chronically cannulated male rats. In addition to the classic diabetic symptoms (hyperphagia, polydipsia, polyuria, glycosuria and hyperglycaemia), the slow body weight gain (0·95 ± 0·5 compared with 2·63 ± 0·5 g/day in non-diabetic controls) was associated with a reduction in bone growth (from 162 ± 9 to 48 ± 4 μm/day) and a reduced pituitary GH content (from 1·5 ± 0·2 to 0·6 ± 0·06 mg/gland). Serial blood sampling during the day or overnight showed that the normal male episodic GH secretory pattern was obliterated in the diabetic animals. The constant osmotic stimulation of hyperglycaemia and high fluid turnover was reflected in a significant reduction in pituitary oxytocin and arginine vasopressin (AVP) stores. Intravenous insulin infusions (67–1340 pmol/h for 4 or 7 days) caused a large initial weight gain (> 20 g in 2 days) followed by a slower increase, and stimulated tibial bone growth (to 100 ± 16 and 126 ± 8 μm/day after 4 or 7 days respectively). Insulin infusion for 7 days also increased pituitary GH content (to 1 ± 0·15 mg/gland), and the normal episodic GH secretory pattern returned. Intravenous infusions of insulin which reduced, but did not completely normalize, blood glucose levels, allowed the resumption of growth and pulsatile GH secretion. Continuous infusion of recombinant human insulin-like growth factor-I (hIGF-I) at 1110 pmol/h for 54 h also caused a large initial rise in body weight in diabetic rats (17·1 ± 1·6 compared with 7·5 ± 2·8 g in saline-infused controls) due primarily to increased fluid retention. This effect of hIGF-I occurred without any significant changes in pituitary GH, AVP, oxytocin, blood glucose or bone growth over this short-term infusion, nor was there any obvious effect on spontaneous GH secretion, monitored over the entire infusion period. We conclude that the diabetic rat is not a good model to study growth stimulation by short-term insulin or IGF-I treatments because the insulin-like effects of these peptides obscure their specific growth-promoting activities in this model.

Journal of Endocrinology (1989) 122, 661–670
Proliferation and differentiation of possible Leydig cell precursors after destruction of the existing Leydig cells with ethane dimethyl sulphonate: the role of LH/human chorionic gonadotrophin
Authors: Teerds K. J. , de Rooij D. G. , Rommerts F. F. G. , van den Hurk R. , Wensing C. J. G.
Published: December 2008

The influence of LH levels on the proliferation and differentiation of possible Leydig cell precursors was investigated in adult rats, after the destruction of the existing Leydig cells with the cytotoxic drug ethane dimethyl sulphonate (EDS). In rats bearing a testosterone implant which prevented the rise in plasma LH levels and kept them within the normal range after the destruction of the Leydig cells, the proliferative activity of possible Leydig cell precursors still increased seven- to eightfold 2 days after EDS administration. Apparently, in this situation, locally produced factors, and not LH, may play a role in the stimulation of proliferation. The proliferative activity of the possible precursor cells could be further stimulated by treating rats with daily injections of human chorionic gonadotrophin (hCG) following EDS administration. It was concluded that the proliferative activity of possible Leydig cell precursors is probably regulated by both paracrine and endocrine factors.

Almost no Leydig cells were formed in the rats bearing a testosterone implant during the first 4 weeks after EDS administration. When these rats were treated with hCG, starting 28 days after administration of EDS, a substantial number of Leydig cells was found after 2 days, and these cells also showed 3β-hydroxysteroid dehydrogenase (3β-HSD) and α-naphtyl esterase (α-NE) activity. When hCG treatment was started at 14 or 21 days after EDS administration, some cells with the nuclear characteristics of Leydig cells were present after 2 days, but no 3β-HSD or α-NE activity could be detected. Finally, when hCG treatment was started directly after EDS administration, a considerable number of Leydig cells was found 14 days after EDS, and some of these cells already showed 3β-HSD and α-NE activity. It is concluded that precursor cells are able to develop into advanced precursor cells at normal LH levels, and that the rate of development of new Leydig cells strongly depends upon LH/hCG levels.

Journal of Endocrinology (1989) 122, 689–696
Effect of noradrenaline and γ-aminobutyric acid on the secretion of corticotrophin-releasing factor-41 and arginine vasopressin from the rat hypothalamus in vitro
Authors: Hillhouse E. W. , Milton N. G. N.
Published: December 2008

Much controversy exists concerning the role of catecholamines in the control of ACTH secretion. In this study, noradrenaline (0·1 nmol–0·1 μmol/l) stimulated the release of both immunoreactive corticotrophin-releasing factor-41 (ir-CRF-41) and ir-arginine vasopressin (ir-AVP) from the rat hypothalamus in vitro. The stimulatory effect of noradrenaline on CRF-41 release was blocked by propranolol, whilst that on AVP release was blocked by phentolamine. γ-Aminobutyric acid (GABA; 10 nmol/l) inhibited the acetylcholine-induced release of both AVP and CRF-41 in vitro, and the effect was blocked by picrotoxin (0·1 μmol/l). Neither substance had any effect on the basal secretion of either neuropeptide. The results indicate that noradrenaline stimulates and GABA inhibits the release of both peptides from the rat hypothalamus in vitro.

Journal of Endocrinology (1989) 122, 719–723
Plasma growth hormone pattern and androgens influence the levels of corticosteroid-binding globulin in rat serum
Authors: Jansson J.-O. , Oscarsson J. , Mode A. , Ritzén E. M.
Published: December 2008

The serum concentration of corticosteroid-binding globulin (CBG) is higher in female rats than in males. Combined hypophysectomy and gonadectomy of female rats reduced the serum concentration of CBG as measured by steady-state polyacrylamide gel electrophoresis, whereas hypophysectomy of male rats increased serum CBG. These effects were seen despite replacement therapy with thyroxine and glucocorticoids. Moreover, neither androgen nor oestrogen treatment affected the serum concentrations of CBG in hypophysectomized rats. Continuous infusions of human or bovine GH (1·4 U/kg per day), by means of osmotic minipumps for 1 week, increased serum concentrations of CBG in both hypophysectomized male and female rats. In contrast, intermittent GH replacement therapy by s.c. injections at 12-h intervals either had no effect or suppressed serum CBG levels. In male rats, neonatal (days 1–2) gonadectomy increased CBG levels more than did prepubertal (day 25) gonadectomy, and testosterone replacement therapy reversed these effects.

It is concluded that GH increases the serum CBG levels of hypophysectomized rats when it is given in a continuous manner, but not when given intermittently. The sex difference in serum CBG levels of normal rats may, therefore, be attributed to the more continuous secretory pattern of GH previously observed in female rats.

Journal of Endocrinology (1989) 122, 725–732
Somatotrophic receptors in hepatic tissue of the developing male pig
Authors: Breier B. H. , Gluckman P. D. , Blair H. T. , McCutcheon S. N.
Published: December 2008

The development of hepatic somatotrophic receptors and plasma concentrations of insulin-like growth factor-I (IGF-I) were investigated at five different ages (2, 20, 35, 105 and 165 days) in four male pigs per group. The specific binding of 125I-labelled porcine GH (pGH) to hepatic somatotrophic membranes was very low at 2 days of age (0·53±0·12%), and increased progressively (P <0·01) with advancing age to 3·60 ± 0·95% at 165 days of age. Specific binding of 125I-labelled bovine GH (bGH) to the same membrane preparations was markedly higher than binding of 125I-labelled pGH; it also showed a distinct developmental increase (P <0·01) with age from 4·4 ± 0·55% at 2 days of age to 24·0±1·90% at 165 days of age. Plasma concentrations of IGF-I increased significantly (P <0·01) from 79 ± 14·0 μg/l at 2 days of age to 610 ± 64·0 μg/l at 165 days of age.

Non-linear regression analysis of the competitive binding data using bGH as labelled and unlabelled ligands showed linear Scatchard plots in the three youngest age groups, with an association constant (Ka) of approximately 3·5 litres/nmol. Curvilinear Scatchard plots were observed in the two oldest age groups. The Ka for the higher affinity binding site (approximately 5·0 litres/nmol) was very similar to that for the sole site observed in the younger animals. The Ka of the lower affinity binding site was approximately 0·35 litres/nmol. There was a significant (P <0·01) developmental increase in the capacity of the higher affinity binding site from 12 ± 4·6 pmol/100 mg liver at 2 days of age to 91 ± 23·0 pmol/100 mg liver at 165 days of age.

These studies demonstrate heterogeneity of somatotrophic hepatic membranes in the pig and show that low concentrations of a high affinity binding site are already present in newborn pigs. A considerable developmental increase was observed in the capacity of this binding site which correlated significantly (r = 0·82, P <0·01, n = 20) with plasma concentrations of IGF-I. The role of the lower affinity binding site which was observed in addition to the high affinity binding site in older pigs is less clear. The data from the present study support the hypothesis that the postnatal rise in plasma concentrations of IGF-I is associated with the developmental increase of the capacity of the high affinity somatotrophic receptors.

Journal of Endocrinology (1989) 123, 25–31
Differential secretory rhythm of growth hormone controls the number of hepatic epidermal growth factor receptors in the rat
Authors: Kashimata M. , Hiramatsu M. , Minami N.
Published: December 2008

The effect of human GH (hGH) on hepatic epidermal growth factor (EGF) receptors in the rat was investigated. Continuous administration of hGH through an osmotic minipump, mimicking the female pattern of GH secretion, to normal male rats reduced the binding of 125I-labelled EGF to hepatic membranes to the normal female levels. The same treatment of hGH applied to hypophysectomized males had no apparent effect on EGF binding. Intermittent s.c. administration of hGH twice a day (every 12 h), mimicking the male pattern of GH secretion, to hypophysectomized male and/or normal female rats, caused a significant increase in EGF binding to the levels of normal male rats. Scatchard analysis of the binding data clearly showed that the change in EGF binding was due to a change in the number of EGF receptors. The results on the affinity labelling and phosphorylation of EGF receptors were in good agreement with those showing differences in the number of EGF receptors among the experimental groups. These results indicate that the number of hepatic EGF receptors in the rat is regulated by the differential secretory rhythm of pituitary GH between the sexes.

Journal of Endocrinology (1989) 123, 75–81
Inhibitory effects of amiloride and its analogues on prostaglandin E2-stimulated fluid transport by cultured porcine thyroid cells: evidence for apical membrane Na+ channels
Authors: Matainaho T. , Cragoe E. J. , Manley S. W. , Huxham G. J. , Pearson J. V. , Bourke J. R.
Published: December 2008

Confluent monolayers of cultured porcine thyroid cells transport fluid from the apical to the basal surface, forming circumscribed zones of detachment (domes) from the culture dish substrate. Fluid transport, as measured by increase in dome height, was stimulated by prostaglandin E2 (PGE2; 1 μmol/l) and inhibited by amiloride (0·1–100 μmol/l). Values of the inhibition constant (Ki) with 95% confidence limits for each of a series of amiloride analogues were: 3′,4′-dichlorobenzamil (DCB), 0·090 (0·045–0·18) μmol/l; 2′,4′;-dimethylbenzamil (DMB), 0·14 (0·074–0·27) μmol/l; amiloride, 0·72 (0·33–1·8) μmol/l; 5-(N,N-hexamethylene)amiloride (HMA), 17 (5·9–43) μmol/l; 5-(N-ethyl-N-isopropyl)amiloride (EIPA), 33 (15–71) μmol/l; and 2-guanidinobenzimidazole, 243 (110–570) μmol/l. Triaminopyrimidine was ineffective at concentrations up to 1 mmol/l. Since DCB and DMB are known to have a higher affinity for Na+ channels, while HMA and EIPA show higher affinity for Na+/H+ antiports, it was concluded that PGE2-stimulated fluid transport involved an apical membrane Na+ channel.

Journal of Endocrinology (1989) 123, 93–97
The fate and uptake of murine epidermal growth factor in the sheep
Authors: Tyson L. M. , Browne C. A. , Jenkin G. , Thorburn G. D.
Published: December 2008

125I-Labelled murine epidermal growth factor (EGF) was injected or infused into conscious ewes through the jugular vein. Its disappearance from the circulation and the pattern of its distribution in other body tissues and compartments were observed. Single bolus injections of 125I-labelled EGF resulted in a transient peak of radioactive EGF in the circulation which occurred within 1 min of the injection. This was followed by a very rapid fall in radioactivity in the plasma (t½ ∼ 1 min) and the gradual appearance of 125I-labelled EGF in the urine. Immunoprecipitable 125I-labelled EGF could be detected in urine within 5 min of the start of the experiment. 125I-Labelled EGF accumulated in the urine for several hours following the injection, although with increasing time a substantial amount of non-immunoprecipitable iodide was also found. The rate of disappearance of the 125I-labelled EGF from the plasma of the ewe was found to be faster than the rate of disappearance of free [125I]iodide that had been injected into the ewe.

125I-Labelled EGF was also administered by a continuous infusion following an initial bolus injection. This again resulted in a rapid initial fall in radio-activity in blood, followed by a slow rise throughout the period of the infusion. When the infusion was stopped, there was a 15-min period of rapid readjustment, after which the radioactivity in the blood fell at a much slower rate (t½ ∼70 min) than was seen initially. Again, intact 125I-labelled EGF was transferred to urine throughout the experiment.

At autopsy, 125I-labelled EGF was increased in bile, liver, thyroid and kidney. Although most of the 125I found in the thyroid was free iodide, some EGF-like material was also present. There was also EGF-like material found in both the kidney cortex and the kidney medulla. These results indicate that complex multi-compartment pathways for the uptake, distribution and clearance of 125I-labelled EGF exist in the sheep.

Journal of Endocrinology (1989) 123, 121–130
The angiotensin II receptor of rabbit liver: characterization in isolated hepatocytes and effect of GTP
Authors: Vandekerckhove A. , Keppens S. , De Wulf H.
Published: December 2008

A homologous population of specific angiotensin II receptors is present on the cell surface of isolated rabbit hepatocytes. The binding characteristics of [3H]angiotensin II to the cells were: association rate constant (K+1) 0·08 l/nmol per min and dissociation rate constant (K−1) 1·9/min, yielding a dissociation constant (Kd) of 24 nmol/l. A very similar Kd (32 nmol/l) has been derived from saturation binding data which indicate a maximal binding capacity of about 200 000 sites/cell.

Analysis of the association binding data to purified liver plasma membranes indicated a Kd of 6 nmol/l in the absence and 30 nmol/l in the presence of GTP. Dissociation was clearly dependent upon the presence of the nucleotide, which shifted the K−1 from 0·12/min to 0·42/min.

The studied binding sites are very likely to be involved in the glycogenolytic action of angiotensin II, since a highly significant correlation was established between the biological activity (activation of glycogen phosphorylase) and the binding affinity of a series of agonistic analogues.

The reported characteristics of the rabbit hepatic angiotensin II receptors show much similarity with those of rat liver.

Journal of Endocrinology (1989) 123, 131–136
Kinetic analysis of the uptake of thyroid hormones by the human erythrocyte
Authors: Ball S. G. , Ramsden D. B.
Published: December 2008

The uptake of 3,5,3′-tri-iodothyronine (T3) by human erythrocytes was shown to occur by two saturable processes. System I (Michaelis constant (Km) 109 ± 40 (mean ± s.d.) nmol/l; maximal velocity (Vmax) = 43 ± 16 pmol/min per 109 cells) was observed over the range 1–50 nmol T3/l whilst system II (Km = 8·8 ± 8·0 pmol/l; Vmax = 29 ± 6 fmol/min per 109 cells) was observed over the range 1–500 pmol T3/l. At low concentrations (1–500 pmol/l) uptake processes equivalent to system II could be demonstrated for thyroxine and 3,3′,5′-tri-iodothyronine also. System II had the properties of a carrier-mediated process.

Journal of Endocrinology (1989) 123, 143–148