bluuebunny/crossref_metadata_2025 huggingface dataset:
The route of secretion of inhibin from the rat testis
Authors: Maddocks S. , Sharpe R.M.
Published: December 2008

Levels of immunoactive and bioactive inhibin were measured in venous blood collected at a point just before (testicular venous) and after (spermatic venous) its passage through the mediastinal venous plexus over the anterior pole of the rete testis, and compared with levels in peripheral venous blood and testicular interstitial fluid (IF). In 15 control rats, levels of inhibin were highest in IF (8900 ± 432 ng/l; mean ± SEM) and lowest in peripheral (290 ± 32 ng/l) and testicular (288 ± 34 ng/l) venous blood, whilst levels in spermatic venous blood (633 ± 99 ng/l) were always higher (P<0.002) than the levels in testicular venous blood. The latter difference was either reduced or abolished after disruption of spermatogenesis by local heating of the testes 8, 14, or 21 days previously, and by ligation of the efferent ducts for 6 h or more, but was not affected by acute removal of the epididymis. It is concluded that inhibin secreted into seminiferous tubule fluid may be reabsorbed from the rete testis and this may be the major route by which it reaches the peripheral bloodstream in rats with normal spermatogenesis.
GROWTH HORMONE INCREASES PLASMA LEVELS OF INSULIN-LIKE GROWTH FACTOR (IGF-I) IN A TELEOST, THE GILTHEAD SEABREAM (SPARUS AURATA)
Authors: Funkenstein B. , Silbergeld A. , Cavari B. , Laron Z.
Published: December 2008

A heterologous radioimmunoassay (RIA) was applied for the determination of immunoreactive (IR)insulin-like growth factor (IGF-I) in a teleost, the gilthead seabream (Sparus aurata). Serial dilutions of the fish plasma gave a linear curve when added to constant amounts of 125I-labelled human IGF-I(53-70) and antiserum prepared against this fragment. The RIA was used to study the effect of GH on plasma levels of IR-IGF-I in S. aurata. A single injection of human recombinant GH (1 μg/g) resulted in a significant increase in IR-IGF-I at 29, 48 and 72 h, when compared with saline-injected fish. This novel observation suggests that in fish, as in mammals, circulating IGF-I levels are modulated by GH.
Differential effects of FSH and testosterone on the maintenance of spermatogenesis in the adult hypophysectomized rat
Authors: Bartlett J. M. S. , Weinbauer G. F. , Nieschlag E.
Published: December 2008

In order to clarify further the role of FSH in the maintenance of spermatogenesis, adult rats were treated with purified human FSH (2 × 5 IU/day per rat), testosterone (1·5 cm silicone elastomer implant) or a combination of both hormones for 2 weeks following hypophysectomy.

After hypophysectomy alone, no elongate spermatids were observed and the numbers of pachytene spermatocytes and round spermatids observed were reduced when compared with untreated controls. Testosterone supplementation alone qualitatively maintained the formation of elongate spermatids in most seminiferous tubules, whilst in FSH-treated rats increased numbers of round spermatids and pachytene spermatocytes were observed when compared with hypophysectomized animals. Formation of elongate spermatids, however, did not occur under FSH treatment alone. A combination of FSH and testosterone treatment maintained spermatogenesis in an almost quantitative fashion. Numbers of pachytene spermatocytes and round spermatids were maintained at about 80% of levels seen in intact control animals. Treatment with FSH or testosterone alone maintained testis weights at significantly higher levels than those seen in hypophysectomized controls (FSH, 0·79 ± 0·05 g; testosterone, 0·81 ± 0·07 g; hypophysectomized, 0·50 ± 0·04 g). Animals treated with FSH and testosterone showed testis weights 20% below control values (1·22 ± 0·05 vs 1·51 ± 0·06 g; P <0·05). No increases in intratesticular or intratubular androgen concentrations or in testosterone: dihydrotestosterone ratios were observed in any of the hormone-treated groups when compared with hypophysectomized controls. In all hypophysectomized animals testicular androgen concentrations were reduced to <5% of control values.

The results obtained in this study suggest that FSH is involved in the maintenance of spermatogenesis in the adult rat and that the effects of FSH are not mediated through changes in intratesticular androgens. Low levels of testosterone in combination with FSH can almost quantitatively maintain spermatogenesis in adult rats.

Journal of Endocrinology (1989) 121, 49–58
Vitamin A deficiency and tri-iodothyronine action at the cellular level in the rat
Authors: Higueret P. , Pailler I. , Garcin H.
Published: December 2008

Young rats were given a diet with or without vitamin A for a period of 7 weeks. In rats on the vitamin A-deficient diet, the formation of tri-iodothyronine (T3) complexes with nuclear proteins was reduced, and Scatchard analysis showed a decreased capacity of nuclear T3 receptors. In these rats, a decrease in growth and a decrease in the activity of lipogenic enzymes (glucose-6-phosphate dehydrogenase, phosphogluconate dehydrogenase and l-malate dehydrogenase) occurred, which could be related to the decreased formation of T3–nuclear protein complexes.

Journal of Endocrinology (1989) 121, 75–79
Oxytocin receptors in the ovine corpus luteum
Authors: Sernia C. , Gemmell R. T. , Thomas W. G.
Published: December 2008

There is inconclusive evidence that oxytocin acts directly on the corpus luteum and affects steroidogenesis. Since any such action would probably be mediated by oxytocin receptors, these should be present in luteal tissue. In this study, homogenates of corpora lutea from both pregnant and non-pregnant ewes were examined for oxytocin receptors by radio-receptor assay. Specific oxytocin binding was not observed in luteal tissue during the oestrous cycle. However specific binding was found in the corpora lutea of pregnant ewes; appearing at a fetal head length of approximately 0·65 cm (about 30 days of pregnancy) and persisting to a head size of 11 cm, the largest size examined in this study. The affinity (Kd) of the receptor was calculated as 2·9 ± 0·3 nmol/l (s.e.m.; n = 9), a value similar to that obtained for the uterus. The receptor number ranged from a low of 8·7± 3·2 fmol/mg protein (n = 6) at a head size of <0·65 cm, to a maximum of 40·1 ± 6·5 fmol/mg protein (n = 25) at a head size of 2·5–3·75 cm. These values were lower than our estimate of 588 ± 39 fmol/mg protein (n = 5) for the uterus. It is concluded that a direct action of oxytocin on the corpus luteum is possible but only after the first month of pregnancy and not in the corpus luteum of the oestrous cycle.

Journal of Endocrinology (1989) 121, 117–123
The interaction of arginine vasopressin and oxytocin with bovine adrenal medulla cells
Authors: Taylor A. H. , Whitley G. St J. , Nussey S. S.
Published: December 2008

Binding of [3H]arginine vasopressin (AVP) and [3H]oxytocin to primary monolayer cultures of bovine adrenal chromaffin cells was time-dependent, and the binding sites for each peptide were specific and saturable. Studies with the V1 AVP antagonist d(CH2)5Tyr(Me)2-AVP, the V2 agonist 1-deamino-8-d-AVP and the V2 antagonist d(CH2)5d-Leu2,Val4-AVP indicated that the AVP receptor was V1 in specificity. Scatchard plots showed that each ligand interacted with a single high-affinity, low-capacity binding site: oxytocin dissociation constant (Kd) 0·29 ± 0·02 nmol/l, maximum binding capacity (Bmax) 7·6 ± 0·2 fmol/106 cells (or 4500 ± 102 sites/cell) (n = 3); AVP Kd 0·09±0·02 nmol/l, Bmax 5·1±0·63 fmol/106 cells (or 3050 ± 318 sites/cell) (n = 3). Although forskolin in concentrations from 1 nmol/l to 1 mmol/l stimulated cyclic AMP (cAMP) production in isolated chromaffin cells, this did not result in detectable catecholamine release. Neither AVP nor oxytocin in concentrations between 10 pmol/l and 10 μmol/l stimulated cAMP production in these cells. Vasopressin in concentrations as low as 10 pmol/l stimulated a sixfold increase in total inositol phosphates; the dose–response curve was triphasic. Oxytocin had little effect on total inositol phosphate accumulation at low concentrations, but concentrations above micromolar stimulated total inositol phosphate production approximately fourfold. There was no measurable release of catecholamines in response to either peptide. The physiological consequences of these AVP-induced changes in inositol phosphate concentrations remain to be elucidated.

Journal of Endocrinology (1989) 121, 133–139
Steroid hormone receptors and the sexual phenotype of the Harderian gland in hamsters
Authors: Vilchis F. , Pérez-Palacios G.
Published: December 2008

To investigate the participation of intracellular steroid hormone receptors in the sexual transformation process of the Harderian gland, a series of experiments were undertaken in adult golden hamsters. The in-vitro labelling of cytosolic steroid-binding sites with appropriate radioligands revealed the presence of androgen, oestrogen and glucocorticoid but not progestin receptors in the glands from animals of both sexes. The androgen receptor of the female gland was further characterized because it was found to be the predominant intracellular steroid receptor. Studies of binding kinetics using [3H]7α,17α-dimethyl-17β-hydroxy-4-oestren-3-one (DMNT) as ligand, demonstrated a high affinity androgen-binding site with an apparent dissociation constant (Kd) of 0·7 nmol/l and maximal saturation binding capacity of 84·0 ± 3·0 (s.d.) fmol/mg protein. Specificity of the androgen receptor was assessed by displacement analysis; DMNT, 5α-dihydrotestosterone, testosterone and 3α-androstanediol were efficient competitors for the androgen-binding site, while oestradiol-17β, progesterone and dexamethasone exhibited very little, if any, competitive potency. The sedimentation coefficient of the androgen receptor in sucrose density gradients was 8–9 S. These data indicate that the physicochemical characteristics of the androgen receptor from the female gland are similar to those previously described in the male gland. The striking observation of a complete lack of oestrogen-inducible and oestrogen-insensitive progestin receptors in glands cytosol, even after stimulation with cholera toxin, adds further support to the concept that the androgen receptor is the key molecule mediating the hormone-induced sexual transformation of the Harderian gland in this species.

Journal of Endocrinology (1989) 121, 149–156
International Standards for human Prolactin: calibration by international collaborative study
Authors: Schulster D. , Gaines Das R. E. , Jeffcoate S. L.
Published: December 2008

Three ampouled preparations of purified human prolactin were assessed by 20 laboratories in eight countries for their suitability to serve as International Standards for the estimation of human prolactin in serum. Bioassays (pigeon crop sac assays and NB2 cell assays) were carried out in two laboratories, radioreceptor assays by one laboratory and radioimmunoassays by 17 laboratories.

By physicochemical analysis the preparations appeared similar. Each preparation contained small amounts of contaminants and/or prolactin variants. No major differences among the three preparations were detected by immunoassay although, in one radioreceptor assay system, one of the preparations was found to differ from the other two.

On the basis of all the available information, the Expert Committee on Biological Standardization of the World Health Organization (ECBS) in 1986 established the preparation in ampoules coded 83/562 as the Second International Standard for Prolactin and in October 1988 established the preparation in ampoules coded 84/500 as the Third International Standard for Prolactin. A value of 0·053 IU (53 mIU) prolactin activity/ampoule was assigned to both the Second and Third IS on the basis that this unitage would, insofar as possible, maintain continuity of the IU defined by the First International Reference Preparation of Prolactin, human, for Immunoassay (coded 75/504).

Journal of Endocrinology (1989) 121, 157–166
The measurement and effect of growth hormone in the presence of growth hormone-binding antibodies
Authors: Pringle P. J. , Hindmarsh P. C. , Di Silvio L. , Teale J. D. , Kurtz A. B. , Brook C. G. D.
Published: December 2008

We have developed methods for measuring the concentrations of free GH in plasma using a polyethylene glycol (PEG) separation procedure to remove antibody-bound GH within 1 h of collection. Total GH concentrations were obtained by acidification of the GH–antibody complex to release the GH followed by PEG precipitation of the antibody. The plasma GH assay had a within-assay coefficient of variation (C.V.) of 6·8% at 4·6 mU/l and a between-assay C.V. of 9·2% at 4·0 mU/l. The PEG-modified assay had a within-assay C.V. of 4·3% at 6·3 mU/l and a between-assay C.V. of 10·9% at 5·3 mU/l. Both assays had a sensitivity of 1·3 mU/l. There was good correlation between plasma and free GH concentrations in 24-h profiles in two tall children (r = 0·98; P < 0·001) and between total and free GH in the same profiles (r = 0·97; P < 0·001).

GH antibodies were measured using a highly sensitive radioimmunoassay. In children who did not develop GH antibodies there was no difference between total, plasma and free GH concentrations. In contrast, in those who developed GH antibodies both total and plasma GH concentrations were markedly increased compared with free GH concentrations. The presence of GH antibodies did not affect the growth, plasma insulin-like growth factor-I concentrations or fasting serum insulin concentration responses to 1 year of therapy with biosynthetic human GH.

Journal of Endocrinology (1989) 121, 193–199
Somatotrophin increases thyroxine-5′-monodeiodinase activity in lactating mammary tissue of the cow
Authors: Capuco A. V. , Keys J. E. , Smith J. J.
Published: December 2008

The effect of administration of bovine somatotrophin (bST) on peripheral conversion of thyroxine (T4) to tri-iodothyronine (T3) was studied in non-pregnant lactating Holstein cows. Six cows were injected daily for 5 days with 40 mg recombinantly derived bST, while six control cows received excipient alone. Blood samples were collected hourly from 08.00 to 19.00 h on a single day the week before treatment and on days 4–5 of treatment. All other tissue samples were obtained at slaughter, 20–23 h after the last injection.

Administration of bST increased milk production and caused a 9% increase in hepatic DNA. Consumption of feed did not differ between control and bST-treated cows. Treatment did not alter serum concentrations of T4 or T3, although concentrations of thyroid hormones in the serum increased from 08.00 to 19.00 h. Activity of thyroxine-5′-monodeiodinase (5′-D) in liver and kidney was similarly unaffected. However, activity of 5′-D in mammary tissue increased approximately twofold in response to bST administration. We suggest that an increase in mammary conversion of T4 to the more biologically potent thyroid hormone T3 plays a role in mediating the galactopoietic response of dairy cattle to bST.

Journal of Endocrinology (1989) 121, 205–211
Adrenal mast cells modulate vascular and secretory responses in the intact adrenal gland of the rat
Authors: Hinson J. P. , Vinson G. P. , Pudney J. , Whitehouse B. J.
Published: December 2008

Mast cells were identified in the rat adrenal gland, located in the walls of arterioles at the point at which they penetrate the connective tissue capsule. The mast cell products, histamine and serotonin, both caused dose-dependent increases in rates of perfusion medium flow and steroid secretion in the isolated, perfused rat adrenal gland in situ. Compound 48–80, a mast cell degranulator, caused a significant increase in perfusion medium flow rate and steroid secretion by the in-situ perfused rat adrenal. Administration of disodium cromoglycate, a mast cell stabilizer, before administration of ACTH(1–24) virtually abolished the normal flow rate increment and significantly attenuated the corticosterone secretory response to ACTH(1–24).

These observations strongly suggest that adrenal mast cells modulate both vascular and secretory responses in the intact adrenal gland of the rat.

Journal of Endocrinology (1989) 121, 253–260
Testosterone concentrations in testicular interstitial fluid collected with a push–pull cannula or by drip-collection from adult rats given testosterone or aminoglutethimide
Authors: Maddocks S. , Setchell B. P.
Published: December 2008

This study was designed to investigate the differences in testosterone concentrations measured in testicular extracellular interstitial fluid obtained with a push–pull cannula or by post-mortem drip-collection. In the first experiment, testosterone-filled silicone elastomer capsules (2–16 cm lengths) or empty 2 cm capsules were implanted s.c. in adult male rats for 1 week. Animals were then anaesthetized and interstitial fluid was collected with a push–pull cannula for 1 h from one testis in each animal. Testicular and peripheral venous blood were then sampled and supernatant fluid was collected from the dispersed cells of the same testis. The contralateral testis in each animal was removed, and post-mortem interstitial fluid obtained by drip-collection for 20 h at 4 °C. In animals given empty capsules, testosterone concentrations in drip-collected interstitial fluid were significantly (P < 0·01) greater than testicular and peripheral venous blood levels, testicular fluid levels, and levels in interstitial fluid calculated from push–pull cannula samples. The concentrations of testosterone calculated in interstitial fluid collected with a push-pull cannula were never significantly greater than testicular venous blood levels. In animals with testosterone-filled capsules, testosterone concentrations measured in drip-collected interstitial fluid were similar to those calculated from push–pull cannulae samples, and to testicular venous blood levels. In a second experiment, a group of adult male rats was pretreated with aminoglutethimide to block steroidogenesis. Two hours later, interstitial fluid was drip-collected from the testes of these animals and from a group of vehicle-treated controls. For each animal, one testis was placed on ice and interstitial fluid collected for only 10 min, while the contralateral testis was kept at 4 °C and interstitial fluid collected for 20 h. In control animals, testosterone concentrations in interstitial fluid collected for 20 h were significantly (P < 0·01) greater than those in interstitial fluid collected for 10 min. In animals pretreated with aminoglutethimide, testosterone concentrations at 10 min and 20 h were not significantly different, and did not differ from those in control samples collected for 10 min.

We believe these results support earlier suggestions that the post-mortem drip-collection technique may give misleading results, and indicate that testosterone synthesis and secretion continues after isolation of the testis. This may also be the case with other Leydig cell products.

Journal of Endocrinology (1989) 121, 303–309
Levels of the small insulin-like growth factor-binding protein are strongly related to those of insulin in prepubertal and pubertal children but only weakly so after puberty
Authors: Holly J. M. P. , Smith C. P. , Dunger D. B. , Edge J. A. , Biddlecombe R. A. , Williams A. J. K. , Howell R. , Chard T. , Savage M. O. , Rees L. H. , Wass J. A. H.
Published: December 2008

We have looked at the relationship between fasting levels of insulin and a small insulin-like growth factor (IGF)-binding protein (IBP-1) in a cross-sectional study of 116 normal subjects aged 5–48 years. The relationship between IBP-1 and insulin was also examined within individual normal children in over-night profiles of IBP-1 and insulin obtained from two children at each stage of puberty (Tanner stages 1–5). In the cross-sectional study high levels of IBP-1 were found in early childhood and these fell throughout puberty as fasting levels of insulin rose. Multiple regression analysis revealed that both these changes were predominantly due to pubertal development rather than to age. After the age of 16 IBP-1 levels remained low despite fasting insulin levels returning to prepubertal levels. A strong negative correlation was obtained between IBP-1 and insulin in children of 5–16 years (r = −0·63; n = 60; P <0·001), no such relationship being found after the age of 16. In the second study, IBP-1 underwent a marked circadian variation in all cases and an inverse correlation with insulin, measured at the same time, was obtained at pubertal stages 1 to 4, but not at stage 5 (pooled data stages 1–4, r = −0·69; n = 53; P <0·001). We have demonstrated that a potential inhibitor of IGF-activity is inversely related to insulin throughout the period of active GH-related growth and that this relationship weakens after puberty.

Journal of Endocrinology (1989) 121, 383–387
Augmentation by epidermal growth factor of basal and stimulated progesterone production by human luteinized granulosa cells
Authors: Richardson M. C. , Gadd S. C. , Masson G. M.
Published: December 2008

Human granulosa cells were prepared from follicular aspirates obtained during oocyte collection for in-vitro fertilization. Following several days in culture, cells were washed and then progesterone output was measured in 2-h incubations. After culture for 3 days, incubated cells responded well to human chorionic gonadotrophin (hCG) and prostaglandin (PG) E2 with similar levels of maximum response. Exposure of cultured cells to epidermal growth factor (EGF) for 2 days (days 3–5) led to substantial increases both in basal production and in responses to hCG and PGE2 during subsequent incubations. These effects of EGF were not accompanied by measurable increases in DNA levels in cultures over this time. Results may point to a possible paracrine role for EGF-like factors modulating the activity of cells forming the early corpus luteum.

Journal of Endocrinology (1989) 121, 397–402
The in-vitro transformation of [3H]dehydroepiandrosterone into its principal metabolites in the adrenal cortex of adult castrated male rats and following steroid treatment
Authors: Canonaco M. , Andò S. , Valenti A. , Tavolaro R. , Panno M. L. , Maggiolini M. , Dessì-Fulgheri F.
Published: December 2008

The adrenal gland of castrated adult male rats metabolized [3H]dehydroepiandrosterone in vitro to Δ4-androsten-3,17-dione (4AD), testosterone, dihydrotestosterone (DHT) and 5α-androstane-3,17-dione (5αAD). Despite the low testosterone values, DHT and 5αAD were higher 30 and especially 60 days after castration, with raised 4AD:testosterone and decreased testosterone:DHT ratios. The 5α-reductase activity thus appears to increase with time after castration. Fourteen days after castration, 4AD was the only metabolite that was raised compared with intact animals, and testosterone was comparable in sham-operated and castrated rats.

The administration of testosterone propionate to castrated rats restored testosterone values to those of intact rat adrenals, whereas 4AD values were greater. The administration of dihydrotestosterone propionate also yielded higher levels of 4AD, in the presence of a lower testosterone value. After administration of oestradiol benzoate, 4AD values were lower especially compared with the other hormone-treated groups, and there was an unexpectedly high testosterone value.

These data indicate that the adrenal gland contributes to the production of androgens, as previously noted by Andò, Canonaco, Beraldi et al. (1988) who showed increased plasma 4AD and testosterone levels in adult male rats 30 days after castration. Furthermore, adrenal androgen production in castrated animals is differentially regulated by sex steroids.

Journal of Endocrinology (1989) 121, 419–424
Heterogeneity in the luteal population following superovulation with pregnant mare serum gonadotrophin and human chorionic gonadotrophin in the sheep
Authors: Hunter M. G. , Southee J. A.
Published: December 2008

In order to investigate the development and possible heterogeneity in the luteal population following superovulation, anoestrous ewes were induced to ovulate using progestagen priming followed by injections of pregnant mare serum gonadotrophin (1000 IU) and hCG (1000 IU). Ovaries were recovered from ewes on each of days 2, 4, 6, 8, 10, 12 and 15, and the weight, progesterone content, 125I-labelled hCG binding and progesterone synthesis in vitro of the individual corpora lutea measured. The results obtained showed that plasma progesterone concentrations on the day of slaughter were significantly correlated with time (P<0·05), total weight of luteal tissue (P< 0·001) and number of corpora lutea (P<0·05). The number of corpora lutea recovered per animal ranged from two to 12 and was significantly (P<0·05) correlated with the day after hCG injection until day 10. There was much variation between individual corpora lutea, particularly in terms of weight and progesterone content, although both were significantly (P<0·001) correlated with day of recovery until day 10. 125I-Labelled hCG binding was significantly (P<0·001) correlated with time until day 15. There was a significant (P<0·001) effect of age of the tissue on progesterone production in vitro, with output declining throughout the luteal phase.

These results show that the number of corpora lutea induced by superovulation in anoestrous ewes was very variable, and suggest that ovulation may have continued to occur during the luteal phase. Moreover, there was much variation between individual corpora lutea recovered from the same animal, and thus it should not be assumed that an individual corpus luteum is representative of the entire population.

Journal of Endocrinology (1989) 121, 459–465
Dexamethasone induces an increased biosynthesis of chromogranin A in rat pituitary gland
Authors: Fischer-Colbrie R. , Wohlfarter T. , Schmid K. W. , Grino M. , Winkler H.
Published: December 2008

The influence of dexamethasone on the regulation of the biosynthesis of chromogranins A and B, two proteins originally described in adrenal chromaffin granules, was investigated in the rat pituitary gland. Dexamethasone increased chromogranin A levels and its corresponding mRNA in a dose-dependent manner. Induction of chromogranin A occurred with a time-lag of 24 h. As shown by immunohistochemistry, chromogranin A was present in control and dexamethasone-treated rats in gonadotrophic cells only. Chromogranin B, secretogranin II and the respective pituitary hormones co-stored with chromogranin A were not changed by dexamethasone. Thus dexamethasone selectively increased chromogranin A levels in gonadotrophic cells.

Journal of Endocrinology (1989) 121, 487–494
The effect of 5-methoxytryptamine on golden hamster gonads is not a consequence of its acetylation into melatonin
Authors: Raynaud F. , Miguel J. L. , Vivien-Roels B. , Masson-Pévet M. , Pévet P.
Published: December 2008

Radioimmunoassay and high performance liquid chromatography were used to determine if the gonadal atrophy induced by late afternoon injections of 5-methoxytryptamine (5-MT) in golden hamsters kept under long photoperiod could be due to the acetylation of this compound into melatonin. An increase in plasma concentrations of melatonin (10–13 nmol/l) was detected 15 min after injection of 130 nmol 5-MT. An injection of 4·3 nmol melatonin generated a similar plasma concentration of melatonin. 5-MT (130 nmol) and melatonin (4·3 nmol) were then injected daily in the late afternoon to golden hamsters kept under long photoperiod. After 8 weeks, 5-MT induced total testicular regression, while melatonin induced partial atrophy only. Thus under these experimental conditions, 5-MT had a physiological activity independent of that of melatonin.

Journal of Endocrinology (1989) 121, 507–512
Growth hormone and testosterone can independently stimulate the growth of hypophysectomized prepubertal lambs without any alteration in circulating concentrations of insulin-like growth factors
Authors: Young I. R. , Mesiano S. , Hintz R. , Caddy D. J. , Ralph M. M. , Browne C. A. , Thorburn G. D.
Published: December 2008

Castrated prepubertal lambs were hypophysectomized and then treated with GH and testosterone either alone or in combination over a series of 3-week treatment periods. Hypophysectomy resulted in a rapid reduction in skeletal growth rate which could be reversed by the administration of either GH (4IU three times a week for 3 weeks) or testosterone propionate (10 mg daily for 3 weeks). When GH or testosterone treatment was withdrawn, skeletal growth fell to the post-operative rate. Combined treatment with both GH and testosterone was no more or less effective than either hormone given singly. The order of administration did not have any effect on the growth rate. Circulating concentrations of insulin-like growth factor-I (IGF-I) were reduced by hypophysectomy, but neither GH nor testosterone treatment, alone or in combination, had any effect on IGF-I concentrations. Concentrations of IGF-II rose following hypophysectomy, and again were not affected by any of the hormonal replacement treatments.

In conclusion, both GH and testosterone could stimulate skeletal growth in the hypophysectomized lamb without any alteration of circulating IGF concentrations, and testosterone can clearly stimulate skeletal growth in the complete absence of GH.

Journal of Endocrinology (1989) 121, 563–570
The role of hypothalamic vasoactive intestinal polypeptide in the maintenance of prolactin secretion in incubating bantam hens: observations using passive immunization, radioimmunoassay and immunohistochemistry
Authors: Sharp P. J. , Sterling R. J. , Talbot R. T. , Huskisson N. S.
Published: December 2008

The role of chicken vasoactive intestinal polypeptide (cVIP) as a prolactin-releasing factor was investigated in incubating bantam hens. Specific antibodies were raised against cVIP (anti-cVIP) for passive immunization studies, to develop a radioimmunoassay and to localize VIP neurones immunohistochemically in the hypothalamus. The concentration of plasma prolactin decreased after i.v. injection of anti-cVIP: this low concentration being maintained by daily injection of anti-cVIP. Incubating hens injected daily with anti-cVIP deserted their nests after 4·5 ± 0·6 days and returned to lay after 20 ± 1 days. This disruption of incubation behaviour with anti-cVIP was prevented by concomitant, twice daily, injections of 30IU ovine prolactin. The concentration of plasma LH was not immediately affected after injection of anti-cVIP but increased when the hens deserted their nests.

The amount of cVIP, measured by radioimmunoassay, was significantly higher in the median eminence (P < 0·01) and medial basal hypothalamus (P = 0·05) in incubating than in laying hens. No differences were seen in the amounts of cVIP in the preoptic hypothalamus or in a part of the forebrain including the nucleus accumbens, between laying and incubating hens.

Morphological observations were made on immunohistochemically identified cVIP cell bodies in the medial basal hypothalamus. These showed that cVIP cell number, cell area and density of immunoreactive product were significantly (P < 0·05) greater in incubating than in laying hens. Further, the density of cVIP reaction product in the anterior median eminence was also significantly (P < 0·01) greater in incubating than in laying hens.

These observations are consistent with the view that hypothalamic VIP is a physiologically important prolactin-releasing hormone in incubating bantam hens.

Journal of Endocrinology (1989) 122, 5–13
Gonadotrophin-releasing activity of neurohypophysial hormones: II. The pituitary oxytocin receptor mediating gonadotrophin release differs from that of corticotrophs
Authors: Evans J. J. , Catt K. J.
Published: December 2008

Neurohypophysial hormones stimulate gonadotrophin release from dispersed rat anterior pituitary cells in vitro, acting through receptors distinct from those which mediate the secretory response to gonadotrophin-releasing hormone (GnRH). The LH response to oxytocin was not affected by the presence of the phosphodiesterase inhibitor, methyl isobutylxanthine, but was diminished in the absence of extracellular calcium and was progressively increased as the calcium concentration in the medium was raised to normal. In addition, the calcium channel antagonist, nifedipine, suppressed oxytocin-stimulated secretion of LH. It is likely that the mechanisms of LH release induced by GnRH and neurohypophysial hormones are similar, although stimulation of gonadotrophin secretion is mediated by separate receptor systems. Oxytocin was more active than vasopressin in releasing LH, but less active in releasing ACTH. The highly selective oxytocin agonist, [Thr4,Gly7]oxytocin, elicited concentration-dependent secretion of LH but had little effect on corticotrophin secretion. The neurohypophysial hormone antagonist analogues, [d(CH2)5Tyr(Me)2]-vasopressin, [d(CH2)5Tyr(Me)2,Orn8]vasotocin and [d(CH2)5d-Tyr(Et)2Val4,Cit8]vasopressin, inhibited the LH response to both oxytocin and vasopressin. However, [d(CH2)5Tyr(Me)2]vasopressin was much less effective in inhibiting the ACTH response to the neurohypophysial hormones, and [d(CH2)5Tyr-(Me)2,Orn8]vasotocin and [d(CH2)5d-Tyr(Et)2,Val4, Cit8]vasopressin exhibited no inhibitory activity against ACTH release. Thus, agonist and antagonist analogues of neurophypophysial hormones display divergent activities with regard to LH and ACTH responses, and the neuropeptide receptor mediating gonadotroph activation is clearly different from that on the corticotroph. Whereas the corticotroph receptor is a vasopressin-type receptor an oxytocin-type receptor is responsible for gonadotrophin release by neurohypophysial hormones.

Journal of Endocrinology (1989) 122, 107–116
Effect of serotonin on α-melanocyte-stimulating hormone secretion from perifused frog neurointermediate lobe: evidence for the presence of serotonin-containing cells in the frog pars intermedia
Authors: Lamacz M. , Tonon M. C. , Leboulenger F. , Héry F. , Idres S. , Verhofstad A. J. , Pelletier G. , Vaudry H.
Published: December 2008

We have examined the presence of 5-hydroxytryptamine (serotonin; 5-HT) in the intermediate lobe of the frog pituitary and investigated the effect of exogenous 5-HT on α-melanocyte-stimulating hormone (α-MSH) release from the perifused neurointermediate lobe (NIL). Using a specific antiserum against 5-HT, the indirect immunofluorescence technique revealed the presence of 5-HT-like immunoreactivity (5-HT-LI) in discrete cells, generally gathered in small clusters among parenchymal cells, and in numerous neurites surrounding melanotrophic cells. At the electron microscopic level, using a silver-gold intensification procedure, 5-HT-LI was localized in dense-core secretory vesicles within specific pituitary cells which appear to be different from pituitary melanotrophs. Dense accumulation of gold particles was also observed in nerve fibres running between parenchymal cells. A combination of high-performance liquid chromatography analysis and electrochemical detection showed the presence of both 5-HT and its metabolite 5-hydroxyindol acetic acid (5-HIAA) in frog NIL extracts (534 ± 40 and 1245 ± 65 (s.e.m.) pg/mg wet tissue respectively). Administration of graded doses of 5-HT (from 1 to 30 μmol/l) to perifused frog NIL induced a dose-dependent inhibition of α-MSH release. Repeated pulses of 5-HT (10 μmol/l each) induced a reproducible inhibition of α-MSH without any desensitization phenomena. The inhibitory effect of 5-HT was partially blocked by the serotonergic antagonists methysergide and ICS-205-930 (10 μmol/l each). Concomitant administration of methysergide and ICS-205-930 (10 μmol/l each) totally abolished 5-HT-evoked inhibition of α-MSH. Fenfluramine, a releaser of 5-HT, induced a slight but significant reduction of α-MSH secretion. While 5-HT caused a marked inhibition of α-MSH release from intact NIL, 5-HT was devoid of effect on acutely dispersed pars intermedia cells suggesting that 5-HT does not exert a direct action on pituitary melanotrophs. We have examined the effect of specific dopaminergic, GABAergic and α-adrenergic antagonists on 5-HT-induced α-MSH inhibition. We observed that sulpiride and SR 95531 (10 μmol/l each) did not affect the response of NIL to 5-HT while yohimbine (10 μmol/l) suppressed the inhibitory action of 5-HT.

Taken together, our results indicate that discrete cells of the frog pars intermedia contain the neurotransmitter 5-HT which may act locally to inhibit α-MSH release. Our data also suggest that the inhibitory effect of 5-HT is mediated via presynaptic stimulation of catecholamine (possibly norepinephrine) release from adrenergic nerve endings terminating in the intermediate lobe of the frog pituitary.

Journal of Endocrinology (1989) 122, 135–146
Towards an antigenic map of human thyroglobulin: identification of ten epitope-bearing sequences within the primary structure of thyroglobulin
Authors: Dong Q. , Ludgate M. , Vassart G.
Published: December 2008

The aim of this study was to define, at the molecular level, epitopes of thyroglobulin recognized by heterologous antibodies and autoantisera.

One hundred thousand clones from a λgt11 human thyroid cDNA library were screened using a rabbit antibody to human thyroglobulin at a 1:2000 dilution. Twenty clones were plaque-purified to homogeneity, and characterization and sequencing of their cDNA inserts showed that they represented four distinct regions of the thyroglobulin molecule, one of them being the 22 carboxyl-terminal amino acids. Rescreening of the library with the same rabbit antibody to human thyroglobulin absorbed with peptides encoded by the carboxyl-terminal clone, led to the definition of six further epitope-bearing fragments of thyroglobulin. The ten regions that we have identified were recognized by ten further rabbit antibodies to human thyroglobulin, showing that they are representative of the repertoire of heterologous epitopes.

In contrast, none of the ten heteroepitope-bearing fragments was recognized by sera from ten patients with autoimmune thyroid disease with various titres of thyroglobulin antibodies. Screening of 2 × 106 clones from the library using a pool of ten autoantisera (individual sera diluted to 1:1000), and of 1 × 106 clones using a single autoantiserum of very high antithyroglobulin titre (diluted 1:400) resulted in no thyroglobulin clones being isolated. The significance of these results to the immune process is discussed.

Journal of Endocrinology (1989) 122, 169–176
Expression of an intercellular adhesion molecule, ICAM-1, by human thyroid cells
Authors: Weetman A. P. , Cohen S. , Makgoba M. W. , Borysiewicz L. K.
Published: December 2008

Intercellular adhesion molecule-1 (ICAM-1), hitherto identified on activated B cells, macrophages, dendritic cells, endothelia and certain epithelial cells, serves as a ligand for the lymphocyte function-associated antigen-1 (LFA-1). ICAM-1 binding by LFA-1 enhances the efficiency of lymphocyte-target cell and lymphocyte-accessory cell interactions. We have investigated the in-vitro expression of ICAM-1 by cultured thyroid cells from five patients with Graves' disease using indirect immunofluorescence analysis, and found that 30 ± 11% (mean ± s.d.) of cells were ICAM-1 positive under basal conditions. The proportion of cells which were ICAM-1 positive and the amount of ICAM-1 per cell (assessed by fluorescence intensity) were both increased in all cases by the cytokines γ-interferon, interleukin-1 and tumour necrosis factor. Immunohistochemical analysis of frozen sections from thyroidectomy specimens demonstrated ICAM-1 on thyroid follicular cells in areas of lymphocytic infiltration in patients with Graves' disease (n = 2) or Hashimoto's thyroiditis (n = 2). ICAM-1 was not found in specimens from a patient with a toxic multinodular goitre or a patient with Graves' disease without focal lymphocytic accumulation. These results suggest that the thyroid epithelium may express ICAM-1 as well as major histocompatibility complex class II antigens, such as HLA-DR, in response to locally synthesized cytokines. The enhanced expression of ICAM-1 may render these cells more susceptible as targets for lymphocytemediated cytotoxicity, and together with HLA-DR antigen expression may increase the accessory cell capability of the thyroid follicular cells.

Journal of Endocrinology (1989) 122, 185–191
Regulation of the onset of the breeding season of the ewe: importance of long days and of an endogenous reproductive rhythm
Authors: Malpaux B. , Robinson J. E. , Wayne N. L. , Karsch F. J.
Published: December 2008

Three experiments were conducted to evaluate the role that the increasing day lengths of late winter and spring play in timing the breeding season of the Suffolk ewe. In the first experiment, ewes were denied their normal complement of increasing day length by maintaining them on the photoperiod experienced at the winter solstice. This prevented the breeding season from occurring the subsequent autumn. In the second experiment, ewes were exposed to increases in day length at different time-intervals after the winter solstice: the normal time, later than normal or earlier than normal. Once the summer solstice photoperiod was reached, it was maintained until the end of the study. When increasing photoperiod was provided early, the breeding season was advanced; when it was provided late, reproduction was delayed. In the third experiment, ewes were exposed to a continuously increasing photoperiod matching the maximal rate of rise in natural conditions; this treatment was begun on the spring equinox and continued until mid-autumn. The steadily increasing photoperiod did not alter the time of reproductive onset in the autumn.

These findings support the following conclusions for timing of the breeding season of the Suffolk ewe. (1) The lengthening photoperiod between the winter and summer solstices is required for the occurrence of the breeding season in the autumn. (2) The time of initial exposure to this lengthening photoperiod provides an important cue for determining when the reproductive period occurs. (3) The time of onset of the breeding season does not depend upon the decreasing photoperiod after the summer solstice, nor does it require the photoperiod to stop increasing as the summer solstice approaches. These findings have been incorporated into a conceptual model for temporal regulation of the annual reproductive cycle of the ewe. An important component of this model is a critical role for increasing photoperiod to initiate a process in the late winter–spring which ultimately leads to an obligatory reproductive onset in the autumn.

Journal of Endocrinology (1989) 122, 269–278
Dynamics of testosterone secretion by the rat testis: implications for measurement of the intratesticular levels of testosterone
Authors: Maddocks S. , Sharpe R. M.
Published: December 2008

Testosterone concentrations have been measured in testicular interstitial fluid (IF), and in blood plasma sampled from various parts of the rat testis and spermatic cord, to assess (1) the most accurate method for determination of the intratesticular levels of testosterone, and (2) the route of secretion of testosterone from the testis. In untreated adult rats, testosterone concentrations were highest in blood collected from veins on the surface of the testis (269·50 ± 30·63 (s.e.m.) nmol/l), but were reduced by 56% on average in blood collected from veins at the proximal end of the spermatic cord (123·06±24·75 nmol/l), and were reduced considerably in peripheral venous blood (4·55 ± 0·55 nmol/l). Similar changes occurred in adult rats in which steroidogenesis was either stimulated (by treatment with human chorionic gonadotrophin; hCG) or inhibited (by treatment with aminoglutethimide; AMG), and in rats of various ages during sexual maturation. The reduction in testosterone levels during passage of blood from the testis up the spermatic cord is probably due mainly to dilution by incoming arterial blood which transfers to venous blood via anastomoses in the spermatic cord. Venous-arterial transfer of testosterone in the cord contributed to this in only a minor way. Concentrations of testosterone in testicular IF were always greater than testicular venous concentrations in control, developing and hCG-stimulated rats, but were comparable in rats treated with AMG to suppress Leydig cell steroidogenesis. These and other results demonstrate that the method of drip-collection of IF results in over-estimation of the actual intratesticular levels of testosterone. However, the present study also suggests that testicular venous blood probably provides the most accurate indication of the concentrations of testosterone in IF (and therefore in the testis) at any given time.

Journal of Endocrinology (1989) 122, 323–329
Endocrine regulation of cytochrome P-450 in the rat brain and pituitary gland
Authors: Warner M. , Tollet P. , Strömstedt M. , Carlström K. , Gustafsson J.-Å.
Published: December 2008

In an effort to understand the physiological functions of cytochrome P-450 in the central nervous system and pituitary gland, we evaluated changes in the level of the enzyme as a function of the endocrine status of rats and the ability of these tissues to synthesize or degrade steroids. The P-450 content of microsomes prepared from the hypothalamic preoptic area (HPOA), the olfactory lobes and the cerebrum was 0·040 ± 0·009 and in the pituitary gland 2·2 ± 0·6 (s.d.) nmol/g tissue. The P-450 content of the HPOA and olfactory lobes, but not of the rest of the cerebrum, was influenced by the endocrine status of rats. In microsomes it increased five- to tenfold over control levels during late pregnancy in the olfactory lobes and during lactation in the HPOA, and in both brain regions treatment of rats with 5α-dihydrotestosterone (DHT) caused an eight- to tenfold increase in the P-450 content. Androstenedione was not a good substrate for brain P-450. The level of androstenedione 19-hydroxylase in the olfactory lobe microsomal fraction was 0·50± 0·06 nmol 19-hydroxyandrostenedione formed/g tissue per h. This activity was tenfold lower in other brain areas and was not detectable in the pituitary gland. The rate of aromatization of androstenedione to oestradiol in the HPOA and olfactory lobe of lactating rats was 0·46 ± 0·14 and 0·38 ± 0·05 pmol/oestradiol formed/g tissue per h respectively. 5α-Androstane-3β,17β-diol (A-5α-3β,17β-diol) was a much better substrate for P-450 throughout the brain and pituitary gland. Catalytic activity was 125 ± 46 and 307 ±108 nmol triols formed/g tissue per h in the brain and pituitary gland respectively. The P-450 responsible for this catalytic activity was isolated and its substrate specificity examined. In addition to A-5α-3β,17β-diol, 5-androstene-3β,17β-diol, dehydroepiandrosterone and DHT were also substrates, with turnover numbers of 27, 8, 12 and 1 mol product/mol P-450 per min respectively. None of these catalytic activities was induced in the rat brain during pregnancy, lactation or DHT treatment. The enzyme was also present in the brains of mice but not guinea-pigs.

The yield of P-450 from the mitochondrial fraction of the HPOA and olfactory lobes in control rats was 0·01–0·02 nmol/g tissue. This increased tenfold during pregnancy. Immunological evidence for the presence of the cholesterol side-chain cleavage enzyme P-450 SCC was found in the HPOA and olfactory lobes of pregnant but not of control rats. However, no SCC catalytic activity was detectable in these brain mitochondrial P-450 fractions. From these studies we conclude that there is a major influence of the endocrine system on the content and quality of P-450 in the brain. However, the function and substrate specificities of these P-450s as well as of those in the pituitary gland remain to be characterized.

Journal of Endocrinology (1989) 122, 341–349