bluuebunny/crossref_metadata_2025 huggingface dataset:
The transport of plasma thyroxine in White storks (Ciconia ciconia) and the association of high levels of plasma transthyretin (thyroxine-binding prealbumin) with moult
Authors: Cookson E. J. , Hall M. R. , Glover J.
Published: December 2008

Antisera raised against domestic fowl transthyretin (thyroxine-binding prealbumin) and quail albumin were used to identify thyroxine-binding proteins in the plasma of White storks (Ciconia ciconia) and to measure seasonal changes in these proteins. Three plasma proteins were shown to bind thyroxine (T4). They were albumin, transthyretin and a protein with an electrophoretic mobility on non-denaturing poly-acrylamide gel electrophoresis intermediate between albumin and transthyretin. There was no thyroxine-binding globulin.

The plasma concentrations of transthyretin and albumin were measured in male and female storks exposed to the photoperiodic and climatic conditions experienced during an annual cycle at 46°N,11°E. The storks were in four age groups, fledglings and 1-, 2- and 3-year-old birds. A strong correlation (P < 0·001) was found between daylength and the concentration of transthyretin during the period from February to August in the 1-, 2- and 3-year-old male and female storks, with peak values of 150–210 mg/l in May, June and July coinciding with the annual moult. Between September and January the concentration of plasma transthyretin was low, ranging between 80 and 100 mg/l. The concentration of plasma albumin was not correlated with seasonal changes in daylength and did not change during moult. It is suggested that seasonally high concentrations of plasma transthyretin may have a physiological role in moult.

J. Endocr. (1988) 117, 75–84
Thyrotrophin and prostaglandin E2 increase calmodulin levels and cyclic AMP phosphodiesterase activity in cultured porcine thyroid cells
Authors: Takasu N. , Yamada T. , Shimizu Y.
Published: December 2008

Thyrotrophin (TSH) and prostaglandin E2 (PGE2) increased cellular cyclic AMP (cAMP), calmodulin levels and cAMP phosphodiesterase activity in cultured porcine thyroid cells. Dibutyryl cAMP (dbcAMP), a stable analogue of cAMP, increased calmodulin levels and cAMP phosphodiesterase activity. These results indicate that TSH- and PGE2-stimulated increases in calmodulin are mediated by cAMP. This increased concentration of calmodulin in turn stimulates cAMP phosphodiesterase. Double reciprocal plots of cAMP hydrolysis yielded two apparent Michaelis constants (Km); the lower in the 1 μmol/l and the higher in the 10 μmol/l range. Thyrotrophin, PGE2 and dbcAMP increased the values of maximal velocity without changing the Km values.

J. Endocr. (1988) 117, 109–114
Release of prolactin is independent of the secretion of thyrotrophin-releasing hormone into hypophysial portal blood of sheep
Authors: Thomas G. B. , Cummins J. T. , Yao B. , Gordon K. , Clarke I. J.
Published: December 2008

In order to determine whether pituitary prolactin release was directly related to the secretion of TRH into hypophysial portal blood, serial portal and jugular venous blood samples were collected from seven lactating and three non-lactating ewes. In another experiment, samples were collected from five ovariectomized ewes while being exposed to an audio-visual stress and then later administered with chlorpromazine. Secretion of TRH was pulsatile in all ewes and independent of prolactin secretion; TRH pulses coincided with significant increases in prolactin secretion in only 15% of cases and only 29% of prolactin pulses were associated with TRH pulses. Sixty-seven per cent of suckling bouts were associated with increases in prolactin secretion, but only 22% of these were associated with significant increases in TRH secretion. Chlorpromazine increased prolactin levels fourfold but did not affect portal concentrations of TRH. Audio-visual stress was not a reliable method of causing prolactin release in this model. Mean portal concentrations of TRH and jugular concentrations of prolactin were not significantly correlated. These results show that hypothalamic TRH and pituitary prolactin are secreted independently in the sheep, implying that increases in prolactin release caused by suckling or chlorpromazine are not the direct result of increased TRH secretion.

J. Endocr. (1988) 117, 115–122
Steroid sequestration within the rat adrenal zona glomerulosa
Authors: Laird S. M. , Vinson G. P. , Whitehouse B. J.
Published: December 2008

Accumulated data from in-vitro experiments have suggested that 18-hydroxysteroids may be stored within the intact rat adrenal zona glomerulosa. The phenomenon was further investigated by comparing the amount of steroid remaining in the zona glomerulosa tissue with that secreted into the media during incubation in vitro. The results showed that 18-hydroxydeoxycorticosterone (18-OH-DOC) and 18-hydroxycorticosterone (18-OH-B) were retained within the tissue against a considerable concentration gradient, with smaller amounts of aldosterone and corticosterone. Lysis of the intact zona glomerulosa, by preincubation in distilled water, yielded an enriched plasma membrane preparation. After subsequent incubation in Krebs–Ringer bicarbonate this preparation contained significantly more 18-OH-DOC than did the intact tissue, suggesting that tissuesequestered 18-OH-DOC is normally metabolized to other products. These may include 18-OH-B and aldosterone.

Fractionation of homogenized intact zona glomerulosa and the enriched plasma membrane preparation by density gradient centrifugation showed that tissue 18-OH-DOC banded in fractions of density 1·063– 1·21 g/ml and that its distribution was highly correlated with protein. Corticosterone, 18-OH-B and aldosterone banded like added free [3H]18-OH-DOC in fractions of density < 1·006 g/ml.

The results suggest that 18-OH-DOC is the major sequestered steroid within the rat adrenal zona glomerulosa and that this sequestration is attributable to the association of 18-OH-DOC with a high-density component of the plasma membrane.

J. Endocr. (1988) 117, 191–196
Osmoregulation in rats with long-term enhanced cerebrospinal fluid levels of vasopressin or vasoactive intestinal polypeptide
Authors: Kruisbrink J. , van Heerikhuize J. J. , Boer G. J.
Published: December 2008

The cerebrospinal fluid level of arginine vasopressin (AVP) or vasoactive intestinal polypeptide (VIP) was enhanced chronically by implantation of a device for controlled drug delivery in the lateral ventricle of the rat. Urine production, water consumption, urine osmolality as well as urinary AVP excretion were then measured for a period of 26 days. During this period the rats were studied under normal hydration and under conditions of osmotic stress induced by water deprivation (2 days) and the drinking of 2% (w/v) NaCl (6 days), in order to see whether the capacity of central systems to react adequately to osmotic stimuli was affected by high central peptide levels.

Immediately after the central AVP treatment was started, a temporary increase was found in urinary AVP levels which was not accompanied by a change in any of the other parameters but which decreased again to control levels within 10 days. After this burst of AVP excretion, AVP-treated rats showed a tendency during periods of normal hydration for a lower urine osmolality, combined with a higher water intake and urine production without changes in urinary AVP excretion. Since there was no clear-cut correlation between urinary AVP excretion and body water turnover, this could still indicate a slowly acquired and slight inhibition of pituitary AVP release by long-term centrally administered AVP. However, the capacity of these rats to respond to osmotic stimuli was not different from the controls.

In the VIP-treated rats a slight but significant reduction in urine production was found in all three periods of normal hydration. During the osmotic stress induced by the drinking of 2% NaCl the VIP-treated rats showed a lower increase in urine production and fluid intake and a lower decrease in urine osmolality when compared with the response of the control rats. This has tentatively been interpreted as a potentiation by VIP of the activation of pituitary AVP release under these conditions.

J. Endocr. (1988) 117, 207–214
Evidence for a novel parathyroid hormone-related protein in fetal lamb parathyroid glands and sheep placenta: comparisons with a similar protein implicated in humoral hypercalcaemia of malignancy
Authors: Rodda C. P. , Kubota M. , Heath J. A. , Ebeling P. R. , Moseley J. M. , Care A. D. , Caple I. W. , Martin T. J.
Published: December 2008

Parathyroid hormone (PTH)-like bioactivity, assayed as adenylate cyclase response in UMR 106-01 osteogenic sarcoma cells, was present in extracts of sheep fetal and maternal parathyroid glands and placenta. Preincubation of extracts with PTH(1–34) antiserum inhibited approximately 40% of the bioactivity in fetal parathyroid extracts, 50% in maternal parathyroid extracts, but only 10% of the bioactivity in the placental extract. Partial purification of placental extracts by chromatography yielded fractions containing PTH-like bioactivity which were similar in behaviour to that of PTH-related protein (PTHrP) from a human lung cancer cell line (BEN). An antiserum against synthetic PTHrP(1–16) partially inhibited the bioactivity of the placental extract and synthetic PTHrP(1–34), but had no effect on the bioactivity of bovine PTH(1–34) or bovine PTH(1– 84). The placental PTH-like bioactivity was higher in mid- than in late gestation. Fetal parathyroid glands contained the highest PTH-like bioactivity.

Thyroparathyroidectomy of one fetal twin lamb in each of 16 ewes between 110 and 125 days of gestation resulted in decreases of the plasma calcium concentration and reversal of the placental calcium gradient that existed between the ewe and the intact fetus. Perfusion of the placenta of each twin in anaesthetized ewes was carried out sequentially with autologous fetal blood in the absence of the exsanguinated fetus. The plasma calcium concentration in the blood perfusing the placenta of each twin increased, but reached a plateau at a lower concentration in the perfusing blood of thyroparathyroidectomized fetuses than in that of the intact fetuses. Addition of extracts of fetal parathyroid glands or of partially purified PTHrP resulted in further increases in plasma calcium in the autologous blood perfusing the placentae of thyroparathyroidectomized fetuses, but addition of bovine PTH(1–84) or rat PTH(1–34) had no effect. The presence of this PTH-like protein in the fetal parathyroid gland and placenta may contribute to the relative hypercalcaemia of the fetal lamb. This protein, which is similar to PTHrP associated with humoral hypercalcaemia of malignancy, stimulates the placental calcium pump responsible for maintaining a relative fetal hypercalcaemia during gestation.

J. Endocr. (1988) 117, 261–271
Specific binding of atrial natriuretic peptide increases cyclic GMP levels in human astrocytoma cells
Authors: Lyall F. , Balmforth A. J. , Morton J. J.
Published: December 2008

Specific high-affinity binding sites (dissociation constant 100 pmol/l) for atrial natriuretic peptide (ANP) have been identified in the clone D384 derived from the human astrocytoma cell line G-CCM. Unrelated peptides such as angiotensin II, vasopressin and bradykinin did not compete for these sites. Of the atrial natriuretic peptides studied, both the human and rat ANP competed equally, while peptides with either C-or N-terminal residue missing or with no internal -S-S-bond either competed less effectively or did not compete at all. Human ANP stimulated the cells to increase their intracellular level of cyclic GMP in a time- and dose-dependent manner with maximum stimulation being approached but not reached at concentrations of 1 μmol/l. These results support both the notion that ANP has an important functional role within the brain and the concept of neurotransmitter/neuromodulator communication between neurones and glia.

J. Endocr. (1988) 117, 315–321
Social status controls LH secretion and ovulation in female marmoset monkeys (Callithrix jacchus)
Authors: Abbott D. H. , Hodges J. K. , George L. M.
Published: December 2008

The suppression of ovulation in subordinate female marmosets was associated with suppressed pituitary LH secretion and reduced pituitary LH response to gonadotrophin-releasing hormone (GnRH). In subordinate females, basal plasma LH concentrations were commonly below 2 IU/1 (n = 5) (maximum 10·7 IU/1). Plasma oestrogen concentrations were similarly low (maximum 0·62 nmol/l) and plasma progesterone concentrations of below 30 nmol/l confirmed the anovulatory condition. This infertility condition was rapidly reversed when subordinate females (n = 5) were removed from their social groups and housed singly, when plasma LH (maximum 140·0 IU/1) and oestrogen (maximum 7·84 nmol/l) concentrations increased preceding ovulation. Infertility was rapidly reimposed when these singly housed females were re-introduced to subordinate status in new social groups, when plasma LH concentrations fell to their previous low values within 4 days; no ovulation occurred thereafter. Plasma oestrogen levels also fell, but less dramatically. The luteal phases of three of the subordinate females were shortened following the re-instatement of subordinate status. The maximum LH response of subordinate females to the highest dose of GnRH (200 ng) was only 19·1±6·7 IU/1 (mean ± s.e.m.; n=8): this contrasted with that in dominant females in either the follicular phase (40·0±13·3 IU/1; n=6) or the luteal phase (126·7±24·9 IU/1; n=10) of the ovarian cycle.

These results suggest that the social suppression of fertility in subordinate female marmosets is mediated by impaired hypothalamic GnRH secretion. Such an immediate and precise behavioural control of LH secretion and ovulation is without equal in anthropoid primates.

J. Endocr. (1988) 117, 329–339
Mesotocin and luteal function in macropodid marsupials
Authors: Curlewis J. D. , Renfree M. B. , Sheldrick E. L. , Flint A. P. F.
Published: December 2008

Pituitary glands and corpora lutea collected at various stages of the reproductive cycle of the tammar wallaby (Macropus eugenii), were extracted and fractionated by high-performance liquid chromatography, and specific radioimmunoassays were used to measure mesotocin ([Ile8]-oxytocin) and oxytocin. Mesotocin, but not oxytocin, was identified in extracts of pituitary; the mean concentration of mesotocin in this tissue was 0·75 nmol/g wet weight. Neither mesotocin nor oxytocin was detected in extracts of corpus luteum. In female Bennett's wallabies passively immunized against mesotocin during seasonal reproductive quiescence, there was no significant effect on peripheral progesterone concentrations and there were no births, matings or changes in vaginal smears in the 2 months following treatment. Thus mesotocin is unlikely to act as a systemic luteostatic agent during seasonal quiescence.

J. Endocr. (1988) 117, 367–372
Pituitary and plasma vasopressin concentrations and fluid balance throughout the oestrous cycle of the rat
Authors: Forsling M. L. , Peysner K.
Published: December 2008

Observations were made of fluid balance and vasopressin concentrations throughout the oestrous cycle of normally cyclic female rats housed under a 12 h light: 12 h darkness regime. Plasma vasopressin concentrations were found to increase progressively during the light period, falling again during the night on all days of the cycle except pro-oestrus. On this day, peak vasopressin concentrations of 3·32 ± 0·8 pmol/l were seen between 10.00 and 12.00 h, with lower concentrations of 1·74± 0·22 pmol/l being seen between 18.00 and 19.00 h. Urine flow appeared to reflect the changes in plasma vasopressin concentrations, being significantly lower during the light phase, with a small increase being seen over this period on pro-oestrus. Pituitary vasopressin concentrations were highest between 09.00 and 10.00 h and fell progressively over each of the 4 days of the oestrous cycle. The changes in pituitary content were greater than could be accounted for in terms of the alterations in the plasma concentrations of vasopressin.

J. Endocr. (1988) 117, 397–402
Evidence for luteotrophic and antiluteolytic actions of prolactin in rats with 5-day oestrous cycles
Authors: Sánchez-Criado J. E. , van der Schoot P. , Uilenbroek J. Th. J.
Published: December 2008

Injection of 1 mg bromocriptine at either 08.00 or 16.00 h on the day of oestrus in rats with 5-day oestrous cycles caused a reduction in the duration of progesterone secretion by the corpus luteum during dioestrous, and a shortening of the ovarian cycle by 1 day. These effects were not present when bromocriptine was injected at 08.00 h on the day of metoestrus. The effect of bromocriptine on progesterone secretion by the corpus luteum was reversed by neutralization of the biological activity of LH at dioestrus by injection of 0·5 ml anti-LH serum at 08.00 h at metoestrus. Injection of the antiserum alone prolonged progesterone secretion by the corpus luteum, but had no effect on the length of dioestrus.

These results are interpreted as suggesting (1) that prolactin secretion on the afternoon of oestrus protects the corpus luteum of the rat ovarian cycle against the luteolytic effects of LH secretion during early dioestrus and (2) that prolactin stimulates progesterone secretion in the absence of such a luteolytic action. This response of the corpus luteum of the rat ovarian reproductive cycle to prolactin results in 5-day oestrous cycles.

J. Endocr. (1988) 117, 455–460
DEVELOPMENT OF A RADIOIMMUNOASSAY FOR OVINE TROPHOBLAST PROTEIN-1, THE ANTILUTEOLYTIC PROTEIN FROM THE SHEEP CONCEPTUS
Authors: Vallet J. L. , Bazer F. W. , Ashworth C.J. , Johnson H. M. , Pontzer C. H.
Published: December 2008

A radioimmunoassay has been developed for quantitation of ovine trophoblast protein-1 (oTP-1), a sheep conceptus secretory protein which allows for maintenance of the corpus luteum during early pregnancy. The assay was validated for dialysed and undialysed culture medium and pregnant uterine flushings ranging from no dilution (neat) to dilutions of 1:2500 for dialysed media, 1:100-1:1000 for undialysed media and 1:50-1:1000 for pregnant uterine flushings. The assay accurately measured oTP-1 added to undiluted and diluted dialysed and undialysed culture media and pregnant uterine flushings. No cross-reaction was detectable for bovine α or γ interferon, bovine calmodulin, feline conceptus secretory proteins, equine conceptus secretory proteins, porcine conceptus secretory proteins, bovine conceptus secretory proteins and proteins in a uterine flushing collected from a non-pregnant ewe. Immunoreactivity in the assay matched that for oTP-1 throughout oTP-1 purification. This assay is the first validated assay which may be used to quantitate production of oTP-1 in culture or content of oTP-1 in uterine flushings.
Attenuation of preovulatory gonadotrophin surges by epostane: a new inhibitor of 3β-hydroxysteroid dehydrogenase
Authors: DePaolo L. V.
Published: December 2008

Epostane, an inhibitor of 3β-hydroxysteroid dehydrogenase, was administered orally to pro-oestrous rats to evaluate further a possible role for preovulatory progesterone secretion in eliciting surges of LH and FSH. Whereas a dose of 10 mg epostane/kg had essentially no effects on preovulatory gonadotrophin surges and ovulation, 200 mg epostane/kg markedly attenuated LH and FSH surges and blocked ovulation. A dose of 50 mg epostane/kg exerted effects on LH and FSH surges and ovulation intermediate between those of doses of 10 and 200 mg/kg. Plasma concentrations of progesterone were significantly lower in all anovulatory epostane-treated rats at 18.00 and 22.00 h on pro-oestrus than those measured in vehicle-treated rats. Concurrent injection of 2 mg progesterone in rats given 200 mg epostane/kg restored gonadotrophin surges to normal, but consistently failed to reverse the inhibitory effects of epostane on ovulation. Peak plasma progesterone levels produced by the progesterone injections were eight- to tenfold higher than the highest levels measured in vehicle-treated rats during the afternoon of pro-oestrus. Insertion of progesterone capsules was less effective than injections of progesterone in restoring gonadotrophin surges to normal, even though peak plasma progesterone concentrations achieved after insertion of two 20 mm long progesterone capsules were double the peak progesterone concentrations measured in control rats. Nevertheless, taken together with recent reports showing attenuation of preovulatory gonadotrophin surges by the progesterone antagonist RU 486 (17β-hydroxy-11β-[4-dimethyl-aminophenyl]-17α-[prop-1-ynl]estra-4,9-diene-3-one), the present results provide support for a role of preovulatory progesterone secretion in enhancing oestrogen-dependent LH/FSH surges on pro-oestrus.

J. Endocr. (1988) 118, 59–68
Efficacy and specificity of monoclonal antibodies to progesterone in preventing the establishment of pregnancy in the mouse
Authors: Ellis S. T. , Heap R. B. , Butchart A. R. , Rider V. , Richardson N. E. , Wang M.-W. , Taussig M. J.
Published: December 2008

Anti-progesterone monoclonal antibody prevents the establishment of pregnancy in BALB/c mice by the prevention of implantation when injected i.p. 32 h after mating. To determine the specificity of this effect, mice were injected with immune and non-immune purified mouse immunoglobulins. The results show that anti-implantation efficacy was due to high-affinity antibody which bound progesterone since two further mouse immunoglobulin (Ig) G1 preparations, mouse IgA and mouse IgM which failed to bind the steroid, had no effect on pregnancy rates. From a panel of anti-progesterone monoclonal antibodies, six with a high affinity (affinity constant, 0·24–0·80 litres/nmol) and specificity for progesterone were selected for additional studies. Anti-implantation efficacy for five antibodies was similar, with a 50% effective dose within the range of 0·8–2·0 nmol. Antibody reached high concentrations in plasma within 12 h after i.p. injection, and declined with a half-life of about 80 h. Purified F(ab′)2 fragments of antibody also bound progesterone, but were less effective than the native molecule in blocking pregnancy. The results show that implantation in the mouse can be blocked by a high-affinity antibody that binds progesterone and which is removed from the blood at a slow rate.

J. Endocr. (1988) 118, 69–80
Baroreceptor-independent medullary mechanism for release of vasopressin during hypotension in rats
Authors: Hashemzadeh-Gargari H. , Baertschi A. J. , Guyenet P. G.
Published: December 2008

Plasma vasopressin (AVP) levels were measured at rest (mean arterial pressure 80–85 mmHg) and during hypotension (mean arterial pressure 38–45 mmHg) induced by ganglionic blockade (trimethaphan) in halothane-anaesthetized respirated rats with end-tidal pCO2 maintained at 34–40 mmHg. Hypotension (15 min) produced a 310% increase in plasma AVP (±60% s.e.m.) which was not reduced significantly by prior baro- and chemoreceptor denervation. The hypotension-induced rise in AVP was blocked by bilateral microinjections (40 nl) of the GABA-mimetic agent muscimol (151 pmol) into the ventrolateral medulla at obex level and significantly attenuated by injections of the same amount in the nucleus tractus solitarius. The rise in AVP was unaffected by microinjections in the pontine locus coeruleus. It was also blocked by bilateral microinjections of the glutamate-receptor antagonist kynurenate (40 nl, 1·8 nmol) into the ventrolateral medulla but unaffected by microinjections of the inactive analogue xanthurenic acid (40 nl, 1·8 nmol). A significantly smaller rise in plasma AVP (88%) was observed following bilateral nephrectomy. It is concluded that, in this preparation, hypotension produces the release of AVP via a mechanism largely independent of baro- and chemoreceptors, which requires the activation of neurones located in the caudal medulla oblongata. The same or closely related neurones may be activated by a neural or hormonal signal generated by the kidney.

J. Endocr. (1988) 118, 101–111
Changes in serum concentrations of steroids during embryonic and post-hatching development of male and female Japanese quail (Coturnix coturnix japonica)
Authors: Schumacher M. , Sulon J. , Balthazart J.
Published: December 2008

Serum concentrations of testosterone, 5α-dihydrotestosterone, oestradiol and progesterone were measured by radioimmunoassay combined with Celite chromatography in male and female Japanese quail (Coturnix coturnix japonica) during the second half of embryonic life (days 9–17 of incubation) and during the first 5 weeks after hatching. The mean level of each of the four steroids was significantly affected by the age of the birds. An overall effect of sex was detected by analysis of variance only on oestradiol concentrations, with females having higher serum concentrations than males during most of the age range studied. Significant peaks of testosterone and progesterone were also detected around hatching time. These results are consistent with the view that oestradiol is the major hormone implicated in the sexual differentiation of reproductive behaviour in the quail. The relationships between the circulating concentrations of oestradiol during ontogeny and the critical period of differentiation as postulated by currently accepted models is also discussed.

J. Endocr. (1988) 118, 127–134
Hypothalamic hormones that release growth hormone stimulate hepatic 5′-monodeiodination activity in the chick embryo
Authors: Kühn E. R. , Vanderpooten A. , Huybrechts L. M. , Decuypere E. , Darras V. , Sharp P. J.
Published: December 2008

Plasma GH, tri-iodothyronine (T3), thyroxine (T4) and liver 5′-monodeiodination (5′-D) activity were measured in 18-day-old chick embryos injected with thyrotrophin-releasing hormone (TRH) and human pancreatic growth hormone releasing factor (hpGRF). Injections of 0·1 and 1 μg TRH and 1·5 μg hpGRF increased the concentration of plasma GH while injection of 15 μg hpGRF had no effect. Concentrations of plasma T3 were raised after injection of TRH or hpGRF. Injections of TRH but not of hpGRF raised the concentration of plasma T4. The increases in concentration of plasma T3 after injection of TRH or hpGRF were parallelled by increases in liver 5′-D activity. An injection of 0·25 μg T4 significantly raised the concentration of T4 in plasma but had no effect on plasma T3 or liver 5′-D activity. It is concluded that the release of chicken GH by TRH or hpGRF is responsible for the observed increases in plasma concentration of T3 and liver 5′-D activity.

J. Endocr. (1988) 118, 233–236
Effects of acute stress on the patterns of LH secretion in the common marmoset (Callithrix jacchus)
Authors: O'Byrne K. T. , Lunn S. F. , Dixson A. F.
Published: December 2008

Stressful stimuli associated with aggressive encounters and low social rank may affect female fertility in a variety of mammalian species. In these experiments we examined the effects of aggressive encounters and physical restraint in a primate chair on the patterns of LH secretion in ovariectomized, oestrogen-primed female marmosets. Receipt of aggression from a female conspecific, followed by physical restraint for collection of blood samples (at 10-min intervals for 4 h), resulted in marked declines in LH concentrations during oestradiol-induced LH surges in five animals (from 112 ± 24 μg/l to 45±12 μg/l; group means ± s.e.m.; P<0·05). This was due to reductions in LH pulse amplitude rather than to changes in pulse frequency. Decreases in plasma concentrations of LH were reversed by treating females with exogenous LH-releasing hormone (LHRH). Cortisol treatment had no effect on LH levels during oestrogen-induced LH surges. Effects of aggressive encounters and physical restraint on plasma LH were not therefore due to reduced pituitary responsiveness to LHRH or to increased plasma concentrations of cortisol. In separate experiments it was found that physical restraint alone had no effect on plasma LH in habituated subjects, and that decreases in plasma LH after receipt of aggression only occurred if animals were subsequently placed in the restraint chair. A summation of stressful effects is therefore required to produce the fall in circulating LH. A summation of social and other environmental stressors may also underlie the reduced fertility seen in free-living animals.

J. Endocr. (1988) 118, 259–264
Involvement of the cytoskeleton in the steroidogenic response of frog adrenal glands to angiotensin II, acetylcholine and serotonin
Authors: Feuilloley M. , Netchitaïlo P. , Delarue C. , Leboulenger F. , Benyamina M. , Pelletier G. , Vaudry H.
Published: December 2008

In order to determine the role of the cytoskeleton in adrenal steroidogenesis, we have studied the effect of cytochalasin B (a microfilament-disrupting agent) and vinblastine (an antimicrotubular drug) on corticosteroid secretion by frog interrenal tissue in vitro. Perifusion of interrenal fragments with cytochalasin B (50 μmol/l) induced a marked inhibition of basal corticosteroid output. In addition, stimulation of corticosteroidogenesis by all corticotrophic factors was also inhibited by cytochalasin B. Using an immunohistochemical technique and specific anti-tubulin antiserum, we verified that vinblastine (10 μmol/l) was responsible for the disappearance of the microtubular network in adrenocortical cells. Administration of vinblastine (10 μmol/l) did not affect the spontaneous secretion of corticosterone and aldosterone and had no effect on the steroidogenic response of interrenal glands to angiotensin II and acetylcholine. In contrast, vinblastine was responsible for a marked decrease in serotonin-induced stimulation of corticosteroid production. On the other hand, data from high-performance liquid chromatography showed that infusion of cytochalasin B or vinblastine was not associated with the production of any new steroid which could interfere in the radioimmunoassays. Taken together, these data suggest that microfilaments are involved in a late and common step of corticosteroidogenesis while microtubules are only required for the coupling of the secretory response to certain corticotrophic factors such as ACTH and serotonin.

J. Endocr. (1988) 118, 365–374
Glucocorticoids in the blood plasma of the platypus Ornithorynchus anatinus
Authors: McDonald I. R. , Than K. A. , Evans B.
Published: December 2008

Blood samples were obtained from two male and two female platypuses at various times after capture and anaesthesia for other experimental purposes. In samples obtained during ketamine–xylazine or pregnanediol anaesthesia 15–24 h after capture, the concentration of total glucocorticoids, measured as 'cortisol equivalent' in a radioligand assay, was 207– 620 nmol/l. In samples taken 14–35 h after injection of dexamethasone (0·2 mg/kg) total glucocorticoid concentration was 79–88 nmol/l.

Individual glucocorticoids were isolated on columns of Sephadex LH-20 and measured separately against appropriate standards. In all except two haemolysed samples obtained from a male that died 25 h after capture, the major glucocorticoid behaved as cortisol, contributing 77–94% of the total. The remainder was made up of varying proportions of substances behaving as corticosterone, 11-deoxycortisol and cortisone. In the haemolysed samples from the moribund animal the major reactive substance, contributing 52–54% of the total, behaved as cortisone. The total adrenal gland weight of this animal was 747 mg, compared with 200–286 mg in two others, suggesting preceding exposure to stress.

Equilibrium dialysis and polyacrylamide gel electrophoresis (PAGE) revealed no evidence for a transcortin-like glucocorticoid- and progesterone-binding protein in platypus plasma. However, as in the echidna, there was a heat-labile, high-capacity binding system migrating with albumin on PAGE.

Glucose was undetectable in the plasma of the moribund animal and only 1·7–2·8 mmol/l in the initial plasma samples from the others. In two animals, injection of glucose i.p and dexamethasone i.m. was followed by an increase in the plasma concentration of glucose to the range 3·8–9·9 mmol/l and commencement of normal swimming and feeding activity for the next 36–48 h.

J. Endocr. (1988) 118, 407–415
Stimulation of aromatase activity in the fetal rat testis by cyclic AMP and FSH
Authors: Weniger J.-P. , Zeis A.
Published: December 2008

The effect of dibutyryl cyclic AMP and FSH on oestrogen biosynthesis was investigated in testes from 18- to 21-day-old fetal rats cultured in vitro in the presence of tritiated testosterone. Oestrone and oestradiol concentrations were measured by determination of constant specific activity after isotopic dilution.

Dibutyryl cyclic AMP and FSH markedly stimulated the conversion of testosterone into both oestrone and oestradiol at all stages studied. Oestradiol synthesis was stimulated by two- to sevenfold, while stimulation of oestrone synthesis was even greater. The results demonstrate that the aromatase enzyme system of the fetal rat testis responds to cyclic AMP and FSH.

J. Endocr. (1988) 118, 485–489
Extracellular calmodulin and its association with epidermal growth factor in normal human body fluids
Authors: Mac Neil S. , Dawson R. A. , Crocker G. , Barton C. H. , Hanford L. , Metcalfe R. , McGurk M. , Munro D. S.
Published: December 2008

In this study we describe the occurrence of a calmodulin-like protein in normal human biological fluids. Extraction of the calmodulin-like protein from breast milk, saliva, serum and urine provided an extract with enhanced calmodulin immunoreactivity which, in the case of milk and saliva, showed a protein band co-migrating with authentic calmodulin (Mr 17 000) on sodium dodecylsulphate-polyacrylamide gel electrophoresis. However, in milk, saliva and serum a major protein band of Mr 14 000–15 000 was always observed, which we speculate may be related to calmodulin, possibly as a partially degraded form.

Estimates of biologically active calmodulin in most normal extracellular fluids were of the order which we have found will stimulate cell division when added to the extracellular medium of cells in culture. Levels ranged from 0·03 nmol/l in urine to 18·6 nmol/l in breast milk, and exhibited a quantitative relationship (r = 0·79, P < 0·01) to epidermal growth factor (EGF) levels in fluids. Where EGF concentrations varied from normal (increased in saliva 24 h after oral surgery and reduced in the urine of patients with renal failure) calmodulin concentrations were similarly affected. The presence of calmodulin in serum may in part be attributable to its release from platelets which are particularly rich in calmodulin. Release of calmodulin from the platelet was associated with that of EGF and other platelet products.

J. Endocr. (1988) 118, 501–509
Cellular distribution of somatogenic receptors and insulin-like growth factor-I mRNA in the rat liver
Authors: van Neste L. , Husman B. , Möller C. , Andersson G. , Norstedt G.
Published: December 2008

Parenchymal and non-parenchymal cell fractions isolated from rat liver were analysed for the distribution of somatogenic receptors and for insulin-like growth factor-I (IGF-I) mRNA. The effect of hypophysectomy either alone or in combination with a single injection of human GH (hGH) on the levels of IGF-I mRNA was also studied in the two cell fractions.

The contamination of parenchymal cells in the non-parenchymal cell fraction was in the range of 2– 3%. Somatogenic receptors were found only in the parenchymal cell fraction. IGF-I mRNA was detected in both cell fractions, although the level of this mRNA was about fivefold higher in parenchymal cells. When RNA was studied by Northern gel analysis no major differences were observed in the size distribution of IGF-I transcripts in the two cell fractions.

In hypophysectomized animals, the IGF-I mRNA level was decreased to 10 and 30% of control values in the parenchymal and non-parenchymal cell fractions respectively. Treatment of hypophysectomized animals with a single dose of hGH restored IGF-I mRNA in parenchymal and in non-parenchymal cells to the extent found in intact animals.

In conclusion, our data indicate that somatogenic receptors are exclusively, and IGF-I mRNA predominantly, expressed in liver parenchymal cells compared with a total non-parenchymal cell fraction. The most marked effect of GH, correlating with the presence of somatogenic receptors, was seen in the parenchymal cells. The minor non-GH dependent expression of IGF-I mRNA suggests that this peptide may elicit effects in addition to the well-established GH-dependent activity.

J. Endocr. (1988) 119, 69–74
Growth hormone (GH) secretion in the conscious rat: negative feedback of GH on its own release
Authors: Clark R. G. , Carlsson L. M. S. , Robinson I. C. A. F.
Published: December 2008

The negative-feedback effects of GH on its own secretion were studied in conscious male and female rats bearing indwelling double-bore venous cannulae. Intravenous infusions of human GH (hGH; 20–60 μg/h) or somatostatin (SS; 10 μg/h) were given while frequent serial microsamples of blood were withdrawn using an automatic blood-sampling system. In both sexes, i.v. infusions of hGH for 6 h inhibited endogenous GH secretory pulses, with a slow onset of the inhibition. There was no rebound GH secretion immediately following the removal of the hGH infusion, but spontaneous GH secretion gradually returned to normal. Infusions of hGH did not inhibit the pituitary GH response to repeated GH-releasing factor (GRF) injections (1 μg) given i.v. every 40 min to female rats. By contrast, infusions of SS, which also blocked spontaneous GH release, dramatically reduced the GH responses to serial GRF injections. When SS Infusions were stopped, the subsequent GRF-induced GH secretory responses were enhanced. These results show that GH can inhibit its own release when given by i.v. infusion to conscious male and female rats. Since GH responses to GRF are maintained during a GH infusion, the feedback effect of GH is unlikely to be exerted directly on the pituitary or by increasing SS release. Our results are consistent with the idea that GH feedback in the conscious rat involves an inhibition of GRF release.

J. Endocr. (1988) 119, 201–209
Neuroendocrine responsiveness to light during the neonatal period in the sheep
Authors: Ebling F. J. P. , Claypool L. E. , Foster D. L.
Published: December 2008

Circulating prolactin concentrations were monitored during the early postnatal period in sheep to evaluate their response to photoperiod. In the first experiment, male and female lambs were exposed from 1 week of age, with their mothers, to either long days (16 h light:8 h darkness; n = 15) or short days (8 h light:16 h darkness; n = 16) to test whether they could discriminate different day lengths. In both sexes, serum prolactin concentrations were higher on long than on short days during the first 7 weeks after birth. In the second experiment, female lambs (n = 21) were raised on long days from 2 weeks of age. The superior cervical ganglia were removed bilaterally at 4 weeks of age from 14 lambs to lesion the sympathetic innervation to the pineal gland, and thus ablate the nocturnal increase in pineal melatonin secretion. After surgery, serum prolactin concentrations on long days were significantly lower in ganglionectomized lambs than in the intact controls. In the third experiment, the amplitude of the night-time melatonin rise was artificially increased in female lambs (n = 8) between 2 and 7 weeks of age to adult levels. Unrestrained lambs were infused during the 8-h dark phase of each day with melatonin by means of a self-contained, computerized syringe-pump. Concentrations of circulating prolactin did not differ from those in uninfused lambs (n = 8) with lower endogenous nocturnal melatonin. These results reveal that the sheep can discriminate photoperiod cues during the early postnatal period, and suggest that the low-amplitude melatonin rhythm in the neonatal lamb is sufficient to mediate this response.

J. Endocr. (1988) 119, 211–218
In-vitro secretion of prolactin and growth hormone in the presence of melatonin by pituitary gland from mink kept under long or short days
Authors: Meunier M. , Brebion P. , Chêne N. , Servely J.-L. , Martinet L.
Published: December 2008

Mink anterior pituitaries were incubated in Medium 199 for up to 9 or 13 days. Biological activity of prolactin and GH was determined. Daily concentrations of prolactin and GH in the incubation medium were also measured by radioimmunoassay and radioreceptor assay. When females were kept under short days for several weeks before the experiment, a significant decrease in prolactin secretion by the anterior pituitary was observed as compared with that in females maintained under long days. In contrast, secretion of GH was not modified by the photoperiodic history of the animals. Pineal gland denervation by ablation of the superior cervical ganglia a few months before the experiment, or addition of melatonin to the incubation medium of anterior pituitaries from intact or ganglionectomized females, did not modify the secretion of prolactin and GH. The pituitary gland does not therefore seem to be a direct target site for melatonin in transducing the duration of daylength on the hypothalamo-pituitary axis.

J. Endocr. (1988) 119, 287–292
Hormonal regulation of DNA polymerase-β activity in the rat thyroid gland
Authors: Nagasaka A. , Yoshida S. , Nakai A. , Ohyama T. , Iwase K. , Ohtani S. , Nakagawa H. , Masunaga R. , Kato S. , Kawabe T. , Kataoka K.
Published: December 2008

Using hypophysectomized rats, it has been shown that DNA polymerase-β activity in the adrenal gland and testis is largely influenced by pituitary trophic hormones. Sucrose gradient centrifugation of thyroid extracts revealed three peaks of DNA polymerase-β activity sedimenting at 3·3S, 7·3S and 12S. Of these, hypophysectomy induced a decrease in the 3·3S DNA polymerase-β, whereas other molecular forms were affected only slightly. DNA polymerase-α and -γ activities were unaffected by hypophysectomy. These changes in DNA polymerase-β caused by hypophysectomy were reversed by daily i.p. injection of TSH. Furthermore, stimulation of the thyroid by excess TSH induced by the administration of 1-methyl-2-mercaptoimidazole resulted in an increase of all forms of thyroid DNA polymerase-β.

These results show that the level of DNA polymerase is relatively constant after hypophysectomy but that DNA polymerase-β in the rat thyroid gland is also modulated by TSH mainly through the change of activity of the polymerase-β which sediments at 3·3S.

J. Endocr. (1988) 119, 303–308
The rebound release of growth hormone (GH) following somatostatin infusion in rats involves hypothalamic GH-releasing factor release
Authors: Clark R. G. , Carlsson L. M. S. , Rafferty B. , Robinson I. C. A. F.
Published: December 2008

We have studied the rebound secretion of GH following short-term somatostatin (SS) infusions in conscious rats, using an automatic sampling system for withdrawing frequent microsamples of blood. Intravenous infusions of SS (5–50 μg/h per rat) inhibited spontaneous GH secretion, but when SS was withdrawn there was a large burst of rebound GH secretion. A sub-anaesthetic dose of urethane reduced such rebound bursts of GH, suggesting a hypothalamic involvement in rebound GH secretion. Passive immunization with an antibody against rat GH-releasing factor (GRF) attenuated the rebound GH secretory response to the withdrawal of an SS infusion (GH concentration during rebound secretion was 26±21 μg/l vs 475 ± 127 μg/l (mean ± s.e.m.), after 0·5 ml anti-GRF serum or non-immune serum respectively). The inhibition of GH rebound secretion was related to the dose of anti-GRF serum administered. Intravenous infusions of human GH (20– 100 μg/h per rat) also reduced the size of the rebound GH secretion following SS withdrawal, in both male and female rats. We suggest that the rebound GH secretion that follows SS withdrawal in vivo is caused mainly by a hypothalamic release of GRF. Exogenous GH inhibits SS-induced rebound GH secretion in the conscious rat, possibly by inhibiting hypothalamic GRF release.

J. Endocr. (1988) 119, 397–404
Inhibitory effect of neurohypophysial peptides on cyclic AMP accumulation by isolated hepatocytes of the trout
Authors: Lahlou B. , Fossat B. , Porthé-Nibelle J. , Bianchini L. , Guibbolini M.
Published: December 2008

Cyclic AMP levels were measured in freshly isolated hepatocytes of the rainbow trout. Compared with basal values, the average levels were increased up to 60 times in a dose-dependent manner either by mammalian glucagon (concentration range 1 nmol– 1 μmol/l; dose giving half maximum response (EC50) 0· 18 μmol/l) or by forskolin (concentration range 0·1–100 μmol/l; EC50 about 10 μmol/l). These stimulatory effects were partially inhibited by fish or mammalian neurohypophysial hormones used at relatively high concentrations (1–5 μmol/l). It is suggested that these results are evidence for the presence of V1-type receptors in fish hepatocytes. Together with previous results obtained with gills on the hormonal inhibition of adenylate cyclase activity, they suggest that teleost fish may possess only V1-type receptors (or two V1-related types), while the V2 receptors have evolved (or have become functional) in higher vertebrates.

J. Endocr. (1988) 119, 439–445
Influence of neonatal opioid blockade or injections of gonadotrophin-releasing hormone on the timing of puberty in female rats: correlation of opioid effects with occupation of hypothalamic μ-opioid receptors
Authors: Landymore K. M. , Wilkinson M.
Published: December 2008

Hypothalamic opioid peptides have been implicated in the timing of sexual maturation in several species. We have examined the effects of neonatal opioid blockade on the timing of puberty in the female rat and have compared these with the effects of neonatal GnRH injection. Intermittent naloxone (2·5 mg/kg) or GnRH (200 ng/100 g body wt) injected s.c. at 6-h intervals for the first 10 days of life only slightly advanced the mean day of vaginal opening (VO). However, the degree of precocity was significantly more marked in a subgroup of drug-injected rats. In contrast, injections of the long-acting opioid antagonist naltrexone (50 mg/kg) had no effect on the timing of VO.

The results suggested that the duration of opioid receptor blockade is critical in determining the degree of opioid antagonist effect. Therefore, additional studies were performed to compare receptor occupancy of naloxone and naltrexone in 9-day-old rat pups. An exvivo binding assay was utilized to determine the availability of hypothalamic opioid-binding sites at various intervals following a single s.c. injection of antagonist. The time-course of inhibition of tritium labelled [d-Ala2-N-Me-Phe4,Gly5-ol]-enkephalin ([3H]DAGO) binding (μ-opioid sites) revealed that naloxone occupies the μ-receptor for a relatively short period of time. Naloxone (2·5 and 50 mg/kg) produced extensive inhibition of [3H]DAGO binding at 30 min following injection but binding was 100% of control at 1 h and 3·5 h respectively. In contrast, hypothalamic binding in naltrexone-treated (50 mg/kg) pups did not reach 50% of control values until 12 h and 100% of control values at 20 h after injection. The results are consistent with naltrexone's role as a long-acting antagonist but indicate that this blockade is not sustained for 24 h as previously indicated by indirect assessment with opiate challenge tests. Our data demonstrate that the duration of μ-opioid receptor occupancy is an important consideration in attempting to understand why intermittent, but not more continuous, blockade of opioid receptors induces some degree of sexual precocity in female rats. Also of interest is the observation that anterior pituitary stimulation with GnRH, during the first 10 days of life, can modify the timing of sexual maturation.

J. Endocr. (1988) 119, 447–452