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L. Shukovski, J. K. Findlay, and A. I. Smith

ABSTRACT

Acid extracts of bovine preovulatory granulosa cells and corpora lutea (CL) were subjected to high-performance liquid chromatography (HPLC) and found to contain two peaks of immunoreactive (ir) oxytocin (OT), one corresponding to authentic OT and the second eluting 8 min later. The second peak was more abundant than authentic irOT in preovulatory follicles and in the early CL, but became less abundant as the CL matured (mid luteal) and was close to the limit of detection in the late CL. This peak could be detected only by an OT antiserum which recognized both the biologically active form of OT, as well as the post-translational processing intermediate Gly10-extended oxytocin. A second more specific OT antiserum (OT-933) did not recognize the second peak as strongly. Further analysis of the second peak revealed a complex of OT bound to its neurophysin (NP-I) which could be dissociated under denaturing conditions. Furthermore, we were able to create this complex in vitro by combining the two materials together under acid conditions, similar to the pH predicted in secretory granules, but not under neutral conditions. Measuring irNP-I by radioimmunoassay showed a single peak with a similar retention time to the OT/NP-I complex, confirming the identity of the unknown peak. Incubation of CL slices in culture showed a time-related release of both OT and NP-I, with OT having a greater rate of release in the mid luteal CL.

These data suggest the presence of an OT/NP-I complex in the bovine preovulatory granulosa cells and CL, as well as the unbound peptide presumably within the secretory granules. The ratio of OT/NP-I complex and free peptide changes with ageing of the the CL, perhaps indicating regulated differences in the post-translational processing of the prohormone.

Journal of Endocrinology (1991) 128, 305–314

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P. VAN DER SCHOOT

In rats with 4- or 5-day reproductive cycles various responses to the blockade of ovulation with sodium pentobarbitone at pro-oestrus (or at pro-oestrus and on the next day) were compared. The blood concentrations of oestradiol decreased rapidly during the 24 h period after injection of sodium pentobarbitone at pro-oestrus in rats with 5-day cycles. In those with 4-day cycles this response took almost a day to develop. Injection of sodium pentobarbitone at pro-oestrus and on the next day interfered with ovulation of the present crop of large follicles in rats with 5-day cycles. In 4-day cyclic rats this procedure delayed ovulation of these follicles by 48 h. Receptive behaviour was absent on the day after the second injection of sodium pentobarbitone in rats with 5-day cycles; some receptivity was, however, induced by the injection of gonadotrophin. The latter injection resulted in the release of a low number of eggs; fertilization of these eggs, however, did not occur. In 4-day cyclic rats receptive behaviour was recorded on the day after the second injection of sodium pentobarbitone: fertilization of delayed ovulated eggs took place normally but pregnancy was seen only rarely.

The results indicated clear differences in responses to blockade of ovulation with sodium pentobarbitone between rats with 4- or 5-day cycles. The differences most probably result from a more advanced 'age' of preovulatory follicles at pro-oestrus of 5-day cycles compared with those of 4-day cycles. Experimental delay of ovulation reveals ageing changes and the probable onset of atresia at an earlier time after blockade in 5-day cyclic than in 4-day cyclic rats.

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T. A. Yarney, J. Z. Kendall, and G. C. B. Randall

ABSTRACT

The ontogeny of fetal lung glucocorticoid receptors and their regulation by the fetal pituitary, adrenal and thyroid gland during lung maturation were investigated. Sites with a specificity typical of glucocorticoid receptors were detectable in lung cytosol, with the order of potency of steroids being dexamethasone > cortisol > corticosterone > 11- -deoxycorticosterone > progesterone > 17α-hydroxyprogesterone > oestradiol -17β ≃ testosterone ≃ androstenedione ≃ oestrone. The binding affinity for [3H]dexamethasone was high (K d = 0·23–0·60 nmol/l) and showed an age-related decrease during the perinatal period when cortisol levels were high. After charcoal treatment of the cytosol, however, a decrease in binding affinity was not as clearly evident. The K d decreased following hypophysectomy of fetuses; thyroidectomy had no significant effect. The concentration of glucocorticoid receptors was high from day 82 to day 100 of gestation (1437 fmol/mg protein) and declined progressively to a lower value at term and following birth (660 fmol/mg protein). Hypophysectomy, but not thyroidectomy, prevented the age-related decline in receptor concentration. Lung glycogen content declined with fetal ageing in association with increases in plasma concentrations of cortisol and thyroxine and with changes in K d and B max, but appeared to be more closely associated with concentrations of thyroxine. Hypophysectomy of fetuses decreased concentrations of both cortisol and thyroxine and prevented the depletion of lung glycogen content. Preliminary results from thyroidectomized fetuses showed decreases in plasma thyroxine and lung glycogen content compared with day-82 fetuses. Plasma cortisol levels, however, were consistent with a fetal age of 113 days. The effects of thyroxine on lung glycogen depletion, therefore, appear to occur, at least in part, through a pathway independent of glucocorticoid receptors.

Journal of Endocrinology (1990) 127, 341–349

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F. T. A. Fitzpatrick and B. D. Greenstein

ABSTRACT

The effects of several steroids on the regenerating thymus in ageing male rats have been studied. Rats aged from 12 to 15 months were orchidectomized and 7 days later implanted s.c. with silicone elastomer tubing containing 25 mg testosterone, 5α-dihydrotestosterone (DHT), oestradiol, progesterone or corticosterone. One group of rats received an empty implant. Thirty days later the rats were killed and the thymus, spleen, ventral prostate and seminal vesicles weighed and retained for histology. Whole blood was taken for total and differential white cell counts; plasma was prepared for radioimmunoassay of testosterone, oestradiol, progesterone and corticosterone.

After orchidectomy only, a multilobular thymus was present, and histologically the tissue appeared healthy. In testosterone- and oestradiol-treated rats, thymus weight was reduced to about 50% of that in untreated animals. Histologically, much of the thymus taken at autopsy was fat and what remained was poorly organized and contained a much lower density of thymocytes. The total white cell count was significantly reduced in these animals, the effect appearing to be predominantly on lymphocytes. Although treatment with DHT also resulted in a lower mean thymus weight than that of orchidectomized animals, histologically the tissue appeared similar to that of the untreated castrated animals. In rats treated with DHT, the total white cell count was significantly higher than in testosterone-implanted rats. Both progesterone and corticosterone implants resulted in significantly smaller mean thymus weights, although these steroids were not as potent as testosterone or oestradiol. Corticosterone, but not progesterone, appeared to cause a significant reduction in circulating lymphocytes. Dihydrotestosterone possessed only half the potency of testosterone in restoring the weights of the accessory sex organs. Serum concentrations of testosterone in orchidectomized old rats were 0·33 ± 0·02 nmol/l and in testosterone-implanted rats 4·8 ± 0·4 nmol/l. These results raise the possibility that testosterone and oestradiol may have caused atrophy of the thymus, while DHT may have retarded regeneration of the thymus without any atrophic effect. It remains to be seen whether the different responses between testosterone and DHT, in both the thymus and accessory sex organs, are due to differences in intrinsic action or differences in the metabolism of the steroids.

J. Endocr. (1987) 113, 51–55

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H. J. CHEN and P. G. WALFISH

SUMMARY

Old male rats of 22–24 months and young ones of 3–5 months were studied to find the effects of ageing, of orchidectomy and of orchidectomy and treatment with testosterone propionate (TP) on the basal serum concentrations of thyrotrophin (TSH) and on the total and free concentrations of tri-iodothyronine (T3) and thyroxine (T4) in the serum. The changes in TSH after treatment with thyrotrophin releasing hormone (TRH) were also observed. Intact old rats had significantly (P < 0·001) lower basal T4 and T3 as well as lower (P < 0·05) testosterone concentrations than were present in young rats. They also had higher basal TSH and per cent free T4 but lower absolute free T3 concentrations than had young rats. Two weeks after orchidectomy, basal TSH concentrations were slightly but significantly (P < 0·05) decreased in both young and old rats while T4 decreased significantly (P < 0·05) only in the young. The responses of TSH to TRH were also reduced by orchidectomy in both age groups with the old rats being less responsive than the young. Orchidectomy and treatment with pharmacological doses of TP produced similar effects on the pituitary-thyrotrophic response for both old and young rats but a greater effect occurred in the basal T4 response in young rats. In all groups basal TSH was influenced by orchidectomy or by treatment with TP but was always higher in the aged rat. Tri-iodothyronine concentration was always lower in the older rat and was not altered by orchidectomy or by treatment with TP in either young or old rats.

These results indicate that (1) in the male rat these age-specific effects on the thyroid–pituitary system are probably due, not only to a reduction in thyroid gland function and plasma T4 protein-binding, but also to a concomitant hyporesponsiveness of the aged male rat pituitary thyrotroph to TRH stimulation and (2) there is probably a significant influence of testicular function on the pituitary–thyroid system of the male rat.

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A M Solomon and P M G Bouloux

biology has become increasingly sophisticated – of special interest are the influence of mechanical stimulation, ageing and endocrine factors ( Dhawan & Rando 2005 , Wagers & Conboy 2005 , Wozniak et al. 2005 , Scime & Rudnicki 2006 , Sherwood

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Zuzana Saidak, Carole Le Henaff, Sofia Azzi, Caroline Marty, and Pierre J Marie

Introduction Bone loss associated with ageing is characterised by decreased bone formation relative to bone resorption, resulting in altered bone microarchitecture, osteoporosis and increased risk of fractures ( Khosla et al . 2011 ). The age

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Cheryl A Conover, Megan A Mason, James A Levine, and Colleen M Novak

and genetic factors influence tissue-specific decline in ageing C. elegans . Nature 419 808 – 814 . Holzenberger M Dupont J Ducos B Leneuve P Geloen A Even PC Cervera P Le Bouc Y 2003 IGF-1 receptor regulates lifespan and

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Mathis Grossmann

Introduction: significance of the clinical problem Around 50% of ageing, obese men presenting to the diabetes clinic have lowered testosterone levels relative to reference ranges based on healthy young men ( Grossmann 2011 ). Many have symptoms

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Lucie E Bourne, Caroline PD Wheeler-Jones, and Isabel R Orriss

and the heart valves. It is a common consequence of atherosclerosis, diabetes, chronic kidney disease (CKD) and ageing and is associated with an increased risk of having a future adverse cardiovascular event (e.g. heart attack, stroke) ( Zhu et al