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J. D. Curlewis
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G. M. Stone
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ABSTRACT

Uterine weight, RNA, DNA, protein content, in-vitro rate of protein synthesis, cytosol oestrogen and progesterone receptors were examined after administration of oestradiol to ovariectomized animals and on days 0, 5, 9 and 13 of the non-pregnant cycle and day 13 of pregnancy. In ovariectomized animals, oestradiol increased uterine weight, RNA: DNA and protein: DNA ratios and the concentration of cytosol receptors for oestradiol and progesterone. During the oestrous cycle there was a linear increase in uterine weight and a significant effect of the corpus luteum on the weight of the ipsilateral uterus. Changes in RNA, DNA and protein content between days 0 and 5 were not observed, but between days 5 and 13 RNA: DNA and protein: DNA ratios increased and the DNA: tissue weight ratio decreased. Thus, cellular hypertrophy and/or increased metabolic activity rather than hyperplasia occur over this period, which is coincident with the known rise in plasma progesterone levels. The rate of in-vitro protein synthesis (per unit tissue protein) during the non-pregnant cycle was greatest at day 0. These changes in uterine metabolic activity were associated with alterations in cytosol receptor concentrations for both steroids. Cytosol progesterone receptor concentrations were highest at day 0 after which they declined to a minimum at day 13. Cytosol oestradiol receptor concentrations, however, rose between days 0 and 5 and then declined. Although lutectomy on day 8 of the cycle does not interfere with the development of a histologically normal luteal phase, high peripheral progesterone levels which occur after day 8 in intact animals are associated with major increases in uterine metabolic activity.

The unilateral effect of the corpus luteum on uterine weight was associated with a decrease in DNA: g tissue ratio and an increase in rate of in-vitro protein synthesis indicating hypertrophy and/or extracellular accumulation of secreted material as well as enhanced metabolic activity. There was a significant effect of pregnancy on uterine weight at day 13 and this was associated with an increase in DNA content of both uteri. There was a unilateral effect of pregnancy on RNA: DNA ratio and in-vitro rate of protein synthesis, but not on uterine weight.

J. Endocr. (1986) 108, 201–210

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J. D. Curlewis
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G. M. Stone
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ABSTRACT

Radioimmunoassays were established for the measurement of total androgens and the specific measurement of testosterone and 5α-dihydrotestosterone in peripheral plasma of the brush-tail possum. Androgen concentrations were measured in blood collected from indwelling jugular cannulae (i) to determine whether the normal pattern of androgen secretion in this species was episodic and (ii) to attempt to relate total androgen and the pattern of testosterone secretion to the changes previously reported in prostatic, but not epididymal, weight in the breeding season. Blood was collected from restrained animals at varying time-intervals during daylight hours and darkness. Despite an apparent good adaptation to the sampling procedure there was generally a progressive decline in plasma androgen level during the collection period. This was true for animals bled during or out of the breeding season. There was no significant seasonal effect on the androgen concentration in the initial blood sample. When less restraint was used, two of three animals showed fluctuations in androgen levels over the 7-h sampling period. Testosterone levels in blood obtained by cardiac puncture were four- to nine-fold higher than those of 5α-dihydrotestosterone but levels of these androgens in samples obtained during the breeding season were not significantly different from those obtained out of season. The results do not argue for a pulsatile release of testosterone in the possum but do demonstrate a marked capacity for changes in the peripheral androgen concentration. There was a poor correlation between testosterone and 5α-dihydrotestosterone levels and prostatic weight.

J. Endocr. (1985) 105, 63–70

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J. D. Curlewis
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M. Axelson
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G. M. Stone
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ABSTRACT

The quantitatively major steroid hormones in ovarian and adrenal venous plasma of the female brush-tail possum were identified by gas chromatography–mass spectrometry. The ovarian vein plasma samples all contained oestradiol and its concentration was highest during the pro-oestrous phase of the reproductive cycle. During this phase the concentration of progesterone was below the limit of detection but at day 13 of the oestrous cycle and pregnancy, the concentration of progesterone exceeded that of oestradiol. Cortisol and corticosterone were the major steroid hormones found in all adrenal vein samples with cortisol predominant. Androgens with a 3-oxo structure, if present, were below the limits of detection in all plasma samples. Radioimmunoassays for the measurement of progesterone and oestradiol in peripheral plasma were used to follow changes in the concentrations of these steroids during the reproductive cycle.

Progesterone in serial blood samples was low at oestrus, rose gradually until day 7 and then increased more rapidly to reach a peak level of 21–29 nmol/l at around day 13. Any differences between the pregnant and non-pregnant cycles were minor. Oestradiol was only detected around oestrus when levels were variable (53·3±20·92 (s.e.m.) pmol/l; n = 4). The results indicate that the reproductive cycle of the brush-tail possum is characterized by a single peak of oestradiol at around pro-oestrus followed by gradually increasing levels of progesterone. Pregnancy appears to have no influence on the circulating concentrations of oestradiol or progesterone.

J. Endocr. (1985) 105, 53–62

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D. DEXTER
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H. B. STONER
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H. N. GREEN
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SUMMARY

The antidiuretic effect of small doses of adenosine triphosphate has been confirmed in both the rat and the rabbit after intravenous, intraperitoneal and intramuscular injection. This action of adenosine triphosphate, which closely resembles that of 'pitressin', was shared, to a lesser degree, by related compounds possessing either a —6NH2 or a —P2O7 group.

The effect could not be explained in terms of pre-renal deviation of water, either by failure of absorption, or by the formation of an exudate at the site of injection. Nor could it be explained by an alteration in renal haemodynamics, since the blood pressure and glomerular filtration rate were unaffected by the intraperitoneal doses used. Noxious afferent stimuli were also excluded as causative factors.

The antidiuretic action of adenosine triphosphate was inhibited by removal of the neurohypophysis and was accompanied by a fall in the antidiuretic hormone content of the posterior lobe of the pituitary.

The plasma and urine of the adenosine triphosphate-treated rats contained an antidiuretic substance which behaved like 'pitressin'.

In the rabbit, doses of adenosine triphosphate below the threshold for intravenous injection, caused marked changes in the rate of water excretion on intracarotid injection. The direction of the response depended on the state of hydration in the same way as the response to 'pitressin'.

The antidiuretic effect of small doses of adenosine triphosphate is thought to be due to the liberation of antidiuretic hormone from the posterior lobe of the pituitary. After large doses of adenosine triphosphate the cardiovascular and the other changes accompanying the state of shock will contribute to the effect.

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D Stone
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A David
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F Bolognani
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PR Lowenstein
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MG Castro
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The transfer of genetic material into endocrine cells and tissues, both in vitro and in vivo, has been identified as critical for the study of endocrine mechanisms and the future treatment of endocrine disorders. Classical methods of gene transfer, such as transfection, are inefficient and limited mainly to delivery into actively proliferating cells in vitro. The development of viral vector gene delivery systems is beginning to circumvent these initial setbacks. Several kinds of viruses, including retrovirus, adenovirus, adeno-associated virus, and herpes simplex virus, have been manipulated for use in gene transfer and gene therapy applications. As different viral vector systems have their own unique advantages and disadvantages, they each have applications for which they are best suited. This review will discuss viral vector systems that have been used for gene transfer into the endocrine system, and recent developments in viral vector technology that may improve their use for endocrine applications - chimeric vectors, viral vector targeting and transcriptional regulation of transgene expression.

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