In some fields of steroid endocrinology, it has long been accepted that the potency of a hormone may depend not only on its secretion rate and rate of hepatic clearance but also on specific metabolic transformation at the site of the target cell. The crucial role of tissue 5α reductase activity on the action of testosterone is a spectacular example (Peterson, ImperatoMcGinley, Gautier & Sturla, 1977). Curiously—it is easy to be wise after the event—this line of thought seems rarely to have been exercized in explanations of corticosteroid action. Clearly, the severity of diseases of corticosteroid excess, of which Cushing's and Conn's syndromes are the best known examples, is strongly correlated with the secretion rate of cortisol and aldosterone respectively. However, in some forms of hypertension where deranged corticosteroid action might be expected to provide an acceptable explanation (e.g. increased mineralocorticoid secretion in low-renin essential hypertension), no abnormalities of secretion
Dopamine is capable of modulating zona glomerulosa function. Of this there now seems little doubt. However, whether varying dopamine levels in vivo forms the basis of a realistic normal physiological control mechanism for aldosterone secretion is far from clear. Reviewers have been cautious (Ganguly, 1984) or enthusiastic (Sowers, 1984) depending on the choice of evidence and the weight given to individual studies, but some resolution of the uncertainty is pressing since aberrations in this as yet unproven relationship have been suggested as basic abnormalities in a number of forms of hypertensive disease.
Evidence for and against the dopamine–aldosterone relationship has been obtained using dopamine itself and antagonists or agonists of its action in whole animals and in tissue preparations. Initial impetus for the dopamine hypothesis came from the observation that the dopamine agonist, bromocriptine, inhibited the response of aldosterone to frusemide-induced sodium loss (Edwards, Thorner, Miall et al. 1975) although
J. COENEGRACHT and R. FRASER
1. A method is described of measuring the early thyroid 'clearance' by the technique of Berson, Yalow, Sorrentino & Roswit , with the results expressed as the percentage of extracellular fluid cleared per half hour. This measurement has satisfactorily segregated from a normal group: 10 out of 10 definitely and 11 out of 11 probably thyrotoxic, 11 out of 12 definitely myxoedematous and 1 out of 4 probably myxoedematous patients. The technique is relatively simple and rapid.
2. Measurement of the early thyroid clearance is closely correlated with the urinary 'T' index of thyroid uptake (for log values of each, r=0·90).
A. WILSON and R. FRASER
A method of estimating the peripheral plasma concentration of 11-deoxycorticosterone (DOC) in man by means of gas-liquid chromatography with electron capture detection is described. Purification requirements for samples and chromatographic media have been simplified by the use of a detector bypass valve which reduces the risk of detector contamination. For this reason the method is relatively short. There was no measurable blank using either plasma from an adrenalectomized subject or water. The normal range was found to lie between 4·1 and 17·2 ng/100 ml (mean 9·8). Dexamethasone administration to one subject resulted in a subnormal plasma DOC concentration.
P. A. MASON and R. FRASER
A method for determining the plasma concentrations of six major corticosteroids, aldosterone, 18-hydroxy-11-deoxycorticosterone (18-OH-DOC), corticosterone, deoxycorticosterone (DOC), cortisol and 11-deoxycortisol using gas–liquid chromatography with electron capture detection is described. Esterification of suitable derivatives of these compounds with heptafluorobutyric anhydride (HFB) allowed detection of quantities of steroid, ranging from 0·3 pg for androstenetrione HFB (from cortisol) to 2·3 pg for corticosterone HFB. No detectable reagent blank was obtained for any compound when water was used instead of plasma and this was also the case when plasma from an adrenalectomized subject was analysed, with the exception of 18-OH-DOC where a reproducible but negligibly small blank occurred. Coefficients of variation for replicate determinations ranged from 8% for corticosterone to 17% for aldosterone. Concentrations in a series of normal human plasma samples were as follows: aldosterone, 4·0–18·0 ng/ 100 ml; 18-OH-DOC, 20–160 ng/ml; corticosterone, 0·08–0·80 μg/100 ml; DOC, 2·8–16·0 ng/100 ml; cortisol, 2·5–10·0 μg/100 ml;and 11-deoxycortisol, 40·0–400·0 ng/100 ml. When seven normal subjects were treated with dexamethasone, concentrations of DOC, cortisol and 11-deoxycortisol fell to below the limit of the normal range, those of 18-OH-DOC and corticosterone were at the lower end of the normal range while the concentration of aldosterone was not significantly affected.
S. Harvey and R. A. Fraser
The refractoriness of guinea-pigs to the growth-promoting actions of exogenous GH has been suggested to be due to a deficiency or defect in tissue GH receptors or in GH-receptor gene expression. GH-receptor mRNA was, however, demonstrated by Northern blot analysis and by the polymerase chain reaction in extracts of guinea-pig liver, adipose tissue, brain, hypothalamus and pituitary gland. High-affinity, low-capacity binding sites for radio-labelled ovine GH were also demonstrated on the plasma membranes of guinea-pig liver and were similar to those in rat liver. These results demonstrate that the unresponsiveness of guinea-pigs to exogenous GH is not due to the absence of GH receptors.
Journal of Endocrinology (1992) 133, 357–362
S. Harvey and R. A. Fraser
'... by analogy to the situation with calcitonin, it appears worthwhile to look for PTH in the brain and for physiological and behavioral effects of the hormone in the central nervous system' (Gennari, 1988)
Parathyroid hormone (PTH)-like peptides, mRNA and degradative enzymes are present in hypophysiotropic regions of the hypothalamus, in which PTHbinding sites are located on neural membranes. Since exogenous PTH stimulates hypothalamic dopamine metabolism and the release of pituitary prolactin, PTH-like peptides in the hypothalamus may have neuroendocrine roles in the regulation of pituitary function. However, as PTH is produced peripherally and neurological disorders are symptomatic of hyperparathyroid disease states, parathyroidal PTH may also participate in the neuroendocrine control of the hypothalamo-pituitary axis.
PTH in the hypothalamo-pituitary axis
Unlike other peptides of the 'diffuse neuroendocrine system', PTH production was believed to be solely by the parathyroid gland (Rosenblatt et al. 1989), from which PTH
W. R. PITNEY and T. RUSSELL FRASER
1. Two in vitro tests are described for measuring the inhibitory potency of antithyroid drugs on enzymic and non-enzymic oxidative iodination of protein; one, a milk enzymic iodination test, and the other an enzyme-free peroxide iodination test.
2. Five recognized antithyroid drugs have been tested: 2-thiouracil, 2-carbethoxythio-methyl-glyoxaline, potassium thiocyanate, resorcinol and sulphathiazole. By means of the milk enzymic test they could be ranged in order of potency, as indicated by the molar concentrations required for 50 % inhibition. They could also be separated into different types by the speed with which they pass from minimal to maximal inhibition with rising molar concentration.
3. With the enzyme-free peroxide test, thiouracil, but not resorcinol, was found to be inert; with the enzymic test, both were nearly equivalent in potency.
SE Dickson, R Bicknell and HM Fraser
Vascular endothelial growth factor (VEGF) is essential for the angiogenesis required for the formation of the corpus luteum; however, its role in ongoing luteal angiogenesis and in the maintenance of the established vascular network is unknown. The aim of this study was to determine whether VEGF inhibition could intervene in ongoing luteal angiogenesis using immunoneutralisation of VEGF starting in the mid-luteal phase. In addition, the effects on endothelial cell survival and the recruitment of periendothelial support cells were examined. Treatment with a monoclonal antibody to VEGF, or mouse gamma globulin for control animals, commenced on day 7 after ovulation and continued for 3 days. Bromodeoxyuridine (BrdU), used to label proliferating cells to obtain a proliferation index, was administered one hour before collecting ovaries from control and treated animals. Ovarian sections were stained using antibodies to BrdU, the endothelial cell marker, CD31, the pericyte marker, alpha-smooth muscle actin, and 3' end DNA fragments as a marker for apoptosis. VEGF immunoneutralisation significantly suppressed endothelial cell proliferation and the area occupied by endothelial cells while increasing pericyte coverage and the incidence of endothelial cell apoptosis. Luteal function was markedly compromised by anti-VEGF treatment as judged by a 50% reduction in plasma progesterone concentration. It is concluded that ongoing angiogenesis in the mid-luteal phase is primarily driven by VEGF, and that a proportion of endothelial cells of the mid-luteal phase vasculature are dependent on VEGF support.
R. FRASER and V. H. T. JAMES
A double isotope derivative assay technique for the simultaneous estimation of aldosterone, corticosterone and cortisol in human peripheral plasma is described. With this method, concentrations of the corticosteroids in plasma from adrenalectomized subjects were not significantly different from zero. Further evidence of specificity for aldosterone was obtained by measuring the disappearance rate of a single intravenous injection of unlabelled aldosterone from peripheral plasma, by demonstrating a rapid rise in aldosterone concentration in normal subjects acutely depleted of sodium, and by a comparison of aldosterone levels in patients with primary aldosteronism before and after surgical treatment. A study has also been made of the variation of the ratio of cortisol to corticosterone in normal subjects and of the effect upon this ratio of corticotrophin administration and haemorrhage.