A Rapid and Sensitive Method for Bioassay of the Adrenal Cortical Hormone

Selye, Hans; Schenker, Victor

doi:10.3181/00379727-39-10257

Cited by 49 publications

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“…The venous blood from the suprarenals was collected as previously described [Vogt, 1943]. The plasma prepared from this blood was assayed for cortical activity by hypodermic injection into groups of suprarenalectomized rats exposed to cold [method of Selye & Schenker, 1938]. The prolongation of the mean survival time of such groups of rats by the administration of plasma measures its content of cortical hormone.…”

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Observations on some conditions affecting the rate of hormone output by the suprarenal cortex

Vogt¹

1944

The Journal of Physiology

142

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Considerable amounts of cortical hormone are continuously released into the circulation by the suprarenals, and can be detected and assayed in the blood collected from the glands [Vogt, 1943]. The present paper deals with the effect of various factors on the rate of this secretion of cortical hormone. The experiments were mainly concerned with the influence of adrenaline and of splanchnic stimulation. Since both these conditions are liable to cause changes in arterial blood pressure and in blood flow through the glands, preliminary experiments had to ascertain the effect, if any, of these two factors on the output of cortical hormone. METHODSThe experiments were done on heparinized, eviscerated dogs, except for two instances, in which cats were used. Anaesthesia was induced with ether and completed with intravenous chloralose. The venous blood from the suprarenals was collected as previously described [Vogt, 1943]. The plasma prepared from this blood was assayed for cortical activity by hypodermic injection into groups of suprarenalectomized rats exposed to cold [method of Selye & Schenker, 1938]. The prolongation of the mean survival time of such groups of rats by the administration of plasma measures its content of cortical hormone. Details will be found in the earlier publication.There are two ways of following the minute output of hormone throughout an experiment. The first is to collect consecutive samples of equal volume.If appreciable changes in blood flow take,place, the periods of collection will be of different length and the output per minute will have to be found by calculation. Since the assay is not strictly quantitative, this introduces an error, and the method, accordingly, was only used in a few preliminary experiments. The alternative is to make the collection times equal, irrespective of the volumes of the samples. In this case, the assay measures the output without any corrections. Since, however, injection into the rats of unequal PH. CIII. 21

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Observations on some conditions affecting the rate of hormone output by the suprarenal cortex

Vogt¹

1944

The Journal of Physiology

142

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“…This fact determined the minimum amount of adrenal plasma required per assay; it was of the order of 10 ml. and thus no smaller than the require¬ ments of the assay according to Selye & Schenker [1938, see also Vogt, 1943 where young adrenalectomized rats are exposed to low environmental temperature and their survival times measured ('cold test').…”

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Parallel Biological Assays of the Cortisone Equivalent of the Dog's Adrenal Vein Blood

Bibile¹,

Vogt²

1953

J Endocrinol

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The rate of secretion of the adrenal cortex of the anaesthetized dog as measured by the mouse eosinophil test is equivalent to about 11 \ g=m\ g cortisone/g fresh tissue/min.When the cortex of the dog's perfused adrenal is stimulated by ACTH, the increased rate of secretion is registered by the eosinophil test in the same way as by the 'cold test'.When the perfused adrenal was subjected to an increased blood concentration of potassium, only small fluctuations in hormone output were produced. These also were registered by both tests simultaneously.On the assumption that mineralocorticoids are not oxygenated in the C-11 position, these results may be interpreted to mean that, within the experimental errors of the assays, there is no variation in the production of (hypothetical) mineralocorticoids under the conditions of these perfusions.When Speirs & Meyer [1949] published their paper on the use of eosinophil counts in mice for the estimation of adrenocortical hormones, it appeared that their test would be much more sensitive than any known biological assay and permit the estimation of corticoids in arterial blood. This hope, however, has been disappointed, and our work had to be hmited to the estimation, in terms of cortisone, of the cortical hormone in extracts of dog adrenal venous blood under varying conditions. Furthermore, tests were carried out in parallel with a second biological assay, the so-called 'cold test'. The object of parallel assays is to get evidence for or against the identity of a biologically active substance. In this instance, it is known that eosinophils respond exclusively to corticoids with an oxygen atom in the C-l 1 position, whereas the response in the cold test is less specific and can be ehcited by substances without that grouping, for instance, by esters of deoxycorticosterone (DOC) [Vogt, 1951]. For a long time, it has been assumed that the adrenal gland secretes specific 'mineralocorticoids', and though it is no longer considered likely that DOC is a natural mineralocorticoid, the work of Spencer [1950] and of Tait, Simpson & Grundy [1952] and Simpson, Tait & Bush [1952] suggests the secretion of a mineralocorticoid of as yet unknown constitution. Provided this new compound or any other active secretory products of the adrenal cortex were not oxygenated in the C-ll position, they might influence survival time in the ' cold test ', while leaving the eosinophils un¬ affected. The secretion of such compounds might then find expression in discrepancies in parallel biological assays by the two methods. The test for discrepancy was made by comparing the results for each individual sample of plasma in terms of cortisone, further, by altering the rate of secretion and observing whether direction and magni¬ tude of change was reflected in both tests in an identical fashion. In order to alter the secretion rate in a way which might be expected to favour the formation of mineralo-

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“…These findings have been used by Selye & Schenker [1938] and others [Vogt, 1943;Dorfman, Shipley, Schiller & Horwitt, 1946] in developing bioassay procedures for the determination of adrenocortical preparations. These findings have been used by Selye & Schenker [1938] and others [Vogt, 1943;Dorfman, Shipley, Schiller & Horwitt, 1946] in developing bioassay procedures for the determination of adrenocortical preparations.…”

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A Method for Screening Compounds for Adrenocortical Activity Using the Resistance of Mice to the Effect of Cold

Pf¹

1957

Journal of Endocrinology

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1. Adrenalectomy in the mouse causes a decrease in its resistance to the effects of cold; this resistance can be completely restored by the administration of cortisone acetate and other 11-oxygenated corticosteroids. A linear relationship exists between the survival time of adrenalectomized mice exposed to cold and the logarithm of the dose of cortisone acetate or other corticosteroids injected.2. The assay depending upon the resistance of mice to cold is simple and easier to perform than assays using either the response of the eosinophils or of liver glycogen in mice. The sensitivity of the method is less than that of the latter assays. Its precision is less than that of the glycogen assay, but is of the same order as that of the eosinophil assay. Compared with the similar assay using rats it is cheaper to perform, requires less time and has a similar order of precision and sensitivity.3. The assay depending upon the resistance of mice to cold provides a simple and rapid means of screening compounds for adrenocortical activity.It has been well established that adrenalectomy reduces the resistance of rats to high or low temperatures, and that the resistance can be restored by the administration of adrenocorticoids. These findings have been used by Selye & Schenker [1938] and others [Vogt, 1943;Dorfman, Shipley, Schiller & Horwitt, 1946] in developing bioassay procedures for the determination of adrenocortical preparations. Publica¬ tions on the use of the adrenalectomized mouse in experiments involving exposure to cold have been very few, and on the basis of the evidence so far available it has been considered that mice are less suitable than rats [Widström, 1935 ;Dorfman et al. 1946].However, in other assays of adrenocortical hormones the mouse has proved to be more suitable than the rat; for example, Speirs & Meyer [1949, 1951], Bibile [1953] and Bibile & Vogt [1953] used the adrenalectomized mouse with success in an assay based on the fall in the number of eosinophils in the circulation. Moreover, Venning, Kazmin & Bell [1946] and Eggleston, Johnston & Dobriner [1946] replaced the rat by the mouse in the assay using liver glycogen and greatly increased its sensitivity. As the mouse was successfully used for the two latter assays it was considered that its suitability for the assay depending upon resistance to cold should be re-investigated.

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A Rapid and Sensitive Method for Bioassay of the Adrenal Cortical Hormone

Cited by 49 publications

References 0 publications

Observations on some conditions affecting the rate of hormone output by the suprarenal cortex

Observations on some conditions affecting the rate of hormone output by the suprarenal cortex

Parallel Biological Assays of the Cortisone Equivalent of the Dog's Adrenal Vein Blood

A Method for Screening Compounds for Adrenocortical Activity Using the Resistance of Mice to the Effect of Cold

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