Localization of Adrenal Cortical Hormones in the Adrenal Cortex of the Cat

Abstract

Recently Reichstein and his collaborators (Reichstein, 1 , 2 Reichstein and von Euw 3 ) have isolated 3 ketone steroid compounds— corticosterone, desoxycorticosterone, and dehydrocorticosterone, any one of which will, in pure form, maintain the life of adrenalectomized animals. They have shown that all the biological activity in an alcoholic extract of beef adrenals is confined to the ketone fraction of the extract, whereas the ketone-free fraction is without specific activity in adrenalectomized animals (Reichstein 4 ). All the active compounds so far isolated have been shown by Reichstein to possess 2 or 3 keto groups. They can be separated, with other keto compounds, from a crude extract by means of their reaction with semi-carbazide, and they will reduce ammoniacal silver solution. This latter reaction is due to the presence in all three compounds of a side chain bearing an α-hydroxy keto grouping.

In the present study these reactions, with others, have been applied to frozen sections of fresh or formol-fixed cat adrenals in an attempt to localize the hormones in the cortex. The ascorbic acid was removed from the sections by treating with iodine, indophenol, or oxygenated glycine-sodium carbonate buffer at pH 8.4. The sections were then placed in a solution of phenylhydrazine hydrochloride in deoxygenated sodium acetate-acetic acid buffer at pH 6.5-6 for a few hours. A distinct yellow band appeared in the outer portion of the fasciculata in the zone occupied by the large lipoid-rich cells called “spongiocytes” by Guieysse. 5 This yellow band appears to be due to the formation of phenylhydrazones with the carbonyl groups in the cortex. The phenylhydrazone formation was prevented by extracting the sections with acetone or alcohol, or by treating the sections with semicarbazide before immersing in phenylhydrazine.