Differential stain for the nuclei of the suprarenal cortical cells

Abstract

Since the production of an active cortical extract by Pfiffiier and Swingle ( '29)' several substances have been recovered from the suprarenal cortex. Wintersteiner and Pfiffner ('35, '36) have isolated seven physiologically inactive crystalline substances, and Mason, Meyers and Kendall ('36) five, one of which corresponds very closely with compound D of the Wintersteiner and Pfiffner group.Studies covering many years have revealed several important facts concerning the functions and relationships of the suprarenal cortex. Thus Jaffe ('24) found a possible relationship between 'it and the thymus. Klein ( '31) was able to promote growth of the male sex organs and retard the development of the female sex organs by the use of a cortical extract. Zalesky ( '34) found a definite relationship between the suprarenal cortex and the reproductive cycle in the thirteen-lined ground squirrel. Bourne ( '34) observed two distinct types of Golgi apparatus in the suprarenal cortical cells, one of which he regarded as the hypertrophied form of the other. Pottinger and Simonsen ('38, '38 a ) , working with the benzene discard of the Pfiffner and Swingle method of extraction, obtained a male sex-stimulating, and a female sex-repressing fraction, and orally active sex-maturation fractions, as well as a gonadotropic substance which was effective in restoring physiologic balance in allergic children.I n view of the finding of various substances in the suprarenal cortex, one might expect to find differential staining of 401 THE INATOXICAL RECORD, VOL. 74, NO. 4 402 D A N DYSART BAKERits cells also. After many efforts directed toward the investigation of this point, a staining method has been developed which does differentiate the nuclei of these cells, although it may not indicate a U the different cells in the cortex. This method also differentiates the cytoplasm to some extent .Many different fixatives were used without success before it was found that human, bat, dog and bovine suprarenal glands could be differentially stained after fixation in a mixture containing 9 parts saturated mercuric chloride to 1 part formalin, which need not be neutral. Human material stains well after fixation in the mercury-form01 mixture for 3 to 4 hours, followed by fixation in 10% formalin for 10 to 12 hours. Zcnlter-form01 was found to be the best kxative for general use.The best staining reactions were obtained by using acid fuchsin, orange G and aniline blue (National Aniline and Chemical Co., Inc.) after Ehrlich hematoxylin and potassium bichromate. Many other stains, including water and alcoholsoluble eosin, Bismarck brown Y, malachite green, carmine, aurantia, methylene blue, toluidin blue and methyl orange were first used, but without success. The nuclei of the cells of the suprarenal cortex of mice, rats, bats, cats, dogs, hogs, guinea pigs, rabbits and cattle have all been differentially stained by this method, as has the suprarenal cortex of man. Similar but not the same staining reactions were obtained in all the cortices examined. The nu...