Histochemical and immunohistochemical properties of the CRF-granules and other "Gomori-positive" substances of the rat.

In adrenalectomized rats, histochemical and immunohistochemical properties of the following secretion products have been investigated: 1. CRF-granules in the outer layer of the median eminence; 2. neurosecretory material (NSM) in the supraoptico-hypophysial system of the hypothalamus; 3. secretory granules in the TSH-cells of the anterior lobe of the hypophysis; 4. secretory granules in the ependymal cells of the subcommissural organ (SCO); 5. beta-cell-granules in the islets of Langerhans in the pancreas. All these substances are characterized by their stainability with the so-called "Gomori method". The experiments have included studies into: a) the extractability of the substances by various solvents; b) the digestability of the substances by pepsin or trypsin; c) their histochemically detectable content of disulfide groups, arginine and periodic acid-Schiff (PAS) reactive carbohydrates; d) their reaction with porcine-neurophysin-II-antibodies. All substances exhibited a positive reaction for disulfide groups. Based on their solubility properties, their resistance to pepsin or trypsin, their respective content of PAS-reactive carbohydrates and their failure to react with anti-neurophysin serum the "Gomori-positive" granules in TSH-, SCO- and pancreatic beta-cells can be distinguished from one another and from CRF- and neurosecretory granules. In contrast, CRF-granules and NSM showed identical properties. Taking into consideration data from the biochemical and histochemical literature, the present findings suggest that CRF-granules and NSM consist of closely related biochemical substances.