The immunological and cytological specificity of antiserum to adrenocorticotrophic hormone was established and the corticotroph defined at the light microscopic level by immunocytochemistry. Our observations substantiated recent reports by several investigators. By comparing thin methacrylate sections with adjacent 1 p-methacrylate sections immunochemically stained with anti-ACTH, the ACTH cell was identified at the ultrastructural level and its morphology studied. Using the criteria established at light and electron microscopic levels, the corticotroph was readily found in tissue which had been fixed in paraformaldehyde-osmium mixture and embedded in epoxy resin. Ultrastructural changes in this cell in response to experimentally designed alterations in t h e pituitary ACTH levels confirmed this identification. The ACT11 cell was likewise identified in glutaraldehyde-fixed material. Thyrotrophs were tentatively defined. Several ultrastructural studies responsible for conflicting reports in the literature were investigated. It was concluded that the corticotroph is a unique cell type and that granule size and morphology remain important criteria for adenohypophyseal cell identification regardless of the fixation used. The need for a definitive immunocytochemical study of thyrotrophs at the ultrastructural level was recognized.

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The 2A8 clone, a normal diploid rat anterior pituitary cell strain, was investigated by immunocytochemistry to determine the cell types into which the clonal cells differentiated in vivo and in vitro. The in vivo study was carried out by injecting the 2A8 clone preparation either into the hypothalamic region or under the kidney capsule. After thirty days the implants were removed and studied by immunocytochemistry. In vitro, many prolactin cells and a few growth hormone cells were found. In vivo, however, prolactin, growth hormone, ACTH and TSH cells and gonadotrophs were identified. We concluded that the 2A8 clone was multipotential. Since the gonadotrophs of the implants made in the hypothalamic region were larger and more plentiful than those in the kidney implants, and since gonadotrophs were lacking in the in vitro system, it appeared that the hypophysiotrophic environment was the most conducive to gonadotrophic differentiation and maintenance, and that the factor or factors necessary for cyto-differentiation were apparently present in the general circulation of the rat but absent in the growth medium of our culture cells.

Two rabbits and two guinea pigs were immunized with arginine vasotocin (AVT) conjugated to bovine albumin with glutaraldehyde. Only one preparation of antiserum (anti-G 7) was of value. Anti-G 7 immunochemically defined the same rat pineal cells previously reported as presumptive AVT cells. However, absorption of anti-G 7 with bovine albumin inhibited the staining of the pineal cells demonstrating that they contained an albumin-like substance. Positive immunochemical staining of the rat pars nervosa suggested that anti-G 7 contained antibodies able to react with arginine vasopressin (AVP). Loss of a positive reaction in the posterior lobe on absorption of anti-G 7 with AVT or AVP confirmed this. However, the addition of AVT to anti-G 7 failed to inhibit the immunochemical staining of the pineal cells. This study reports the presence of an albumin-like substance in pineal cells previously described as presumptive AVT cells, and discusses possible explanations for the inability of anti-G 7 to recognize immunocytochemically the native AVT molecule. Confirmation of AVT in the pineal gland by immunocytochemistry must await the availability of more specific antisera.