AN APPLICATION OF IN SITU HYBRIDIZATION AND ANTISENSE OLIGONUCLEOTIDE METHOD TO VISUALIZE AND MANIPULATE GENE EXPRESSION OF STEROID HORMONE RECEPTOR PROTEINS IN THE NERVOUS SYSTEM

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Chromaffin cells were found to contain glucocorticoid receptor-immunoreactivity in their nucleus. When dexamethasone was added into the culture medium, the extension of the processes, intensity of immunoreactivity of MAP-2 were significantly diminished. These results indicated that the differentiation of chromaffin cells to neurons is determined, at least in part, by humoral factor of glucocorticoid through its receptor. The addition of antisense oligos for glucocorticoid receptor cleary caused the significant extension of processes of chromaffin cells, indicating that the protein synthesis of steroid receptor was blocked by the antisense inhibitory action to its mRNA.In the sexually dimorphic area of the hypothalamus, immunocytochemical studies show that estrogen induces a significant changes of numbers of immunoreactive never fibers containing neurotensin, calcitonin-gene related peptide (CGRP), Met-enkephalin, cholecystokinin-8 (CCK-8) and dopamine, indicating that axonal sprouting … More of neurons containing these chemical messengers occur in response to estrogen. The relationship between estrogen-receptor and gene expression of these biochemical messengers in concomitant with the process formation or elongation has been discussed, particularly in the context of male and female differences. Estrogen-receptor was found to localize in the cell nucleus by immunocytochemistry in conjunction with the use of estrogen-receptor antibody. Colchicine treatment blocked immunpositive reaction in the cell nucleus, suggesting that estrogen-receptor protein was synthesized in the cytoplasm followed by the transport to the nucleus through nuclear pore in association with microtubules. Monolayr cultures from male and female rat hypothalami were developed and the differences of neurite extension was investigated. In this point, it should be noted that developmental and estrogen-induced changes in mRNAs coding specific proteins for the cytoskeleton, including tubulin-like molecules, have been indentified in the hypothalamic preoptic area. Less