Functional implication of autophagy in steroid-secreting cells of the rat.

Abstract

Autophagy, while frequently observed in embryonic cells undergoing differentiation and in pathologically altered cells, appears to occur less commonly in normal, fully differentiated cells. Our previous work revealed that the frequency of autophagic activity was rather high in the Leydig cells of rat testes, but the functional significance of autophagy in Leydig cells remains obscure. The purpose of the present study is to investigate the possible role of autophagy in steroid-secreting cells. The autophagic activity was investigated in two steroid-secreting cells, e.g., Leydig cells and adrenocortical fasciculata cells of rats. Cytidine monophosphatase (CMPase) cytochemistry was utilized to show the activity of lysosomal enzymes in autophagosomes. Electron microscopic morphometry was employed to analyze the frequencies of autophagy in the cells of the rats intact or treated with related hormones resulting in a hyper- or hypo-secretion of testosterone and corticosterone. Autophagy took place in normal steroid-secreting cells with higher frequencies than in many other cells including the tubular cells of kidney and hepatocytes. The large number of autophagosomes or autophagic vacuoles allowed to outline the autophagic process in these cells. The C-shaped double-membrane profiles tending to demarcate a portion of cytoplasm were referred to as pre-autophagosomes. So-called early autophagosomes were the vacuoles enclosed completely by double delimiting membranes, containing normal-looking cellular components. The majority of sequestered organelles appeared to be mitochondria and smooth endoplasmic reticulum. The autophagosomes starting digestion were considered as late autophagosomes or autophagic vacuoles, the indications of which were the destruction of their contents or the presence of lysosomal enzymes demonstrated by a positive CMPase reaction. Residual bodies were frequently observed to be exocytosed. The quantitative assay revealed an alteration of autophagic activity in close relation with steroid-secreting states. The number of autophagosomes was one-fold higher in hyposecreting Leydig cells after 2 days testosterone administration, and three-fold higher in hyposecreting adrenocortical fasciculata cells after one dosage of dexamethasone administration. In addition, the autophagosomes showed a four-fold decrease in hypersecreting Leydig cells stimulated by LRH for 2 days. Considering that most of the autophagocytosed organelles were steroid-producing apparatus, we may conclude that, by removing part of steroid-producing organelles, autophagy might play a role in adapting to or even regulating the secretory activity. This hypothesis was strongly supported by the fact that the intensity of autophagy varied in company with the fluctuation of steroid secretion.