Open AccessConditionally immortalized adrenocortical cell lines at␣undifferentiated␣states exhibit inducible expression of␣glucocorticoid‐synthesizing genes
Author(s) -
Mukai Kuniaki,
Nagasawa Hideko,
AgakeSuzuki Reiko,
Mitani Fumiko,
Totani Keiko,
Yanai Nobuaki,
Obinata Masuo,
Suematsu Makoto,
Ishimura Yuzuru
Publication year - 2002
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.0014-2956.2002.02624.x
Subject(s) - zona glomerulosa , zona fasciculata , steroid 11 beta hydroxylase , aldosterone , aldosterone synthase , biology , corticosterone , adrenal cortex , steroidogenic acute regulatory protein , cell culture , cholesterol side chain cleavage enzyme , endocrinology , medicine , cellular differentiation , microbiology and biotechnology , gene expression , gene , messenger rna , steroid , renin–angiotensin system , angiotensin ii , biochemistry , hormone , genetics , blood pressure
To facilitate studies on differentiation of adrenocortical cells and regulation of steroidogenic genes, we established cell lines from adrenals of adult transgenic mice harboring a temperature‐sensitive large T‐antigen gene of simian virus 40. Adrenal glands of the mice exhibited normal cortical zonation including a functionally undifferentiated cell‐layer between the aldosterone‐synthesizing zona glomerulosa cells and the corticosterone‐synthesizing zona fasciculata cells. At a permissive temperature (33 °C), established cell lines AcA201, AcE60 and AcA101 expressed steroidogenic genes encoding steroidogenic factor‐1, cholesterol side‐chain cleavage P450scc, and steroidogenic acute regulatory protein, which are expressed throughout adrenal cortices and gonads. Genes encoding 3β‐hydroxysteroid dehydrogenase and steroid 21‐hydroxylase P450c21, which catalyze the intermediate steps for syntheses of both aldosterone and corticosterone, were inducible in the three cell lines in temperature‐ and/or dibutyryl cAMP‐dependent manners. Notably, these cell lines displayed distinct expression patterns of the steroid 11β‐hydroxylase P45011β gene responsible for the zone‐specific synthesis of corticosterone. AcA201 cells expressed the P45011β gene at 33 °C, showing the property of the zona fasciculata cells, while AcE60 cells expressed it upon a shift to a nonpermissive temperature (39 °C). On the other hand, AcA101 expressed the P45011β gene at 39 °C synergistically with exposure to dibutyryl cAMP. None of these clones express the zona glomerulosa‐specific aldosterone synthase P450aldo gene under the conditions we tested. These results show that AcE60 and AcA101 cells display a pattern of the steroidogenic gene expression similar to that of the undifferentiated cell‐layer and are capable of differentiating into the zona fasciculata‐like cells in vitro .