Document Detail


Long-lived cytoplasmic factors that suppress adrenal steroidogenesis.
MedLine Citation:
PMID:  6280168     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Adrenal cells secrete steroids after stimulation with corticotropin (ACTH), whereas cells reconstructed by fusing adrenal cell nuclei to fibroblast cytoplasms are temporarily (2-3 wk) unresponsive to ACTH. In this report, we characterize this inhibition by using "cybrids" (cytoplasmic hybrids) isolated by either genetic selection or a new procedure that utilizes the fluorescence-activated cell sorter and the vital mitochondrial dye rhodamine 123. Such cybrids, which contain both adrenal and fibroblast cytoplasmic components, are unable to produce steroids, suggesting the existence of cytoplasmic inhibitory factors. In order to elucidate this cytoplasmic inhibition of steroidogenesis, techniques are described that test the contribution of fibroblast mitochondria to this phenomenon. The first technique utilizes purified mitochondria, isolated from chloramphenicol (CAP)-resistant fibroblasts, to confer CAP resistance on an otherwise sensitive adrenal cell. The resulting CAPr cells, termed mitochondrial transformants, are responsive to ACTH. The second technique utilizes a procedure for isolating small fragments of cytoplasm (microcytospheres) from fibroblasts. Microcytospheres, which do not contain mitochondria, are stained with rhodamine 18, a vital membrane dye, and then fused to unstained adrenal cells. The fusion products are then isolated with the fluorescence-activated cell sorter. Approximately 30% of the fusion products are inhibited in their ability to respond to ACTH. These results suggest that the fibroblast cytoplasm contains nonmitochondrial long-lived inhibitory factors that temporarily suppress steroidogenic function in adrenal cells.
Authors:
M A Clark; J W Shay
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  79     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1982 Feb 
Date Detail:
Created Date:  1982-06-14     Completed Date:  1982-06-14     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1144-8     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Adrenal Gland Neoplasms
Adrenal Glands / metabolism*,  ultrastructure
Adrenocorticotropic Hormone / pharmacology*
Cell Fusion
Cell Line
Cytoplasm / physiology*
Fibroblasts
Mitochondria / physiology
Steroids / biosynthesis*
Time Factors
Grant Support
ID/Acronym/Agency:
GM 29261/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Steroids; 9002-60-2/Adrenocorticotropic Hormone
Comments/Corrections

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