Document Detail


Coculturing posterior pituitary and GH3 cells: dramatic stimulation of prolactin gene expression.
MedLine Citation:
PMID:  8419149     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Recent evidence suggests that the posterior pituitary (PP), also called the neurointermediate lobe, regulates PRL release. We previously reported that cocultures of anterior pituitary and PP cells resulted in a 2- to 3-fold increase in PRL content and release. For this study we chose GH3 cells (a somatomammotroph tumor cell line) to determine whether coculturing GH3 with PP cells: 1) stimulates the release and cell content of PRL as compared with GH; 2) increases GH3 cell proliferation; and 3) affects PRL messenger RNA (mRNA) levels. Exp 1. GH3 cells (25,000 cells per well; 25K) were cocultured with PP cells (0K, 12.5K, or 25K) from male rats in serum-free media for 1, 2, 4, and 7 days; hormones were measured by RIA. Coculturing resulted in 5- to 10-fold increases in both media and cell PRL that were linear with time and dependent on the number of PP cells. In contrast, media GH increased only 1.5- to 2-fold, and GH cell content reduced by 75%. Exp 2. GH3, PP, and GH3 + PP cells were cultured for 1, 2, and 4 days and then incubated with [3H]thymidine for 5 h. The incorporation of [3H]thymidine in GH3 cells remained constant over time and showed a small, early increase in cocultures. In contrast, incubation of PP cells alone resulted in a 50- to 60-fold rise in [3H]thymidine incorporation from days 1-4 in culture. Exp 3. Cytoplasmic mRNA was determined by slot blot hybridization with 32P-labeled complementary DNA probes for PRL and GH. After coculturing 25K GH3 cells with 12.5K and 25K PP cells for 4 days, PRL mRNA levels increased 15- and 30-fold, respectively, whereas GH mRNA levels rose less than 2-fold. Neither PRL nor GH mRNA were detected in PP cells. Conclusions: 1) coculturing GH3 with PP cells dramatically stimulates PRL gene expression, synthesis, and release; 2) this response is specific for PRL, has little effect on GH, and is not due to increased GH3 cell proliferation; and 3) we speculate that a subpopulation of intermediate lobe cells, possibly with a proliferative capacity, is responsible for inducing these effects.
Authors:
A Corcia; R Steinmetz; J W Liu; N Ben-Jonathan
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Endocrinology     Volume:  132     ISSN:  0013-7227     ISO Abbreviation:  Endocrinology     Publication Date:  1993 Jan 
Date Detail:
Created Date:  1993-02-05     Completed Date:  1993-02-05     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0375040     Medline TA:  Endocrinology     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  80-5     Citation Subset:  AIM; IM    
Affiliation:
Department of Physiology and Biophysics, Indiana University School of Medicine, Indianapolis 46202.
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MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Northern
Cells, Cultured
DNA / biosynthesis
Gene Expression*
Growth Hormone / genetics,  metabolism
Male
Nucleic Acid Hybridization
Pituitary Gland, Posterior / physiology*
Prolactin / genetics*,  metabolism
RNA, Messenger / analysis,  metabolism
Rats
Rats, Wistar
Tumor Cells, Cultured
Grant Support
ID/Acronym/Agency:
DK-39551/DK/NIDDK NIH HHS; NS-13243/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/RNA, Messenger; 9002-62-4/Prolactin; 9002-72-6/Growth Hormone; 9007-49-2/DNA

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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