Stimulation by atrial natriuretic factor of cyclic GMP production in cultured anterior and intermediate pituitary tissues: evidence for a major contribution of proliferating nonendocrine cells.
Primary cultures of anterior and intermediate pituitary tissues were monitored immunocytochemically for the presence of endocrine and nonendocrine cells and simultaneously tested for their ability to produce cyclic GMP in response to atrial natriuretic factor (ANF). Cells cultured for 3 days and 6 days, in which nonendocrine (vimentin-positive) cells were found to rapidly overgrow the endocrine cells, showed a dramatic elevation in cyclic GMP production stimulated by ANF, with maximum stimulation 300-700% that seen in 1-day cultured cells. Also, ANF-induced accumulation of cyclic GMP in an enriched population of vimentin-positive cells appeared to closely match that triggered in a 3-day culture of anterior pituitary cells, emphasizing the major role played by nonendocrine cells and their ability to synthesize cyclic GMP. In contrast, in the homogeneous population of tumor corticotrophs AtT-20, there was a close relationship between cyclic GMP formation and cell density. It thus appears that contamination of primary cultures of anterior and intermediate pituitary tissues by proliferating nonendocrine cells (mainly fibroblasts), in which ANF-induced accumulation of cyclic GMP may be confused with that of the very secretory cells, leads to overestimation and masking of guanylate cyclase activity of endocrine cells.
Lutz-Bucher
B
B
Institut de Physiologie, URA CNRS 98, Strasbourg, France.
Schimchowitsch
S
S
Félix
J M
JM
Stoeckel
M E
ME
Koch
B
B
D000324:Adrenocorticotropic Hormone / Q000032:analysis; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000502:physiology*; D002452:Cell Count; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000096:biosynthesis*; D005455:Fluorescent Antibody Technique; D008297:Male; D010902:Pituitary Gland / Q000166:cytology / Q000378:metabolism*; D010903:Pituitary Gland, Anterior / Q000166:cytology / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013997:Time Factors; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000032:analysis; D000521:alpha-MSH / Q000032:analysis
D014746:Vimentin; D000521:alpha-MSH; D009320:Atrial Natriuretic Factor; D000324:Adrenocorticotropic Hormone; D006152:Cyclic GMP