Detection of membrane-bound guanylate cyclase activity in rat C6 glioma cells at different growth states following activation by natriuretic peptides.
We studied the activity and the ultracytochemical localization of membrane-bound guanylate cyclase (GC) after stimulation with rat atrial natriuretic peptide (rANP), porcine brain natriuretic peptide (pBNP), rat brain natriuretic peptide (rBNP), or porcine C-type natriuretic peptide (CNP) in rat C6 glioma cells during proliferation or following exposure of confluent cells to dibutyryl cyclic AMP (db-cAMP) or retinoic acid (RA). Under our experimental conditions all peptides were activators of GC as demonstrated by the accumulation of cGMP within cells. During proliferation of C6 cells, the amounts of cGMP remained approximately constant. However, at subconfluency, confluency and postconfluency, the GC reaction product was located at different sites in C6 cells. At subconfluency, GC reaction product was on membranes of protoplasmic extensions, at postconfluency, GC reaction product was in association with membranes of cell bodies, and at confluency, both localizations of GC reaction product were detected. Incubation of confluent cells in culture medium containing db-cAMP or RA induced the appearance of long and slender protoplasmic extensions. Under these conditions, the GC reaction product was localized exclusively to these processes. These data suggest that GC is differentially located depending on the state of growth of glial cells, and that in differentiating glial cells GC is preferentially located in cell processes.
Sorci
G
G
Department of Experimental Medicine and Biochemical Sciences, University of Perugia, Italy.
Spreca
A
A
Donato
R
R
Rambotti
M G
MG
D000818:Animals; D009320:Atrial Natriuretic Factor / Q000494:pharmacology*; D003994:Bucladesine / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D003470:Culture Media; D006152:Cyclic GMP / Q000378:metabolism; D005910:Glioma / Q000201:enzymology* / Q000648:ultrastructure; D006162:Guanylate Cyclase / Q000378:metabolism*; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D020097:Natriuretic Peptide, Brain; D020098:Natriuretic Peptide, C-Type; D009419:Nerve Tissue Proteins / Q000494:pharmacology*; D011506:Proteins / Q000494:pharmacology*; D051381:Rats; D013552:Swine; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D003470:Culture Media; D009419:Nerve Tissue Proteins; D011506:Proteins; D020097:Natriuretic Peptide, Brain; D020098:Natriuretic Peptide, C-Type; D006165:Guanylyl Imidodiphosphate; D014212:Tretinoin; D003994:Bucladesine; D009320:Atrial Natriuretic Factor; D006162:Guanylate Cyclase; D006152:Cyclic GMP