Attenuation of parasite cAMP levels in T. cruzi-host cell membrane interactions in vitro.
Previous investigations have shown that the adhesion of T. cruzi plasma membrane vesicles (PMV) to monolayers of host cell myoblasts and to immobilized heart muscle sarcolemma membranes (PAM) on polyacrylamide beads is mediated by the interaction of T. cruzi attachment sites with the muscarinic cholinergic and beta-adrenergic receptors of the host cell membrane. It has also been shown that this interaction is blunted by the specific antagonists of the mammalian receptors atropine and propranol, respectively. In the studies reported here, PAM also rapidly attached to swimming T. cruzi trypomastigotes in a complex, concentration-dependent fashion and binding isotherms showed that the equilibrium between free and bound PAM is rapidly reached within 2 minutes of incubation in physiologically balanced salt solutions. In this time frame, trypomastigote cAMP levels are significantly reduced from steady state values within 30 seconds of the addition of PAM in a buffer system containing a diesterase inhibitor. Maximal attenuation of cAMP levels was measured between 1 and 2 minutes of the addition of PAM to T. cruzi trypomastigotes. The degree of cAMP level attenuation was reduced by blocking PAM attachment with either atropine or propranol. On the basis of these results we propose that a likely pathway for the negative parasite signal generated upon adhesion of host muscle cell membranes to the surface of the flagellates is from the parasite's surface attachment sites directly to a Pertussis toxin sensitive inhibitory protein Gi, thereby blunting adenyl cyclase activity and cAMP formation.
Von Kreuter
B F
BF
Department of Pathology, Cornell University Medical College, New York, New York 10021.
Walton
B L
BL
Santos-Buch
C A
CA
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D037361:Adenylate Cyclase Toxin; D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D001285:Atropine / Q000494:pharmacology; D002448:Cell Adhesion / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism*; D002463:Cell Membrane Permeability; D002772:Cholera Toxin / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000096:biosynthesis*; D005033:Ethylmaleimide / Q000494:pharmacology; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000031:analogs & derivatives / Q000494:pharmacology; D006153:Guanosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D006325:Heart Atria / Q000166:cytology / Q000469:parasitology; D006790:Host-Parasite Interactions; D007545:Isoproterenol / Q000494:pharmacology; D037342:Pertussis Toxin; D011433:Propranolol / Q000494:pharmacology; D012508:Sarcolemma / Q000469:parasitology*; D013552:Swine; D013873:Thionucleotides / Q000494:pharmacology; D014349:Trypanosoma cruzi / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D037361:Adenylate Cyclase Toxin; D000067956:Adenylyl Cyclase Inhibitors; D013873:Thionucleotides; D010566:Virulence Factors, Bordetella; C034882:guanosine 5'-O-(1-thiotriphosphate); D006153:Guanosine Diphosphate; D005576:Colforsin; D016244:Guanosine 5'-O-(3-Thiotriphosphate); C023427:guanosine 5'-O-(2-thiodiphosphate); D001285:Atropine; D002772:Cholera Toxin; D011433:Propranolol; D000242:Cyclic AMP; D037342:Pertussis Toxin; D000262:Adenylyl Cyclases; D007545:Isoproterenol; D005033:Ethylmaleimide; D015056:1-Methyl-3-isobutylxanthine