Crotoxin, a phospholipase A2 neurotoxin from snake venom, interacts with epithelial mammary cells, is internalized and induces secretion.
Prolactin (PRL) induces liberation of arachidonic acid (AA) from phospholipids of lactating mammary epithelial cells and stimulates casein secretion. In order to investigate the possible involvement of phospholipase A2 (PLA2) activity in the hormonal control of casein secretion by PRL, we examined the effects of crotoxin, a PLA2 neurotoxin from snake venom, on mammary epithelial cells. Crotoxin is made of two subunits: a basic PLA2 with low toxicity (component B, CB) and an acidic, non-toxic and enzymatically inactive component A (CA) which enhances the pharmacological action of CB. While CA is inactive, the PLA2 subunit (CB) induces an accumulation of secretory products in the lumen of mammary acini, an extensive development of the Golgi apparatus. The secretion of newly synthesized casein is increased in the presence of CB and this effect is inhibited by nordihydroguaiaretic acid (NDGA) and caffeic acid, two inhibitors of the lipoxygenase pathway which also prevent stimulation of secretion by PRL. Further, CB transiently induces the release of radiolabelled AA from mammary tissues previously labelled with [14C]AA, the highest release being observed between 15 s and 5 min of contact with CB and CA. Immunofluorescence labelling by anti-CB antibodies of epithelial mammary tissues previously incubated with CA, CB or a combination of CA and CB indicates that CB binds to epithelial cells and is internalized, at least in part, and that CA enhances both CB binding and its internalization. These observations emphasize the involvement of PLA2 in the control of casein secretion and suggest that PLA2 acts intracellularly.
Ollivier-Bousquet
M
M
Laboratoire de Biologie Cellulaire et Moléculaire, INRA, Jouy-en-Josas, France.
Radvanyi
F
F
Bon
C
C
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D002364:Caseins / Q000378:metabolism*; D003439:Crotoxin; D005260:Female; D005455:Fluorescent Antibody Technique; D008321:Mammary Glands, Animal / Q000378:metabolism* / Q000648:ultrastructure; D009498:Neurotoxins; D010741:Phospholipases A / Q000502:physiology*; D054467:Phospholipases A2; D011388:Prolactin / Q000502:physiology; D011817:Rabbits; D051381:Rats; D011919:Rats, Inbred Strains
D002364:Caseins; D009498:Neurotoxins; D016718:Arachidonic Acid; D011388:Prolactin; D003439:Crotoxin; D010741:Phospholipases A; D054467:Phospholipases A2