Phospholipid hydrolysis and loss of membrane integrity following treatment of rat brain synaptosomes with beta-bungarotoxin, notexin, and Naja naja atra and Naja nigricollis phospholipase A2.
The effects of the phospholipase A2 (PLA2) toxins, beta-bungarotoxin and notexin, and the PLA2 enzymes from Naja naja atra and Naja nigricollis snake venoms on the plasma membrane integrity of synaptosomes were examined. Synaptosomes were isolated from rat brain cerebral cortex, corpus striatum and hippocampus. Osmotic activity, lactate dehydrogenase leakage, and leakage of 2-deoxy-D-(1-3H)-glucose-6-phosphate were monitored (37 degrees C, 10-120 min) following incubation with 0.5, 5 and 50 nM concentrations of toxins and enzymes. Damage to the synaptosomal plasma membrane was time and concentration but not tissue dependent. The potencies of the treatments were as follows: N. n. atra PLA2 greater than or equal to N. nigricollis PLA2 greater than notexin greater than beta-bungarotoxin. Chelation of Ca2+ with 5 mM EDTA completely inhibited plasma membrane disruption caused by beta-bungarotoxin and N. n. atra PLA2. One mg/ml of bovine serum albumin also blocked the disruptive action of N. n. atra PLA2, while 8 mg/ml was required to antagonize beta-bungarotoxin. A correlation between phospholipid hydrolysis and loss of membrane integrity was also observed. The generation of phospholipid hydrolytic products may be critical in the permeabilization of synaptic plasma membranes by these toxins and enzymes, however, they do not explain the presynaptic specificity and potency of beta-bungarotoxin and notexin.
Yates
S L
SL
Section of Pharmacology and Toxicology, University of Connecticut, School of Pharmacy U-92, Storrs 06269.
Burns
M
M
Condrea
E
E
Ghassemi
A
A
Shina
R
R
Rosenberg
P
P
D000818:Animals; D001921:Brain / Q000187:drug effects / Q000648:ultrastructure*; D002038:Bungarotoxins / Q000633:toxicity*; D002118:Calcium / Q000502:physiology; D002462:Cell Membrane / Q000187:drug effects*; D003840:Deoxycholic Acid / Q000494:pharmacology; D004546:Elapid Venoms / Q000633:toxicity*; D006868:Hydrolysis; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D009994:Osmolar Concentration; D010741:Phospholipases A / Q000633:toxicity; D054467:Phospholipases A2; D010743:Phospholipids / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013574:Synaptosomes / Q000187:drug effects*
D002038:Bungarotoxins; D004546:Elapid Venoms; D010743:Phospholipids; D003840:Deoxycholic Acid; C005082:notexin; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium