Release of secretory phospholipase A2 from rat neuronal cells and its possible function in the regulation of catecholamine secretion.
Here we show that secretory phospholipase A2 (sPLA2) that is immunochemically indistinguishable from type II sPLA2 is (i) stored in neuroendocrine cells, (ii) released in response to neurotransmitters or depolarization, and (iii) involved in the regulation of catecholamine secretion by these cells. Rat brain synaptic vesicle fractions contained PLA2 activity, which was neutralized completely by an antibody raised against rat type II sPLA2. sPLA2 immunoreactive with anti-(type II sPLA2) antibody was released from synaptosomes in response to depolarization evoked by a high concentration of potassium in the presence of Ca2+. Rat pheochromocytoma PC12 cells, which differentiated into adherent cells similar to sympathetic neurons in response to nerve growth factor, were used for the detailed analysis of the dynamics and function of sPLA2 in neuronal cells. Antibody against rat type II sPLA2 precipitated approximately 80% of the PLA2 activity in PC12 cell lysates. Transcript for type II sPLA2 was detected in PC12 cells by reverse transcriptase-PCR. When neuronally differentiated PC12 cells were stimulated with carbamylcholine or potassium, sPLA2 was released into the medium and reached a maximal approximately 40% release by 15 min. Inhibitors specific to type II sPLA2 suppressed catecholamine secretion by PC12 cells which had been activated by carbamylcholine. Furthermore, treatment of PC12 cells with exogenous type II sPLA2 alone elicited catecholamine secretion. These observations indicate that sPLA2 released from neuronal cells may regulate the degranulation process leading to release of neurotransmitters and are compatible with our earlier finding that this enzyme is involved in the degranulation of rat mast cells.
Matsuzawa
A
A
Faculty of Pharmaceutical Science, University of Tokyo, Japan.
Murakami
M
M
Atsumi
G
G
Imai
K
K
Prados
P
P
Inoue
K
K
Kudo
I
I
D000310:Adrenal Gland Neoplasms; D000818:Animals; D000906:Antibodies; D001921:Brain / Q000201:enzymology*; D002118:Calcium / Q000494:pharmacology; D002454:Cell Differentiation; D002458:Cell Fractionation; D002846:Chromatography, Affinity; D017931:DNA Primers; D005071:Evoked Potentials / Q000187:drug effects; D006706:Homeostasis; D015151:Immunoblotting; D007150:Immunohistochemistry; D007700:Kinetics; D008297:Male; D009414:Nerve Growth Factors / Q000494:pharmacology; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D009638:Norepinephrine / Q000378:metabolism*; D016716:PC12 Cells; D010673:Pheochromocytoma; D010741:Phospholipases A / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D054467:Phospholipases A2; D016133:Polymerase Chain Reaction; D051381:Rats; D017208:Rats, Wistar; D013572:Synaptic Vesicles / Q000201:enzymology*; D013574:Synaptosomes / Q000201:enzymology*; D014158:Transcription, Genetic
D000906:Antibodies; D017931:DNA Primers; D009414:Nerve Growth Factors; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium; D009638:Norepinephrine