Cultured rat microglial cells synthesize the endocannabinoid 2-arachidonylglycerol, which increases proliferation via a CB2 receptor-dependent mechanism.
Microglia, as phagocytes and antigen-presenting cells in the central nervous system, are activated in such disease processes as stroke and multiple sclerosis. Because peripheral macrophages are capable of producing endocannabinoids, we have examined endocannabinoid production in a macrophage-colony stimulating factor (M-CSF)-dependent rat microglial cell line (RTMGL1) using reversed phase high-pressure liquid chromatography and liquid chromatography-mass spectroscopy. We determined that cultured microglial cells produce the endocannabinoid 2-arachidonylglycerol (2-AG) as well as anandamide in smaller quantities. When 2-AG, but not anandamide, is added exogenously, RTMGL1 microglia increase their proliferation. This increased proliferation is blocked by an antagonist of the CB(2) receptor N-[(1S)endo-1,3,3-trimethyl bicyclo heptan-2-yl]-5-(4-chloro-3-methylphenyl)-1-(4-methylbenzyl)-pyrazole-3-carboxamide (SR144528) and mimicked by the CB(2) receptor-specific agonist 1,1-dimethylbutyl-1-deoxy-Delta(9)-tetrahydrocannabinol (JWH133). Accompanying the increase in proliferation seen with 2-AG is an increase in active ERK1 that is also blocked with SR144528. The RTMGL1 microglial cells, which exist in a primed state, express the CB(1) and CB(2) receptors as demonstrated by reverse transcription-polymerase chain reaction and immunostaining. The CB(2) receptor in untreated cells is expressed both at the cell surface and internally, and exposure of the cells to 2-AG significantly increases receptor internalization. These data suggest that 2-AG activation of CB(2) receptors may contribute to the proliferative response of microglial cells, as occurs in neurodegenerative disorders.
Carrier
Erica J
EJ
Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, WI 53226-0509, USA.
Kearn
Christopher S
CS
Barkmeier
Andrew J
AJ
Breese
Nicole M
NM
Yang
Wenqi
W
Nithipatikom
Kasem
K
Pfister
Sandra L
SL
Campbell
William B
WB
Hillard
Cecilia J
CJ
D000704:Analysis of Variance; D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism*; D063385:Cannabinoid Receptor Modulators / Q000378:metabolism*; D002455:Cell Division; D002478:Cells, Cultured; D063388:Endocannabinoids; D004789:Enzyme Activation; D005434:Flow Cytometry; D005989:Glycerides / Q000378:metabolism*; D016173:Macrophage Colony-Stimulating Factor / Q000378:metabolism; D017628:Microglia / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D051381:Rats; D043885:Receptor, Cannabinoid, CB2 / Q000819:agonists / Q000378:metabolism
D001095:Arachidonic Acids; D063385:Cannabinoid Receptor Modulators; D063388:Endocannabinoids; D005989:Glycerides; D043885:Receptor, Cannabinoid, CB2; D016173:Macrophage Colony-Stimulating Factor; C094503:glyceryl 2-arachidonate; D020928:Mitogen-Activated Protein Kinases