Characterization of CB1 cannabinoid receptor immunoreactivity in postmortem human brain homogenates.
The CB1 cannabinoid receptor (CB1) is the predominant type of cannabinoid receptor in the CNS, in which it displays a unique anatomical distribution and is present at higher densities than most other known seven transmembrane domain receptors. Nevertheless, as with almost all seven transmembrane domain receptors, the tertiary and quaternary structure of this receptor is still unknown. Studies of CB1 in rat cerebral tissue are scarce, and even less is known regarding the expression of CB1 in the human brain. Thus, the aim of the present work was to characterize CB1 expression in membranes from postmortem human brain using specific antisera raised against this protein. Western blot analysis of P1 and P2 fractions, and crude plasma membrane preparations from the prefrontal cortex showed that CB1 migrated as a 60 kDa monomer under reducing conditions. These data were confirmed by blotting experiments carried out with human U373MG astrocytoma cells as a positive control for CB1 expression and wild-type CHO cells as negative control. In addition, when proteins were solubilized in the absence of dithiothreitol, the anti-human CB1 antiserum detected a new band migrating at around 120 kDa corresponding in size to a putative CB1 dimer. This band was sensitive to reducing agents (50 mM dithiothreitol) and showed sodium dodecylsulphate stability, suggesting the existence of disulfide-linked CB1 dimers in the membrane preparations. Important differences in the anatomical distribution of CB1 were observed with regard to that described previously in monkey and rat; in the human brain, CB1 levels were higher in cortex and caudate than in the cerebellum.
Department of Pharmacology, Faculty of Pharmacy, University of the Basque Country, C/Paseo de la Universidad 7, E-01006 Vitoria-Gasteiz, Spain. kfblodem@vc.ehu.es
M López
ML
De Jesús
J
J
Sallés
J J
JJ
Meana
L F
LF
Callado
D000367:Age Factors; D000918:Antibody Specificity; D001921:Brain / Q000276:immunology* / Q000378:metabolism; D001923:Brain Chemistry / Q000276:immunology*; D063385:Cannabinoid Receptor Modulators / Q000378:metabolism*; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D005260:Female; D006031:Glycosylation; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D043884:Receptor, Cannabinoid, CB1 / Q000276:immunology* / Q000378:metabolism; D013347:Subcellular Fractions
D063385:Cannabinoid Receptor Modulators; D043884:Receptor, Cannabinoid, CB1