2014 Apr 2;3(4):495-504. doi: 10.1016/j.molmet.2014.03.007. eCollection 2014.
Cannabinoid control of brain bioenergetics: Exploring the subcellular localization of the CB1 receptor.
Hebert-Chatelain E1, Reguero L2, Puente N2, Lutz B3, Chaouloff F1, Rossignol R4, Piazza PV1, Benard G1, Grandes P2,Marsicano G1.
Abstract
Brain mitochondrial activity is centrally involved in the central control of energy balance. When studying mitochondrial functions in the brain, however, discrepant results might be obtained, depending on the experimental approaches. For instance, immunostaining experiments and biochemical isolation of organelles expose investigators to risks of false positive and/or false negative results. As an example, the functional presence ofcannabinoid type 1 (CB1) receptors on brain mitochondrial membranes (mtCB1) was recently reported and rapidly challenged, claiming that the original observation was likely due to artifact results. Here, we addressed this issue by directly comparing the procedures used in the two studies. Our results show that the use of appropriate controls and quantifications allows detecting mtCB1 receptor with CB1 receptor antibodies, and that, if mitochondrial fractions are enriched and purified, CB1 receptor agonists reliably decrease respiration in brain mitochondria. These data further underline the importance of adapted experimental procedures to study brain mitochondrial functions.
KEYWORDS:
BSA, bovine serum albumin; Brain bioenergetics; CB1 receptor; CB1, cannabinoid type 1 receptor; DAB–Ni, Ni-intensified 3,3ʹ-diaminobenzidine–4HCl; DMSO, dimethyl sulfoxide; KO, knock-out; LDHa, lactate dehydrogenase a; SDHA, succinate dehydrogenase a; Slp2, stomatin-like protein 2; WIN, WIN55,212-2; WT, wild-type; antibodies; electron microscopy; mitochondria; organelle purification
- PMID:
- 24944910
- [PubMed]
