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 of cannabinoid 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.