| Popis: |
Obesity is a worldwide health problem; the number of obese and extremely obese individuals in the US increased rapidly between 1962 to 2000, from 14-36 % (NIH). There are many factors that relate obesity to diseases such as diabetes mellitus, cardiovascular diseases, and infertility, highlighting the need to find a way to reverse this trend. In fact, obese parents can directly affect their offspring health. One of the most important factors that contributes to obesity is lack of voluntary physical activity. A key motivational element of controlling physical activity is the involvement of the endocannabinoid system (ECS). This system consists of two receptors, cannabinoid receptor-1 and 2 (CB1-R and CB2-R) which are G-protein-coupled receptors, and their ligands. Overeating can alter the expression of CB1 receptors in the brain. Previous research has shown that offspring mice of high fat diet (HFD) fed-fathers run more than offspring mice of low fat diet (LFD) fed-fathers (Zhang et al.). In the present study, brain tissues of offspring mice from both groups were flash frozen, post-fixed and sectioned to be used to compare the expression of CB1-R after optimizing the protocol. I hypothesize that alterations in expression of CB1-R within the CNS reward circuitry in hippocampus and hypothalamus are related to different levels of voluntary physical activity in offspring of obese HFD-fed fathers versus LFD-fed fathers. We found that of three primary anti- CB1-R antibodies tested, only one showed specific binding, and that both blocking solution composition and degree of dilution of the primary antibody affect the staining and apparent CB1-R immunoreactivity in cerebellum and hippocampus. In conclusion, CB1-R was expressed and specifically detected in both cerebellum, which was used as a positive control, and hippocampus. The protocol developed will be used in the proposed studies of offspring CB1-R. |
