A separate set of rats (N = 32) that were either socially isolated or group reared and concomitantly handled or non-handled (n = 8/group) were decapitated on postnatal day 70. Brains were rapidly dissected, and snap frozen in precooled isopentane (−30°C). Brains were stored at low temperature (−30°C and −80°C) until use. The right hemisphere of the rat brain was used to measure cannabinoid receptor densities and distribution using [3H]CP55,940 binding and quantitative autoradiography. The left hemisphere was used to obtain tissue punches for quantification of endocannabinoid content using liquid chromatography/mass spectrometry (LC/MS).
