The effects of stress due to brief (4–5 min) ether and pentobarbital anesthesia vs. decapitation on assays of seven enzymes in homogenates of synovium, articular and epiphyseal cartilage, and metaphyseal and cortical bone were compared. Etherization caused twofold changes in synovial and articular cartilage G-6-PDH, LDH, CPK, glutamic DH, and ICDH based on tissue dry weight and DNA content. Pentobarbital anesthesia produced only slightly lower activities, per gram DNA, of LDH, acid phosphatase, and glutamic-DH in cortical bone. Epiphyseal cartilage metabolism was unaffected by either mode of anesthesia. No differences could be detected between the levels of enzyme activities of the several tissues taken from rats that had been decapitated or anesthetized with pentobarbital. The changes in enzyme activities suggested that pentobarbital was non-stressful and appropriate to metabolic studies in the skeleton.