1. Rat liver beta-N-acetylhexosaminidase was separated into several different molecular forms by DEAE-cellulose chromatography. 2. The subunit composition of the isoenzymes, as well as the similarities to human hexosaminidases, were determined by using the specific active alpha subunit substrate 4-methylumbelliferyl-beta-N-acetylglucosamine-6-sulphate. 3. As in human tissues, the intermediate form lacked the active alpha subunit and resembled hexosaminidase B rather than A. 4. The intermediate form was markedly increased in foetal liver and in regenerating liver after partial hepatectomy. 5. The variations in isoenzyme expression were accompanied by variations in specific activity of hexosaminidase. 6. Sulphated substrate analysis and thermal stability experiments indicated that the rapid cell proliferation had a greater effect on the formation of beta-subunit of hexosaminidase than on that of alpha-subunit.