The expression of CD45 isoforms and of CDw29 has been analyzed as a function of time by correlating cell surface phenotype with mRNA synthesis. After activation, T cells lose CD45R and acquire a high density of CD45 p180 and of CDw29. Throughout this transition the density of CD45 common determinants steadily increases, resulting in a net gain of surface CD45. The gradual loss of CD45R could reflect a rapid switch in CD45 mRNA splicing patterns followed by a slow loss of surface CD45R. Alternatively it could reflect a delayed activation of splicing to produce the 4.8-kb CD45 mRNA, or long lived 5.4-kb CD45 mRNA. Analysis of CD45 mRNA indicated that at 24 h postactivation, 5.4-kb CD45 mRNA is lost and only 4.8-kb mRNA is detectable. The amount of 4.8-kb mRNA increases until day 3 and then decreases somewhat. Thus our results support the interpretation that transitions in CD45 isoform mRNA expression occur within the first 24 h after activation and that the persistence of CD45R+ T cells until day 3 to 4 of culture results from slow turnover of surface CD45R glycoprotein.