Temporal regulation of six crystallin transcripts during mouse lens development

Using the polymerase chain reaction (PCR) and RNA blot analysis, we have examined the differential expression patterns of the gamma-crystallins during lens development. Since only four of these genes had been previously characterized, the cDNAs for the remaining two genes, gamma C and gamma F, were isolated and sequenced. The steady-state mRNA profiles were then determined by RNA blot analysis of samples from embryonic stages to 180 days after birth, with gene-specific probes for gamma A, gamma B, gamma C, and gamma D, and a common probe for gamma E and gamma F. Due to the paucity of mismatches between the gamma E and gamma F-crystallin genes, the PCR technique was exploited to determine their relative abundance. The data showed that while all six gamma-crystallin genes were expressed in the embryonic lens, they were differentially regulated during development. At early stages, the levels of gamma B and gamma C mRNAs were found to be relatively low in comparison to those for gamma A, gamma D, gamma E and gamma F. After 30-40 days, however, the levels of gamma A, gamma E, and gamma F mRNAs declined rapidly, and the gamma B, gamma C and gamma D transcripts became the major gamma-crystallin mRNA species. The utility of the PCR technique in studying the relative abundance of steady-state gamma-crystallin mRNAs was also investigated.