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.
Goring
D R
DR
Genetics Department, Hospital for Sick Children, Toronto, Canada.
Breitman
M L
ML
Tsui
L C
LC
D000818:Animals; D001483:Base Sequence; D003459:Crystallins / Q000235:genetics*; D004247:DNA / Q000737:chemistry; D005786:Gene Expression Regulation / Q000502:physiology*; D015151:Immunoblotting; D007908:Lens, Crystalline / Q000254:growth & development* / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D014158:Transcription, Genetic / Q000502:physiology
D003459:Crystallins; D012333:RNA, Messenger; D004247:DNA