A SET OF ANTI-CRYSTALLIN MONOCLONAL-ANTIBODIES FOR DETECTING LENS SPECIFICITIES - BETA-CRYSTALLIN AS A SPECIFIC MARKER FOR DETECTING LENTOIDOGENESIS IN CULTURES OF CHICKEN LENS EPITHELIAL-CELLS

Seven hybridoma lines which produced monoclonal antibodies against len s crystallins were established. They could detect alphaA-, alphaB-, ga mma-, delta-, and several beta-crystallins of different species of ani mals with high avidities. Although immunohistological analysis of chic ken and mouse lenses showed the typical distributions of each crystall in as have been reported so far, the ectopic expression of alphaB or d elta-crystallin was observed in the brain or the kidney of chicken, re spectively. Subsequently, crystallin production during the process of lentoidogenesis in cultures of chicken lens epithelial cells was exami ned with these antibodies. Western blot analysis revealed that beta-cr ystallins were not detected in cultured cells before lentoidogenesis, although all other crystallins were detected throughout the culture pe riod. Immunostaining of cultures indicated clearly that expression of beta-crystallins was restricted to lentoid bodies. These data confirme d the lens fiber specificity of beta-crystallins as previously reporte d in the messenger RNA level. In addition, we found that small-size de lta-crystallin (48 kDa) accumulated before the onset of lentoidogenesi s. These results strongly suggest that the differentiated state of len s cells in vitro could be classified by examination of the expression pattern of crystallins. In addition, the anti-crystallin monoclonal an tibodies produced in this study could be useful for detecting lens spe cificities.