Tgfbeta receptor expression in lens: implications for differentiation and cataractogenesis.
TGFbeta induces changes characteristic of some forms of cataract. However, the responsiveness of lens epithelial cells to TGFbeta is age-dependent; weanling and adult, but not neonatal, lens epithelial cells respond. This study investigated TGFbeta receptor (TbetaRI and TbetaRII) expression during rat lens development and the effects of FGF-2 on TGFbeta responsiveness and TbetaR expression. Immunofluorescence, immunoblotting, RT-PCR and in situ hybridization were used to examine the spatio-temporal expression patterns of TbetaR. Lens explants were used to investigate the effects of FGF-2 on TGFbeta responsiveness and TbetaR expression. In the lens epithelium, little or no immunoreactivity was detected at P3 but at P21 there was distinct reactivity for TbetaRI and TbetaRII. Reactivity for both receptors was also found in the differentiating fibers in the transitional zone and cortex at both ages. Western blotting of lens membrane extracts identified multiple molecular weight forms of TbetaRI (30, 50, 90 kDa) and TbetaRII (70-120 kDa). In situ hybridization with a rat probe for Alk5 (TbetaRI) showed that the lens expresses Alk5 mRNA in epithelium and fibers throughout development. A rat TbetaRII probe revealed distinct expression of a TbetaRII mRNA in lens fibers throughout development and in the lens epithelium at P21 but not at P3. In vitro studies showed that lens epithelial explants from P9 rats did not undergo cataractous changes in response to TGFbeta but P13 explants did. Addition of FGF-2 to P9 explants induced increased TbetaR immunoreactivity and enhanced the competency of lens epithelial cells to respond to TGFbeta. These data indicate that the overall increased expression of TGFbeta receptors in lens epithelium during postnatal development (P3-P21) underlies an age-related change in TGFbeta responsiveness. The results also suggest that lens cells may express multiple forms of TbetaR. Expression of TbetaR in lens fibers throughout lens development and the induction of enhanced TbetaR expression by FGF suggest a role for TGFbeta signaling during FGF-induced responses and fiber differentiation.
de Iongh
R U
RU
Department of Anatomy and Histology and Institute for Biomedical Research (F13), The University of Sydney, Sydney, NSW 2006, Australia. robbdei@anatomy.usyd.edu.au
Gordon-Thomson
C
C
Chamberlain
C G
CG
Hales
A M
AM
McAvoy
J W
JW
D000375:Aging / Q000502:physiology; D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D002386:Cataract / Q000209:etiology / Q000378:metabolism*; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000032:analysis; D016222:Fibroblast Growth Factor 2 / Q000502:physiology; D005455:Fluorescent Antibody Technique; D017403:In Situ Hybridization; D007908:Lens, Crystalline / Q000378:metabolism*; D051379:Mice; D008970:Molecular Weight; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D018125:Receptors, Transforming Growth Factor beta / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016212:Transforming Growth Factor beta / Q000502:physiology
D018076:DNA, Complementary; D012333:RNA, Messenger; D018125:Receptors, Transforming Growth Factor beta; D016212:Transforming Growth Factor beta; D016222:Fibroblast Growth Factor 2