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L. A. Williams, M. E. Boulton, M. A. Wride; Identification of Tumour Necrosis Factors and Expression of TRAIL and TRAF 3 During Chick Lens Development. Invest. Ophthalmol. Vis. Sci. 2007;48(13):2908.
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The Tumour Necrosis Factor (TNF) ligand and receptor families are involved in a number of diverse processes including cell proliferation, apoptosis and differentiation. However, their roles in development have not been well characterised. Previous work has suggested a role for TNFs in lens development. Therefore, the purpose of the current study was to examine the spatio-temporal patterns of expression of selected TNF family members and related signalling molecules in the chick lens during development using RT-PCR and immunohistochemistry.
Lenses were collected from White Leghorn chickens at embryonic days (E) 6, 8, 10, 12, 14 and 16 and RNA isolated using TRIzol®. Primers were designed for a number of chick TNF family members from the NCBI database and semi-quantitative RT-PCR was carried out. Band intensities were determined using Scion Image and compared between time points. Immunohistochemistry was carried out to examine the spatio-temporal patterns of expression of TNF Related Apoptosis Inducing Ligand (TRAIL) and TNF Receptor Associated Factor 3 (TRAF 3) during lens development.
Examples of members of the TNF family shown to be expressed in the embryonic chick lens using RT-PCR are TRAF 1,Tumour Necrosis Factor Receptor 1 (TNFR1), TRAIL and TRAIL-like which were all most highly expressed at E8, TRAF2 peaked at E12, Tumour Necrosis Factor-Alpha-Converting enzyme (TACE) peaked at E16, while TRAF 3 was most highly expressed at E10. Immunohistochemistry showed TRAIL to be highly expressed in the nuclei of the lens fibres up to E14 at which time the staining disappeared from central fibres. TRAF 3 showed cytoplasmic staining in the epithelium and differentiating secondary fibre cells. This cytoplasmic staining increased with embryonic age.
A number of TNF family members, previously unidentified in the lens, were shown to be expressed. These results suggest a role for TNF signalling in regulation of proliferation, differentiation and/or denucleation of fibre cells during lens development.
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