Abstract
Purpose: :
The expression of Math5 associated with the down-regulation of Notch-1. Whether Notch-1 can change the role of Math5 on the retinal ganglion cell ( RGC ) expression-patterns in retinal progenitor cells ( RPCs ) was investigated in this study.
Methods: :
RPCs were cultured,transfected by recombinant Math5 plasmid with internal ribosome entry site and enhanced green fluorescent protein ( pIRES2-EGFP-Math5 ), and then induced differentiation in the culture medium with Notch-1 antisense oligonucleotides ( group A ) or missense oligonucleotides ( group B ) for 14d, with pIRES2-EGFP transfected ( group C ) and no plasmid transfected ( group D ) induced differentiation in the culture medium with missense oligonucleotides as control. RGCs were identified by Thy1.1 immunocytochemistry methods and analyzed by Leica Qwin V3.1 system.
Results: :
pIRES2-EGFP-Math5 could transfect RPCs and the transfection rate was 24.68 %. After plating, different groups of RPCs could differentiate and express retina-specific markers, including RGC marker Thy1.1. The percentage of Thy1.1-positive cells was 31.22 % ± 6.73 % in group A, 29.51 % ± 6.31 % in group B, 15.51 % ± 3.60 % in group C, 16.62 % ± 3.56 % in group D. The differences between group A and C or D were statistically significant ( F = 97.387, P <0.001 ). The differences between group B and C or D were statistically significant ( F = 83.178, P <0.001 ). However, the differences between group A and B were not statistically significant ( F = 1.029, P = 0.315 ).
Conclusions: :
Math5 may up-regulate RGC expression-patterns in RPCs. However, inhibiting the Notch-1 seems to have no effect on the role of Math5.
Keywords: ganglion cells • retinal culture • gene transfer/gene therapy