Abstract
Purpose: :
The retinal development is complex processes that a single type of migrating neuroblasts differentiates into various cell types. The goal of this study was to analyze the presence and distribution of CD44v6 and ADAM10 in the early developing retina, and to investigate whether CD44v6 shedding and migration of neuroblasts in vitro is mediated by ADAM10.
Methods: :
Indirect immunohistochemistry for CD44v6 and ADAM10 was performed on paraffin-embedded developing chick retinas. Neuroblasts isolated from developing chick retinas at embryonic day 6 (E6) were primary cultured with GM6001 or ADAM10 inhibitor.
Results: :
A prominent CD44v6 immunoreactivities were distributed uniformly on the most neuroblast cells in contact with each other at E5, whereas was confined mainly to the differentiated ganglion cells at E7 and E8. A similar pattern of distribution was also observed for ADAM10. The presence of ADAM10 protein was also detected by Western blot analysis using retinal lysates. Primary cultured neuroblasts from E6 aggregated in small clumps. After 1 day in culture, outgrowing cells started to extend neurites. Neuroblasts outgrowth and CD44v6 shedding was significantly blocked in aggregate cultures incubated with GM6001 or ADAM10 inhibitor in a dose dependent manner.
Conclusions: :
Our data suggest a role for CD44v6 in cell-ECM interaction, ADAM10 plays an important role for CD44v6 shedding, and ADAM10 activity may involved initially in dissociation of cell-ECM interactions, and subsequently in the generation of retinal neurons.
Keywords: retinal development • retinal culture • retinal adhesion