Abstract
Purpose:
Nestin, a marker of neural stem cells, is known to be expressed in Müller cells during retinal development. However, the expression of nestin in retinal vascular development is not well established. Thus, we aimed to investigate the expression of nestin in developmental mouse retina, and to identify which retinal cells are related with the expression of nestin during the retinal vascular development.
Methods:
Eyes were enucleated from C57BL/6 mice on postnatal day (P)4, P8, P12, P16, and P26. Paraffin-embedded eyes were sliced into 4 μm thickness sections. Immunohistochemistry was performed to identify the expression pattern of nestin in the developing retina. Immunofluorescence was used to evaluate nestin expression in relation to endothelial cells (isolectin B4, IB4), pericytes (α-smooth muscle actin, α-SMA), and astrocytes (glial fibrillary acidic protein, GFAP).
Results:
Nestin was strongly expressed from ganglion cell layer (GCL) to retinoblast layer at P4. At P8, P12, and P16, the expression of nestin was observed from upper border of GCL where the end feet of Muller cells located, and vertically penetrating to outer border of outer nuclear layer. At P26, the expression of nestin was decreased and confined to GCL and inner nuclear layer (INL). Interestingly, there was strong vascular shape expression of nestin at all stages. Indeed, superficial, deep, and intermediate vascular plexus was completely merged with nestin expression at P4, P8, P12, P16. At P26, nestin expression was partially merged with IB4 expression. In addition, the nestin expression merged with those of α-SMA, but not that of GFAP.
Conclusions:
Nestin was expressed in endothelial cells and pericytes during retinal vascular development in retina. These results suggested that nestin could play an important role in developmental angiogenesis via interplay with endothelial cells and pericytes.