Bit1-a novel regulator of astrocyte function during retinal development: proliferation, migration, and paracrine effects on vascular endothelial cell

AbstractStudies have shown that astrocyte plays an important role in the formation of retinal vasculature during development. For our study, we investigated the role of Bcl2 inhibitor of transcription 1 (Bit1) in regulating astrocyte function from developing retina and its paracrine effects on vascular endothelial cell. Expression pattern of Bit1 was analyzed by immunofluorescent staining of whole mount rat retina. Astrocytes and retinal microvascular endothelial cells (RMECs) were isolated from rat retina for cultural studies. The proliferation and migration of astrocytes and RMECs were evaluated by CCK-8 assay, scratch assay, and transwell migration assay. Cell apoptosis was detected by anoikis assay. Angiogenesis assay was used to measure the ability of RMECs to form tube-like microvascular structure. siRNA knockdown assay was employed to regulate Bit1 expression in astrocytes. Immunofluorescent staining showed Bit1 expression in migrating retinal astrocytes co-localized with the marker glial fibrillary acidic protein (GFAP). Isolated retinal astrocytes from post-natal rat eyes have an elevated expression of Bit1 and show increased cell survival and decreased anoikis as compared with retinal astrocytes from embryo. Suppressing Bit1 by siRNA assay leads to decreased cell proliferation, migration, and increased anoikis of astrocytes. Meanwhile, Bit1 knockdown could decrease the astrocytic vascular endothelial growth factor (VEGF) expression leading to inhibitory paracrine ef...

Source: Human Cell - July 30, 2019 Category: Cytology Source Type: research