Int. J. Dev. Biol. In Press
doi: 10.1387/ijdb.200113fz
© UPV/EHU Press

Photoreceptor progenitor dynamics in the zebrafish embryo retina and its modulation by primary cilia and N-cadherin

Gonzalo Aparicio1,2, Magela Rodao1,2, José L. Badano2 and Flavio R. Zolessi1,2

1Sección Biología Celular, Facultad de Ciencias, Universidad de la República, Uruguay and 2Institut Pasteur Montevideo, Uruguay

ABSTRACT Photoreceptors of the vertebrate neural retina are originated from the neuroepithelium, and like other neurons, must undergo cell body translocation and polarity transitions to acquire their final functional morphology, which includes features of neuronal and epithelial cells. We analyzed this process in detail on zebrafish embryos using in vivo confocal microscopy and electron microscopy. Photoreceptor progenitors were labeled by the transgenic expression of EGFP under the regulation of the photoreceptor-specific promoter crx, and structures of interest were disrupted using morpholino oligomers to knock-down specific genes. Photoreceptor progenitors detached from the basal retina at pre-mitotic stages, rapidly retracting a short basal process as the cell body translocated apically. They remained at an apical position indefinitely to form the outer nuclear layer (ONL), initially extending and retracting highly dynamic neurite-like processes, tangential to the apical surface. Many photoreceptor progenitors presented a short apical primary cilium. The number and length of these cilia was gradually reduced until nearly disappearing around 60 hpf. Their disruption by knocking-down ift88 and elipsa caused a notorious defect on basal process retraction. To assess the role of cell adhesion in the organization of photoreceptor progenitors, we knocked-down cdh2/N-cadherin and observed the cell behavior by time-lapse microscopy. The ectopic photoreceptor progenitors initially migrated in an apparent random manner, profusely extending cell processes, until they encountered other cells to establish cell rosettes in which they stayed acquiring the photoreceptor-like polarity. Altogether, our observations indicate a complex regulation of photoreceptor progenitor dynamics to form the retinal ONL, previous to the post-mitotic maturation stages.

Keywords:

neuroepithelium; cell polarity; cell adhesion; retina lamination; neurogenesis

*Corresponding author e-mail: fzolessi@fcien.edu.uy