Characterization of three zebrafish cell polarity genes, pard3, lin7, and scribble are described here. Anti-rabbit anti-Pard3 antiserum was generated and detected 180 and 150 kDa protein bands on the immunoblots. The Pard3 protein localized to the apical region of the neuroepithelial cells, either co-localizing with the apical adherens junctions or slightly apical to the adheren junctions. The apical subcellular localization of Pard3 protien is dependent on the aPKC and Nok proteins. Anti-Rabbit anti-Lin7 recognizes all three Lin7 protiens on the immunoblots. Lin7 protein localizes to retinal bipolar and horizontal cells. The correct subcellular localization of Lin7 requires the functional Nok protein in the retinal neuroepithelium.The Zebrafish scrib gene encodes a protein of 1724 amino acids that possesses one LRR and four PDZ domains in the same arrangement as in the human and mouse Scrib proteins. Two anti-rabbit anti-Scrib antisera were generated and detected a major protein band (250 kDa) and a minor protein band (235 kDa) on the immunoblot. The Scrib protein localized at the basalateral subdomain of the retinal neuroepithelial cells and expressed in all neural cell in mature retina. Anti-scrib morpholinos knocked down Scrib protein and resulted in a small retina without discernable neural layers. Co-injection of in vitro transcribed full-length scrib mRNA partially rescued the scrib morphant phenotype. TUNEL labeling showed a large amount of cell death throughout the retina. PVNA and BrdU labeling showed that the neuroepithelial cells in the scrib morphants retina failed to exit the cell cycle and continue to proliferation. Specification of retinal neural cells was abolished and the Ath5 differentiation signaling pathway required for ganglion cell differentiation was inhibited in the scrib morphant retina. Loss of Scrib expression resulted in the persistent expression of cyclin D1 but not cyclin B1. Knockdown of Scrib protein did not significantly affect the subcellular localization of the apical proteins like Pard3 and Nok, though the apical Nok-expressing subdomains were significantly expanded. Thus, Scrib contributes to the zebrafish retinal lamination in a different way relative to the apical cell polarity proteins.