AbstractWe identified Rab11‐family interacting protein 4 (Rab11‐FIP4) as a gene strongly expressed in the developing mouse retina. The major transcript encoding a full‐length protein, mRab11‐FIP4A, was expressed predominantly in neural tissues; whereas an alternative transcript encoding an N‐terminally truncated form of the protein, mRab11‐FIP4B, was expressed ubiquitously as a minor form. Gain‐of‐function of mRab11‐FIP4A in retina promoted cell cycle exit and increased subpopulations of retinal cells localized in the inner nuclear layer, such as bipolar cells and Müller glia. Reversal of the phenotype was observed in the loss‐of‐function experiment. Furthermore, Shh signaling was suggested to be involved in these functions. Analysis using truncation mutants revealed the essential role of the N‐terminal region containing a conserved EF‐hand motif for the retinal phenotypes induced by the expression of mRab11‐FIP4A, whereas binding to Rab11 was dispensable, suggesting the involvement of a novel Rab11‐independent mechanism for mRab11‐FIP4A action in the regulation of retinal development. Developmental Dynamics 236:214–225, 2007. © 2006 Wiley‐Liss, Inc.