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Samelia Okpodu, Chunqiao Liu, Alexander Bassuk, Helen May-Simera, Vinit B. Mahajan, Werner Graf, Anand Swaroop, Tiansen Li; Functional Characterization of Prickle 2, a core Planar Cell Polarity protein, in mouse retina. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3961.
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Planar cell polarity (PCP) refers to an asymmetry along the plane of tissue that is perpendicular to the baso-apical axis. PCP is critical for many aspects of development such as convergent extension of cell movement during gastrulation and cochlea duct formation in the inner ear. To understand functions of PCP pathway in retina we focused on functional characterization of Prickle 2, a core player of PCP pathway.
In situ hybridization and qRT-PCR was used to study Prickle 2 gene expression during retinal development. Polyclonal antibodies against Prickle2 were generated to study Prickle 2 protein expression by immunoblotting and Prickle 2 subcellular localization by immunohistochemistry. Cochlea hair cell formation was examined by phalloidin staining for actin filament in wildtype and prickle2 null mice at postnatal day 1. Retinal functions of Prickle 2 knockout mice were assessed by electroretinography (ERG).
Prickle 2 expression in retina is detected as early as at embryonic day 13.5 (E13.5) by in situ hybridization and is present throughout the developing retina. In adult retina, it is moderately expressed in the inner nuclear layer, weakly expressed in ganglion cells, and minimally expressed in photoreceptor layer. Immunoblotting shows Prickle2 to be a single band of 90 kDa which is absent in the KO retina. Prickle 2 protein is upregulated in NRL knockout mouse retina both by immunoblotting and by qRT-PCR. ERG at one month shows that loss of PK2 protein does not lead to a major deficit in photoreceptor development or function. No gross abnormality in stereocilia bundle formation and orientation was detected in the knockout mice. Further work is in progress, which will investigate the inner retinal neurons and circuitry in addition to the PK2 subcellular localization.
Sensory neurons (photoreceptors and cochlear hair cells) appeared to develop normally in mice lacking Prickle 2. Functional redundancy with Prickle1 may offer an explanation. PK2 expression appears to be influenced by NRL, the significance of which is under investigation.
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