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Cryopreservation of Mouse Embryos by Ethylene Glycol-Based Vitrification
JoVE 3155 11/18/2011

May 2013 Regulating gene expression in photoreceptor precursor cells


mouse_of_month_201305

Regulating gene expression in photoreceptor precursor cells

B6.Cg-Tg(Crx-cre/ERT2)1Tfur/TfurRbrc RBRC05559

mn1305_0101

Courtesy of Takahisa Furukawa, M.D., Ph.D., Osaka University

X-gal staining of sections isolated from a Crx-cre/ERT2; CAG-CAT-Z mouse with tamoxifen. Application of tamoxifen induced Cre-mediated recombination of the LacZ reporter transgene in the developing photoreceptor layer of the CAG-CAT-Z reporter mouse (provided by Dr. J. Miyazaki, Osaka University). ONL: outer nuclear layer, INL: inner nuclear layer, GCL: ganglion cell layer.

Cone-rod homeobox containing gene (CRX) is a transcription factor that is predominantly expressed in developing and mature photoreceptor cells in the retina, and plays a crucial role in the terminal differentiation of rods and cones. Crx is one of the earliest expressed genes in photoreceptor precursor cells [1-4]. To obtain the regulation of Cre expression in photoreceptor precursor cells, Furukawa and colleagues established a transgenic mouse line (Crx-CreERT2) that expresses CreERT2 under the control of a 2-kb promoter fragment from the Crx gene [5, 6]. Application of tamoxifen to Crx-CreERT2; CAG-CAT-Z indicator mice induced Cre-mediated recombination of a LacZ reporter transgene in the developing photoreceptor layer. They also inactivated the retina and anterior neural fold homeobox (Rax) gene in early postmitotic photoreceptor precursor cells using the CreERT2 system and revealed that the expression of the transcription factor orthodenticle homolog 2 (Drosophila) (Otx2) was severely attenuated in the Rax-induced conditional knockout retina [6]. Crx-CreERT2 transgenic mice allow us to regulate gene expression in photoreceptor precursor cells from the embryonic to the adult stage.

 

Related strains : 129.B6C3-Tg(Crx-cre)1Tfur/TfurRbrc  RBRC05426
129S6;B6C3-Tg(Crx-EGFP)G3Tfur/TfurRbrc  RBRC05560
129(Cg)-Rax<tm1.1Tfur>/TfurRbrc  RBRC05558
B6.Cg-Tg(Crx-rtTA)G2Tfur/TfurRbrc  RBRC05562
Depositor : Takahisa Furukawa, M.D., Ph.D.
Institute for Protein Research, Osaka University
References :  [1] Furukawa T, Morrow EM, Cepko CL. Crx, a novel otx-like homeobox gene, shows photoreceptor-specific expression and regulates photoreceptor differentiation. Cell; 91(4):531-41, 1997.
 [2] Furukawa T, Morrow EM, Li T, Davis FC, Cepko CL. Retinopathy and attenuated circadian entrainment in Crx-deficient mice. Nat Genet.; 23(4):466-70, 1999.
 [3] Nishida A, Furukawa A, Koike C, Tano Y, Aizawa S, Matsuo I, Furukawa T. Otx2 homeobox gene controls retinal photoreceptor cell fate and pineal gland development. Nat Neurosci.; 6(12):1255-63, 2003.
 [4] Muranishi Y, Sato S, Inoue T, Ueno S, Koyasu T, Kondo M, Furukawa T. Gene expression analysis of embryonic photoreceptor precursor cells using BAC-Crx-EGFP transgenic mouse. Biochem Biophys Res Commun.; 392(3):317-22, 2010.
 [5] Furukawa A, Koike C, Lippincott P, Cepko CL, Furukawa T. The mouse Crx 5’-upstream transgene sequence directs cell-specific and developmentally regulated expression in retinal photoreceptor cells. J Neurosci.; 22(5):1640-7, 2002.
 [6] Muranishi Y, Terada K, Inoue T, Katoh K, Tsujii T, Sanuki R, Kurokawa D, Aizawa S, Tamaki Y, Furukawa T. An essential role for RAX homeoprotein and NOTCH-HES signaling in Otx2 expression in embryonic retinal photoreceptor cell fate determination. J Neurosci.; 31(46):16792-807, 2011.