Engrafted Neural Progenitor Cells Express a Tissue-Restricted Reporter Gene Associated With Differentiated Retinal Photoreceptor Cells
Abstract:Neural progenitor cells (NPCs) have shown ability to repair injured CNS, and might provide precursors to retinal neurons. NPCs were isolated from the brains of 14 day murine embryos of transgenic mice that express -galactosidase (-gal) on the arrestin promoter, which specifically directs expression to retinal photoreceptor cells. NPCs were transferred to adult, syngeneic mice via inoculation into the anterior chamber of the eye, the peritoneal cavity, or the brain. At 14 weeks postgrafting, tissues were collected and examined to determine if differentiated NPC progeny were present in retina based on histochemical detection of -gal. Four of six anterior chamber-inoculated recipients showed Bluo-gal-stained cells in retina, indicating the presence of transferred NPCs or their progeny. Because the progenitor cells do not express -gal, positive staining indicates differentiation leading to activation of the arrestin promoter. Two recipients inoculated by the intraperitoneal route also exhibited Bluo-gal staining in retina. The NPCs did not express -gal if inoculated into brain, but survived and dispersed. Most recipients, regardless of inoculation route, were PCR positive for -gal DNA in extraocular tissues, but no Bluo-gal staining was found outside of the retina. Injury to the retina promoted, but was not required, for progenitor cell engraftment. -Gal-positive cells were concentrated in the outer layers of the retina. In summary, a reporter gene specifically expressed in differentiated retinal photoreceptor cells due to the activity of the arrestin promoter was expressed in recipient mouse retina following transfer of NPCs prepared from the -gal transgenic mice. The presence of -gal DNA, but not Bluo-gal staining, in spleen and other tissues revealed that the cells also migrated elsewhere and took up residence in other organs, but did not undergo differentiation that led to -gal expression.
Keywords: Cell differentiation; Immunofluorescence; Immunohistochemistry; Mice, transgenic; Neural proteins; Neurons; Photoreceptor cells; Progenitor cell transplantation; Retina; Stem cell transplantation
Document Type: Research Article
Publication date: February 1, 2006
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