Notch signaling pathway inhibitor promotes differentiation of Müller cell-derived retinal stem cells into retinal ganglion cells_Chinese Journal of Ocular Fundus Diseases

Authors：

SongWeitao , ZhangXueyong , XiongSiqi , 蒋剑 , 文丹 , 喻一心 ,  夏晓波

Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha 410008, China;

Corresponding author：

夏晓波, Email: xbxia21@163.com

Keywords：

Microglia; Retinal ganglion cells; Stem cells; Receptors, notch; Animal experimentation

DOI：

10.3760/cma.j.issn.1005-1015.2014.04.015

Video：

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Objective To observe the role of Notch signaling pathway inhibitor in differentiation process of stem cells derived from retinal Müller cells into the ganglion cell. Methods Retinas of Sprague Dawley rat at postnatal 10-20 days were dissociated from eye balls. The third passage of Müller cells was used in this experiment, which cultured by repeated incomplete pancreatic enzyme digestion method. The retinal Müller cells were induced in the serum-free dedifferentiation medium. The cell proliferation state was observed under an inverted microscope. The expression of the specific markers Nestin and Ki-67 of retinal stem cells was measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The positive rate of nucleus was detected by Edu. The retinal stem cells was divided into Gamma secretase inhibtor-I (GSI) group and control group, the rate of ganglion cells was counted by using immunofluorescence staining. Results The cell proliferation had gathered to form a sphere. Immunofluorescence staining showed that the expressions of Nestin and Ki-67 were (92.94±6.48%) and (85.96±6.04%) respectively. Edu positive rate of nucleus was (82.80±6.65)%. RT-PCR and Western blot further confirmed the high expression of Nestin and Ki-67 in the cell spheres but not in the Müller cells. The positive rate of ganglion cells were (16.98±2.87)% and (11.17±0.71)% in GSI group and control group respectively, with the significant difference (t=3.210, P=0.002). Conclusion Notch signaling pathway is an important regulatory gene in stem cells differentiated into retinal ganglion cell.

Citation： SongWeitao, ZhangXueyong, XiongSiqi. Notch signaling pathway inhibitor promotes differentiation of Müller cell-derived retinal stem cells into retinal ganglion cells. Chinese Journal of Ocular Fundus Diseases, 2014, 30(4): 395-398. doi: 10.3760/cma.j.issn.1005-1015.2014.04.015 Copy

1.	Fimbel SM, Montgomery JE, Burket CT, et al.Regeneration of inner retinal neurons after intravitreal injection of ouabain in zebrafish[J].J Neurosci, 2007, 27: 1712-1724.
2.	Yurco P, Cameron DA.Responses of Müller glia to retinal injury in adult zebrafish[J].Vision Res, 2005, 45:991-1002.
3.	Fischer AJ, Reh TA.Potential of Müller glia to become neurogenic retinal progenitor cells[J]. Glia, 2003, 43:70-76.
4.	Ooto S, Akagi T, Kageyama R, et al.Potential for neural regeneration after neurotoxic injury in the adult mammalian retina[J]. Proc Natl Acad Sci USA, 2004, 101:13654-13659.
5.	Susan H, Branden RN, Brian B, et al. Notch signaling regulates regeneration in the avian retina[J].Developmental Biology, 2007, 312:300-311.
6.	Song WT, Zhang XY, Xiong SQ, et al. Comparison of two methods used to culture and purify rat retinal Müller cells[J]. Int J Ophthalmol, 2013, 6:778-784.
7.	Giannelli SG, Demontis GC, Pertile G, et al. Adult human Müller glia cells are a highly efficient source of rod photoreceptors[J].Stem Cells, 2011, 29:344-356.
8.	Cho MS, Kim SJ, Ku SY, et al. Generation of retinal pigment epithelial cells from human embryonic stem cell-derived spherical neural masses[J]. Stem Cell Res, 2012, 9:101-109.
9.	Sapkota D, Wu F, Mu X. Focus on molecules: Math5 and retinal ganglion cells[J]. Exp Eye Res, 2011, 93:796-797.
10.	Fischer AJ, McGuire CR, Dierks BD, et al. Insulin and fibroblast growth factor 2 Activate a neurogenic program in Müller glia of the chicken retina[J].J Neurosci, 2002, 22:9387-9398.
11.	Das AV, Mallya KB, Zhao X, et al. Neural stem cell properties of Müller glia in the mammalian retina: regulation by Notch and Wnt signaling[J].Dev Biol, 2006, 299: 283-302.
12.	Song WT, Zhang XY, Xia XB. Atoh7 promotes the differentiation of retinal stem cells derived from Müller cells into retinal ganglion cells by inhibiting Notch signaling[J].Stem Cell Res Ther, 2013, 4:94.

1. Fimbel SM, Montgomery JE, Burket CT, et al.Regeneration of inner retinal neurons after intravitreal injection of ouabain in zebrafish[J].J Neurosci, 2007, 27: 1712-1724.
2. Yurco P, Cameron DA.Responses of Müller glia to retinal injury in adult zebrafish[J].Vision Res, 2005, 45:991-1002.
3. Fischer AJ, Reh TA.Potential of Müller glia to become neurogenic retinal progenitor cells[J]. Glia, 2003, 43:70-76.
4. Ooto S, Akagi T, Kageyama R, et al.Potential for neural regeneration after neurotoxic injury in the adult mammalian retina[J]. Proc Natl Acad Sci USA, 2004, 101:13654-13659.
5. Susan H, Branden RN, Brian B, et al. Notch signaling regulates regeneration in the avian retina[J].Developmental Biology, 2007, 312:300-311.
6. Song WT, Zhang XY, Xiong SQ, et al. Comparison of two methods used to culture and purify rat retinal Müller cells[J]. Int J Ophthalmol, 2013, 6:778-784.
7. Giannelli SG, Demontis GC, Pertile G, et al. Adult human Müller glia cells are a highly efficient source of rod photoreceptors[J].Stem Cells, 2011, 29:344-356.
8. Cho MS, Kim SJ, Ku SY, et al. Generation of retinal pigment epithelial cells from human embryonic stem cell-derived spherical neural masses[J]. Stem Cell Res, 2012, 9:101-109.
9. Sapkota D, Wu F, Mu X. Focus on molecules: Math5 and retinal ganglion cells[J]. Exp Eye Res, 2011, 93:796-797.
10. Fischer AJ, McGuire CR, Dierks BD, et al. Insulin and fibroblast growth factor 2 Activate a neurogenic program in Müller glia of the chicken retina[J].J Neurosci, 2002, 22:9387-9398.
11. Das AV, Mallya KB, Zhao X, et al. Neural stem cell properties of Müller glia in the mammalian retina: regulation by Notch and Wnt signaling[J].Dev Biol, 2006, 299: 283-302.
12. Song WT, Zhang XY, Xia XB. Atoh7 promotes the differentiation of retinal stem cells derived from Müller cells into retinal ganglion cells by inhibiting Notch signaling[J].Stem Cell Res Ther, 2013, 4:94.

Chinese Journal of Ocular Fundus Diseases

Notch signaling pathway inhibitor promotes differentiation of Müller cell-derived retinal stem cells into retinal ganglion cells

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