title: NGF Expression in Reelin-Deprived Retinal Cells: A Potential Neuroprotective Effect.
abstract: We recently reported that increased NGF and p75(NTR) as well as decreased trkA(NGFR) characterized the Reelin-deprived (E-Reeler) retina, prospecting a potential contribution of NGF during E-Reeler retinogenesis. Herein, retinal ganglion cells (RGCs), glial cells and rod bipolar cells (RBCs) were isolated from E-Reeler retinas, and NGF, trkA(NGFR)/p75(NTR) expression and apoptosis were investigated. E-Reeler (n = 28) and E-control (n = 34) retinas were digested, and RGCs, glial cells and RBCs were isolated by the magnetic bead separation. Expression of NGF, trkA(NGFR), p75(NTR), Annexin V/PI and Bcl2/Bax was quantified by flow cytometry and validated by real-time PCR or WB. In E-Reeler retinas, NGF was significantly increased in RGCs and glial cells, p75(NTR) was increased in both RBCs and RGCs, and trkA(NGFR) was unchanged. In E-control retinas, NGF and p75(NTR) were expressed mainly in RBCs and RGCs and faintly in glial cells, while trkA(NGFR) was weakly expressed by RBCs and RGCs. In RBCs and RGCs, Annexin V expression was unchanged, while Bcl2 increased and Bax decreased selectively in E-Reeler RGCs. The data indicate that E-Reeler RBCs and RGCs overexpress NGF and p75(NTR) as a protective endogenous response to Reelin deprivation. The observation is strongly supported by the absence of apoptosis in both cell types.
authors.affiliation: Laboratory of Ophthalmology, Ocular Surface Unit, IRCCS-G.B. Bietti Foundation, via Alvaro del Portillo 21, 00128, Rome, Italy.
authors.forename: Bijorn Omar
authors.initials: BO
authors.lastname: Balzamino
authors.forename: Graziana
authors.initials: G
authors.lastname: Esposito
authors.forename: Ramona
authors.initials: R
authors.lastname: Marino
authors.forename: Flavio
authors.initials: F
authors.lastname: Keller
authors.forename: Alessandra
authors.initials: A
authors.lastname: Micera
mesh_terms: D000818:Animals; D017304:Annexin A5 / Q000096:biosynthesis / Q000235:genetics; D017209:Apoptosis; D015816:Cell Adhesion Molecules, Neuronal / Q000172:deficiency* / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000172:deficiency* / Q000235:genetics; D005136:Eye Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008818:Mice, Neurologic Mutants; D020932:Nerve Growth Factor / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000172:deficiency* / Q000235:genetics; D009457:Neuroglia / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis / Q000235:genetics; D020800:Receptor, Nerve Growth Factor / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D020917:Receptor, trkA / Q000096:biosynthesis / Q000235:genetics; D000091042:Reelin Protein; D012160:Retina / Q000378:metabolism* / Q000473:pathology; D051245:Retinal Bipolar Cells / Q000378:metabolism; D012165:Retinal Ganglion Cells / Q000378:metabolism; D012697:Serine Endopeptidases / Q000172:deficiency* / Q000235:genetics; D015398:Signal Transduction; D051028:bcl-2-Associated X Protein / Q000096:biosynthesis / Q000235:genetics
chemical_list: D017304:Annexin A5; C494827:Bax protein, mouse; D015816:Cell Adhesion Molecules, Neuronal; D016326:Extracellular Matrix Proteins; D005136:Eye Proteins; D009419:Nerve Tissue Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D020800:Receptor, Nerve Growth Factor; D000091042:Reelin Protein; D051028:bcl-2-Associated X Protein; C052617:Bcl2 protein, mouse; D020932:Nerve Growth Factor; D020917:Receptor, trkA; C000717128:Reln protein, mouse; D012697:Serine Endopeptidases