Retinal detachment is a serious cause of visual impairment. Gaining an understanding of its cellular effects can help improve management of the disease and contribute to our basic knowledge of the retina and its relationship to the pigment epithelium. We have recently identified specific cellular and biochemical changes in long-term detachments using an animal model. Gaining a further understanding of these changes using a combination of structural, biochemical, and molecular techniques is a major goal of this project. Specifically we will determine: 1) when the upregulation of intermediate filament proteins and the downregulation of enzymatic and vitamin A binding proteins occurs in Muller cells (by using immunocytochemistry and immunoblot analyses); 2) if the mRNA levels,for one protein in each of these classes, changes in concert with the changes in protein expression (by Northern and slot blot analyses; appropriate cellular expression of the mRNAs will be studied by in situ hybridization); 3) the onset and extent of opsin redistribution in photoreceptors (by immunocytochemistry); 4) if opsin synthesis and outer segment renewal continues (by immunoprecipitation and Lucifer yellow band displacement analyses); 5) if opsin mRNA levels change and if opsin mRNA distribution changes in cells that have redistributed opsin or whose morphology is severely disrupted (by Northern, slot blot and in situ hybridization studies); 6) the extent of the proliferative response and identification of the cell types involved (by continuous delivery of 3H-thymidine, LM and EM autoradiography); 7) if growth factors are involved (by immunocytochemistry) or if they can mimic the effects of detachment (by injection of growth factors into the eye or their addition to cultures of Muller cells); 8) if the changes in protein expression or redistribution or the proliferative response are arrested or modified by retinal reattachment. A greater understanding of the synaptic organization of the human retina will be gained by studying synaptic circuitry by serial-section electron microscopy of adult and developing human retinas and the analysis of Golgi- impregnated whole-mounts. In the latter, we will: 1) study the stained neurons in the 75 existing whole-mounts by standard Golgi descriptive techniques, and 2) analyze the synaptic input into the Golgi stained cells by electron microscopy (by obtaining new postmortem tissue from donor eyes and using a variety of fixation protocols to develop better structural preservation of the retina).

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37EY000888-28
Application #
2856879
Study Section
Special Emphasis Panel (NSS)
Project Start
1976-03-01
Project End
1999-12-31
Budget Start
1999-01-01
Budget End
1999-12-31
Support Year
28
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of California Santa Barbara
Department
Type
Organized Research Units
DUNS #
City
Santa Barbara
State
CA
Country
United States
Zip Code
93106
Mandal, Nakul; Lewis, Geoffrey P; Fisher, Steven K et al. (2011) Protein changes in the retina following experimental retinal detachment in rabbits. Mol Vis 17:2634-48
Lesnik Oberstein, Sarit Y; Lewis, Geoffrey P; Dutra, Thomas et al. (2011) Evidence that neurites in human epiretinal membranes express melanopsin, calretinin, rod opsin and neurofilament protein. Br J Ophthalmol 95:266-72
Oberstein, Sarit Y Lesnik; Byun, Jiyun; Herrera, Diego et al. (2011) Cell proliferation in human epiretinal membranes: characterization of cell types and correlation with disease condition and duration. Mol Vis 17:1794-805
Luna, Gabriel; Kjellstrom, Sten; Verardo, Mark R et al. (2009) The effects of transient retinal detachment on cavity size and glial and neural remodeling in a mouse model of X-linked retinoschisis. Invest Ophthalmol Vis Sci 50:3977-84
Lewis, Geoffrey P; Chapin, Ethan A; Byun, Jiyun et al. (2009) Muller cell reactivity and photoreceptor cell death are reduced after experimental retinal detachment using an inhibitor of the Akt/mTOR pathway. Invest Ophthalmol Vis Sci 50:4429-35
Linberg, Kenneth A; Lewis, Geoffrey P; Fisher, Steven K (2009) Retraction and remodeling of rod spherules are early events following experimental retinal detachment: an ultrastructural study using serial sections. Mol Vis 15:10-25
Eibl, Kirsten H; Fisher, Steven K; Lewis, Geoffrey P (2009) Alkylphosphocholines: a new approach to inhibit cell proliferation in proliferative vitreoretinopathy. Dev Ophthalmol 44:46-55
Verardo, Mark R; Lewis, Geoffrey P; Takeda, Masumi et al. (2008) Abnormal reactivity of muller cells after retinal detachment in mice deficient in GFAP and vimentin. Invest Ophthalmol Vis Sci 49:3659-65
Lewis, Geoffrey P; Betts, Kellen E; Sethi, Charanjit S et al. (2007) Identification of ganglion cell neurites in human subretinal and epiretinal membranes. Br J Ophthalmol 91:1234-8
Nakazawa, Toru; Takeda, Masumi; Lewis, Geoffrey P et al. (2007) Attenuated glial reactions and photoreceptor degeneration after retinal detachment in mice deficient in glial fibrillary acidic protein and vimentin. Invest Ophthalmol Vis Sci 48:2760-8

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