Retinal Degenerative Diseases (eBook)

Laboratory and Therapeutic Investigations

Robert E. Anderson, Joe G. Hollyfield, Matthew M. Lavail (Herausgeber)

eBook Download: PDF

2010 | 2010
XLVII, 714 Seiten
Springer New York (Verlag)
978-1-4419-1399-9 (ISBN)

Infos

Robert E. Anderson, MD, PhD, is George Lynn Cross Research Professor, Dean A. McGee Professor of Ophthalmology, and Adjunct Professor of Biochemistry & Molecular Biology and Geriatric Medicine at The University of Oklahoma Health Sciences Center in Oklahoma City, Oklahoma. He is also Director of Research at the Dean A. McGee Eye Institute. He received his Ph.D. in Biochemistry (1968) from Texas A&M University and his M.D. from Baylor College of Medicine in 1975. In 1968, he was a postdoctoral fellow at Oak Ridge Associated Universities. At Baylor, he was appointed Assistant Professor in 1969, Associate Professor in 1976, and Professor in 1981. He joined the faculty of the University of Oklahoma Health Sciences Center in January of 1995. He has received several honorary appointments including Visiting Professor, West China School of Medicine, Sichuan University, Chengdu, China; Honorary Professorship, Xi'an Jiaotong University, Xi'an, China; and Honorary Professor of Sichuan Medical Science Academy, Sichuan Provincial People's Hospital, Sichuan, China. Dr. Anderson has received the Sam and Bertha Brochstein Award for Outstanding Achievement in Retina Research from the Retina Research Foundation (1980), and the Dolly Green Award (1982) and two Senior Scientific Investigator Awards (1990 and 1997) from Research to Prevent Blindness, Inc. He received an Award for Outstanding Contributions to Vision Research from the Alcon Research Institute (1985), and the Marjorie Margolin Prize (1994). He has served on the editorial boards of Investigative Ophthalmology and Visual Science, Journal of Neuroscience Research, Neurochemistry International, Current Eye Research, and Experimental Eye Research. Dr. Anderson has published extensively in the areas of lipid metabolism in the retina and biochemistry of retinal degenerations. He has edited 14 books, 13 on retinal degenerations and one on the biochemistry of the eye. Dr. Anderson has received grants from the National Institutes of Health, The Retina Research Foundation, the Foundation Fighting Blindness, and Research to Prevent Blindness, Inc. He has been an active participant in the program committees of the Association for Research in Vision and Ophthalmology (ARVO) and was a trustee representing the Biochemistry and Molecular Biology section. He was named a Gold Fellow by ARVO in 2009. He has served on the Vision Research Program Committee and Board of Scientific Counselors of the National Eye Institute and the Board of the Basic and Clinical Science Series of The American Academy of Ophthalmology. Dr. Anderson is a past Councilor, Treasurer, and President of the International Society for Eye Research.

Matthew M. LaVail, PhD, is Professor of Anatomy and Ophthalmology at the University of California, San Francisco School of Medicine. He received his Ph.D. degree in Anatomy (1969) from the University of Texas Medical Branch in Galveston and was subsequently a postdoctoral fellow at Harvard Medical School. Dr. LaVail was appointed Assistant Professor of Neurology-Neuropathology at Harvard Medical School in 1973. In 1976, he moved to UCSF, where he was appointed Associate Professor of Anatomy. He was appointed to his current position in 1982, and in 1988, he also became director of the Retinitis Pigmentosa Research Center at UCSF, later named the Kearn Family Center for the Study of Retinal Degeneration. Dr. LaVail has published extensively in the research areas of photoreceptor-retinal pigment epithelial cell interactions, retinal development, circadian events in the retina, genetics of pigmentation and ocular abnormalities, inherited retinal degenerations, light-induced retinal degeneration, and pharmaceutical and gene therapy for retinal degenerative diseases. He has identified several naturally occurring murine models of human retinal degenerations and has developed transgenic mouse and rat models of others. He is the author of more than 150 research publications and has edited 13 books on inherited and environmentally induced retinal degenerations. Dr. LaVail has received the Fight for Sight Citation (1976); the Sundial Award from the Retina Foundation (1976); the Friedenwald Award from the Association for Research in Vision and Ophthalmology (ARVO, 1981); two Senior Scientific Investigators Awards from Research to Prevent Blindness (1988 and 1998); a MERIT Award from the National Eye Institute (1989); an Award for Outstanding Contributions to Vision Research from the Alcon Research Institute (1990); the Award of Merit from the Retina Research Foundation (1990); the first John A. Moran Prize for Vision Research from the University of Utah (1997); the first Trustee Award from The Foundation Fighting Blindness (1998); and the Llura Liggett Gund Award from the Foundation Fighting Blindness (2007). He has served on the editorial board of Investigative Ophthalmology and Visual Science and as an Executive Editor of Experimental Eye Research. Dr. LaVail has been an active participant in the program committee of ARVO and has served as a Trustee (Retinal Cell Biology Section) of ARVO. He was named a Gold Fellow of ARVO in 2009. He has been a member of the program committee and a Vice President of the International Society for Eye research. He has also served on the Scientific Advisory Board of the Foundation Fighting Blindness since 1973.

Joe G. Hollyfield, PhD, is the Llura and Gordon Gund Professor of Ophthalmology Research in the Cole Eye Institute at The Cleveland Clinic Foundation, Cleveland, Ohio. He received a Ph.D. from the University of Texas at Austin and did postdoctoral work at the Hubrecht Laboratory in Utrecht, The Netherlands. He has held faculty positions at Columbia University College of Physicians and Surgeons in New York City and at Baylor College of Medicine in Houston, Texas. He was Director of the Retinitis Pigmentosa Research Center in The Cullen Eye Institute at Baylor from 1978 until his move to The Cleveland Clinic Foundation in 1995. He is currently Director of the Foundation Fighting Blindness Research Center at The Cleveland Clinic Foundation. Dr. Hollyfield has published over 200 papers in the area of cell and developmental biology of the retina and retinal pigment epithelium in both normal and retinal degenerative tissue. He has edited 14 books, 13 on retinal degenerations and one on the structure of the eye. Dr. Hollyfield received the Marjorie W. Margolin Prize (1981, 1994), the Sam and Bertha Brochstein Award (1985) and the Award of Merit in Retina Research (1998) from the Retina Research Foundation; the Olga Keith Weiss Distinguished Scholars' Award (1981) and two Senior Scientific Investigator Awards (1988, 1994) from Research to Prevent Blindness, Inc.; an award for Outstanding Contributions to Vision Research from the Alcon Research Institute (1987); the Distinguished Alumnus Award (1991) from Hendrix College, Conway, Arkansas; the Endre A. Balazs Prize (1994) from the International Society for Eye Research (ISER); and the Proctor Medal (2009) from the Association for Research in Vision and Ophthalmology (ARVO). He was named a Gold Fellow by ARVO in 2009. He is currently Editor-in-Chief of the journal, Experimental Eye Research published by Elsevier. Dr. Hollyfield has been active in the Association for Research in Vision and Ophthalmology (ARVO) serving on the Program Committee (1976), as Trustee (Retinal Cell Biology, 1989-94), as President (1993-94) and as Immediate Past President (1994-95). He was also President (1988-91) and Secretary (1984-87) of the International Society of Eye Research. He is Chairman of the scientific review panel for the Macular Degeneration program of the American Health Assistance Foundation (Clarksburg, MD), serves on the scientific advisory boards of the Foundation Fighting Blindness (Owings Mills, MD), the Knights Templar Eye Research Foundation (Chicago, IL), the Helen Keller Eye Research Foundation (Birmingham, AL), the South Africa Retinitis Pigmentosa Foundation (Johannesburg, South Africa), and is Co-Chairman of the Medical and Scientific Advisory Board of Retina International (Zurich, Switzerland).

Inhalt

Dedication 6
Preface 10
Contents 14
Contributors 24
Travel Awards 44
About the Editors 46
Part I Basic Science Underlying Retinal Degeneration 49
1 Analysis of Genes Differentially Expressed During Retinal Degeneration in Three Mouse Models 50
1.1 Introduction 50
1.2 Methods 51
1.2.1 RNA Preparation and cDNA Labeling 51
1.2.2 Hybridization of Slides, Image Acquisition and Bioinformatics 51
1.2.3 Real-Time PCR 51
1.3 Results 51
1.3.1 Microarray Analysis of Opsin 02550256 0/0 Model 52
1.3.2 Microarray Analysis of Bouse C Model 54
1.3.3 Microarray Analysis of MOT1 Mouse 57
1.4 Discussion 59
References 59
2 Regulation of Angiogenesis by Macrophages 61
2.1 Macrophage Polarization and Its Role in Angiogenesis 63
References 64
3 Protein Kinase C Regulates Rod Photoreceptor Differentiation Through Modulation of STAT3 Signaling 66
3.1 Introduction 66
3.2 Materials and Methods 67
3.2.1 Reagents 67
3.2.2 Animals and Retina Explant Culture 68
3.2.3 Cell Culture 68
3.2.4 Western Blot Assay 68
3.3 Results 69
3.3.1 Phorbol Esters Increase Rod Generation 69
3.3.2 Expression of PKC Isoforms in Developing Retina 70
3.3.3 Activation of PKC Decreases Phosphorylation of STAT3 70
3.4 Discussion 70
References 72
4 Pigment Epithelium-derived Factor Receptor (PEDF-R): A Plasma Membrane-linked Phospholipase with PEDF Binding Affinity 74
4.1 Introduction 74
4.2 Identification of a PEDF Receptor 75
4.3 In Silico Information 75
4.4 Expression and Distribution in the Retina 76
4.5 Transmembrane Topology 78
4.6 Binding to PEDF Ligands 78
4.7 Phospholipase Activity 78
4.8 PEDF-R Activity in Retinal Cells 79
4.9 Conclusions 79
References 81
5 The Function of Oligomerization-Incompetent RDS in Rods 83
References 89
6 The Association Between Telomere Length and Sensitivity to Apoptosis of HUVEC 91
6.1 Introduction 91
6.2 Methods 92
6.2.1 The Culture of HUVEC and the Construction of Cell Division Model 92
6.2.2 Construction of an Apoptosis Model of HUVEC with Free Hydroxyl Radicals 92
6.2.3 Measurement of Apoptosis Rates and Telomere Lengths 93
6.2.4 Statistics Analysis 93
6.3 Results 93
6.3.1 Relationship Between the Time of Culture and the Telomere Length 93
6.3.2 Relationship Among Apoptosis Rates, Culture Times and Oxidation 93
6.3.3 Oxidation Enhances the Telomere Shortening 94
6.4 Discussion 95
References 96
7 Photoreceptor Guanylate Cyclases and cGMP Phosphodiesterases in Zebrafish 98
7.1 Regulation of cGMP Levels in Photoreceptor Outer Segments 98
7.2 Retinal Disorders Associated with Mutations in RetGCs and PDE6 99
7.3 Analysis of Teleost RetGC and PDEs in Retinal Function and Disorders 100
References 103
8 RDS in Cones Does Not Interact with the Beta Subunit of the Cyclic Nucleotide Gated Channel 105
References 111
9 Increased Expression of TGF-1 and Smad 4 on Oxygen-Induced Retinopathy in Neonatal Mice 113
9.1 Introduction 113
9.2 Material and Methods 114
9.2.1 Animals 114
9.2.2 Methods 114
9.2.2.1 TGF- Immunohistochemistry (IH) and Smad-4 In Situ Hybridization (ISH) 114
9.2.3 Statistical Analysis 114
9.3 Results 115
9.4 Discussion 117
References 118
10 ZBED4, A Novel Retinal Protein Expressed in Cones and Mller Cells 120
10.1 Introduction 120
10.2 Methods and Results 121
10.2.1 ZBED4 mRNA is Expressed in Human Retina 121
10.2.2 ZBED4 mRNA is Expressed in Mouse and Human Cones 121
10.2.3 ZBED4 is Expressed Both in Nuclei and Cytoplasm of Human Cones 124
10.2.3.1 Human ZBED4 is Also Expressed in Müller Cells Endfeet 124
10.2.4 Human ZBED4 is Distributed Between Nuclear and Cytoplasmic Retinal Fractions 124
10.2.5 Subcellular Localization of ZBED4 in Stably Transfected Cells 124
10.2.6 Purification of His-Tagged ZBED4 and Its Dimerization In Vivo 126
10.2.7 Mass Spectrometry Identifies Putative Proteins Interacting with ZBED4 126
10.3 Discussion 127
References 128
11 Tubby-Like Protein 1 (Tulp1) Is Required for Normal Photoreceptor Synaptic Development 129
11.1 Introduction 129
11.2 Methods 130
11.2.1 Animals 130
11.2.2 Immunofluorescent Staining of Retinal Sections 131
11.3 Results 131
11.4 Discussion 134
References 135
12 Growth-Associated Protein43 (GAP43) Is a Biochemical Marker for the Whole Period of Fish Optic Nerve Regeneration 137
12.1 Introduction 137
12.2 Experimental Procedures 138
12.2.1 Animal 138
12.2.2 Immunohistochemistry 138
12.2.3 RT-PCR Analysis 139
12.2.4 Behavioral Analysis 139
12.3 Results 139
12.3.1 Immunohistochemical Studies of GAP43 Protein in the Goldfish Retina After Optic Nerve Transection 139
12.3.2 Time Course of GAP43 mRNA Expression in the Goldfish Retina During Optic Nerve Regeneration 139
12.3.3 Chasing Behavior of Two Goldfish with Treatment of Optic Nerve Transection During Optic Nerve Regeneration 141
12.4 Discussion 142
12.4.1 Termination of Optic Nerve Regeneration in Goldfish 142
12.4.2 GAP43 Is a Good Marker for Monitoring the Long Process of Optic Nerve Regeneration in Fish 142
References 144
13 Multiprotein Complexes of Retinitis Pigmentosa GTPase Regulator (RPGR), a Ciliary Protein Mutated in X-Linked Retinitis Pigmentosa (XLRP) 145
13.1 X-Linked RP (XLRP) 145
13.2 Retinitis Pigmentosa GTPase Regulator (RPGR) 146
13.3 RPGR Isoforms in the Retina 147
13.4 Animal Models of RPGR 147
13.5 Sensory Cilia 147
13.6 Retinal Degeneration Caused by Mutations in Ciliary Proteins 148
13.7 Macromolecular Complexes of RPGR ORF15 148
13.8 Dissection of RPGR ORF15 Complexes 149
13.9 Conclusion 150
References 151
14 Misfolded Proteins and Retinal Dystrophies 155
14.1 Endoplasmic Reticulum Stress and Retinal Degeneration 155
14.2 Misfolded Proteins in Photoreceptors 156
14.3 Misfolded Proteins in Retinal Pigment Epithelial Cells 158
14.4 Pharmacologic Targeting of Protein Misfolding to Prevent Retinal Degeneration 159
References 159
15 Neural Retina and MerTK-Independent Apical Polarity of v5 Integrin Receptors in the Retinal Pigment Epithelium 162
15.1 Introduction 163
15.2 Functions of Apical v5 Integrin Receptors in Retinal Phagocytosis and Adhesion 163
15.3 Apical Polarity of v5 Integrin Receptors is Independent of the Neural Retina 164
15.4 Apical Polarity of v5 Receptors is Independent of the Essential Engulfment Receptor MerTK 167
15.5 Motifs of the 5 Integrin Subunit Cytoplasmic Domain that May Promote Apical Polarity of v5 Integrin Receptors 168
15.6 Perspective 169
References 170
16 Mertk in Daily Retinal Phagocytosis: A History in the Making 171
16.1 Introduction 171
16.2 RCS Rat and MerTK Receptor: An Intimate Story 172
16.3 Changes Associated with Absence of MerTK in the Rat Retina 173
16.4 Daily Rhythmic Activation of Mertk: The Intracellular Way 174
16.5 The Debate About MerTK Ligands In Vivo 175
16.6 Perspectives 176
References 176
17 The Interphotoreceptor Retinoid Binding (IRBP)Is Essential for Normal Retinoid Processing in ConePhotoreceptors 179
17.1 Introduction 179
17.2 The Cone Population in Irbp/Mice 181
17.3 Implications for IRBP and Cone Function 184
17.4 The Cone Visual Cycle 184
References 186
18 Aseptic Injury to Epithelial Cells Alters Cell Surface Complement Regulation in a Tissue Specific Fashion 188
18.1 Introduction 188
18.2 Material and Methods 189
18.2.1 Reagents 189
18.2.2 Cell Culture 190
18.2.3 Flow Cytometry 190
18.3 Results 191
18.3.1 Oxidative Stress, but Not Chemical Hypoxia, Alters the Expression of Complement Regulatory Proteins on ARPE-19 Cells 191
18.3.2 Oxidative Stress of Renal Tubular Epithelial Cells Does Not Alter Surface Expression of Crry by the Cells 192
18.3.3 Expression of MCP, CD55 and CD59 on the Surface of Lung Epithelial Cells Increases After Oxidative Stress, but This Does Not Prevent Complement-Activation on the Cell Surface 192
18.4 Discussion 192
References 195
19 Role of Metalloproteases in Retinal Degeneration Induced by Violet and Blue Light 196
19.1 Introduction 197
19.2 Objective 198
19.3 Materials and Methods 198
19.4 Results 199
19.5 Conclusion 200
References 200
20 Mitochondrial Decay and Impairment of Antioxidant Defenses in Aging RPE Cells 202
20.1 Summary 202
20.2 Introduction 203
20.3 Materials and Methods 204
20.3.1 Primary Human RPE Cell Culture 204
20.3.2 Hydrogen Peroxide Toxicity -- PI Assays 204
20.3.3 Mitochondrial Morphometrics 204
20.3.4 Protein and Weight Estimation of RPE Cells and Mitochondria 205
20.3.5 Measurement of ROS, ATP and Mitochondrial Membrane Potential (00 m ) 205
20.3.6 Measurement of ([Ca2+]c) and ([Ca2+ ]m) 206
20.3.7 Expression of Mitochondrial Associated Genes 206
20.4 Results 206
20.4.1 Age Related Sensitivity of RPE Cells to Oxidative Stress 206
20.4.2 Variation in Mitochondrial Number, Structure, and Size 207
20.4.3 ROS and ATP Production, and 00 m Decrease in RPE Cells with Aging 209
20.4.4 Age-Related Variations in ([Ca2+]c) and ([Ca 2+] m ) in RPE Cells 211
20.4.5 Expression of Genes Associated with Mitochondrial Function 212
20.5 Discussion 214
References 217
21 Ciliary Transport of Opsin 221
21.1 Introduction 221
21.2 Methods 222
21.3 Results 223
21.4 Discussion 223
References 226
22 Effect of Hesperidin on Expression of Inducible Nitric Oxide Synthase in Cultured Rabbit Retinal Pigment Epithelial Cells 228
22.1 Introduction 229
22.2 Materials and Methods 230
22.2.1 Preparing Hesperidin Extract of Pericarpium Citri Reticulatae 230
22.2.2 Identification of Hesperidin by High Performance Liquid Chromatogram (HPLC) 231
22.2.3 Cell Culture 231
22.2.4 MTT Cell Viability Assay 232
22.2.5 Assay of NO Production 232
22.2.6 Cellular Immunohistochemistry of iNOS 232
22.2.7 Statistical Analysis 233
22.3 Results 233
22.3.1 Identification of Hesperidin by HPLC 233
22.3.2 RPE Cells Morphology 233
22.3.3 Influence of Hesperin on RPE Cell Proliferation Under the Condition of High Glucose 233
22.3.4 Assay of NO and iNOS 233
22.4 Discussion 234
References 235
23 Profiling MicroRNAs Differentially Expressed in Rabbit Retina 237
23.1 Introduction 237
23.2 Materials and Methods 238
23.2.1 Rabbit Retina Tissues 238
23.2.2 RNA Extraction 239
23.2.3 miRNA Microarray Analysis 239
23.2.4 Data Analysis 239
23.2.5 Bioinformatics Analysis of the Selected Mirnas 239
23.3 Results and Discussion 240
23.3.1 miRNA Microarray Analysis 240
23.3.2 Putative miRNA Target Gene Prediction 241
References 242
24 Unexpected Transcriptional Activity of the Human VMD2 Promoter in Retinal Development 244
24.1 Introduction 244
24.2 Materials and Methods 245
24.2.1 Experiment with Animals 245
24.2.2 -Galactosidase Assay 245
24.3 Results 245
24.3.1 Generation of Transgenic Mice 245
24.3.2 Localization of Cre Function in Transgenic Mice 246
24.4 Discussion 246
References 249
25 Microarray Analysis of Hyperoxia Stressed Mouse Retina: Differential Gene Expression in the Inferior and SuperiorRegion 250
25.1 Introduction 251
25.2 Methods 251
25.3 Result 252
25.4 Conclusions 255
References 255
26 Photoreceptor Sensory Cilia and Inherited Retinal Degeneration 256
26.1 PSC Proteins Involved in Inherited Retinal Degenerations 256
26.2 Structure of Photoreceptor Sensory Cilium Complex 257
26.3 Protein Components of Photoreceptor Sensory Cilium: PSC Proteome 258
26.4 Novel Photoreceptor Cilia Proteins in PSC Proteome 259
26.4.1 Subcellular Locations of Candidate Novel PSC Proteins 259
26.4.2 Functional Analysis of Novel PSC Proteins in Photoreceptor and Renal Cilia 260
26.4.2.1 shRNAs Against Novel PSC Genes 260
26.4.2.2 Evaluation of Phenotypes of shRNA Knockdown in mIMCD3 Cells and PSCs 260
26.5 TTC21B Protein in Photoreceptor Sensory Cilia and Renal Primary Cilia 261
26.5.1 TTC21B Localizes to the Basal Bodies and Transition Zone of Primary and Photoreceptor Sensory Cilia 261
26.5.2 TTC21B is Required for Primary Cilia and Photoreceptor Sensory Cilia Formation 262
26.6 Future Direction: Screening Novel PSC Genes for Mutations that Cause IRDs 263
References 263
27 Role of Elovl4 Protein in the Biosynthesisof Docosahexaenoic Acid 266
27.1 Introduction 266
27.2 Materials and Methods 267
27.2.1 RNA Interference 267
27.2.2 Construction of Mouse Anti Elovl4 Gene shRNA 267
27.2.3 Tissue Culture 268
27.2.4 Fatty Acid Analysis 268
27.3 Results 268
27.3.1 661W Cells Express Elovl4 and Can Elongate 18:3n3 and 22:5n3 to Longer Chain Fatty Acids 268
27.3.2 Knock-Down of Endogenous Elovl4 Does Not Affect C18--C24 PUFA Synthesis 269
27.4 Discussion 269
References 273
Part II Molecular Genetics and Candidate Genes 276
28 Molecular Pathogenesis of Achromatopsia Associated with Mutations in the Cone Cyclic Nucleotide-Gated Channel CNGA3 Subunit 277
28.1 Introduction 277
28.2 Materials and Methods 279
28.2.1 Constructs, Cell Culture and Transfection 279
28.2.2 Ratiometric Measurement of Intracellular Ca2+ Concentration 279
28.2.3 Electrophysiological Recordings 279
28.2.4 SDS-PAGE and Western Blot Analysis 280
28.2.5 Immunofluorescence Labeling and Confocal Microscopy 280
28.3 Results 280
28.3.1 The R218C and R224W Mutations Cause Loss of Channel Function 280
28.3.2 The R218C and R224W Mutations Cause Channel Mis-Localization 282
28.3.3 Co-Expression of The R218C and R224W Mutants with the Wild Type Channel Does Not Affect the Channel Activity 282
28.4 Discussion 283
References 284
29 Mutation Spectra in Autosomal Dominant and Recessive Retinitis Pigmentosa in Northern Sweden 286
29.1 Introduction 286
29.2 Materials and Methods 287
29.2.1 Patients and Ophthalmologic Examinations 287
29.2.2 Molecular Genetic Analysis 287
29.3 Results and Discussion 288
29.3.1 adRP 288
29.3.2 Bothnia Dystrophy 291
29.4 Conclusions 292
References 293
30 1 Rhodopsin Mutations in Congenital Night Blindness 294
30.1 Introduction 294
30.2 Properties of Rhodopsin CSNB Mutants 295
30.2.1 Spectral and Photochemical Properties 295
30.2.2 Retinal Binding Kinetics of Rhodopsin CSNB Mutants 296
30.2.3 Activity of CSNB Mutants 297
30.2.3.1 In Vitro Assays of CSNB Mutants 297
30.2.3.2 Electrophysiological Studies on Transgenic Animal Models 298
30.3 Proposed Mechanisms of CSNB Mutations 300
30.3.1 Desensitization Due to Mutant Opsin Activity in Xenopus 300
30.3.2 Proposed Dark-Active Rhodopsin in Mouse 301
30.4 Future Studies 302
References 302
31 GCAP1 Mutations Associated with Autosomal Dominant Cone Dystrophy 304
31.1 Heterogeneity of Autosomal Dominant Cone and Cone-Rod Dystrophies 305
31.2 Guanylate Cyclase 1 (GC1) and GCAP1 305
31.3 The EF Hand Motifs of GCAP1 308
31.4 GUCA1A Mutations Associated with adCD and adCRD 308
31.5 EF3: The GCAP1(Y99C) and GCAP1(N104K) Mutations 309
31.6 EF4: The GCAP1(I143NT), GCAP1(L151F) and GCAP1(E155G) Mutations 310
31.7 Conclusion 311
References 311
32 Genotypic Analysis of X-linked Retinoschisis in Western Australia 314
32.1 Introduction 314
32.2 Methodology 315
32.2.1 Molecular Genetic Studies 315
32.2.2 Electrophysiological Studies 316
32.3 Results 316
32.3.1 RS1 Mutations in Western Australian Families 316
32.3.2 Compromised Full-Field and mfERG in an Obligate Carrier with 52+1G 0 T Mutation 317
32.3.3 Likely Pathogenicity of the Novel 289TG Genetic Variant 317
32.3.3.1 Family Information 317
32.3.3.2 Patient Information 317
32.3.3.3 Genetic Information 317
32.4 Discussion 320
References 321
33 Mutation Frequency of IMPDH1 Gene of Han Population in Ganzhou City 323
33.1 Introduction 323
33.2 Materials and Methods 324
33.2.1 Subjects 324
33.2.2 DNA Extraction 325
33.2.3 Amplification of IMPDH1 Genes 325
33.2.4 RFLP Analysis 325
33.2.5 Statistical Analysis 325
33.3 Results 326
33.4 Discussion 326
References 327
Part III Diagnostic, Clinical, Cytopathological and Physiologic Aspects of Retinal Degeneration 328
34 Reversible and Size-Selective Opening of the Inner Blood-Retina Barrier: A Novel Therapeutic Strategy 329
34.1 Introduction 329
34.2 Materials and Methods 331
34.2.1 Animal Experiments and Experimental Groups 331
34.2.2 Web-Based siRNA Design Protocols Targeting Claudin-5 332
34.2.3 In Vivo Delivery of siRNA to Murine Retinal Capillary Endothelial Cells by Large Volume Hydrodynamic Injection 332
34.2.4 Indirect Immunostaining of Retinal Flatmounts 332
34.2.5 Assessment of BRB Integrity by Perfusion of Hoechst (H33342) 333
34.2.6 Magnetic Resonance Imaging (MRI) 333
34.3 Results 333
34.3.1 Claudin-5 Levels in Retinal Flatmounts 333
34.3.2 Perfusion of Hoechst 33342 (562 Da) in Mice Post-Delivery of Claudin-5 Sirna 333
34.3.3 MRI Analysis of Ibrb Integrity Following Rnai of Claudin-5 334
34.4 Discussion 334
References 336
35 Spectral Domain Optical Coherence Tomography and Adaptive Optics: Imaging Photoreceptor Layer Morphology to Interpret Preclinical Phenotypes 337
35.1 Introduction 337
35.2 Materials and Methods 338
35.2.1 Subjects 338
35.2.2 Adaptive Optics Retinal Imaging 339
35.2.3 Spectral Domain Optical Coherence Tomography 340
35.3 Results 341
35.3.1 Cone Photoreceptor Mosaic Topography 341
35.3.2 Outer Nuclear Layer Thickness 342
35.4 Discussion 343
References 343
36 Pharmacological Manipulation of Rhodopsin RetinitisPigmentosa 345
36.1 Introduction 345
36.2 Pharmacological Strategies for Misfolding Mutant Rod Opsin 346
36.2.1 Pharmacological Chaperones 346
36.2.2 Kosmotropes 347
36.2.3 Molecular Chaperone Inducers 348
36.2.4 Autophagy Inducers 349
36.3 Conclusion 349
References 350
37 Targeted High-Throughput DNA Sequencing for Gene Discovery in Retinitis Pigmentosa 352
37.1 Introduction 352
37.2 Methods 354
37.2.1 Selection of Families 354
37.2.2 VisionCHIP Gene Selection 354
37.2.3 VisionCHIP Validation 355
37.2.4 Evaluating Potentially Pathogenic Variants 356
37.3 Conclusion 357
References 358
38 Advances in Imaging of Stargardt Disease 359
38.1 Introduction 359
38.2 Fundus Autofluorescence 360
38.3 OCT 361
38.4 Adaptive Optics Scanning Laser Ophthalmoscope 361
38.5 Conclusion 364
References 365
39 Protamine Sulfate Downregulates Vascular Endothelial Growth Factor (VEGF) Expression and Inhibits VEGF and Its Receptor Binding in Vitro 367
39.1 Materials and Methods 368
39.1.1 Cell Culture 368
39.1.2 Semi-Quantitative Assay of VEGF Expression in the Culture Cells by ICC 368
39.1.3 VEGF Expression was Determined by ELISA 369
39.1.4 Statistical Analysis 369
39.2 Results 369
39.2.1 The Maximum Inhibition of VEGF Expression by Protamine Sulfate 369
39.2.2 Protamine Sulfate Inhibits the RF/6A Cell VEGF Expression at the Hypoxic Condition 369
39.2.3 Protamine Sulfate Inhibits the Binding of VEGF to Its Receptor 370
39.3 Discussions 371
39.3.1 The Inhibition Effect of Protamine Sulfate on VEGF 372
39.3.2 Inhibition of the Binding Between VEGF and Its Receptor 372
39.3.3 The Potential Use of Protamine Sulfate Inhibition of Angiogenic Eye Diseases 373
References 373
40 Computer-Assisted Semi-Quantitative Analysis of Mouse Choroidal Density 374
40.1 Introduction 374
40.2 Methods 375
40.2.1 Immunohistochemial Staining of Choroidal Endothelia 375
40.2.2 Analysis of Choriodal Density with Photoshop 8.0 375
40.3 Results and Discussion 376
40.3.1 Analysis Of Choroidal Density 376
40.3.2 Usefulness of the Methodology 377
40.3.3 Summary 377
References 378
41 Thioredoxins 1 and 2 Protect Retinal Ganglion Cells from Pharmacologically Induced Oxidative Stress, Optic Nerve Transection and Ocular Hypertension 379
41.1 Introduction 379
41.2 Methods 380
41.2.1 Animals 380
41.2.2 RGC Counting 381
41.2.3 RGC Isolation 381
41.2.4 Western Blot Analysis 381
41.2.5 RGC-5 Culture and Transfection 381
41.2.6 Cell Viability Assay 382
41.2.7 In Vivo Electroporation (ELP) 382
41.2.8 Statistical Analysis 382
41.3 Results 382
41.3.1 Expression of TRX1, TRX2 and TXNIP in the Retina After ONT and IOP Elevation and in RGC-5 Cells with Induced Oxidative Stress 382
41.3.1.1 TRX Expression in RGC-5 Cells in Response to Oxidative Stress 382
41.3.1.2 The Levels of TRX Proteins After ONT 383
41.3.1.3 The Levels of TRX Proteins After IOP Elevation 383
41.3.2 The Effect of TRX1 and TRX2 Overexpression on RGC Survival 383
41.3.2.1 TRX1 and TRX2 Overexpression Protects RGC-5 cells Against Oxidative Stress 383
41.3.2.2 TRX1 and TRX2 Overexpression Increases RGC Survival After ONT 384
41.3.2.3 TRX1 and TRX2 Overexpression Increases RGC Survival After IOP Elevation 384
41.4 Discussion 385
References 386
42 Near-Infrared Light Protect the Photoreceptor from Light-Induced Damage in Rats 388
42.1 Introduction 389
42.2 Material and Methods 390
42.2.1 Animal 390
42.2.2 Light Damage 390
42.2.3 670 nm LED Treatment 390
42.2.4 Evaluation of Photoreceptor Cell Function by Electroretinography 390
42.2.5 Morphological Evaluation of Photoreceptor Rescue by Quantitative Histology 391
42.2.6 Statistical Analysis 391
42.3 Results 391
42.3.1 LED Attenuated the Light Damage Area in Retinas 391
42.3.2 LED Protected the Morphology of Light Damage Retina 391
42.3.3 LED Protected the Function of Light Damage Retina 393
42.4 Discussions 394
References 396
43 BDNF Improves the Efficacy ERG Amplitude Maintenance by Transplantation of Retinal Stem Cells in RCS Rats 398
43.1 Introduction 398
43.2 Methods 399
43.2.1 Animals 399
43.2.2 Cell Preparation and Subretinal Transplantation 399
43.2.3 Flash-Electroretinogram (F-ERG) Recordings 400
43.2.4 Histology and Quantification 400
43.2.5 Data Analysis 401
43.3 Results 401
43.3.1 ERG Amplitudes and Latencies 401
43.3.2 ONL Thickness 402
43.3.3 Graft Cells Survival After Subretinal Transplantation 402
43.4 Discussion 402
References 406
44 The Role of Purinergic Receptors in Retinal Functionand Disease 408
44.1 Introduction 408
44.2 Mechanisms of ATP Release and Degradation 409
44.2.1 ATP Release 409
44.2.2 Degradation of ATP 409
44.3 Purinergic Signaling in the Retina 410
44.3.1 Purinergic Modulation of Neuronal Signaling 410
44.3.2 ATP and Glial Transmission 411
44.4 The Role of Purinergic Receptors in Retinal Disease 411
44.5 Concluding Remarks 412
References 412
Part IV Macular Degeneration 415
45 Fundus Autofluorescence Imaging in Age-Related Macular Degeneration and Geographic Atrophy 416
45.1 Background 416
45.2 Fundus Autofluorescence Overview 417
45.3 FAF Findings in Early AMD with Drusen Only 419
45.4 FAF Findings in Late AMD with Geographic Atrophy 419
45.5 Progression of Geographic Atrophy 420
45.6 Mechanisms of Progression 420
45.7 Research to Prevent Progression 421
45.8 Discussion 422
References 422
46 Endoplasmic Reticulum Stress as a Primary Pathogenic Mechanism Leading to Age-Related Macular Degeneration 424
46.1 Age Related Macular Degeneration Is a Leading Cause of Vision Loss 424
46.2 Oxidative Stress and Complement Activation are Common Pathways in End-Stage Disease 425
46.3 ER Stress and Oxidative Stress Interact 426
46.4 ER and Oxidative Stress as Triggers for Inflammation and Disease 426
46.5 Future Experimental Approaches 427
References 428
47 Proteomic and Genomic Biomarkers for Age-Related Macular Degeneration 431
47.1 Introduction 431
47.2 Methods 432
47.3 Results 433
47.3.1 CEP Adducts and Autoantibodies Are Elevated in AMD Plasma 433
47.3.2 AMD Risk Based on CEP Biomarkers and Genotype 433
47.3.3 The Association Between CEP Biomarkers and AMD Risk Genotypes 434
47.4 Discussion 436
References 436
48 Impaired Intracellular Signaling May AllowUp-Regulation of CTGF-Synthesis and Secondary Peri-Retinal Fibrosis in Human Retinal Pigment Epithelial Cells from Patients with Age-Related Macular Degeneration 438
48.1 Introduction 438
48.2 Methods 439
48.2.1 Chemicals 439
48.2.2 Establishment and Maintenance of hRPE Cell Cultures 440
48.2.3 Cellular Proliferation 440
48.2.4 Immunoprecipitation Assay 440
48.2.5 Statistical Analysis 441
48.3 Results 441
48.3.1 Effect of Glucose on 14C-CTGF Synthesis in hRPE Cells 441
48.3.2 Effect of IGF-1 on 14C-CTGF Synthesis in hRPE cells 441
48.3.3 Effect of PD98059 on Glucose Stimulated 14C-CTGF Synthesis in hRPE Cells 442
48.3.4 Effect of PD98059 on IGF-1 Stimulated 14C-CTGF Synthesis in hRPE Cells 442
48.4 Discussion 445
References 446
49 PPAR Nuclear Receptors and Altered RPE Lipid Metabolism in Age-Related Macular Degeneration 448
49.1 Introduction 448
49.1.1 Current Hypotheses Surrounding Sub-RPE Deposit Formation 449
49.1.2 Long Chain Poly-Unsaturated Fatty Acids (LCPUFA) are Associated with ARMD Risk 449
49.1.3 Peroxisome Proliferator Activated Receptors (PPARs) are Expressed in ARPE19 Cells 450
49.2 LcPUFA Regulates Gene Expression in ARPE19 Cells 451
49.2.1 Purpose and Methods 451
49.2.2 Results 451
49.2.3 Discussion 452
References 453
50 The Pathophysiology of Cigarette Smoking and Age-Related Macular Degeneration 456
50.1 Introduction 456
50.2 Cigarette Smoking as a Risk Factor for AMD 457
50.2.1 AMD and Cigarette Smoke 457
50.2.2 Cigarette Smoke Constituents 457
50.3 Oxidative Stress 457
50.3.1 Oxidative Damage in AMD 457
50.3.2 Reactive Oxygen Species in Cigarette Smoke 458
50.3.3 Acrolein-Induced Oxidative Stress 458
50.3.4 Cadmium-Induced Oxidative Stress 458
50.4 Cigarette Smoke Depletion of Antioxidant Protection 459
50.4.1 Systemic Antioxidant Mechanisms 459
50.4.2 Local Ocular Antioxidants 459
50.5 Non-oxidative Chemical Damage by Cigarette Smoke 460
50.5.1 Nicotine 460
50.5.2 Polycyclic Aromatic Hydrocarbons 460
50.6 Inflammation 460
50.6.1 Inflammation and AMD 460
50.6.2 Cigarette Smoke and Complement Pathway 461
50.6.3 Cigarette Smoke and Other Inflammatory Mediators 461
50.7 Vascular Changes 461
50.8 Conclusions 461
References 462
51 Oxidative Stress and the Ubiquitin Proteolytic System in Age-Related Macular Degeneration 466
51.1 Oxidative Stress and Age-Related Macular Degeneration 466
51.2 The Ubiquitin Proteolytic System (UPS) and Oxidative Stress in the Retina 467
51.3 The UPS and the Cytoprotective Transcription Factor, Nrf2 471
References 473
52 Slit-Robo Signaling in Ocular Angiogenesis 476
52.1 Ocular Angiogenesis 476
52.2 Slit-Robo Signaling in Axon Guidance 477
52.3 Slit-Robo Signaling in Angiogenesis 478
52.4 Slit-Robo Signaling in Ocular Angiogenesis 479
52.5 Signaling Pathway of Slit-Robo System in Angiogenesis 480
52.6 Perspective 481
References 481
Part V Animal Models of Retinal Degeneration 483
53 Evaluation of Retinal Degeneration in P27KIP1 Null Mouse 484
53.1 Introduction 485
53.2 Materials and Methods 485
53.2.1 Animals and Biosafety 485
53.2.2 MNU-Induced Retinal Degeneration 485
53.2.3 Electroretinography 485
53.2.4 Histological Examination and Immunohistochemistry 486
53.3 Results 486
53.3.1 Fundus Examination and Histology of the Retina 486
53.3.2 ERG 486
53.3.3 BrdU Incorporation 487
53.3.4 Immunohistology of Nestin 487
53.4 Discussion 487
References 488
54 Differences in Photoreceptor Sensitivity to Oxygen Stress Between Long Evans and Sprague-Dawley Rats 489
54.1 Introduction 489
54.2 Methods 490
54.2.1 Animal Strains and Oxygen Exposure 490
54.2.2 Electroretinography 490
54.2.3 Immunohistochemistry and TUNEL Labeling 491
54.3 Results 491
54.3.1 Rod and Cone Components of the ERG after Hyperoxia 491
54.3.2 Impact of Hyperoxia on the Rate of Photo receptor Death 491
54.3.3 Impact of Hyperoxia on GFAP Expression 491
54.4 Discussion 493
References 494
55 Retinal Degeneration in a Rat Model of Smith-Lemli-Opitz Syndrome: Thinking Beyond Cholesterol Deficiency 496
55.1 Introduction 496
55.2 The AY9944 Rat Model of SLOS: Biochemical Findings 498
55.3 Retinal Degeneration in the SLOS Rat Model: Histology and Ultrastructure 499
55.4 Retinal Degeneration in the SLOS Rat Model: Electrophysiological Deficits 501
55.5 Effects of Feeding a High-Cholesterol Diet 501
55.6 Perspective: Thinking Beyond the Cholesterol Deficiency in SLOS 502
References 503
56 Do Calcium Channel Blockers Rescue Dying Photoreceptors in the Pde6brd1 Mouse? 505
56.1 Introduction 505
56.1.1 The Pde6brd1 Mouse and Increased [cGMP] 506
56.1.2 Calcium Regulation and Overload in the Photoreceptor Inner Segment 507
56.2 D-cis-diltiazem and Neuroprotection in the Retina 508
56.2.1 Criticism of the Frasson Study 508
56.2.2 Subsequent Evidence Shows L-Type Channels Are Involved in Degeneration 509
56.3 Other Players May Be Involved 510
References 511
57 Effect of PBNA on the NO Content and NOS Activityin Ischemia/Reperfusion Injury in the Rat Retina 514
57.1 Introduction 515
57.2 Materials and Methods 515
57.2.1 Animals and Reagents 515
57.2.2 Induction of Retinal I/R 515
57.2.3 Detection of MDA and NO Concentration, SOD and GSH-PX Activity 516
57.2.4 Statistical Analysis 516
57.3 Results 516
57.3.1 Effect of PBNA on Serum NO Content in Retinal I/R Injury 516
57.3.2 Effect of PBNA on T-NOS Activity in Retinal I/R Injury 516
57.3.3 Effect of PBNA on iNOS Activity in Retinal I/R Injury 517
57.3.4 Effect of PBNA on Serum eNOS Activity in Retinal I/R Injury 518
57.4 Discussion 519
References 519
58 Recent Insights into the Mechanisms Underlying Light-Dependent Retinal Degeneration from X. Laevis Models of Retinitis Pigmentosa 521
References 526
59 A Hypoplastic Retinal Lamination in the Purpurin Knock Down Embryo in Zebrafish 528
59.1 Introduction 528
59.2 Materials and Methods 529
59.2.1 Experimental Animals 529
59.2.2 Screening of Genomic DNA for Zebrafish Purpurin 529
59.2.3 Construction of the pur-GFP Reporter Vector 530
59.2.4 Morpholino and Microinjections 530
59.2.5 In Situ Hybridization 530
59.2.6 RNA Isolation, RT-PCR and mRNA Synthesis 530
59.3 Results 531
59.3.1 Isolation and Characterization of Zebrafish Purpurin Gene 531
59.3.2 Similar Phenotypes of Purpurin and Crx Morphant 531
59.4 Rescuing Effect of Purpurin mRNA to the Crx Morphant 532
59.5 Discussion 533
References 534
60 Functional Changes in Inner Retinal Neurons in Animal Models of Photoreceptor Degeneration 536
60.1 Introduction 536
60.2 Bipolar Cell Function in Retinal Degeneration 537
60.2.1 Glutamate Receptors of Bipolar Cells in the Normal and Degenerating Retina 537
60.2.2 Evidence for Bipolar Cell Dysfunction 538
60.2.2.1 Rod Bipolar Cells 538
60.2.2.2 Cone Bipolar Cells 540
60.3 Ganglion Cell Function in Retinal Degeneration 540
References 542
61 Photoreceptor Cell Degeneration in Abcr--/-- Mice 544
61.1 Introduction 544
61.2 Methods 545
61.2.1 Animals and Rearing 545
61.2.2 Measurement of Outer Nuclear Layer Thickness 546
61.2.3 Counting Photoreceptor Nuclei 546
61.3 Results 546
61.4 Discussion 548
References 549
62 Investigating the Mechanism of Disease in the RP10 Form of Retinitis Pigmentosa 551
62.1 Introduction 551
62.2 Retinitis Pigmentosa 552
62.3 RP10 Disease Caused by Mutations in IMPDH1 552
62.4 IMPDH Structure and Function 553
62.5 IMPDH Binds Single Stranded Nucleic Acids 554
62.6 Retinal Isoforms of IMPDH1 554
62.7 Kinetic and Nucleic Acid Binding Properties of Retinal IMPDH1 556
62.8 Conclusion 556
References 557
63 Congenital Stationary Night Blindness in Mice A Tale of Two Cacna1f Mutants 559
63.1 Introduction 560
63.2 Methods 560
63.3 Results 561
63.4 Discussion 564
63.5 Conclusion 566
References 566
64 Protection of Photoreceptors in a Mouse Model of RP10 569
64.1 Introduction 569
64.2 Results 570
64.2.1 Evaluation of Optimal IMPDH1 Suppressors 570
64.2.2 RP10 Mouse Model 571
64.2.3 Rescue of Photoreceptor Cells by rAAV-Mediated Downregulation of Mutant IMPDH1 572
64.3 Discussion 573
References 574
65 Correlation Between Tissue Docosahexaenoic Acid Levels and Susceptibility to Light-Induced Retinal Degeneration 576
65.1 Introduction 576
65.2 Methods 577
65.3 Results 578
65.4 Discussion 580
References 581
66 Activation of Mller Cells Occurs During Retinal Degeneration in RCS Rats 583
66.1 Introduction 583
66.2 Materials and Methods 584
66.2.1 Animal 584
66.2.2 Immunohistochemical Staining 584
66.2.3 Western Blot Test 585
66.2.4 Müller Cell Cultures 585
66.2.5 Data Analysis 585
66.3 Results 586
66.3.1 Morphology and Quantity Changes of Müller Cells 586
66.3.2 Expression of GFAP and ERK in RCS Rat Müller Cells 586
66.3.3 Effect of Mixed Retinal Cells of RCS Rats on Normal Müller Cells 587
66.4 Discussion 587
References 590
67 Effect of 3-Daidzein Sulfonic Sodium on theAnti-oxidation of Retinal Ischemia/Reperfusion Injury in Rats 592
67.1 Introduction 593
67.2 Materials and Methods 593
67.2.1 Animals and Reagents 593
67.2.2 Induction of RI/R 593
67.2.3 Detection of MDA and NO Concentration, SOD and GSH-PX Activity 594
67.2.4 Statistical Analysis 594
67.3 Results 594
67.3.1 The Effect of DSS on the Concentration of MDA in Serum After RI/R Injury 594
67.3.2 The Effect of DSS on the Activity of SOD in Serum After RI/R Injury 595
67.3.3 The Effect of DSS on the Activity of Serum GSH-PX After RI/R Injury 595
67.3.4 The Effect of DSS on the Concentration of Serum NO After RI/R Injury 596
67.4 Discussion 597
References 597
68 Structural and Functional Phenotyping in theCone-Specific Photoreceptor Function Loss 1 (cpfl1) Mouse Mutant -- A Model of Cone Dystrophies 599
68.1 Introduction 600
68.2 Materials and Methods 600
68.2.1 Animals 600
68.2.2 Functional Testing 600
68.2.3 In Vivo Imaging 601
68.3 Results 601
68.3.1 Function 601
68.3.2 Morphology 603
68.4 Discussion 603
References 605
69 The Differential Role of Jak/Stat Signaling in Retinal Degeneration 606
69.1 Introduction 606
69.2 Materials and Methods 607
69.2.1 Mice and Light Exposure 607
69.2.2 Semi-Quantitative Real Time Polymerase Chain Reaction (PCR) 607
69.3 Results 608
69.3.1 STATs Are Induced Differently in Retinas of Light-Exposed and rd1 Mice 608
69.3.2 Shp-1 Is Induced After Light Exposure But Not in the rd1 Mouse 609
69.3.3 Jak3 mRNA Is Induced Similarly in the Model of Light Induced Photoreceptor Cell Death and the rd1 Mouse Model 609
69.4 Discussion 611
References 612
Part VI Neuroprotection and Gene Therapy 613
70 Gene Therapy in the Retinal Degeneration Slow Model of Retinitis Pigmentosa 614
70.1 Introduction 614
70.2 Diseases Associated with RDS Mutations 615
70.3 Current Animal Models 615
70.4 Gene Therapy in rds Models 616
70.5 Viral Gene Therapy Approaches 616
70.6 Non-viral Approaches 618
References 620
71 PEDF Promotes Retinal Neurosphere Formation and Expansion In Vitro 623
71.1 Introduction 623
71.2 Materials and Methods 625
71.2.1 Retinal Stem Cell Isolation and Culture 625
71.2.2 Single Sphere Passaging 625
71.2.3 Bromodeoxyuridine Labeling 625
71.2.4 Retinal Stem Cell Differentiation 626
71.2.5 Immunofluorescence 626
71.3 Results 626
71.3.1 PEDF Promotes Retinal Neurospheres Growth and Self-Renewal 626
71.3.2 Retinal Neurosphere Proliferation 628
71.3.3 Differentiation of Retinal Cells Precursors from RSCs 629
71.4 Discussion 630
References 631
72 A Multi-Stage Color Model Revisited: Implications for a Gene Therapy Cure for Red-Green Colorblindness 633
72.1 Introduction 633
72.2 A Brief History of Color Vision Theory 634
72.3 Color Vision from an Evolutionary Perspective 635
72.4 Evolutionary Constraints Lead to an Extension of Devalois Model 636
72.5 The Possibility of Gene Therapy to Cure Red-Green Colorblindness 639
References 640
73 Achromatopsia as a Potential Candidate for Gene Therapy 641
73.1 Human Achromatopsia 641
73.1.1 Clinical Manifestations 642
73.1.2 Current Achromatopsia Treatments 642
73.2 Genetics of Human Achromatopsia 642
73.2.1 GNAT2 Achromatopsia 643
73.2.2 CNG Achromatopsia 644
73.2.3 Achromatopsia Gene Therapy 644
73.3 The Mutant Gnat2 Mouse and Gene Therapy 644
73.3.1 The Cnga3 Mutant Mouse and Gene Therapy 645
73.3.2 The Cngb3 Mutant Dog and Gene Therapy 647
73.4 Prospects for Achromatopsia Gene Therapy 647
References 647
74 Function and Mechanism of CNTF/LIF Signalingin Retinogenesis 649
74.1 Introduction 649
74.2 Effects of CNTF/LIF on Photoreceptor and Bipolar Neuron Differentiation 650
74.3 Effects of CNTF/LIF on Muller Glia Genesis and Late Progenitor Proliferation 651
74.4 Effects of LIF Misexpression on Retinal Vasculature Development 651
74.5 Expression of CNTF/LIF Signaling Components in the Developing Retina 652
74.6 Signaling Events Triggered by CNTF/LIF During Retinogenesis 652
74.7 CNTF/LIF Regulate Numerous Genes Involved in Retinogenesis 653
74.8 Perspective 654
References 654
75 gp130 Activation in Muller Cells is Not Essentialfor Photoreceptor Protection from Light Damage 657
75.1 Introduction 657
75.2 Conditional gp130 Knockout in the Retinal Mller Cells 658
75.3 Effect of Impaired gp130 Activation in Mller Cells on LIF-Induced Photoreceptor Protection 659
75.4 Discussion 660
References 662
76 Neuroprotectin D1 Modulates the Induction of Pro-Inflammatory Signaling and Promotes Retinal Pigment Epithelial Cell Survival During Oxidative Stress 664
76.1 The Importance of RPE Cell Function and Integrity for Photoreceptor Survival 664
76.2 The Loss of RPE Cells in Retinal Degeneration 667
76.3 DHA and NPD1 Properties and Neuroprotection 668
76.4 NPD1 Modulates the Expression of Survival and Apoptotic-Related Proteins 669
References 669
77 Adeno-Associated Virus Serotype-9 Mediated Retinal Outer Plexiform Layer Transduction is Mainly Through the Photoreceptors 672
77.1 Introduction 673
77.2 AAV9-Mediated Gene Transfer in the Retina 673
77.3 The Sub-Cellular Location of AAV9 Transduction in the OPL 675
77.4 AAV9-Mediated Retinal Gene Transfer in mdx 3cv Mice 675
77.5 Subretinal Injection of AAV9 Vector Did Not Cause Acute Retinal Damage 677
77.6 Conclusions 677
References 677
Index 680

Produkt-Details

• Erscheint lt. Verlag: 17.3.2010
• Reihe/Serie: Advances in Experimental Medicine and Biology
• Reihe/Serie: Advances in Experimental Medicine and Biology
• Zusatzinfo: XLVII, 808 p. 100 illus.
• Verlagsort: New York
• Sprache: englisch
• Themenwelt: Medizin / Pharmazie ► Medizinische Fachgebiete ► Augenheilkunde
• Schlagworte: Adaptive optics • Hypertension • macular degeneration • Morphology • pathogenesis • retina
• ISBN-10: 1-4419-1399-8 / 1441913998
• ISBN-13: 978-1-4419-1399-9 / 9781441913999