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Effects of AQP4 Deficiency on Light-induced Retinal Damage

Author: ZhangWenJing
Tutor: JiangQin；HuGang
School: Nanjing Medical University
Course: Pharmacology
Keywords: Aquaporin 4 Photodamage ERG Degeneration of photoreceptor cells Müller cells
CLC: R774.1
Type: Master's thesis
Year: 2009
Downloads: 38
Quote: 0
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Abstract


Aquaporin (aquaporin, AQP) are a class of widely present in the eukaryotic cell membrane selectively efficient transporter specific channels of the water molecules, and on the body of liquid water balance and stability of the cellular microenvironment play an important regulatory role. Aquaporin family of 13 members (AQP0-12) so far, have been found in mammals. Aquaporin 4 (aquaporin-4, AQP4) is mainly distributed in the central nervous system (central nervous system, CNS), followed by the body tissue of the spinal cord, eyes, lungs, kidneys and gastrointestinal tract also a certain amount of expression . In the eye, AQP4 expression in Müller cells of the retina and the optic nerve astrocytes. The former accounted for 90% of the retina glial component is only across the full thickness of the cells of the retina, the steady-state environment on the maintenance of the retina, uptake of neurotransmitters and fight free radicals damage important role, while Müller cells are widely involved in a variety of acute the occurrence of chronic retinal diseases, development. Environment and artificial light sources may be a potential threat to retinal light damage. Continued to glare as easily lead to retinal photochemical damage, injury early retinal dysfunction, loss of photoreceptor cells, the latter part of the inner nuclear layer cell necrosis, which eventually led to the loss of visual function. In addition, there are many similarities in age-related macular degeneration, retinal pigment degeneration and retinal light damage. Retinal light damage mechanisms and drug prevention and treatment has been an important research topic in the field of ophthalmology. Evidence has been made in recent years show that the expression of AQP4 Müller cells play an important role in ophthalmic physiology and pathology, physiological state Müller cells missing the AQP4 slightly reduced the ERG (electroretinogram, ERG) b-wave amplitude; in rat retinal light damage model AQP4 protein expression increased. In addition, in a mouse model of retinal ischemia AQP4 gene knockdown in addition to significantly slow down the process of degeneration of the retinal cells of the inner nuclear layer. AQP4 role in the study of retinal photochemical damage due to the current lack of a specific inhibitor, requires the use of knockout mice of this effective means. To carry out a study of the role of AQP4 in retinal light damage, will face groping CD1 genetic background of colored mouse retinal light damage light source intensity conditions. Therefore, application of the AQP4 gene knockout mice, the AQP4 gene knockout of retinal light damage before and after the function and morphology in the establishment of retinal light damage model based on the study, to further explore its mechanism for retinal light damage The mechanism of accumulation of the academic foundation and provide a new strategy for the treatment of human ophthalmic diseases. Objective: To study AQP4 gene knockout retinal light damage. Methods: 2-month-old adult wild-type (AQP4 / ) and AQP4 gene deletion (AQP4 - / - ) female mice, 1) retinal light damage model making : mouse dark adaptation 24h, 1% tropicamide faint red light under dilate, 5min after exposure to a white fluorescent light, the light intensity of 16,000 ± 2,000 lux, every day to 12h (8:00 am-20: 00pm) irradiation night's rest 12h, six days in a row, the establishment of the retinal light damage model. 2) ERG (ERG) detection: ERG detection AQP4 / and of AQP4 of - / - the mouse basal state (control, Con) light damage after the first day ( post-light 1 day, PL1), on the third day (post-light day, PL3) and the seventh day (post-light 7 day, PL7) retinal function. 3) of pathology paraffin HE staining optical Con with light damage before the injury PL3 PL7 three groups within the retina (inner nuclear layer, INL), the morphology of the thickness of the outer nuclear layer (outer nuclear layer, ONL) qualitative and quantitative evaluation . 4) immunofluorescence method: observation Müller cell-specific marker glial fibrillary acidic protein (glial fibrillary acidic protein, GFAP) Con, PL3 groups express strength and morphological changes. Results: 1) the the physiological state of AQP4 under / and AQP4 in - / - mouse ERG and retinal morphology were not significantly different. 2) light after the third day (PL3) of AQP4 / mouse retina electroretinogram a, b wave amplitude decreased to a light level before in PL7 when recovery; of AQP4 - / - < / sup> mouse retina after light damage the retina a, b wave in PL3 disappear and PL7 when a, b amplitude does not improve. AQP4 / mice in the light before and after the normal retinal morphology clear hierarchy structure, the inner and outer nuclear layer neat rules, outer nuclear layer cells arranged in layers 9-11; of AQP4 - / - PL3 when mice illumination retinal structure was destroyed, the performance for the thinning of the outer nuclear layer cell arrangement sparse cell layers is reduced to 2-3 layers disappear to the PL7 when the outer nuclear layer inner and outer sections shorter, inner nuclear layer cells disorganized, and cell loss. 3) immunofluorescence observed GFAP expression of Müller cells of AQP4 / mice PL3 cell body larger; AQP4 - / - mice Müller cell body larger and synaptic extending outward activation compared of AQP4 / mice significantly. CONCLUSION: AQP4 gene deletion leads CD1 mouse retina outer nuclear layer more vulnerable to light due to degenerative changes; photodamage, missing the AQP4 The Müller cell GFAP expression increased, and extended the branch outward nuclear layer (ONL) extends to show that the light injury reactive gliosis, and the occurrence of retinal damage in this reaction may be accelerated. In summary, the research innovation lies: Application AQP4 gene knock addition to mice to examine the role of AQP4 in retinal light damage, to obtain direct evidence of AQP4 play a neuroprotective; findings suggest that AQP4 might regulate Müller cells \

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CLC: > Medicine, health > Ophthalmology > Retina and optic nerve diseases > Retinal disease
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