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Mesencephalic Astrocyte-derived Neurotrophic Factor (MANF) Protects Rod and Cone Photoreceptors from Degeneration in Transgenic Rats Carrying the S334ter Rhodopsin Mutation
Presentation Start/End Time:
Monday, May 07, 2012, 3:45 PM - 5:30 PM
Life and Death of Photoreceptors
280 photoreceptor degeneration/apoptosis: cell biology and pathology - RC
Rong Wen, Lingyu Luo, Dequang Huang, Xin Xia, Zhengying Wang, Pingping Chen, Yiwen Li.
Bascom Palmer Eye Inst, University of Miami, Miami, FL.
612 neuroprotection, 645 photoreceptors, 692 retinal degenerations: cell biology
Mesencephalic astrocyte-derived neurotrophic factor (MANF) is a member of a novel evolutionarily conserved protein family with neurotrophic capabilities. It was identified from the conditional medium of a rat type-1 astrocyte cell line, the ventral mesencephalic cell line 1 (VMCL1) to be a factor that promotes the survival of cultured embrayonic dopaminergic neurons. It also significantly reduces the infarction in the ischemic cortex in a rat model of stroke and promotes the survival of cultured heart muscle cells. In the present study, we examined the neuroprotective effects of MANF on photoreceptors.
Recombinant human MANF was expressed in E. coli and purified. To examine the neuroprotective effect of MANF on rod photoreceptors, recombinant human MANF (6 µg) was intravitreally injected to the left eyes of transgenic rat S334ter rats at PD9, and the right eyes were injected with PBS as controls. Eyes were collected at PD21 examined by light microscopy. To examine effect of MANF on cone photoreceptors, MANF (6 µg) was injected intravitreally to the left eyes of the transgenic rats at PD20, and the right eyes were injected with PBS as controls. Retinas were collected at PD30 and cone outer segments were stained with Alexa 488 conjugated peanut agglutinin (PNA). Flat-mounted retinas were examined by confocal microscopy.
MANF significant protected rod photoreceptors. In MANF-treated retinas, three to four rows of nuclei remained in the outer nuclear layer (ONL), as compare to only one row of nuclei in PBS-treated fellow eyes. The thickness of ONL in MANF-treated retinas (superior region) (17.47±3.96 µm, mean±SD, n=5) is significantly greater than that in PBS-treated retinas (7.07±1.12, n=5) (
< 0.001, student
-test). Loss of cone outer segments was characterized by many PNA-negative areas distributed across the retina in PBS treated retinas, as reported previously,. In RdCVF treated eye, however, the PNA negative areas became much smaller or in many cases completely disappeared. Quantitative analysis showed that PNA-positive cells are significantly more in MANF treated retinas (569.5±46.5/0.1 mm
, mean±SD, n=6) than in PBS-treated retinas (398.7±25.4, n=6) (
< 0.001, student
Intravitreal injection of recombinant human MANF protein protects both rod and cone from degeneration.
NIH Grant R01EY018586, P30EY14801; Hope for Vision, the James and Esther King Biomedical Research Program of the State of Florida JEK 08-KN-09; the Department of Defense Grant W81XWH-09-1-0674; RPB.
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