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M. J. Seiler, B. B. Thomas, Z. Chen, Q. Peng, S. R. Sadda, R. B. Aramant; Transplanted Cells Extend Multiple Neuronal Processes Into Degenerate Host Retina - Supporting Results of Restored Visual Function. Invest. Ophthalmol. Vis. Sci. 2007;48(13):5081.
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To further investigate the interactions between degenerating retina and retinal progenitor sheet transplants.
Donor retinal sheets were isolated from E19 fetuses of transgenic rats expressing human alkaline phosphatase (hPAP), and transplanted to the subretinal space of 3.5-5 week old S334ter-3 rat recipients with fast retinal degeneration. Recipients were sacrificed at the age of 7-32 weeks. Five of 8 rats were recorded electrophysiologically in the superior colliculus (SC) at the age of 23 to 32 weeks. Vibratome slices were stained by immunohistochemistry for the donor marker hPAP, with an ABC kit with DAB as a substrate. In some experiments, staining was enhanced by silver-gold toning.
All recorded transplanted rats showed visual responses in the SC in an area corresponding to the placement of the transplant in the retina, with visual thresholds between -2.8 and -3.4 log cd/m2. Control age-matched no-surgery S334ter-3 rats had no such responses. Donor cells and processes could easily be identified in the host by light and electron microscopy. The hPAP antibody penetrated approximately 10 µm of the vibratome slice surface. Silver-gold toning improved the staining for the EM level. Although glial barriers were present in some areas between transplant and host, transplant processes penetrated the inner host retina. Labeled neuronal processes were found in the host, especially in the inner plexiform layer, sometimes forming synapses with unlabeled presumably host cells.
These data suggest that synaptic connectivity between retinal sheet transplants and degenerating host retinas contributes to visual responses in the superior colliculus, supporting previous results of restored visual function and synaptic connectivity shown by trans-synaptic tracing.
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