A Model of Human RP and LCA

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Profiling Retinal Biochemistry in the MPDZ Mutant
Retinal Dysplasia and Degeneration Chick: A Model of
Human RP and LCA
From  James R. Beattie,1,2 Sorcha Finnegan,1 Ross W. Hamilton,1 Manir Ali,3
Christopher F. Inglehearn,3 Alan W. Stitt,1 John J. McGarvey,1,2 Paul M. Hocking,4
and William J. Curry1
PURPOSE. Raman microscopy, a rapid nondestructive technique
that profiles the composition of biological samples, was used to
characterize retinal biochemistry in the retinal dysplasia and
degeneration (rdd) and wild-type (wt) chick retina during retinogenesis
and at hatching.
METHODS. Embryonic day (E)13 and posthatch day (P)1 rdd and
wt retinal cross-sections (n
3 of each line at each age) were
profiled using 633 helium–neon laser excitation. The biochemical
composition was determined using computational analysis
of the Raman spectra. In parallel histology, TUNEL and glial
fibrillary acidic protein (GFAP) immunostaining were used to
visualize retinal dysfunction.
RESULTS. Principal component (PC) analysis of the Raman spectra
identified 50 major biochemical profiles, but only PCs that
made significant contributions to variation within rdd and wt
retina were mapped. These significant PCs were shown to arise
from DNA, various fatty acids, melanin, and a number of proteins.
Distinct patterns of GFAP immunostaining and a larger
population of TUNEL-positive nuclei were observed in the rdd
versus wt retina.
CONCLUSIONS. This study has demonstrated that Raman microscopy
can discriminate between major retinal biomolecules,
thus providing an unbiased account of how their composition
varies due to the impact of the MPDZ null mutation in the rdd
chick relative to expression in the normal wt retina. (Invest
Ophthalmol Vis Sci. 2012;53:413– 420) DOI:10.1167/