Initiation of impaired outer segment degradation in vivo using an antisense oligonucleotide.
This paper describes the first successful in vivo application of antisense DNA technology to induce the accumulation of photoreceptor outer segment derived debris in the retina. An antisense oligonucleotide (CatSC), which was previously demonstrated to be an effective tool to induce debris accumulation in vitro, was injected into the vitreous of pigmented and non-pigmented rats. The animals were euthanased 7 days after the injections. The number of inclusions significantly increased in the RPE layer of Long Evans and RCS-rdy + rats injected with 66 ug of CatSC to 96.2 +/- 13.6 (SD) (p < 0.0003) and 204.2 +/- 39.3 (SD) (p < 0.0001), respectively. The difference between the number of phagosome-like inclusions present in control saline, 6.6 ug of CatSC or 66 ug of sense oligonucleotide (S1) injected animals was not statistically significant. There were no abnormalities observed in the inner layers of the retina but the accumulation of phagosome-like inclusions was accompanied by disorganisation in the apices of outer segments. The large number of inclusions found in CatSC treated animals showed the characteristics of phagosomes containing stacks of undigested photoreceptor outer segment membranes which suggest that the lysosomal digestion process was halted or at least slowed down by the antisense oligonucleotide.
Rakoczy
P E
PE
Centre for Ophthalmology and Visual Science, University of Western Australia, Nedlands, Australia.
Lai
M C
MC
Vijayasekaran
S
S
Robertson
T
T
Rapp
L
L
Papadimitriou
J
J
Constable
I
I
D000818:Animals; D005260:Female; D008854:Microscopy, Electron; D016376:Oligonucleotides, Antisense; D010588:Phagosomes / Q000502:physiology; D010857:Pigment Epithelium of Eye / Q000502:physiology / Q000648:ultrastructure; D051381:Rats; D011919:Rats, Inbred Strains; D012160:Retina / Q000648:ultrastructure; D012374:Rod Cell Outer Segment / Q000502:physiology* / Q000648:ultrastructure
D016376:Oligonucleotides, Antisense