Figure 1.
Visualization of the transplanted tissue.
(A) A retinal detachment (RD, white line) was detectable immediately after transplantation from fundus of the transplanted eye in which the graft was placed just under the neural retina. (B–E) Photos of angiograph from P23H rat retinas. (B and E) Fundus photos from a transplanted eye after surgery. (C and D) Fundus photos from a control eye without transplantation. (B and C) Fundus photos taken with red-free light at 1 week after operation (Fundus of the transplanted retina was similar as that of the control, showing no detachment and hemorrhage). (D and E) Autofluorescence images taken at 2 months after operation. There was no hyper autofluorescence in the control retina (D), while it was observed in the transplanted retina (E), from transplant stained by PKH26 fluorescent cell linker in the superior of the optic nerve (ON, indicated by a white head arrow). (F) Vertical sections 6 months after transplantation showing the transplant (t) labeled with anti-rhodopsin antibody (green) apposed to the neural retina of the host (h).
Figure 2.
Effect of transplantation to P23H retinas at 9 months of age.
(A) Immunolabeling of the flat-mounted degenerating retina (Rho-4D2 antibody, red) with retinal transplant. Due to degeneration, almost no rods were detected except the transplant in the dorsal host retina. Images1 and 2 were taken respectively in the nasal-ventral, temporal-ventral quadrants, both far from and opposite to the transplant. D: dorsal, V: ventral, N: nasal, T: temporal, PNA lectin: green, Rho-4D2 antibody: red. (B) Immunolabeling of cones with PNA lectin in the control and photoreceptor transplanted retinas from the same animal. The two images were taken in the same areas from the nasal-ventral quadrant, showing that cone outer segments are longer in the transplanted retina than in the control retina.
Figure 3.
The distribution of cones after retinal transplantation and the size of tip area of cone outer segments.
(A) This image of cone counting with 193 fields by stereological approach shows the surviving cones distributed in the whole retina, not only localized in the area of transplant. The transplanted area was enclosed by a black line. This image corresponds to Figure 2A. (B) Comparison of the size of tip areas of cone outer segments. (C) The distribution of the size of tip area of cone outer segments in three experimental groups (photoreceptor transplanted, contralateral control, sham control), corresponding to B.
Figure 4.
Morphology of cones in the control and photoreceptor transplanted retinas.
(A) The cones in the untreated control P23H rat retina present large areas of tip sheath and short outer segments. (B) While, small tip areas associated with long cone outer segments are observed in the photoreceptor transplanted P23H rat retina. A1–A3, B1–B3, images with high magnification corresponding to the small parts in A and B.
Table 1.
Size of tip area of cone outer segments in different experimental groups.
Figure 5.
Comparison in the different groups by cell counting and photopic ERG b-wave amplitude.
CO: control eye, SM: sham control eye, RT: retinal transplanted eye, PT: photoreceptor transplanted eye. (A) Scatter plot shows normalization of changing cones from each rat by comparison of treated eye and its corresponding contralateral control eye. 100% for each control eye. Means of percentage in treated groups, SM: 98%, P = 0.3; RT: 111%, * P<0.01; PT: 111%, * P<0.01. (B and C) Comparison of photopic ERG average of 5 single flashes from P23H cases after transplantation. b-wave amplitude: B, control eye- 33 µV, operated eye- 72 µV. C, control eye- 27 µV, operated eye- 86 µV. (D) Difference of photopic ERG b-wave amplitudes in each group between the control and treated eyes.