No Isl1-immunoreactive elements were distinguishable before St29-

No Isl1-immunoreactive elements were distinguishable before St29-30. From the onset of the appearance of Isl1 expression in the retina (St29-30), immunoreactivity appeared progressively following central-to-peripheral and vitreal-to-scleral gradients. From St42 Carfilzomib manufacturer onwards all retinal cell types were fully differentiated and the Isl1 expression pattern did not change significantly.3.1. Isl1 Expression in the X. laevis Differentiated RetinaThe Isl1 expression pattern in fully differentiated retinas will be reported first so as to establish a set of referents with which to compare developmental stages. Thus, at St53, the retina is almost fully developed, exhibiting complete lamination as determined from DAPI-stained cryosections (Figure 2(a)).

Anti-Isl1 antibody stained most of the cells located in the ganglion cell layer (GCL) (Figures 2(b), 2(c), 2(e), 2(f), 2(h), and 2(i)). Furthermore, different populations of Isl1-containing cells located in the inner nuclear layer (INL) could be distinguished by both morphological and topographical features: (i) a small population of cells with the major axis oriented parallel to the vitreal surface, lying at the border between the outer plexiform layer (OPL) and the INL, in the horizontal cell layer (Figures 2(b) and 2(c)); (ii) a population of cells located in the middle of the INL, in the bipolar cell layer (Figures 2(b), 2(c), 2(e), 2(f), 2(h), and 2(i)); and (iii) a small population of cells located at the inner edge of the INL, in the region where the amacrine cells are normally found (Figures 2(b), 2(c), 2(e), 2(f), 2(h), and 2(i)).

To further characterize Isl1-expressing cells, a double labeling with antibodies against calbindin-D28k (CB) and calretinin (CR) was also performed. In the developing and adult X. laevis retina, CB labels cones and subpopulations of bipolar, amacrine, and ganglion cells [35, 45]. Some of the CB-immunoreactive bipolar cells also expressed Isl1 (Figures 2(d)�C2(f)). CR labels subsets of horizontal, bipolar, amacrine, and ganglion cells [35]. Some of the CR-expressing horizontal, bipolar, and ganglion cells also expressed Isl1 (Figures 2(g)�C2(i)).Figure 2Morphological features and expression patterns of Isl1 and other cell differentiation markers in the St53 Xenopus laevis central retina. (a)�C(c) DAPI fluorescence combined with Isl1 immunofluorescence.

DAPI staining showed well-organized retinal …3.2. Isl1 Expression in the X. laevis Developing RetinaBy St29-30, although the lumen of the ventricle was still observed, the most distal part of the vesicle wall had invaginated to Brefeldin_A form a two-layered optic cup (Figure 3(a)). The neural retina was composed of a neuroblastic layer (NbL) (Figure 3(a)), and weak Isl1 immunoreactivity was first observed in sparse nuclei located near the vitreal surface of the central retina (Figure 3(b)). Similar morphological features and staining patterns were observed at St31 (Figures 3(c) and 3(d)).

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