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Fig. 2. In vivo visualization of IPL formation and sublamination in
Pax6-DF4:mGFP line 220. (A) GFP expression at different stages of
development in line 220. At 37 hpf, expression is present in neuroblasts with
processes spanning the thickness of the retina. By 54 hpf, GFP expression
becomes gradually confined to ACs whose neurites take part in forming a
continuous inner plexiform layer (IPL), located approximately four or five
cell bodies away from the internal limiting membrane (ILM). GFP expression
becomes almost exclusively confined to ACs and their neurites by 71 hpf. Note
that anterior retina (left) develops before posterior. L, lens; F, choroid
fissure (marks ventral retina). (B) Time-lapse confocal images showing
progressive addition of GFP+ ACs over time. Each image is a
z-projection of a 19 µm confocal image stack. Arrowheads indicate
the same cells in all time frames. Arrow marks the approximate locations of
emerging GFP-expression in ACs and neurites that contribute to the forming
IPL. (C) Appearance of sublamination in the IPL. Each image is a
z-projection of a 3 µm stack of confocal planes. Arrowheads
indicate the same region across time points. For (B,C), dorsal retina is
towards the right. Time-lapse movies of B and C (Movies 1 and 2) can be found
at
http://dev.biologists.org/supplemental).
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