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Fig. 5. Impairment of retinal development in ntn mutant embryos. (A)
Confocal optical section images through the retina of Bodipy-ceramide stained
wild-type (wt) and mutant (ntn) embryos at 27 hpf, 36 hpf and 48 hpf.
An arrow indicates cell-free spaces. (B) TUNEL staining of retinal sections of
wild-type (wt) and mutant (ntn) embryos at 30 hpf, 36 hpf and 48 hpf.
(C) Immunostaining with zn5 antibody (green) of retinal sections of wild-type
(wt) and mutant (ntn) embryos at 30 hpf, 36 hpf and 48 hpf. Nuclei of
retinal cells were counterstained with Sytox (red). (D) Confocal composite
images of nAChRß3 gene promoter-driven EGFP signals in wild-type (wt) and
mutant (ntn) embryos at 30 hpf, 36 hpf and 48 hpf in coronal view and
those at 48 hpf in lateral view. Wild-type embryos have RGCs over the entire
retina, whereas ntn mutants have sparse RGCs. Note that ntn
mutants show EGFP signals in the trigeminal ganglion and Rohon-Beard sensory
neurons and the pituitary gland. dlf, dorsal longitudinal fasciculus; on,
optic nerve; p, pituitary gland; rb, Rohon-Beard neurons; tg, trigeminal
ganglion. (E) Confocal composite images of immunostaining with anti-acetylated
tubulin of retinal sections of wild-type (wt) and mutant (ntn)
embryos at 30 hpf, 36 hpf and 48 hpf. Arrows indicate RGC axons. (F) In situ
hybridization of ath5 mRNA in wild-type (wt) and mutant
(ntn) embryos at 33 hpf. Ventral view. (G) In situ hybridization of
brn3b mRNA in wild-type (wt) and mutant (ntn) embryos at 36
hpf and 48 hpf. Ventral view. Arrowheads indicate the RGC layer. Scale bars:
50 µm in A-C; 100 µm in D-G.
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