|
Fig. 2
Simultaneous Removal of BC Partner Neurons (RGCs and ACs) Does Not Prevent BC Axons from Forming a Sublaminated Neuropil
(A and B) Despite the loss of ACs, a phalloidin-rich IPL forms in the ptf1a−/−;ptf1aMO retina.
(C) WT retina with the actin-rich IPL shown by phalloidin (arrow) and BC terminals labeled with vsx1:GFP.
(D) Phalloidin staining demonstrates that in the AC/RGC-free retina (ath5−/−;ptf1a−/− ;ptf1aMO), the IPL is actin rich and positioned along the basal surface of the retina (arrow).
(E) The thickness of the IPL was measured for each genotype at 5 dpf, demonstrating it is significantly thinner after AC removal (ptf1a−/−;ptf1aMOs), and thicker after RGC removal (ath5−/−). One-way ANOVA and Tukey post hoc tests; ∗∗p < 0.01; error bars ± 1 SEM. The number of retinas measured is given within the bars.
(F) In the WT retina, Q16-labeled BCs stratify basally to Q19-labeled BCs in the IPL.
(G) After the removal of many ACs and RGCs by ptf1aMO and ath5MO injections (G), the typical Q16-basal, Q19-apical pattern is apparent in many areas.
(H) Inset in (F).
(I) Inset in (G).
(J) Line intensity profile measurements from 45 regions (three different retinas/animals) demonstrates the basal enrichment of Q16 signal and apical enrichment of Q19 signal in ath5;ptf1a morphants. Scale bars, 20 μm.
See also Figures S2 and S3.