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Fig. 5
(A–B) Immunofluorescence images of (A) control heat-shocked (control-hs), and (B) LMNA overexpressing heat-shocked (LMNA OE-hs) at 90 hours post fertilization (hpf). Tg(lhx-1:eGFP) embryos were used for ctl-hs, and Tg(lhx-1:eGFP) × Tg(hsp70:LMNA-mKate2) double-transgene retinae for LMNA OE-hs experiments. Tg(lhx-1:eGFP) marks horizontal cells (HCs) (green), Tg(hsp70:LMNA-mKate2) is expressed in nuclear envelopes upon heat-shock (blue), DAPI labels nuclei (magenta), and phalloidin marks actin (yellow). Red arrowheads: ectopically positioned HCs, dashed circle: lens. Scale bar: 50 μm. (C) Position of HCs relative to outer plexiform layer (OPL) in control heat-shocked (ctl-hs) (n=27, N=5), and LMNA OE-hs (n=283, N=11) at 92 hpf. See Figure 5—source data 1. (D) Stills from a 20 hr time-lapse of Tg(lhx-1:eGFP) × Tg(hsp70:LMNA-mKate2) double-transgene retinae after heat-shock (LMNA OE-hs). Arrowheads: tracked HCs. Scale bar: 20 μm. (E) Migration trajectory of tracked HCs from (D). HC-2, HC-3, and HC-4 remained ectopically positioned in the basal part of the retina, while HC-1 successfully reached the HC layer. See Figure 5—source data 1. See also Figure 5—figure supplement 1.
Interference with nuclear properties impairs efficient horizontal cell migration and layer formation.