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. 2020 Apr 27;9:e56301. doi: 10.7554/eLife.56301

Figure 1. Initial analysis of sagd1.

(A) Lateral views of isl2b:Gfp+ SAG neurons in live wild-type (wt) embryos and sagd1 mutants at the indicated times. The anterior/vestibular portion of the SAG (outlined in white) is deficient in sagd1 mutants, whereas the posterior SAG (outlined in red) appears normal. (B) Number (mean and s.d.) of isl2b:Gfp+ anterior neurons and posterior neurons in wild-type and sagd1 embryos at the times indicated. Sample sizes are indicated. Asterisks, here and in subsequent figures, indicate significant differences (p<0.05) from wild-type controls. (C–F) Cross-sections through the anterior/vestibular portion of the otic vesicle (outlined) showing expression of ngn1 in wild-type embryos and sagd1 mutants at 24 and 30 hpf. (G, H) Mean and standard deviation of ngn1+ cells in the floor of the otic vesicle (G) and in recently delaminated SAG neuroblasts outside the otic vesicle (H), as counted from serial sections. (I–P) Cross-sections through the anterior/vestibular and posterior/auditory regions of the otic vesicle showing expression of neurod in transit-amplifying SAG neuroblasts at 36 and 48 hpf. (Q) Mean and standard deviation of neurod+ SAG neuroblasts at 30 and 48 hpf counted from serial sections. Sample sizes are indicated (B, G, Q).

Figure 1.

Figure 1—figure supplement 1. Quantitation of mature Isl1/2+ SAG neurons and TUNEL in sagd1 mutants.

Figure 1—figure supplement 1.

(A–H) Cross sections (dorsal up, medial to the right) through the anterior/vestibular portion of the otic vesicle in control embryos (A–D) and sagd1 mutants (E–H) showing anti-Isl1/2 stained SAG neurons (A,C, E, G) and co-staining for TUNEL (B, D, F, H). (I) Number of Isl1/2+ SAG neurons at 30 hpf counted from whole mount preparations. Anterior/vestibular SAG neurons are under-produced in sagd1 mutants, whereas posterior/auditory neurons develop normally. (J) Number of TUNEL+ apoptotic cells at the indicated times. sagd1 mutants show fewer apoptotic cells than normal at 24 and 30 hpf, indicating that the deficiency in vestibular SAG neurons is not due to cell death. Sample sizes are indicated (I, J).