Fig. 1 - Supplemental 1

Phenotypes of testes and early gonads of the moto-/- mutant. (A) PH3 and TdT-mediated nick-end labeling (TUNEL) assays of wild-type and moto-/- testes. PH3-positive spermatogonia were detected in moto+/+ and moto-/- testes. TUNEL-positive cells were barely detected in moto+/+ testes, while a considerable number of cells were detected in moto-/- testes. Black arrowheads indicate positive spermatogonia. (B) Histology of early gonads of moto+/- and moto-/-. Growing oocytes (arrowheads) were observed in heterozygous larvae but not in mutant larvae. Developmental stages are indicated by days post-fertilization (dpf). Scale bar: 20 µm. (C) Comparison of mouse and zebrafish Meioc protein sequences. The sequence of zebrafish Meioc (ENSDARG00000090664) was aligned to that of mouse MEIOC (ENSMUSG00000051455). Alignment analyses were performed using EMBOSS NEEDLE, an online software program (https://www.ebi.ac.uk/Tools/psa/emboss_needle/). A coiled-coil domain composed of four helices, PF15189 (previously DUF4582), is conserved in animals. The two mutations predicted to disrupt the zebrafish meioc gene are shown, and the approximate position of the premature stop codon is indicated relative to the total length of the Meioc protein. (D) HE-stained sections of wild-type testes with all stages of spermatogonia (arrowheads), spermatocytes, and spermatozoa present; the meioct31533 mutant, an additional nonsense allele meiocsa13122, and transheterozygote meioct31533/sa13122 exhibited testes containing single spermatogonia and spermatogonia in small clusters. Scale bar: 20 µm. (E) Western blot analysis of meioc+/+, meioc+/mo, and meiocmo/mo testis extracts. (F) Immunostaining of zebrafish wild-type testes. Absorbed anti-Meioc IgG was treated with Meioc recombinant protein. Scale bar: 10 µm. (G) Immunostaining of meiocmo/mo single spermatogonia. Note that Meioc was not detected in meiocmo/mo spermatogonia. Scale bar: 10 µm.