Fig. 1: The inactivation of gdf6Y in Nothobranchius furzeri leads to a full male-to-female sex reversal. | Nature Communications

Fig. 1: The inactivation of gdf6Y in Nothobranchius furzeri leads to a full male-to-female sex reversal.

From: The master male sex determinant Gdf6Y of the turquoise killifish arose through allelic neofunctionalization

Fig. 1

a Schematic of Gdf6X/Y protein domains (SP – signal peptide), exonal distribution, the coding sequence comparison between gdf6X and gdf6Y, and the mutation strategy with two gdf6Y-specific sgRNAs facilitating DNA double-strand breaks at a 109 bp distance. Coding sequence differences between gdf6X and gdf6Y are indicated in gdf6Y’s exons. SNVs: Dark gray – non-synonymous, light gray – synonymous. White – 9 bp deletion. b Phenotype of mosaic F0 GRZ-gdf6Y− animals (XY*, phenofemales) compared to unaltered F0 females (XX) and wild-type males (XY) at about 3 months of age. c One exemplarily HE stained F0 XX, F0 XY*, and XY gonad at about 5 months of age. Oocytes: P – previtellogenic, V – vitellogenic, M – mature. SC – spermatocytes, ST – spermatids, SZ – spermatozoa. d Sexually mature offspring (age: 1 month) of phenofemales with males consists of females, phenofemales, and males in equal parts missing YY* animals expected due to Mendelian ratios. Schematic created in BioRender. Richter, A. (2024) https://BioRender.com/n01l529. e Gdf6Y mutant variants causing full male-to-female sex reversal in F1 (frameshifts at either sgRNA position causing early stop codons (asterisks), in-frame deletions of 9 bp at the sgRNA Y2 site or between both sgRNAs). Two mutations were propagated in fish lines. RT-qPCR of (f) gdf6Y separately and together with gdf6X and (g) male and female marker genes in F1 gonads (Supplementary Table 1, XY*: n = 3; XX, XY: n = 4, n = 3 for foxl2) at 2.4 months of age (mean with standard deviation). Statistical testing by a two-tailed Welch’s t-test (gdf6Y) or Welch’s ANOVA and Dunnett’s T3 multiple comparisons test (others). P-values < 0.05 are displayed. h One-quarter of a phenofemale’s (GRZ-gdf6Ydel9) offspring with males develops malformations and carries two Y chromosomes (YY*). i Reflected light (RL) and brightfield (BF) microscopy images of YY* embryos (RL, n = 13; BF, n = 31) within the egg compared to normally developed animals (RL, n = 45; BF, n = 94; here: XX at 0 dph) from independent samplings from two phenofemales (RL, GRZ-gdf6Ydel6, del8; BF, GRZ-gdf6Ydel9 from h). Arrows: Black – eye, white – tail. d, f–h Source data are provided as a Source Data file.

Back to article page