Molecular Sexing of<i>Nothobranchius furzeri</i>Embryos and Larvae

Richter, Annekatrin; Krug, Johannes; Englert, Christoph

doi:10.1101/pdb.prot107782

Cited by 2 publications

(3 citation statements)

References 11 publications

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“…Dechorionation of embryos and animals at 0 dph was performed as described previously (Richter et al 2022). Phenotypes of embryos and larvae up to 2 dph were documented using reflective or transmission light with the Zeiss SteREO Discovery.V8 equipped with a Zeiss AxioCam MRc and a Volpi dual gooseneck pole mount light or the Zeiss Axio Zoom.V16 equipped with a Zeiss AxioCam HRc and an HXP 200 C light source, respectively.…”

Section: Discussionmentioning

confidence: 99%

“…Genomic DNA samples were derived from caudal fin biopsies of adult animals or tail tips from embryos and larvae as described previously by Krug et al (2023) and Richter et al (2022), respectively. For sexing and genotyping PCRs, DreamTaq DNA Polymerase (Thermo Fisher Scientific) was used according to the manufacturer's instructions with an annealing temperature of 60°C, if not indicated otherwise.…”

Section: Molecular Sexing and Genotyping Of Animalsmentioning

confidence: 99%

“…S1 B) or as described previously (see Supplemental Fig. S2 B; Richter et al (2022)). For the gdf6Yspecific PCR, an annealing temperature of 64°C was applied.…”

Section: Molecular Sexing and Genotyping Of Animalsmentioning

confidence: 99%

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The Tgf-β family member Gdf6Y determines the male sex inNothobranchius furzeriby suppressing oogenesis-inducing genes

Richter,

Mörl,

Thielemann

et al. 2023

Preprint

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The short-lived African killifish Nothobranchius furzeri lives in seasonal freshwater ponds and has evolved remarkable traits to survive in this limited environment. One of those traits is a genetic XX/XY sex-determination system, which ensures an equal distribution of both sexes. Comparisons of female and male genomic sequences identified the Y-chromosomal copy of the TGF-β family member gdf6 as the candidate male sex-determining (SD) gene, which was named gdf6Y in contrast to the X-chromosomal allele gdf6X. CRISPR/Cas9-mediated inactivation of gdf6Y in N. furzeri led to a complete male-to-female sex reversal in XY animals. The homozygous inactivation of gdf6X on the other hand led to a detrimental phenotype post-hatching. This phenotype was compensated by gdf6Y, revealing that the latter became the SD gene while retaining at least some of its original gdf6 function. Gdf6Y is expressed in testicular somatic cells already prior to hatching, where it represses the germ cell-intrinsic feminizing gene foxl2l. We have identified components of the TGF-β signaling pathway, especially the inhibitor of DNA binding genes id1/2/3, and the mRNA decay activator zfp36l2, as Gdf6Y targets. We conclude that Gdf6Y exerts its function as the male sex-determining gene by suppressing female-specific genes in the developing gonad of male N. furzeri.

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Section: Discussionmentioning

confidence: 99%

Section: Molecular Sexing and Genotyping Of Animalsmentioning

confidence: 99%

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The Tgf-β family member Gdf6Y determines the male sex inNothobranchius furzeriby suppressing oogenesis-inducing genes

Richter,

Mörl,

Thielemann

et al. 2023

Preprint

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Generation of a transparent killifish line through multiplex CRISPR/Cas9mediated gene inactivation

Krug

Perner

Albertz

et al. 2023

eLife

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Body pigmentation is a limitation for in vivo imaging and thus for the performance of longitudinal studies in biomedicine. A possibility to circumvent this obstacle is the employment of pigmentation mutants, which are used in fish species like zebrafish and medaka. To address the basis of aging, the short-lived African killifish Nothobranchius furzeri has recently been established as a model organism. Despite its short lifespan, N. furzeri shows typical signs of mammalian aging including telomere shortening, accumulation of senescent cells and loss of regenerative capacity. Here, we report the generation of a transparent N. furzeri line by simultaneous inactivation of three key loci responsible for pigmentation. We demonstrate that this stable line, named klara, can serve as a tool for different applications including behavioral experiments and the establishment of a senescence reporter by integration of a fluorophore into the cdkn1a (p21) locus and in vivo microscopy of the resulting line.

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Molecular Sexing ofNothobranchius furzeriEmbryos and Larvae

Cited by 2 publications

References 11 publications

The Tgf-β family member Gdf6Y determines the male sex inNothobranchius furzeriby suppressing oogenesis-inducing genes

The Tgf-β family member Gdf6Y determines the male sex inNothobranchius furzeriby suppressing oogenesis-inducing genes

Generation of a transparent killifish line through multiplex CRISPR/Cas9mediated gene inactivation

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