Evolutionarily conserved sperm factors, DCST1 and DCST2, are required for gamete fusion
- Department of Cell Science, Institute of Biomedical Sciences, School of Medicine, Fukushima Medical University, Japan
- Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Japan
Figures
Figure 1
Alignments of dendrocyte expressed seven transmembrane protein domain-containing 1 (DCST1) and dendrocyte expressed seven transmembrane protein domain-containing 2 (DCST2) amino acid sequences, and mRNA expression profiles.
(A) Sequence alignments of DCST1 in Homo sapiens (human; GenBank: NP_689707.2), Mus musculus (mouse; GenBank: NP_084250.1), Caenorhabditis elegans (nematode; GenBank: NP_001343858.1), and Drosophila …
Figure 2
Strategy for Dcst1 and Dcst2 genes disruption and its validation.
(A) Targeted disruption of the Dcst1/2, Dcst1, Dcst2, and Spaca6 genes, and constructs for Dcst1 and Dcst2 transgenic mice. Complete structures of the wild-type mouse Dcst1/2 alleles are shown. …
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Figure 2—source data 1
IVF in SPACA6 heterozygous and homozygous KO spermatozoa.
- https://cdn.elifesciences.org/articles/66313/elife-66313-fig2-data1-v1.xlsx
Figure 3
Identification of new gamete fusion-related factors dendrocyte expressed seven transmembrane protein domain-containing 1 (DCST1) and dendrocyte expressed seven transmembrane protein domain-containing 2 (DCST2).
(A) Accumulation of many acrosome-reacted (AR) IZUMO1 positive spermatozoa (red) in the perivitelline space of eggs recovered from the oviducts of females 8 hr after coitus with a Dcst1/2-/- male. …
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Figure 3—source data 1
Gamete fusion assay, IVF and litter size in DCST1/2-related mutants.
- https://cdn.elifesciences.org/articles/66313/elife-66313-fig3-data1-v1.xlsx
Videos
Video 1
6 hr after insemination of Dcst1/2-disrupted spermatozoa.
The first and second half of the video show Dcst1/2 heterozygous and homozygous KO spermatozoa insemination, respectively.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus) | Dcst1/2 KO | This article | Deposited in RIKEN BRC (ID RBRC05733) | |
| Strain, strain background (Mus musculus) | Dcst1 KO | This article | Deposited in RIKEN BRC (ID RBRC10275) | |
| Strain, strain background (Mus musculus) | Dcst2 KO | This article | Deposited in RIKEN BRC (ID RBRC10366) | |
| Strain, strain background (Mus musculus) | Spaca6 KO | This article | Deposited in RIKEN BRC (ID RBRC10367) | |
| Strain, strain background (Mus musculus) | Izumo1 KO | Inoue et al., 2005 | RRID MGI_3576518 | |
| Strain, strain background (Mus musculus) | Dcst1 TG | This article | Deposited in RIKEN BRC (ID RBRC05856) | |
| Strain, strain background (Mus musculus) | Dcst2 TG | This article | ||
| Strain, strain background (Mus musculus) | Izumo1 TG | Inoue et al., 2005 | ||
| Antibody | Anti-mouse SPACA6 (rabbit polyclonal) | This article | (WB: 1:500) | |
| Antibody | Anti-mouse IZUMO1 (rat monoclonal) | Inoue et al., 2013 | Mab125 | Gift from Dr. Ikawa (IF: 0.5 µg ml−1) |
| Antibody | Anti-mouse IZUMO1 (rat monoclonal) | Inoue et al., 2015 | Mab18 | (WB: 1 µg ml−1) |
| Antibody | Anti-mouse FR4 (JUNO) (rat monoclonal) | BioLegend | TH6, RRID AB_1027724 | (IF: 0.25 µg ml−1) |
| Antibody | Anti-mouse CD9 (rat monoclonal) | BioLegend | MZ3, RRID AB_1279321 | (IF: 0.5 µg ml−1) |
| Antibody | Anti-mouse EMMPRIN (BASIGIN) (goat polyclonal) | Santa Cruz Biotechnology | G-19, RRID AB_2066959 | (WB: 0.2 µg ml−1) |
| Recombinant DNA reagent | Plasmid pX330 vector | Addgene | RRID Addgene_42230 | |
| Sequence-based reagent | Primers | This article | Detailed in 'Materials and methods' | |
| Peptide, recombinant protein | SPACA646-139 | This article | Detailed in 'Materials and methods' |
