CCDC113 stabilizes sperm axoneme and head-tail coupling apparatus to ensure male fertility
- Guangzhou Women and Children's Medical Center, Guangzhou Medical University, China
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Stem Cell and Regenerative Medicine Innovation Institute, Chinese Academy of Sciences, China
- University of Chinese Academy of Sciences, China
- Department of Urology, Department of Reproductive Medicine Center, Peking University Third Hospital, China
- Department of Urology, Peking University First Hospital Institute of Urology, Peking University, China
Figures
Figure 1 with 1 supplement
CCDC113 is an evolutionarily conserved axoneme-associated protein.
(A) Multiple species phylogenetic tree of CCDC113. Structural similarity scores (Z scores) of CCDC113 orthologs in H. sapiens, Mus musculus, Phascolarctos cinereus, Danio rerio, Chlamydomonas …
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Figure 1—source data 1
Original files for western blot in Figure 1C-H.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig1-data1-v1.zip
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Figure 1—source data 2
Labelled files for western blot in Figure 1C-H.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig1-data2-v1.zip
Figure 1—figure supplement 1
CCDC113 could not bind to ODF1 and ODF2.
(A–B) CCDC113 was expressed alone or co-expressed with ODF1 (A) or ODF2 (B) in HEK293T cells, and the interactions between CCDC113 and ODF1 or ODF2 were examined by co-immunoprecipitation. IB: …
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Figure 1—figure supplement 1—source data 1
Original files for western blot in Figure 1—figure supplement 1A-B.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig1-figsupp1-data1-v1.zip
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Figure 1—figure supplement 1—source data 2
Labelled files for western blot in Figure 1—figure supplement 1A-B.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig1-figsupp1-data2-v1.zip
Figure 2 with 1 supplement
Ccdc113 knockout leads to male infertility.
(A) The CRISPR-Cas9 strategy for generating the Ccdc113 knockout mice. (B) Genotyping to identify Ccdc113 knockout mice. (C) Immunoblotting of CCDC113 in Ccdc113+/+ and Ccdc113–/– testes. TUBULIN …
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Figure 2—source data 1
Original file for gel in Figure 2B.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig2-data1-v1.zip
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Figure 2—source data 2
Labelled file for gel in Figure 2B.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig2-data2-v1.zip
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Figure 2—source data 3
Original files for western blot in Figure 2C.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig2-data3-v1.zip
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Figure 2—source data 4
Labelled file for western blot in Figure 2C.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig2-data4-v1.zip
Figure 2—figure supplement 1
Ccdc113–/– mice did not exhibit the ciliopathies, such as hydrocephalus, situs inversus, and abnormal ciliogenesis of tracheal cilia.
(A) Immunoblotting of CCDC113 in Ccdc113+/+ and Ccdc113–/– testes, with TUBULIN serving as the loading control. An asterisk indicates the CCDC113 band. IB: immunoblotting. (B) The Ccdc113–/– mice …
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Figure 2—figure supplement 1—source data 1
Original files for western blot in Figure 2—figure supplement 1A.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig2-figsupp1-data1-v1.zip
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Figure 2—figure supplement 1—source data 2
Labelled file for western blot in Figure 2—figure supplement 1A.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig2-figsupp1-data2-v1.zip
Figure 3 with 1 supplement
Ccdc113 knockout results in sperm flagellar defects and sperm head-tail detachment.
(A) Hematoxylin-eosin (H&E) staining of the cauda epididymis from 2-month-old Ccdc113+/+ and Ccdc113–/– male mice. Red circles indicate coiled eosin-stained structures without sperm heads in the …
Figure 3—figure supplement 1
Ccdc113 knockout leads to abnormal sperm head shaping defect.
(A) Periodic acid Schiff (PAS) staining of spermatid at different steps from Ccdc113+/+ and Ccdc113–/– mice. Asterisk indicates abnormal spermatid shapes were found from step 11. (B) Spermatids from …
Figure 4 with 1 supplement
CCDC113 localizes to the head-tail coupling apparatus (HTCA), manchette, and sperm flagellum.
(A) CCDC113 was expressed starting in postnatal day 7 (P7) testes. TUBULIN served as the loading control. An asterisk indicates nonspecific bands. (B) Immunofluorescence of CCDC113 (red) and …
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Figure 4—source data 1
Original files for western blot in Figure 4A.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig4-data1-v1.zip
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Figure 4—source data 2
Labelled file for western blot in Figure 4A.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig4-data2-v1.zip
Figure 4—figure supplement 1
Subcellular localization of CCDC113 in testicular germ cells from Ccdc113+/+ and Ccdc113–/– mice.
(A) RNA expression data from various tissues using RNA-Seq data from Mouse ENCODE (https://www.ncbi.nlm.nih.gov/gene/244608). CCDC113 is highly expressed in the testis, but not significantly in the …
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Figure 4—figure supplement 1—source data 1
Original files for western blot in Figure 4—figure supplement 1B.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig4-figsupp1-data1-v1.zip
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Figure 4—figure supplement 1—source data 2
Labelled file for western blot in Figure 4—figure supplement 1B.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig4-figsupp1-data2-v1.zip
Figure 5
CCDC113 is indispensable for the docking of CFAP91 and DRC2 to the doublet microtubules (DMTs) to maintain the structural integrity of the axoneme.
(A) Transmission electron microscopy (TEM) analysis of spermatozoa from the cauda epididymidis of Ccdc113+/+ and Ccdc113–/– male mice. The flagellar longitudinal sections of Ccdc113–/– spermatozoa …
Figure 6
Ccdc113 knockout spermatids display impaired head-tail coupling apparatus (HTCA).
(A) The proportion of decapitated tails in Ccdc113+/+ and Ccdc113–/– corpus, caput, and cauda epididymis (n=3 independent experiments). At least 200 spermatozoa were analyzed from each mouse. Data …
Figure 7
CCDC113 interacts with SUN5 and CENTLEIN, participating in sperm head-tail linkage.
(A–C, F) Head-tail coupling apparatus (HTCA)-associated proteins (SUN5, CENTLEIN, PMFBP1, SPATA6) were expressed alone or co-expressed with CCDC113 in HEK293T cells, and the interactions between …
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Figure 7—source data 1
Original files for western blot in Figure 7A-F.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig7-data1-v1.zip
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Figure 7—source data 2
Labelled files for western blot in Figure 7A-F.
- https://cdn.elifesciences.org/articles/98016/elife-98016-fig7-data2-v1.zip
