Testis-specific Proteins, TSNAXIP1 and 1700010I14RIK, Are Important for Sperm Motility and Male Fertility in Mice

Overview

Journal Andrology

Specialties Reproductive Medicine
Urology

Date 2023 Jan 4

PMID 36598146

Authors

Yuki Kaneda

Haruhiko Miyata

Keisuke Shimada

Seiya Oura

Masahito Ikawa

Affiliations

Soon will be listed here.

Abstract

Background: TSN (translin), also called testis brain RNA-binding protein, binds to TSNAX (translin-associated factor X) and is suggested to play diverse roles, such as RNA metabolism and DNA damage response. TSNAXIP1 (Translin-associated factor X-interacting protein 1) was identified as a TSNAX-interacting protein using a yeast two-hybrid system, but its function in vivo was unknown.

Objective: To reveal the function of TSNAXIP1 in vivo in mice.

Materials And Methods: We generated Tsnaxip1 knockout mice using the CRISPR/Cas9 system and analyzed their fertility and sperm motility. Further, we generated 1700010I14Rik knockout mice, because 1700010I14RIK is also predominantly expressed in testes and contains the same Pfam (protein families) domain as TSNAXIP1.

Results: Reduced male fertility and impaired sperm motility with asymmetric flagellar waveforms were observed in not only Tsnaxip1 but also 1700010I14Rik knockout mice. Unlike Tsn knockout mice, no abnormalities were found in testicular sections of either Tsnaxip1 or 1700010I14Rik knockout mice. Furthermore, TSNAXIP1 was detected in the sperm tail and fractionated with axonemal proteins.

Discussion And Conclusion: Unlike the TSN-TSNAX complex, whose disruption causes abnormal vacuoles in mouse testes, TSNAXIP1 and 1700010I14RIK may play roles in regulating sperm flagellar beating patterns.

Citing Articles

FBXO24 deletion causes abnormal accumulation of membraneless electron-dense granules in sperm flagella and male infertility.

Kaneda Y, Miyata H, Xu Z, Shimada K, Kamoshita M, Nakagawa T Elife. 2024; 13.

PMID: 39163107 PMC: 11335345. DOI: 10.7554/eLife.92794.

Golgi associated RAB2 interactor protein family contributes to murine male fertility to various extents by assuring correct morphogenesis of sperm heads.

Wang H, Iida-Norita R, Mashiko D, Pham A, Miyata H, Ikawa M PLoS Genet. 2024; 20(6):e1011337.

PMID: 38935810 PMC: 11236154. DOI: 10.1371/journal.pgen.1011337.

FBXO24 deletion causes abnormal accumulation of membraneless electron-dense granules in sperm flagella and male infertility.

Kaneda Y, Miyata H, Xu Z, Shimada K, Kamoshita M, Nakagawa T bioRxiv. 2023; .

PMID: 37986737 PMC: 10659433. DOI: 10.1101/2023.11.10.566635.

CCDC183 is essential for cytoplasmic invagination around the flagellum during spermiogenesis and male fertility.

Shimada K, Ikawa M Development. 2023; 150(21).

PMID: 37882665 PMC: 10629680. DOI: 10.1242/dev.201724.

TSNAXIP1 is required for sperm head formation and male fertility.

Sultana T, Iwamori T, Iwamori N Reprod Med Biol. 2023; 22(1):e12520.

PMID: 37389156 PMC: 10304756. DOI: 10.1002/rmb2.12520.

References

Gupta A, Pillai V, Chittela R . Translin: A multifunctional protein involved in nucleic acid metabolism. J Biosci. 2020; 44(6). View

Fujihara Y, Satouh Y, Inoue N, Isotani A, Ikawa M, Okabe M . SPACA1-deficient male mice are infertile with abnormally shaped sperm heads reminiscent of globozoospermia. Development. 2012; 139(19):3583-9. DOI: 10.1242/dev.081778. View

Oyama Y, Miyata H, Shimada K, Fujihara Y, Tokuhiro K, X Garcia T . CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in mice. Asian J Androl. 2021; 24(3):266-272. PMC: 9226692. DOI: 10.4103/aja.aja_63_21. View

Castaneda J, Hua R, Miyata H, Oji A, Guo Y, Cheng Y . TCTE1 is a conserved component of the dynein regulatory complex and is required for motility and metabolism in mouse spermatozoa. Proc Natl Acad Sci U S A. 2017; 114(27):E5370-E5378. PMC: 5502601. DOI: 10.1073/pnas.1621279114. View

Miyata H, Oura S, Morohoshi A, Shimada K, Mashiko D, Oyama Y . SPATA33 localizes calcineurin to the mitochondria and regulates sperm motility in mice. Proc Natl Acad Sci U S A. 2021; 118(35). PMC: 8536318. DOI: 10.1073/pnas.2106673118. View

Wang J, Boja E, Oubrahim H, Chock P . Testis brain ribonucleic acid-binding protein/translin possesses both single-stranded and double-stranded ribonuclease activities. Biochemistry. 2004; 43(42):13424-31. DOI: 10.1021/bi048847l. View

Miyata H, Satouh Y, Mashiko D, Muto M, Nozawa K, Shiba K . Sperm calcineurin inhibition prevents mouse fertility with implications for male contraceptive. Science. 2015; 350(6259):442-5. DOI: 10.1126/science.aad0836. View

Li L, Gu W, Liang C, Liu Q, Mello C, Liu Y . The translin-TRAX complex (C3PO) is a ribonuclease in tRNA processing. Nat Struct Mol Biol. 2012; 19(8):824-30. PMC: 3414638. DOI: 10.1038/nsmb.2337. View

Naito Y, Hino K, Bono H, Ui-Tei K . CRISPRdirect: software for designing CRISPR/Cas guide RNA with reduced off-target sites. Bioinformatics. 2014; 31(7):1120-3. PMC: 4382898. DOI: 10.1093/bioinformatics/btu743. View

10.

Kluin P, Kramer M, de Rooij D . Spermatogenesis in the immature mouse proceeds faster than in the adult. Int J Androl. 1982; 5(3):282-94. DOI: 10.1111/j.1365-2605.1982.tb00257.x. View

11.

Letunic I, Khedkar S, Bork P . SMART: recent updates, new developments and status in 2020. Nucleic Acids Res. 2020; 49(D1):D458-D460. PMC: 7778883. DOI: 10.1093/nar/gkaa937. View

12.

Niwa H, Yamamura K, Miyazaki J . Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene. 1991; 108(2):193-9. DOI: 10.1016/0378-1119(91)90434-d. View

13.

Eddy E, Toshimori K, OBrien D . Fibrous sheath of mammalian spermatozoa. Microsc Res Tech. 2003; 61(1):103-15. DOI: 10.1002/jemt.10320. View

14.

Bray J, Chennathukuzhi V, Hecht N . Identification and characterization of cDNAs encoding four novel proteins that interact with translin associated factor-X. Genomics. 2002; 79(6):799-808. DOI: 10.1006/geno.2002.6779. View

15.

Tiscornia G, Singer O, Verma I . Production and purification of lentiviral vectors. Nat Protoc. 2007; 1(1):241-5. DOI: 10.1038/nprot.2006.37. View

16.

Wu X, Lefrancois S, Morales C, Hecht N . Protein-protein interactions between the testis brain RNA-binding protein and the transitional endoplasmic reticulum ATPase, a cytoskeletal gamma actin and Trax in male germ cells and the brain. Biochemistry. 1999; 38(35):11261-70. DOI: 10.1021/bi990573s. View

17.

Hwang J, Mannowetz N, Zhang Y, Everley R, Gygi S, Bewersdorf J . Dual Sensing of Physiologic pH and Calcium by EFCAB9 Regulates Sperm Motility. Cell. 2019; 177(6):1480-1494.e19. PMC: 8808721. DOI: 10.1016/j.cell.2019.03.047. View

18.

Abbasi F, Miyata H, Shimada K, Morohoshi A, Nozawa K, Matsumura T . RSPH6A is required for sperm flagellum formation and male fertility in mice. J Cell Sci. 2018; 131(19). PMC: 6198453. DOI: 10.1242/jcs.221648. View

19.

Chennathukuzhi V, Kurihara Y, Bray J, Hecht N . Trax (translin-associated factor X), a primarily cytoplasmic protein, inhibits the binding of TB-RBP (translin) to RNA. J Biol Chem. 2001; 276(16):13256-63. DOI: 10.1074/jbc.M009707200. View

20.

Miyata H, Morohoshi A, Ikawa M . Analysis of the sperm flagellar axoneme using gene-modified mice. Exp Anim. 2020; 69(4):374-381. PMC: 7677079. DOI: 10.1538/expanim.20-0064. View