Animal Reproduction (AR)
https://www.animal-reproduction.org/article/doi/10.1590/1984-3143-AR2020-0521
Animal Reproduction (AR)
Original Article

Septin14, a gene specifically expressed in the testis and seminal vesicle of the Banna mini-pig inbred line (BMI)

Pei Wang; Xia Zhang; Hailong Huo; Shuyan Wang; Xue Song; Jinlong Huo

Downloads: 0
Views: 729

Abstract

Abstract: Septin14 is an important spermatogenesis related gene involved in the pathogenesis of male infertility that has not been well studied. Here, full-length Septin14 cDNA of the Banna mini-pig inbred line (BMI) was cloned using the RACE method and expressed in pig kidney epithelial cells (PK15) and E. coli Rosetta (DE3) cells. Septin14 expression was identified in somatic tissues and testis in different developmental stages. The pig Septin14 CDS is 1,299 bp long, and encodes a peptide (or protein) of 432 amino acids (MW=50.4 kDa). Phylogenetic analysis indicated that pig Septin14 was highly evolutionarily conserved. Subcellular localization of GFP-tagged Septin14 fusion protein revealed that Septin14 was distributed throughout the testicular cells. Among 34 pig tissues, Septin14 mRNA was found specifically in testis and seminal vesicle. In six different postnatal developmental stages, the testicular level of Septin14 mRNA was barely detectable on day 2, while the highest level occurred on day 75. The spatiotemporal expression profile of Septin14, reported herein for the first time in pig, indicated that Septin14 might be involved in the division, development and apoptosis of germ cells. Furthermore, using a pET prokaryotic expression system, we expressed and isolated recombinant 67.9 kDa Septin14 protein.

Keywords

Septin14, pigs, molecular cloning, subcellular localization, gene expression, protein purification

References

Barzi NV, Sabbaghian M, Onsory KH, Hosseinifar H, Modarresi T, Nickhah Z. Comparison of septin14 expression in testes of patients with positive and negative testicular sperm retrieval. Iran J Reprod Med. 2014;12(6):24-5.

Dolat L, Hu Q, Spiliotis ET. Septin functions in organ system physiology and pathology. Biol Chem. 2014;395(2):123-41. http://dx.doi.org/10.1515/hsz-2013-0233. PMid:24114910.

Douglas LM, Alvarez FJ, McCreary C, Konopka JB. Septin function in yeast model systems and pathogenic fungi. Eukaryot Cell. 2005;4(9):1503-12. http://dx.doi.org/10.1128/EC.4.9.1503-1512.2005. PMid:16151244.

Hall PA, Russell SE. The pathobiology of the septin gene family. J Pathol. 2004;204(4):489-505. http://dx.doi.org/10.1002/path.1654. PMid:15495264.

Huo HL, Wang P, Wang SY, Miao YW, Wei HJ, Zhang YY, Zhang QL, Li FQ, Liu LX, Wang R, Li WZ, Zhao Y, Huo JL, Xiao H. cDNA cloning, molecular characterization, and expression analyses of two novel porcine ARRDC genes: ARRDC1 and ARRDC5. Turk J Biol. 2015;39(4):523-31. http://dx.doi.org/10.3906/biy-1404-101.

Ihara M, Kinoshita A, Yamada S, Tanaka H, Tanigaki A, Kitano A, Goto M, Okubo K, Nishiyama H, Ogawa O, Takahashi C, Itohara S, Nishimune Y, Noda M, Kinoshita M. Cortical organization by the septin cytoskeleton is essential for structural and mechanical integrity of mammalian spermatozoa. Dev Cell. 2005;8(3):343-52. http://dx.doi.org/10.1016/j.devcel.2004.12.005. PMid:15737930.

Kinoshita A, Kinoshita M, Akiyama H, Tomimoto H, Akiguchi I, Kumar S, Noda M, Kimura J. Identification of septins in neurofibrillary tangles in Alzheimer’s disease. Am J Pathol. 1998;153(5):1551-60. http://dx.doi.org/10.1016/S0002-9440(10)65743-4. PMid:9811347.

Kinoshita M. Diversity of septin scaffolds. Curr Opin Cell Biol. 2006;18(1):54-60. http://dx.doi.org/10.1016/j.ceb.2005.12.005. PMid:16356703.

Kuhlenbäumer G, Hannibal MC, Nelis E, Schirmacher A, Verpoorten N, Meuleman J, Watts GD, De Vriendt E, Young P, Stögbauer F, Halfter H, Irobi J, Goossens D, Del-Favero J, Betz BG, Hor H, Kurlemann G, Bird TD, Airaksinen E, Mononen T, Serradell AP, Prats JM, Van Broeckhoven C, De Jonghe P, Timmerman V, Ringelstein EB, Chance PF. Mutations in SEPT9 cause hereditary neuralgic amyotrophy. Nat Genet. 2005;37(10):1044-6. http://dx.doi.org/10.1038/ng1649. PMid:16186812.

Kuo YC, Lin YH, Chen HI, Wang YY, Chiou YW, Lin HH, Pan HA, Wu CM, Su SM, Hsu CC, Kuo PL. SEPT12 mutations cause male infertility with defective sperm annulus. Hum Mutat. 2012;33(4):710-9. http://dx.doi.org/10.1002/humu.22028. PMid:22275165.

Mostowy S, Cossart P. Septins: the fourth component of the cytoskeleton. Nat Rev Mol Cell Biol. 2012;13(3):183-94. http://dx.doi.org/10.1038/nrm3284. PMid:22314400.

Peterson EA, Kalikin LM, Steels JD, Estey MP, Trimble WS, Petty EM. Characterization of a SEPT9 interacting protein, SEPT14, a novel testis-specific septin. Mamm Genome. 2007;18(11):796-807. http://dx.doi.org/10.1007/s00335-007-9065-x. PMid:17922164.

Peterson EA, Petty EM. Conquering the complex world of human septins: implications for health and disease. Clin Genet. 2010;77(6):511-24. http://dx.doi.org/10.1111/j.1399-0004.2010.01392.x. PMid:20236126.

Shafipour M, Sabbaghian M, Shahhoseini M, Sadighi Gilani MA. Comparative expression analysis of Septin 14 in testes of infertile men with normal spermatogenesis and spermatogenic failure. Iran J Reprod Med. 2014;12(3):205-8. PMid:24799881.

Shinoda T, Ito H, Sudo K, Iwamoto I, Morishita R, Nagata K. Septin 14 is involved in cortical neuronal migration via interaction with Septin 4. Mol Biol Cell. 2010;21(8):1324-34. http://dx.doi.org/10.1091/mbc.e09-10-0869. PMid:20181826.

Spiliotis ET, Dolat L. Septins: cytoskeletal filaments with structural and regulatory functions. Encycl Cell Biol. 2016;2(3):579-87.

Sugino Y, Ichioka K, Soda T, Ihara M, Kinoshita M, Ogawa O, Nishiyama H. Septins as diagnostic markers for a subset of human asthenozoospermia. J Urol. 2008;180(6):2706-9. http://dx.doi.org/10.1016/j.juro.2008.08.005. PMid:18951558.

Weirich CS, Erzberger JP, Barral Y. The septin family of GTPases: architecture and dynamics. Nat Rev Mol Cell Biol. 2008;9(6):478-89. http://dx.doi.org/10.1038/nrm2407. PMid:18478031.
 


Submitted date:
10/02/2020

Accepted date:
11/30/2020

602ec8c30e8825ab3b7b23c6 animreprod Articles
Links & Downloads

Anim Reprod

Share this page
Page Sections