Expression of Attractin in male reproductive tract of human and mice and its correlation with male reproduction

Abstract

The expression of Attractin mRNA and protein in testis and semen of human and male mice was investigated. Human testis and semen samples were all collected from Reproductive Center of Renmin Hospital, Wuhan University in December, 2012. Testis samples were collected from 7 cases of obstructive azoospermias when they were subjected to diagnosed testis biopsy, and 30 normal human semen samples were obtained from those cases of semen analysis. Adult mice testis tissues were obtained from 10 2-month-old male BALB/c mice, and 60 male mice at different ages were classified into 10 groups (day 1, 5, 10, 15, 21, 28, 35, 42, 56, and 120 respectively, n=6 each). The expression of Attractin mRNA and protein in testis was detected by RT-PCR and Western blotting respectively. Human semen samples were centrifuged into sperm plasma (SP) and sperm extract (SE), and mice sperm samples were collected from the epididymis of 10 adult male BALB/c mice. Western blotting was used to determine the Attractin protein expression level. Attractin mRNA and protein were expressed in the testis of both patients with obstructive azoospermias and adult Bcl/B mice. Quantitative RT-PCR revealed that no Attractin mRNA was detectable in day 1 male BALB/c mice group. The Attractin mRNA and protein levels were low on the day 10, and increased with age until day 56. On the day 120, the expression levels of Attractin were decreased. As for human semen samples, Attractin protein was expressed in both SP and SE, but didn’t exist in samples from the epididymis of male BALB/c mice. It was suggested that Attractin acted as a novel active substance and was involved in male reproduction in both human and BALB/c mice, but it exerted a different expression profile in different mammal species.

Keywords

Attractin / male reproduction / spermatogenesis

References

[1] Duke-CohanJS, GuJ, McLaughlinDF, et al. . Attractin (DPPT-L), a member of the CUB family of cell adhesion and guidance proteins, is secreted by activated human T lymphocytes and modulates immune cell interactions. Proc Natl Acad Sci USA, 1998, 95(19): 11 336-11 341

[2] NakadateK, SakakibaraS, UedaS. Attractin/mahogany protein expression in the rodent central nervous system. J Comp Neurol, 2008, 508(1): 94-111 PMID: 18302151

[3] WjstM. Public data mining shows extended linkage disequilibrium around ADAM33. Allergy, 2007, 62(4): 444-446 PMID: 17362257

[4] FàbregaA, GuyonnetB, DacheuxJL, et al. . Expression, immunolocalization and processing of fertilins ADAM-1 and ADAM-2 in the boar (Sus domesticus) spermatozoa during epididymal maturation. Reprod Biol Endocrinol, 2011, 30(9): 96

[5] RadhakrishnanY, HamilKG, TanJA, et al. . Novel partners of SPAG11B isoform D in the human male reproductive tract. Biol Reprod, 2009, 81(4): 647-56 PMID: 19535787 PMCID: 2754882

[6] LiuJ, XiongC. Immunohistochemical localization of attractin in the testis and epididymis of mature male rats. Zhonghua Nan Ke Xue (Chinese), 2004, 10(7): 515-517

[7] ZhuY, XiongCL, ShenSL, et al. . Distribution of Mahoganoid protein and mRNA in the testes and epididymides of mature male rats. Zhonghua Nan Ke Xue (Chinese), 2005, 11: 601-604

[8] SuiC, XiongCL. The influence on the level of reproductive hormone of male mice after knocking out Attractin gene or Mahoganoind gene. Zhong Guo Ji Hua Sheng Yu Xue Za Zhi (Chinese), 2007, 15(12): 729-731

[9] ChengD, XiongC, LiJ, et al. . The effect of Mahogunin gene mutant on reproduction in male mice: a new sight for infertility?. Andrologia, 2014, 46(2): 98-105 PMID: 23210986

[10] WalkerWP, AradhyaS, HuCL, et al. . Genetic analysis of attractin homologs. Genesis, 2007, 45(12): 744-756 PMID: 18064672

[11] GohmaH, KuramotoT, MatalonR, et al. . Absence-like and tonic seizures in aspartoacylase/attractin double-mutant mice. Exp Anim, 2007, 56(2): 161-165 PMID: 17460362

[12] PazJ, YaoH, LimHS, et al. . The neuroprotective role of attractin in neurodegeneration. Neurobiol Aging, 2007, 28(9): 1446-1456 PMID: 16860906

[13] CotaCD, LiuRR, SumberacTM, et al. . Genetic and phenotypic studies of the dark-like mutant mouse. Genesis, 2008, 46(10): 562-573 PMID: 18821597

[14] Fernandez-FernandezR, MartiniAC, NavarroVM, et al. . Novel signals for the integration of energy balance and reproduction. Mol Cell Endocrinol, 2006, 254–255: 127-132 PMID: 16759792

[15] TokudaS, KuramotoT, SerikawaT. PCR-based genotyping of the rat Atrn (mv) mutation. Exp Anim, 2004, 53(1): 73-76 PMID: 14993747

[16] PantN, PantAB, ChaturvediPK, et al. . Semen quality of environmentally exposed human population: the toxicological consequence. Environ Sci Pollut Res Int, 2013, 20(11): 8274-8281 PMID: 23690079

[17] LiHG, DingXF, LiaoAH, et al. . Expression of CatSper family transcripts in the mouse testis during post-natal development and human ejaculated spermatozoa: relationship to sperm motility. Mol Hum Reprod, 2007, 13(5): 299-306 PMID: 17347248

[18] WrengerS, FaustJ, FriedrichD, et al. . Attractin, a dipeptidyl peptidase IV/CD26-like enzyme, is expressed on human peripheral blood monocytes and potentially influences monocyte function. J Leukoc Biol, 2006, 80(3): 621-629 PMID: 16835316

[19] SakakibaraS, NakadateK, OokawaraS, et al. . Non-cell autonomous impairment of oligodendrocyte differentiation precedes CNS degeneration in the Zitter rat: implications of macrophage/microglial activation in the pathogenesis. BMC Neurosci, 2008, 5(9): 35

[20] TangW, GunnTM, McLaughlinDF, et al. . Secreted and membrane attractin result from alternative splicing of the human ATRN gene. Proc Natl Acad Sci USA, 2000, 97(11): 6025-6030 PMID: 10811918 PMCID: 18552

[21] MutoY, SatoK. Pivotal role of attractin in cell survival under oxidative stress in the zitter rat brain with genetic spongiform encephalopathy. Brain Res Mol Brain Res, 2003, 111(1–2): 111-122 PMID: 12654511

[22] SunK, JohnsonBS, GunnTM. Mitochondrial dysfunction precedes neurodegeneration in mahogunin (Mgrn1) mutant mice. Neurobiol Aging, 2007, 28(12): 1840-1852 PMID: 17720281 PMCID: 2128279