Abstract
It is widely accepted that mature sperm contains RNA. The first hypothesis was that sperm RNAs have no functions of their own but are simply residues of spermatogenesis reflecting the events that occurred during their formation in the testes. More recently new discoveries have essentially expanded these views, showing that sperm mRNAs constitute a population of stable full-length transcripts, many of which are selectively retained during spermatogenesis and delivered to oocytes contributing to early embryo development. It is well known that semen quality can be influenced by occasional physical stress, infection, and variation in temperature and the definition of new markers for evaluation of semen could offer knowledge about the fertility potential of a semen sample. The aim of the present study was to evaluate the presence and the relative quantity of transcripts and protein of heat shock protein 70 (HSP70), 90 (HSP90) and clusterin (CLU) in Percoll-selected spermatozoa collected from seven adult boars of proven fertility routinely employed for artificial insemination. Our results showed the presence of HSP70, HSP90 and CLU transcripts with different level of expression: high for HSPs and low for CLU transcripts. The transcript level of both HSPs are similar among selected spermatozoa derived from high quality sperm with the exception of one boar that showed a reduced content of HSP70 and HSP90 mRNA together with a lower semen quality. At protein level, both HSPs were detected with similar amount among all seven boars whilst no band was evidenced for CLU protein.
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References
Althouse GC (2008) Sanitary procedures for the production of extended semen. Reprod Domest Anim 43:374–378. doi:10.1111/j.1439-0531.2008.01187.x
Althouse GC, Lu KG (2005) Bacteriospermia in extended porcine semen. Theriogenology 63:573–584. doi:10.1016/j.theriogenology.2004.09.031
Althouse GC, Kuster CE, Clark SG, Weisiger RM (2000) Field investigations of bacterial contaminants and their effects on extended porcine semen. Theriogenology 53:1167–1176. doi:10.1016/S0093-691X(00)00261-2
Dadoune JP, Pawlak A, Alfonsi MF, Siffroi JP (2005) Identification of transcripts by macroarrays, RT-PCR and in situ hybridization in human ejaculate spermatozoa. Mol Hum Reprod 11:133–140. doi:10.1093/molehr/gah137
Driver AM, Khatib H (2013) Physiology and endocrinology symposium: heat shock proteins: potentially powerful markers for preimplantation embryonic development and fertility in livestock species. J Anim Sci 91:1154–1161. doi:10.2527/jas.2012-5928
Fantinati P, Zannoni A, Bernardini C et al (2009) Evaluation of swine fertilisation medium (SFM) efficiency in preserving spermatozoa quality during long-term storage in comparison to four commercial swine extenders. Animal 3:269–274. doi:10.1017/S1751731108003443
Feugang JM, Rodriguez-Osorio N, Kaya A et al (2010) Transcriptome analysis of bull spermatozoa: implications for male fertility. Reprod BioMed Online 21:312–324. doi:10.1016/j.rbmo.2010.06.022
Gilbert I, Bissonnette N, Boissonneault G et al (2007) A molecular analysis of the population of mRNA in bovine spermatozoa. Reproduction 133:1073–1086. doi:10.1530/REP-06-0292
Hamatani T (2012) Human spermatozoal RNAs. Fertil Steril 97:275–281. doi:10.1016/j.fertnstert.2011.12.035
Han Z, Wang Z, Cheng G et al (2012) Presence, localization, and origin of clusterin in normal human spermatozoa. J Assist Reprod Genet 29:751–757. doi:10.1007/s10815-012-9779-x
Huang SY, Kuo YH, Lee YP et al (2000a) Association of heat shock protein 70 with semen quality in boars. Anim Reprod Sci 63:231–240
Huang SY, Kuo YH, Tsou HL et al (2000b) The decline of porcine sperm motility by geldanamycin, a specific inhibitor of heat-shock protein 90 (HSP90). Theriogenology 53:1177–1184. doi:10.1016/S0093-691X(00)00262-4
Huszar G, Stone K, Dix D, Vigue L (2000) Putative creatine kinase M-isoform in human sperm is identified as the 70-kilodalton heat shock protein HspA2. Biol Reprod 63:925–932
Hwang JY, Mulligan BP, Kim H-M et al (2013) Quantitative analysis of sperm mRNA in the pig: relationship with early embryo development and capacitation. Reprod Fertil Dev 25:807–817. doi:10.1071/RD12160
Ibrahim NM, Troedesson MHT, Foster DN et al (1999) Reproductive tract secretions and bull spermatozoa contain different clusterin isoforms that cluster cells and inhibit complement induced cytolysis. J Androl 20:230–240
Ibrahim NM, Gilbert GR, Loseth KJ, Crabo BG (2000) Correlation between clusterin-positive spermatozoa determined by flow cytometry in bull semen and fertility. J Androl 21:887–894
Ibrahim NM, Foster DN, Crabo BG (2001) Localization of clusterin on freeze-preserved bull spermatozoa before and after glass wool-sephadex filtration. J Androl 22:891–902. doi:10.1002/j.1939-4640.2001.tb02594.x
Jodar M, Selvaraju S, Sendler E et al (2013) The presence, role and clinical use of spermatozoal RNAs. Hum Reprod Update 19:604–624. doi:10.1093/humupd/dmt031
Kasimanickam V, Kasimanickam R, Arangasamy A et al (2012) Association between mRNA abundance of functional sperm function proteins and fertility of Holstein bulls. Theriogenology 78:2007–2019. doi:10.1016/j.theriogenology.2012.07.016
Kempisty B, Antosik P, Bukowska D et al (2008) Analysis of selected transcript levels in porcine spermatozoa, oocytes, zygotes and two-cell stage embryos. Reprod Fertil Dev 20:513–518
Knox R, Levis D, Safranski T, Singleton W (2008) An update on north American boar stud practices. Theriogenology 70:1202–1208. doi:10.1016/j.theriogenology.2008.06.036
Krawetz SA (2005) Paternal contribution: new insights and future challenges. Nat Rev Genet 6:633–642. doi:10.1038/nrg1654
Kumar S, Deb R, Singh U, Ganguly I, Mandal DK, Tyagi S, Kumar M, Sengar G, Sharma S, Singh R, Singh R (2015) Bovine circadian locomotor output cycles kaput (CLOCK) and clusterin (CLU) mRNA quantitation in ejaculated crossbred bull spermatozoa. Reprod Domest Anim 50(3):505–509. doi:10.1111/rda.12522
Kunavongkrit A, Suriyasomboon A, Lundeheim N et al (2005) Management and sperm production of boars under differing environmental conditions. Theriogenology 63:657–667. doi:10.1016/j.theriogenology.2004.09.039
Lambard S, Galeraud-Denis I, Martin G et al (2004) Analysis and significance of mRNA in human ejaculated sperm from normozoospermic donors: relationship to sperm motility and capacitation. Mol Hum Reprod 10:535–541. doi:10.1093/molehr/gah064
Martínez-Heredia J, de Mateo S, Vidal-Taboada JM et al (2008) Identification of proteomic differences in asthenozoospermic sperm samples. Hum Reprod 23:783–791. doi:10.1093/humrep/den024
Matàs C, Vieira L, Garcìa-Vàzquez FA et al (2011) Effects of centrifugation through three different discontinuous Percoll gradients on boar sperm function. Anim Reprod Sci 127:62–72. doi:10.1016/j.anireprosci.2011.06.009
Muciaccia B, Sette C, Paronetto MP et al (2010) Expression of a truncated form of KIT tyrosine kinase in human spermatozoa correlates with sperm DNA integrity. Hum Reprod 25:2188–2202. doi:10.1093/humrep/deq168
Neuer A, Spandorfer SD, Giraldo P et al (2000) The role of heat shock proteins in reproduction. Hum Reprod Update 6:149–159
O’Bryan MK, Murphy BF, Liu DY et al (1994) The use of anticlusterin monoclonal antibodies for the combined assessment of human sperm morphology and acrosome integrity. Hum Reprod 9:1490–1496
Ostermeier GC, Dix DJ, Miller D et al (2002) Spermatozoal RNA profiles of normal fertile men. Lancet 360:772–777. doi:10.1016/S0140-6736(02)09899-9
Ostermeier GC, Miller D, Huntriss JD et al (2004) Reproductive biology: delivering spermatozoan RNA to the oocyte. Nature 429:154. doi:10.1038/429154a
Spinaci M, Volpe S, Bernardini C et al (2005) Immunolocalization of heat shock protein 70 (Hsp 70) in boar spermatozoa and its role during fertilization. Mol Reprod Dev 72:534–541. doi:10.1002/mrd.20367
Spinaci M, Vallorani C, Bucci D, Bernardini C, Tamanini C, Seren E, Galeati G (2010) Effect of liquid storage on sorted boar spermatozoa. Theriogenology 74:741–748
Sylvester SR, Morales C, Oko R, Griswold MD (1991) Localization of sulfated glycoprotein-2 (clusterin) on spermatozoa and in the reproductive tract of the male rat. Biol Reprod 45:195–207
Turba ME, Fantinati P, Bernardini C et al (2007) Relationships between innovative and traditional parameters to investigate semen quality in pigs. Anim Reprod Sci 99:72–81. doi:10.1016/j.anireprosci.2006.04.045
Volpe S, Galeati G, Bernardini C et al (2008) Comparative immunolocalization of heat shock proteins (Hsp)-60, −70, −90 in boar, stallion, dog and cat spermatozoa. Reprod Domest Anim 43:385–392. doi:10.1111/j.1439-0531.2007.00918.x
Wang P, Wang Y-F, Wang H et al (2014) HSP90 expression correlation with the freezing resistance of bull sperm. Zygote 22:239–245. doi:10.1017/S096719941300004X
Yang CC, Lin YS, Hsu CC et al (2009) Identification and sequencing of remnant messenger RNAs found in domestic swine (Sus scrofa) fresh ejaculated spermatozoa. Anim Reprod Sci 113:143–155. doi:10.1016/j.anireprosci.2008.08.012
Zannoni A, Zaniboni A, Bernardini C et al (2015) Expression fo clusterin isoforms in LPS treated porcine aortic endothelial cells. Pak Vet J 33:236–238
Zhang B, Peñagaricano F, Driver A et al (2011) Differential expression of heat shock protein genes and their splice variants in bovine preimplantation embryos. J Dairy Sci 94:4174–4182. doi:10.3168/jds.2010-4137
Zhang X-G, Hu S, Han C et al (2015) Association of heat shock protein 90 with motility of post-thawed sperm in bulls. Cryobiology 70:164–169. doi:10.1016/j.cryobiol.2014.12.010
Acknowledgements
This study was supported by grant from Bologna University (RFO, Ricerca Fondamentale Orientata). The authors wish to thank Dr. Diego Bucci for his technical support.
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Zannoni, A., Bernardini, C., Zaniboni, A. et al. Relative abundance of heat shock proteins and clusterin transcripts in spermatozoa collected from boar routinely utilised in an artificial insemination centre: preliminary results. Vet Res Commun 41, 233–239 (2017). https://doi.org/10.1007/s11259-017-9689-6
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DOI: https://doi.org/10.1007/s11259-017-9689-6