title: PLCz functional haplotypes modulating promoter transcriptional activity are associated with semen quality traits in Chinese Holstein bulls.
abstract: The sperm-specific phospholipase C zeta (PLCz) is a candidate sperm-borne oocyte-activating factor that triggers a characteristic series of physiological stimuli via cytoplasmic Ca(2+) oscillations during fertilization. The molecular mechanisms involved in the regulation of PLCz gene expression remain largely unknown. To explore the genetic variations in the 5'-flanking region of the PLCz gene and their common haplotypes in Chinese Holstein bulls, as well as to determine whether these variations affect bovine semen quality traits and transcriptional activity, DNA samples were collected from Chinese Holstein bulls and sequenced for the identification of genetic variants in the 5'-flanking region of PLCz. Two genetic variants were identified, and their haplotypic profiles were constructed. The two novel genetic variations (g. -456 G>A and g. +65 T>C) were genotyped in 424 normal Chinese Holstein bulls. Bioinformatics analysis revealed that both loci are in transcription factor binding sites of the core promoter region. The association studies revealed that the two genetic variations and their haplotype combinations significantly affected semen quality traits. Using serially truncated constructs of the bovine PLCz promoters and the luciferase reporter, we found that a 726 bp (-641 nt to +112 nt) fragment constitutes the core promoter region. Furthermore, four haplotypes, H1H1 (GTGT), H2H2 (GCGC), H3H3 (ATAT), and H4H4 (ACAC), were significantly associated with semen quality traits and successfully transfected into MLTC-1 cell lines. The luciferase reporter assay showed that the different haplotypes exhibited distinct promoter activities. Maximal promoter activity was demonstrated by the H2H2 haplotypes, as compared with the other haplotypes. To the best of our knowledge, this study is the first report on genetic variants and their respective haplotypes in the 5'-flanking region of PLCz gene that can influence the semen quality of Chinese Holstein bulls as well as contribute to the transcriptional activity of the PLCz promoter.
authors.affiliation: Dairy Cattle Research Center, Shandong Academy of Agricultural Science, Jinan, PR China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Pan
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Ju
authors.forename: Jinming
authors.initials: J
authors.lastname: Huang
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Chao
authors.initials: C
authors.lastname: Qi
authors.forename: Qin
authors.initials: Q
authors.lastname: Gao
authors.forename: Lei
authors.initials: L
authors.lastname: Zhou
authors.forename: Qiuling
authors.initials: Q
authors.lastname: Li
authors.forename: Lingling
authors.initials: L
authors.lastname: Wang
authors.forename: Jifeng
authors.initials: J
authors.lastname: Zhong
authors.forename: Mei
authors.initials: M
authors.lastname: Liu
authors.forename: Changfa
authors.initials: C
authors.lastname: Wang
mesh_terms: D024506:5' Flanking Region; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D015870:Gene Expression; D014644:Genetic Variation; D060005:Genotyping Techniques; D006239:Haplotypes; D008297:Male; D008969:Molecular Sequence Data; D054801:Phosphoinositide Phospholipase C / Q000235:genetics*; D011401:Promoter Regions, Genetic; D019655:Quantitative Trait, Heritable; D012098:Reproduction / Q000235:genetics; D055101:Semen Analysis; D014158:Transcription, Genetic; D015533:Transcriptional Activation
chemical_list: D054801:Phosphoinositide Phospholipase C
references.citation: Physiol Rev. 2000 Oct;80(4):1291-335
references.pmid: 11015615
references.citation: Reproduction. 2007 Nov;134(5):695-704
references.pmid: 17965260
references.citation: Biol Reprod. 2005 Apr;72(4):992-6
references.pmid: 15601914
references.citation: Dev Biol. 2000 Jul 15;223(2):238-50
references.pmid: 10882513
references.citation: Anim Reprod Sci. 2009 Aug;114(1-3):14-22
references.pmid: 18829190
references.citation: Toxicol Appl Pharmacol. 2005 Sep 1;207(2 Suppl):84-90
references.pmid: 16002116
references.citation: J Immunol. 1995 Sep 15;155(6):3013-20
references.pmid: 7673719
references.citation: Dev Biol. 2000 Jul 15;223(2):251-65
references.pmid: 10882514
references.citation: Biol Reprod. 2001 May;64(5):1338-49
references.pmid: 11319137
references.citation: Anim Reprod Sci. 2002 Mar 15;70(1-2):99-109
references.pmid: 11841910
references.citation: Anim Reprod Sci. 2006 May;92(3-4):349-63
references.pmid: 16242873
references.citation: Biol Reprod. 2011 Jul;85(1):78-88
references.pmid: 21389344
references.citation: Reprod Domest Anim. 2012 Apr;47(2):213-23
references.pmid: 21752105
references.citation: J Biol Chem. 2004 Mar 12;279(11):10408-12
references.pmid: 14701816
references.citation: Immunogenetics. 2008 Apr;60(3-4):157-65
references.pmid: 18330558
references.citation: Reproduction. 2005 Aug;130(2):157-63
references.pmid: 16049153
references.citation: Genome Res. 2001 Jan;11(1):143-51
references.pmid: 11156623
references.citation: J Anim Sci. 2006 Feb;84(2):283-90
references.pmid: 16424254
references.citation: Dev Biol. 2000 Sep 1;225(1):1-12
references.pmid: 10964460
references.citation: DNA Cell Biol. 2012 Jun;31(6):1038-45
references.pmid: 22304649
references.citation: Domest Anim Endocrinol. 1999 Oct;17(2-3):139-48
references.pmid: 10527117
references.citation: Dev Biol. 2004 Oct 15;274(2):370-83
references.pmid: 15385165
references.citation: Cell Res. 2005 Feb;15(2):97-8
references.pmid: 15740637
references.citation: Reprod Domest Anim. 2007 Jun;42(3):326-8
references.pmid: 17506813
references.citation: Genome Res. 1999 Aug;9(8):720-31
references.pmid: 10447507
references.citation: Fertil Steril. 2009 May;91(5 Suppl):2230-42
references.pmid: 18710717
references.citation: Dev Biol. 1993 Jul;158(1):62-78
references.pmid: 8392472
references.citation: Anim Reprod Sci. 2011 Nov;129(1-2):14-21
references.pmid: 22030336
references.citation: J Immunol. 1998 Sep 15;161(6):3169-75
references.pmid: 9743385
references.citation: Dev Biol. 1999 Jul 15;211(2):157-76
references.pmid: 10395780
references.citation: Nucleic Acids Res. 1987 Aug 25;15(16):6643-53
references.pmid: 3628002
references.citation: Hum Reprod. 2009 Oct;24(10):2417-28
references.pmid: 19584136
references.citation: J Clin Invest. 2008 Nov;118(11):3671-81
references.pmid: 18924610
references.citation: Curr Opin Cell Biol. 1997 Apr;9(2):240-6
references.pmid: 9069263
references.citation: Reproduction. 2002 Nov;124(5):611-23
references.pmid: 12416999
references.citation: Biol Reprod. 2008 Jun;78(6):1081-90
references.pmid: 18322275
references.citation: Anim Genet. 2005 Apr;36(2):152-5
references.pmid: 15771727
references.citation: Reproduction. 2006 Sep;132(3):393-401
references.pmid: 16940280
references.citation: Mol Biol Rep. 2011 Jun;38(5):3411-5
references.pmid: 21104139
references.citation: Urol Clin North Am. 2008 May;35(2):211-20, viii
references.pmid: 18423241
references.citation: Science. 2000 Jan 14;287(5451):319-21
references.pmid: 10634790
references.citation: J Dairy Sci. 1998 Dec;81(12):3321-30
references.pmid: 9891279
references.citation: Development. 2002 Aug;129(15):3533-44
references.pmid: 12117804
references.citation: Genet Mol Res. 2011 Apr 26;10(2):731-8
references.pmid: 21523652