title: Differential gene expression of serine protease inhibitors in bovine ovarian follicle: possible involvement in follicular growth and atresia.
abstract: BACKGROUND
  SERPINs (serine protease inhibitors) regulate proteases involving fibrinolysis, coagulation, inflammation, cell mobility, cellular differentiation and apoptosis. This study aimed to investigate differentially expressed genes of members of the SERPIN superfamily between healthy and atretic follicles using a combination of microarray and quantitative real-time PCR (QPCR) analysis. In addition, we further determined mRNA and protein localization of identified SERPINs in estradiol (E2)-active and E2-inactive follicles by in situ hybridization and immunohistochemistry.
  
  
  METHODS
  We performed microarray analysis of healthy (10.7 +/- 0.7 mm) and atretic (7.8 +/- 0.2 mm) follicles using a custom-made bovine oligonucleotide microarray to screen differentially expressed genes encoding SERPIN superfamily members between groups. The expression profiles of six identified SERPIN genes were further confirmed by QPCR analysis. In addition, mRNA and protein localization of four SERPINs was investigated in E2-active and E2-inactive follicles using in situ hybridization and immunohistochemistry.
  
  
  RESULTS
  We have identified 11 SERPIN genes expressed in healthy and atretic follicles by microarray analysis. QPCR analysis confirmed that mRNA expression of four SERPINs (SERPINA5, SERPINB6, SERPINE2 and SERPINF2) was greater in healthy than in atretic follicles, while two SERPINs (SERPINE1 and SERPING1) had greater expression in atretic than in healthy follicles. In situ hybridization showed that SERPINA5, SERPINB6 and SERPINF2 mRNA were localized in GCs of E2-active follicles and weakly expressed in GCs of E2-inactive follicles. SERPING1 mRNA was localized in both GCs and the theca layer (TL) of E2-inactive follicles and a weak hybridization signal was also detected in both GCs and TL of E2-active follicles. Immunohistochemistry showed that SERPINA5, SERPINB6 and SERPINF2 were detected in GCs of E2-active and E2-inactive follicles. SERPING1 protein was localized in both GCs and the TL of E2-active and E2-inactive follicles.
  
  
  CONCLUSIONS
  Our results demonstrate a characteristic expression of SERPIN superfamily member genes in bovine healthy and atretic follicles. The cell-type-and stage-specific expression of SERPINs may be associated with bovine follicular growth and atresia.
authors: Reproductive Biology Research Unit, Division of Animal Science, National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan.
authors: Ken-Go
authors: KG
authors: Hayashi
authors: Koichi
authors: K
authors: Ushizawa
authors: Misa
authors: M
authors: Hosoe
authors: Toru
authors: T
authors: Takahashi
references: J Reprod Fertil. 1995 May;104(1):115-23
references: 7636792
references: Reproduction. 2002 May;123(5):711-9
references: 12006099
references: Biochem Biophys Res Commun. 2005 Jan 14;326(2):435-41
references: 15582596
references: J Thromb Haemost. 2007 Jul;5 Suppl 1:102-15
references: 17635716
references: Reproduction. 2006 Aug;132(2):191-206
references: 16885529
references: Reproduction. 2008 Feb;135(2):253-65
references: 18239053
references: Biol Reprod. 2004 Sep;71(3):887-93
references: 15128599
references: J Reprod Fertil. 1989 Sep;87(1):223-30
references: 2621698
references: Biol Reprod. 2006 Jun;74(6):1051-9
references: 16481595
references: J Biol Chem. 1988 Apr 25;263(12):5797-803
references: 3128548
references: J Reprod Fertil. 1994 Aug;101(3):547-55
references: 7966007
references: J Biol Chem. 1980 May 10;255(9):3931-4
references: 6989830
references: Anim Reprod Sci. 2004 Jul;82-83:109-26
references: 15271447
references: Biol Reprod. 1995 Apr;52(4):745-52
references: 7779996
references: Mol Reprod Dev. 2008 May;75(5):904-14
references: 17948250
references: Mol Immunol. 2008 Oct;45(16):4057-63
references: 18674818
references: Biol Reprod. 2005 Aug;73(2):324-33
references: 15829623
references: Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8989-93
references: 8090757
references: FEBS J. 2005 Oct;272(19):4852-7
references: 16176259
references: J Biol Chem. 1985 Jun 10;260(11):7029-34
references: 3997857
references: Cell Struct Funct. 1996 Oct;21(5):425-30
references: 9118251
references: Biol Reprod. 2000 May;62(5):1160-7
references: 10775162
references: Mol Cell Endocrinol. 2002 Feb 22;187(1-2):29-38
references: 11988309
references: Biol Reprod. 2002 May;66(5):1350-8
references: 11967197
references: Hypertens Res. 2003 Sep;26(9):723-9
references: 14620928
references: Biol Reprod. 2004 May;70(5):1475-84
references: 14736815
references: J Biol Chem. 1998 Jan 23;273(4):1851-4
references: 9442015
references: PLoS One. 2009 Jun 05;4(6):e5814
references: 19503832
references: Reprod Fertil Dev. 2007;19(8):967-75
references: 18076829
references: Biol Reprod. 1996 Dec;55(6):1187-94
references: 8949873
references: J Pharmacol Sci. 2005 Sep;99(1):6-38
references: 16177542
references: J Biochem. 2007 Oct;142(4):435-42
references: 17761692
references: Mol Reprod Dev. 2009 Apr;76(4):351-66
references: 18932212
references: Endocrinology. 1985 Sep;117(3):879-85
references: 3160575
references: Cell. 1980 Aug;21(1):37-45
references: 6157479
references: Biol Reprod. 1995 Dec;53(6):1302-10
references: 8562685
references: Biochem J. 1993 Jul 1;293 ( Pt 1):187-93
references: 7687128
references: J Vet Med Sci. 2004 Mar;66(3):225-9
references: 15107548
references: Reproduction. 2006 Jan;131(1):125-37
references: 16388016
references: Mol Reprod Dev. 2006 Jan;73(1):9-19
references: 16175637
references: FEBS Lett. 2005 Jun 13;579(15):3310-6
references: 15943976
references: Mol Reprod Dev. 2008 Sep;75(9):1457-72
references: 18288646
references: Blood. 1999 Mar 15;93(6):2089-97
references: 10068683
references: Reprod Biol Endocrinol. 2007 Apr 27;5:17
references: 17462098
references: Biol Chem. 2002 Dec;383(12):1917-24
references: 12553728
references: Reprod Biol Endocrinol. 2010 Feb 05;8:11
references: 20132558
references: Br J Haematol. 2005 May;129(3):307-21
references: 15842654
references: Biol Reprod. 2006 Feb;74(2):344-51
references: 16221993
references: Biochim Biophys Acta. 1995 Sep 27;1252(1):28-34
references: 7548163
references: Genome Biol. 2006;7(5):216
references: 16737556
references: Biol Reprod. 1988 Sep;39(2):308-17
references: 3052602
references: J Biol Chem. 1996 Jan 19;271(3):1605-12
references: 8576159
references: Mol Reprod Dev. 2003 Feb;64(2):152-65
references: 12506347
references: Science. 1999 Jul 9;285(5425):245-8
references: 10398599