HDAC1和HDAC2对小鼠卵母细胞组蛋白去乙酰化的作用
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摘要
旨在探索组蛋白去乙酰化酶1 (histone deacetylase 1,HDAC1)和组蛋白去乙酰化酶2 (histone deacetylase 2,HDAC2)在卵母细胞减数分裂期组蛋白去乙酰化过程中的作用。首先利用免疫荧光技术,检测HDAC1与HDAC2在体细胞及卵母细胞不同时期的表达分布,然后分别利用抑制剂抑制HDAC1和HDAC2的活性,观察其对卵母细胞组蛋白去乙酰化的作用。结果:HDAC1与HDAC2分布在间期的体细胞和卵母细胞的细胞核中,但分裂期细胞中只有卵母细胞染色体上存在HDAC1的表达,抑制酶活性后该HDAC1的表达也消失;在小鼠卵母细胞第一次减数分裂双线期(germinal vesicle stage,GV期)卵母细胞体外成熟液中添加TSA (trichostatin A,HDACs广谱抑制剂)、Pyroxamide (HDAC1特异性抑制剂)、Santacruzamate A (HDAC2特异性抑制剂),发现Pyroxamide和Santacruzamate A均能显著抑制卵母细胞减数分裂过程中组蛋白乙酰化修饰的去除,但未能达到广谱抑制剂的抑制效果。研究表明,小鼠卵母细胞在减数分裂组蛋白去乙酰化过程中,HDAC1和HDAC2均具有组蛋白去乙酰化的作用,且与HDAC2相比,HDAC1发挥着主要的去乙酰化作用,这为组蛋白去乙酰化作用机理提供理论参考。
The aim of this research was to explore the regulatory effects of HDAC1 ( histone deacetylase 1,HDAC1) and HDAC2 ( histone deacetylase 2) on histone deacetylation during the oocyte meiophase. Firstly,the immunofluorescence technique was used in the experiment to detect the expression distribution of HDAC1 and HDAC2 in somatic cells and oocytes of different phases. Then,different inhibitors were added to suppress HDAC1 and HDAC2 to further observe their effects on histone deacetylation. The results showed that,during interphase,both HDAC1 and HDAC2 were distributed in the nucleus of somatic cells and oocytes,but only HDAC1 was co-located with chromosome on oocytes during division stage; and when the enzyme activity was inhibited,the co-locat ion disappeared. Adding TSA ( Trichostatin A,HDACs broad-spectrum inhibitor),Pyroxamide ( HDAC1 specific inhibitor),and Santacruzamate A ( HDAC2 specific inhibitor) to mouse in vitro maturation medium,we found that both Pyroxamide and Santacruzamate A significantly inhibited the deacetylation of histone in the process of meiosis in oocytes,but the inhibitory effect was not broad-spectrum. The present study showed that both HDAC1 and HDAC2 had histone deacetylation effects in the meiotic histone deacetylation in mouse oocytes,but they do not play all the roles,and that,compared with HDAC2,HDAC1 played a major role in this process. These results provided a theoretical reference for exploration into histone deacetylation mechanism.
The aim of this research was to explore the regulatory effects of HDAC1 ( histone deacetylase 1,HDAC1) and HDAC2 ( histone deacetylase 2) on histone deacetylation during the oocyte meiophase. Firstly,the immunofluorescence technique was used in the experiment to detect the expression distribution of HDAC1 and HDAC2 in somatic cells and oocytes of different phases. Then,different inhibitors were added to suppress HDAC1 and HDAC2 to further observe their effects on histone deacetylation. The results showed that,during interphase,both HDAC1 and HDAC2 were distributed in the nucleus of somatic cells and oocytes,but only HDAC1 was co-located with chromosome on oocytes during division stage; and when the enzyme activity was inhibited,the co-locat ion disappeared. Adding TSA ( Trichostatin A,HDACs broad-spectrum inhibitor),Pyroxamide ( HDAC1 specific inhibitor),and Santacruzamate A ( HDAC2 specific inhibitor) to mouse in vitro maturation medium,we found that both Pyroxamide and Santacruzamate A significantly inhibited the deacetylation of histone in the process of meiosis in oocytes,but the inhibitory effect was not broad-spectrum. The present study showed that both HDAC1 and HDAC2 had histone deacetylation effects in the meiotic histone deacetylation in mouse oocytes,but they do not play all the roles,and that,compared with HDAC2,HDAC1 played a major role in this process. These results provided a theoretical reference for exploration into histone deacetylation mechanism.
引文
[1]Shen Y,Wei W,Zhou D X.Histone acetylation enzymes coordinate metabolism and gene expression[J].Trends Plant Sci,2015,20(10):614-621.
[2]Saha R N,Pahan K.HATs and HDACs in neurodegeneration:a tale of disconcerted acetylation homeostasis[J].Cell Death Differ,2006,13(4):539-550.
[3]Peserico A,Simone C.Physical and functional HAT/HDAC interplay regulates protein acetylation balance[J].J Biomed Biotechnol,2011,doi:10.1155/2011/371832.
[4]Filgueiras L R,Brandt S L,Ramalho T R D,et al.Imbalance between HDAC and HAT activities drives aberrant STAT1/My D88 expression in macrophages from type 1 diabetic mice[J].J Diabetes Complicat,2017,31(2):334-339.
[5]Byskov A G,Andersen C Y,Nordholm L,et al.Chemical-structure of sterols that activate oocyte meiosis[J].Nature,1995,374(6522):559-562.
[6]Yoshida M,Furumai R,Nishiyama M,et al.Histone deacetylase as a new target for cancer chemotherapy[J].Cancer Chemoth Pharm,2001,48(Suppl 1):S20-S26.
[7]Kroesen M,Gielen P R,Brok I C,et al.HDAC inhibitors and immunotherapy;a double edged sword?[J]Oncotarget,2014,5(16):6558-6572.
[8]Brunmeir R,Lagger S,Seiser C.Histone deacetylase 1 and 2-controlled embryonic development and cell differentiation[J].Int J Dev Biol,2009,53(2-3):275-289.
[9]Ma P,Schultz R M.HDAC1 and HDAC2 in mouse oocytes and preimplantation embryos:Specificity versus compensation[J].Cell Death Differ,2016,23(7):1119-1127.
[10]Akiyama T,Kim J M,Nagata M,et al.Regulation of histone acetylation during meiotic maturation in mouse oocytes[J].Mol Reprod Dev,2004,69(2):222-227.
[11]Kim J M,Liu H L,Tazaki M,et al.Changes in histone acetylation during mouse oocyte meiosis[J].J Cell Biol,2003,162(1):37-46.
[2]Saha R N,Pahan K.HATs and HDACs in neurodegeneration:a tale of disconcerted acetylation homeostasis[J].Cell Death Differ,2006,13(4):539-550.
[3]Peserico A,Simone C.Physical and functional HAT/HDAC interplay regulates protein acetylation balance[J].J Biomed Biotechnol,2011,doi:10.1155/2011/371832.
[4]Filgueiras L R,Brandt S L,Ramalho T R D,et al.Imbalance between HDAC and HAT activities drives aberrant STAT1/My D88 expression in macrophages from type 1 diabetic mice[J].J Diabetes Complicat,2017,31(2):334-339.
[5]Byskov A G,Andersen C Y,Nordholm L,et al.Chemical-structure of sterols that activate oocyte meiosis[J].Nature,1995,374(6522):559-562.
[6]Yoshida M,Furumai R,Nishiyama M,et al.Histone deacetylase as a new target for cancer chemotherapy[J].Cancer Chemoth Pharm,2001,48(Suppl 1):S20-S26.
[7]Kroesen M,Gielen P R,Brok I C,et al.HDAC inhibitors and immunotherapy;a double edged sword?[J]Oncotarget,2014,5(16):6558-6572.
[8]Brunmeir R,Lagger S,Seiser C.Histone deacetylase 1 and 2-controlled embryonic development and cell differentiation[J].Int J Dev Biol,2009,53(2-3):275-289.
[9]Ma P,Schultz R M.HDAC1 and HDAC2 in mouse oocytes and preimplantation embryos:Specificity versus compensation[J].Cell Death Differ,2016,23(7):1119-1127.
[10]Akiyama T,Kim J M,Nagata M,et al.Regulation of histone acetylation during meiotic maturation in mouse oocytes[J].Mol Reprod Dev,2004,69(2):222-227.
[11]Kim J M,Liu H L,Tazaki M,et al.Changes in histone acetylation during mouse oocyte meiosis[J].J Cell Biol,2003,162(1):37-46.