半胱亚磺酸脱羧酶在妊娠小鼠子宫和胚胎中的表达定位研究
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摘要
目的研究半胱亚磺酸脱羧酶(CSD)mRNA和蛋白在妊娠小鼠子宫和胚胎中的表达定位。方法构建CSD原位杂交探针,收集正常妊娠小鼠第1～9天子宫组织和第11天胚胎,采用免疫组织化学染色、原位杂交检测CSD的表达情况。结果小鼠妊娠第1～2天CSD主要定位在子宫腔上皮细胞和腺上皮细胞,基质细胞从第3天开始表达,第6天开始胚胎周围的子宫蜕膜细胞中检测不到CSD蛋白,第8天子宫CSD的表达明显减弱,第9天表达更弱。从囊胚到第9天时胚胎都有CSD表达。CSD在第7天定位于着床点子宫和胚胎中,胚胎周围的子宫蜕膜细胞无表达,在第11天定位于胚胎的脑、心脏、脊髓。结论子宫和胚胎在小鼠妊娠过程中可表达CSD。
Objective To explore the expression and localization of cysteine sulfinate decarboxylase(CSD)in mouse uterus and embryo during pregnancy.Methods CSD in situ hybridization probes were constructed,then collected the uterus of pregnant mice from the first day to the ninth day and the embryos on the eleventh day,the expression of CSD was detected by immunohistochemistry and in situ hybridization.Results The epithelial and gland cells of uterus were positive for CSD from the first day to the second day,the stromal cells became intensely positive for CSD from the third day.CSD protein was not detected in decidual cells around embryo from the sixth day.The CSD response in the uterus was significantly weakened on the eighth day,and the expression was even weaker on the ninth day.The embryo expressed CSD from the blastocyst to the embryo on the ninth day.Uterus and embryo were positive for CSD whereas the decidual cells in the implantation site were unstained on the seventh day,CSD was positive at brain,heart and spinal cord in the embryo on the eleventh day.Conclusion CSD expresses in the uterus and embryo of the pregnant mice.
Objective To explore the expression and localization of cysteine sulfinate decarboxylase(CSD)in mouse uterus and embryo during pregnancy.Methods CSD in situ hybridization probes were constructed,then collected the uterus of pregnant mice from the first day to the ninth day and the embryos on the eleventh day,the expression of CSD was detected by immunohistochemistry and in situ hybridization.Results The epithelial and gland cells of uterus were positive for CSD from the first day to the second day,the stromal cells became intensely positive for CSD from the third day.CSD protein was not detected in decidual cells around embryo from the sixth day.The CSD response in the uterus was significantly weakened on the eighth day,and the expression was even weaker on the ninth day.The embryo expressed CSD from the blastocyst to the embryo on the ninth day.Uterus and embryo were positive for CSD whereas the decidual cells in the implantation site were unstained on the seventh day,CSD was positive at brain,heart and spinal cord in the embryo on the eleventh day.Conclusion CSD expresses in the uterus and embryo of the pregnant mice.
引文
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[14]胡森,王君伟,姜骞等.猪传染性胃肠炎病毒TH-98株S基因5′端部分片段克隆及序列分析[J].中国兽医杂志,2004,4(1):3-7.
[15]CARSON D D,BAGCHI I,DEY S K,et al.Embryo implantationt[J].Dev Biol,2000,223:217-237.
[16]LOBO M V,ALONSO F J,LATORRE A,et al.Immunohistochemical localization of taurine in the rat ovary,oviduct,and uterus[J].J Histochem Cytochem,2001,49(9):1133-1142.
[17]KIM H W,YOON S H,PARK T,et al.Gene expressions of taurine transporter and taurine synthetic enzyme during mouse and chicken embryonic development[J].Adv Exp Med Biol,2006,583:69-77.
[18]STURMAN J A,MESSING J M.Dietary taurine content and feline reproduction and outcome[J].J Nutr,1991,121(8):1195-1203.
[19]WANG Y,LI X W,LIU J,et al.Antenatal taurine supplementation in fetal rats with growth restriction improves neural stem cell proliferation by inhibiting the activities of Rho family factors[J].J Matern Fetal Neonatal Med,2017,31(11):1-15.
[20]ITO T,KIMURA Y,UOZUMI Y,et al.Taurine depletion caused by knocking out the taurine transporter gene leads to cardiomyopathy with cardiac atrophy[J].J Mol Cell Cardiol,2008,44(5):927-937.
[2]LI F,TENG H Y,LIU J,et al.Antenatal taurine supplementation increases taurine content in intrauterine growth restricted fetal rat brain tissue[J].Metab Brain Dis,2014,29(3):867-871.
[3]LIN T,DIAO Y F,KANG J W,et al.Tauroursodeoxycholic acid improves the implantation and live-birth rates of mouse embryos[J].Reprod Biol,2015,15(2):101-105.
[4]HARRIS S E,GOPICHANDRAN N,PICTON H M,et al.Nutrient concentrations in murine follicular fluid and the female reproductive tract[J].Theriogenology,2005,64(4):992-1006.
[5]MILLER J G,SCHULTZ G A.Amino acid content of preimplantation rabbit embryos and fluids of the reproductive tract[J].Biol Reprod,1987,36(1):125-129.
[6]BARNETT D K,BAVISTER B D.Hypotaurine requirement for in vitro development of golden hamster one-cell embryos into morulae and blastocysts,and production of term offspring from in vitro-fertilized ova[J].Biol Reprod,1992,47(2):297-304.
[7]GUYADER-JOLY C,GUERIN P,RENARD J P,et al.Precursors of taurine in female genital tract:Effects on developmental capacity of bovine embryo produced in vitro[J].Amino Acids,1998,15(1):27-42.
[8]NAKAMURA K,MORIMOTO K,SHIMA K,et al.The effect of supplementation of amino acids and taurine to modified KSOM culture medium on rat embryo development[J].Theriogenology,2016,86(8):2083-2090.
[9]STURMAN J A,MESSING J M.Dietary taurine content and feline reproduction and outcome[J].J Nutr,1991,121(8):1195-1203.
[10]STURMAN J A.Taurine in development[J].Physiol Rev,1993,73(1):119-147.
[11]HUXTABLE R J.Physiologicaly actions of taurine[J].Physiol Rev,1992,1(72):101-142.
[12]MA Q W,ZHAO J J,CAO W,et al.Estradiol decreases taurine level by reducing cysteine sulfinic acid decarboxylase via the estrogen receptor-alpha in female mice liver[J].Am J Physiol Gastrointest Liver Physiol,2015,308(4):G277-286.
[13]李春利,王云龙,李晨阳,等.猪瘟病毒E2蛋白A3BCD主要抗原结构域的原核表达[J].生物技术通讯,2009,20(1):55-58.
[14]胡森,王君伟,姜骞等.猪传染性胃肠炎病毒TH-98株S基因5′端部分片段克隆及序列分析[J].中国兽医杂志,2004,4(1):3-7.
[15]CARSON D D,BAGCHI I,DEY S K,et al.Embryo implantationt[J].Dev Biol,2000,223:217-237.
[16]LOBO M V,ALONSO F J,LATORRE A,et al.Immunohistochemical localization of taurine in the rat ovary,oviduct,and uterus[J].J Histochem Cytochem,2001,49(9):1133-1142.
[17]KIM H W,YOON S H,PARK T,et al.Gene expressions of taurine transporter and taurine synthetic enzyme during mouse and chicken embryonic development[J].Adv Exp Med Biol,2006,583:69-77.
[18]STURMAN J A,MESSING J M.Dietary taurine content and feline reproduction and outcome[J].J Nutr,1991,121(8):1195-1203.
[19]WANG Y,LI X W,LIU J,et al.Antenatal taurine supplementation in fetal rats with growth restriction improves neural stem cell proliferation by inhibiting the activities of Rho family factors[J].J Matern Fetal Neonatal Med,2017,31(11):1-15.
[20]ITO T,KIMURA Y,UOZUMI Y,et al.Taurine depletion caused by knocking out the taurine transporter gene leads to cardiomyopathy with cardiac atrophy[J].J Mol Cell Cardiol,2008,44(5):927-937.