The role of nucleotide pyrophosphatase in Mullerian duct regression.
The role of nucleotide pyrophosphatase in Mullerian duct regression.
复制标题
核苷酸焦磷酸酶在苗勒氏管消退中的作用。
DOI：
10.1016/0012-1606(83)90230-0
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发表时间：
1983
影响因子：
2.7
通讯作者：
Donahoe,PK
中科院分区：
文献类型：
--
作者：
Fallat,ME;Hutson,JM;Budzik,GP;Donahoe,PK
苗勒管抑制物质（Mullerian inhibiting substance，MIS）是一种来自胎儿睾丸的糖蛋白，可引起胚胎苗勒管的退行性变。在体外，当Mn 2+存在时，包括GTP、NAD、ATP、AMP在内的多种核苷酸及几种不可水解的合成ATP类似物都能抑制MIS。细胞外核苷酸焦磷酸酶（NPPase）是一种能够水解多种核苷酸和类似物的酶，发现其抑制苗勒管退化，通过组织化学染色（H. Sierakowska和D. Shugar（1963）.生物化学生物物理学Res. Commun. 11，70-74），以确定在消退期间NPP酶是否定位在苗勒管中或周围。取14 ± 12 ~ 17 ± 12日龄大鼠胚胎的尿生殖嵴冰冻切片，用α-萘胸苷-5 ′-磷酸（naphthaltythromidine-5′-phosphate，naphthalTMP）和固红TR孵育。核苷酸焦磷酸酶水解萘TMP，释放萘酚，然后与固红反应，在酶位点产生颜色。核苷酸水解检测周围退化男性（n= 16）苗勒管细胞在16 - 12天的妊娠，但没有水解检测周围女性（n= 17）苗勒管细胞在任何阶段。不含底物孵育的对照（n= 24）未染色。添加外源性ATP（n= 20）的组织化学孵育介质抑制雄性苗勒管核苷酸水解，这表明这种染色是特异性的焦磷酸酶活性。在体内的结果证实了在体外孵育14 - 1 - 2天的雌性大鼠泌尿生殖嵴与MIS为72小时之前，组织化学染色。在MIS中加入睾酮是体外检测染色的必要条件（n= 10）。局部NPPase活性周围的回归苗勒管表明，NPPase可能会出现作为一个结果的管道回归，并可能会采取行动，以控制膜磷酸化的程度，通过降解过量的三核苷酸。
Mullerian inhibiting substance (MIS), a glycoprotein from the fetal testis causing regression of the embryonic Mullerian duct, can be inhibited in vitro in the presence of Mn 2+ by a wide range of nucleotides including GTP, NAD, ATP, AMP, and several nonhydrolyzable synthetic ATP analogs. Extracellular nucleotide pyrophosphatase (NPPase), an enzyme able to hydrolyze the wide variety of the nucleotides and analogs found to inhibit Mullerian duct regression, was studied by histochemical staining (H. Sierakowska and D. Shugar (1963). Biochem. Biophys. Res. Commun. 11, 70–74) to determine if NPPase localized in or around the Mullerian duct during regression. Frozen sections of urogenital ridges from 14 1 2-to 17 1 2-day rat fetuses (n= 77) were incubated with a-naphthyl thymidine-5′-phosphate (naphthyl TMP) and Fast Red TR. Nucleotide pyrophosphatase hydrolyzes naphthyl TMP, releasing naphthol, which then reacts with Fast Red to produce color at the enzyme site. Nucleotide hydrolysis was detected around regressing male (n= 16) Mullerian duct cells at 16 1 2 days of gestation, but no hydrolysis was detected around female (n= 17) Mullerian duct cells at any stage. Controls (n= 24) incubated without substrate did not stain. Addition of exogenous ATP (n= 20) to the histochemical incubation medium inhibited nucleotide hydrolysis on male Mullerian ducts, suggesting that this staining is specific for pyrophosphatase activity. Results in vivo were confirmed in vitro by incubating 14 1 2 day female rat urogenital ridges with MIS for 72 hr prior to histochemical staining. The addition of testosterone to MIS was obligatory to detect staining in vitro (n= 10). The localized NPPase activity around the regressing Mullerian duct suggests that NPPase may appear as a consequence of duct regression and may act to control the degree of membrane phosphorylation by degrading excess trinucleotides.