Purification, measurement, and tissue distribution of a dansyl-derivatized glycopeptide from low-molecular weight follicle-stimulating hormone-inhibitor-containing fractions of porcine follicular fluid.
Purification, measurement, and tissue distribution of a dansyl-derivatized glycopeptide from low-molecular weight follicle-stimulating hormone-inhibitor-containing fractions of porcine follicular fluid.
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从猪卵泡液中含有低分子量卵泡刺激激素抑制剂的级分中纯化、测量和组织分布丹磺酰衍生糖肽。
DOI：
10.1095/biolreprod40.2.407
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发表时间：
1989
影响因子：
3.6
通讯作者：
ReichertJr,LE
中科院分区：
文献类型：
--
作者：
Sluss,PM;Branca,AA;Ford,JJ;Krishnan,KA;ReichertJr,LE
我们用丹磺酰氯(5-二甲氨基-1-萘磺酰氯)在卵泡液或含有与受体结合的低分子量促卵泡激素(FSH)抑制剂(FSH-BI)的高纯度组分中形成含胺化合物的丹磺酰衍生物。这种方法可以根据衍生物的荧光性质对其进行灵敏的检测。利用Dansyl基团的疏水性，从猪卵泡液中纯化了低分子FSH-BI制剂中鉴定的Dansyl衍生物(Rf=0.15)。根据四种不同系统(薄层层析[TLC]和高效液相色谱)的层析标准，纯化的衍生化因子(D15)似乎是均一的。建立了一种从卵泡液或组织提取液中直接测定D15的化学方法。D15在成年大鼠卵巢组织中含量最高(153 ng/mg)，在肾和肝组织中含量较低(分别为93和62 ng/mg)，在横隔膜和心脏组织中含量更低(分别为5和0.5 ng/mg)。在未成熟大鼠的衍生提取物中观察到高浓度的D15，其中在间质细胞中D15的含量(241 ng/mg)大约是生精小管中D15含量(136 ng/mg)的两倍。在猪卵巢组织中，来自大卵泡和黄体的颗粒细胞(分别为69和91 ng/mg)的含量至少是卵泡壁组织(14 ng/ml)的4倍。在先前显示含有低分子FSH-BI的牛卵泡液中，也测定了D15物质的相对浓度。发情周期黄体期奶牛小卵泡液中D15的浓度最高(15.6微克/毫升)。这些卵泡还含有最高浓度的低分子FSH-BI。从妊娠奶牛的大小卵泡液或囊性卵泡液中未检测到D15。这些数据表明，低分子量FSH-BI可以在丹糖化后用TLC进行定量，并提示性腺组织中存在高局部浓度，特别是黄体生成素反应细胞类型。在这些数据的基础上，该因子是FSH作用的局部调节剂(拮抗剂)的假设值得进一步研究。
We have used dansyl chloride (5-dimethylamino-1-naphthalenesulfonyl choloride) to form dansyl derivatives of amine-containing compounds in follicular fluid or highly purified fractions containing a low molecular weight (MW) inhibitor of follicle-stimulating hormone (FSH) binding to receptor (FSH-BI). This approach allowed sensitive detection of the derivatives based on their fluorescent properties. By taking advantage of the hydrophobic nature of the dansyl group, a dansyl derivative (RF= 0.15) identified in low MW FSH-BI preparations was purified from porcine follicular fluid. Based on chromatographic criteria using four different systems (thin-layer chromatography [TLC] and high performance liquid chromatography), the derivatized factor (D15) that was purified appeared to be homogeneous. A direct, chemical assay was developed for quantification of D15 from follicular fluid or tissue extracts. The highest concentration (153 ng/mg) of D15 was found in ovarian tissue of adult rats, lesser amounts were observed in kidney and liver tissues (93 and 62 ng/mg, respectively) and even less in diaphram and heart tissues (5 and 0.5 ng/mg, respectively). High concentrations of D15 were observed in derivatized extracts of tests from immature rats in which approximately twice as much D15 was found in Leydig cells (241 ng/mg) as in seminiferous tubules (136 ng/mg). In porcine ovarian tissue, granulosa cells from large follicles and corpora lutea (69 and 91 ng/mg, respectively) contained at least 4-fold higher concentrations than follicle wall tissue (14 ng/ml). Relative concentrations of D15 material were also determined in pools of bovine follicular fluid previously shown to contain low MW FSH-BI. The concentration of D15 was highest (15.6 µg/ml) in fluid from small follicles obtained from cows during the luteal phase of the estrous cycle. These follicles also contained the highest concentration of low MW FSH-BI. No D15 was detected in fluid from large or small follicles from pregnant cows or from cystic follicles. These data indicate that low MW FSH-BI can be quantified by TLC after dansylation and suggest that high local concentrations exist in gonadal tissues, particularly luteinizing hormone-responsive cell types. On the basis of these data, the hypothesis that this factor is a local modulator (antagonist) of FSH action deserves further study.