HYPOXANTHINE IS THE PRINCIPAL INHIBITOR OF MURINE OOCYTE MATURATION IN A LOW-MOLECULAR WEIGHT FRACTION OF PORCINE FOLLICULAR-FLUID
HYPOXANTHINE IS THE PRINCIPAL INHIBITOR OF MURINE OOCYTE MATURATION IN A LOW-MOLECULAR WEIGHT FRACTION OF PORCINE FOLLICULAR-FLUID
复制标题
DOI：
10.1073/pnas.82.2.454
复制
发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
EPPIG, JJ
中科院分区：
文献类型：
--
作者：
DOWNS, SM;COLEMAN, DL;EPPIG, JJ
研究人员进行了鉴定和定量猪卵泡液 (PFF) 成分的研究，该成分负责抑制小鼠卵母细胞成熟。通过透析制备 PFF 的低 MW 级分 (< 1000) 并用于所有实验。该 PFF 级分含有小鼠卵母细胞成熟抑制剂，在 250-260 nm 的紫外线范围内吸收最大。当 PFF 级分被木炭提取时，导致 250、260 和 280 nm 处吸光度显着损失，这对应于抑制活性的损失。通过离子交换色谱法分离 PFF 的四种主要成分，并根据其紫外光谱特征和抑制活性进行表征。当汇集具有相同光谱的各个级分并通过高效液相色谱进行分析时，第一个汇集的级分 (A) 是不纯的，但腺嘌呤包含 80% 的 UV 吸收材料。级分 B、C 和 D 分别被表征为纯尿嘧啶、次黄嘌呤和 7-甲基肌苷。 PFF 中这些化合物的浓度估计为 0.06 mM 腺嘌呤、0.44 mM 尿嘧啶、1.41 mM 次黄嘌呤和 0.19 mM 7-甲基肌苷。对这些化合物的商业制剂的效力进行比较，确定次黄嘌呤是低分子量 PFF 组分的主要抑制成分。次黄嘌呤的商业制剂模仿了 PFF 对小鼠卵母细胞成熟的作用，它产生了对卵母细胞成熟的短暂抑制，卵泡刺激激素和二丁酰环腺苷单磷酸显着增强了这种抑制作用。当比较嘌呤和嘧啶碱基与核苷的抑制效力时，它们的相对效力降序为嘌呤碱基>嘌呤核苷>嘧啶碱基=嘧啶核苷。次黄嘌呤是 PFF 中主要的低分子量成分，可抑制小鼠卵母细胞成熟，但其他嘌呤/嘧啶也可能在体内维持减数分裂停滞中发挥作用。
Studies were carried out to identify and quantify the porcine follicular fluid (PFF) component(s) responsible for inhibition of murine oocyte maturation. A low MW fraction of PFF (< 1000) was prepared by dialysis and used in all experiments. This PFF fraction contained an inhibitor(s) of mouse oocyte maturation that absorbed maximally in the UV range at 250-260 nm. When the PFF fraction was charcoal-extracted, significant loss of absorbance at 250, 260 and 280 nm resulted, which corresponded to loss of inhibitory activity. Four major components of PFF were separated by ion-exchange chromatography and characterized according to their UV spectral characteristics and inhibitory activity. When individual fractions demonstrating identical spectra were pooled and analyzed by high performance liquid chromatography, the 1st pooled fraction (A) was impure, but adenine comprised 80% of the UV-absorbing material. Fractions B, C, and D were characterized as pure uracil, hypoxanthine and 7-methylinosine, respectively. The concentrations of these compounds in PFF were estimated to be 0.06 mM adenine, 0.44 mM uracil, 1.41 mM hypoxanthine and 0.19 mM 7-methylinosine. Comparison of the potencies of commercial preparations of these compounds established that hypoxanthine is the major inhibitory component of the low MW PFF fraction. A commerical preparation of hypoxanthine mimicked the action of PFF on mouse oocyte maturation in that it produced a transient inhibition of oocyte maturation that was significantly potentiated by follicle-stimulationg hormone and dibutyryl cyclic adenosine monophosphate. When the inhibitory efficacies of purine and pyrimidine bases and nucleosides were compared, their relative potencies in decreasing order were purine bases > purine nucleosides > pyrimidine bases = pyrimidine nucleosides. Hypoxanthine is the predominant low MW component of PFF that inhibits mouse oocyte maturation but that other purines/pyrimidines may also play a role in vivo in maintaining meiotic arrest.