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黑腹果蝇卵子发生和早期胚胎发生过程中特定蛋白质的局部合成。

Localized synthesis of specific proteins during oogenesis and early embryogenesis inDrosophila melanogaster.

作者信息

Gutzeit Herwig O, Gehring Walter J

机构信息

Biozentrum der Universität Basel, Klingelbergstraße 70, CH-4056, Basel, Switzerland.

出版信息

Wilehm Roux Arch Dev Biol. 1979 Jun;187(2):151-165. doi: 10.1007/BF00848268.

Abstract

Protein synthesis in egg follicles and blastoderm embryos ofDrosophila melanogaster has been studied by means of two-dimensional gel electrophoresis. Up to 400 polypeptide spots have been resolved on autoradiographs. Stage 10 follicles (for stages see King, 1970) were labelled in vitro for 10 to 60 min withS-methionine and cut with tungsten needles into an anterior fragment containing the nurse cells and a posterior fragment containing the oocyte and follicle cells. The nurse cells were found to synthesize a complex pattern of proteins. At least two proteins were detected only in nurse cells but not in the oocyte even after a one hour labelling period. Nurse cells isolated from stages 9, 10 and 12 follicles were shown to synthesize stage specific patterns of proteins. Several proteins are synthesized in posterior fragments of stage 10 follicles but not in anterior fragments. These proteins are only found in follicle cells. No oocyte specific proteins have been detected. Striking differences between the protein patterns of anterior and posterior fragments persist until the nurse cells degenerate. In mature stage 14 follicles, labelled in vivo, no significant differences in the protein patterns of isolated anterior and posterior fragments could be detected; this may be due to technical limitations. At the blastoderm stage localized synthesis of specific proteins becomes detectable again. When blastoderm embryos, labelled in vivo, are cut with tungsten needles and the cells are isolated from anterior and posterior halves, differences become apparent. The pole cells located at the posterior pole are highly active in protein synthesis and contribute several specific proteins which are found exclusively in the posterior region of the embryo. In this study synthesis of specific proteins could only be demonstrated at those developmental stages which are characterized by the presence of different cell types within the egg chamber, while no differences were detected when stage 14 follicles were cut and anterior and posterior fragments analyzed separately. The differences in the pattern of protein synthesis by pole cells and blastoderm cells indicate that even the earliest stages of determination are reflected by marked changes at the biochemical level.

摘要

利用二维凝胶电泳技术对黑腹果蝇卵泡和胚盘胚胎中的蛋白质合成进行了研究。在放射自显影片上可分辨出多达400个多肽斑点。用³⁵S-甲硫氨酸对10期卵泡(关于各阶段,见King,1970)进行体外标记10至60分钟,然后用钨针切成一个包含滋养细胞的前部片段和一个包含卵母细胞及卵泡细胞的后部片段。发现滋养细胞合成的蛋白质模式复杂。即使经过一小时的标记期,至少有两种蛋白质仅在滋养细胞中检测到,而在卵母细胞中未检测到。从9期、10期和12期卵泡中分离出的滋养细胞显示出合成阶段特异性的蛋白质模式。10期卵泡的后部片段中合成了几种蛋白质,而前部片段中未合成。这些蛋白质仅在卵泡细胞中发现。未检测到卵母细胞特异性蛋白质。前部和后部片段的蛋白质模式之间的显著差异一直持续到滋养细胞退化。在体内标记的成熟14期卵泡中,未检测到分离的前部和后部片段的蛋白质模式有显著差异;这可能是由于技术限制。在胚盘阶段,又可检测到特定蛋白质的局部合成。当用钨针切割体内标记的胚盘胚胎并从前后两半分离细胞时,差异变得明显。位于后极的极细胞在蛋白质合成方面高度活跃,并贡献了几种仅在胚胎后部区域发现的特定蛋白质。在这项研究中,仅在卵室内存在不同细胞类型的那些发育阶段证明了特定蛋白质的合成,而当切割14期卵泡并分别分析前部和后部片段时未检测到差异。极细胞和胚盘细胞蛋白质合成模式的差异表明,即使是最早的决定阶段也会在生化水平上表现出明显变化。

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