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生长中的小鼠卵母细胞中的核糖体RNA积累和蛋白质合成模式

rRNA accumulation and protein synthetic patterns in growing mouse oocytes.

作者信息

Kaplan G, Abreu S L, Bachvarova R

出版信息

J Exp Zool. 1982 May 1;220(3):361-70. doi: 10.1002/jez.1402200311.

Abstract

The rRNA contents of mouse primordial oocytes, three stages of growing oocytes, full-grown oocytes, and ovulated ova have been measured by hybridization of RNA samples to excess 3H-DNA complementary to rRNA. Since it was known from previous work that rRNA is stable, the results when plotted against days of oocyte growth indicated that rRNA was synthesized at a constant rate over the first 9 days of growth and about 1.5 times faster in the last 5 days. The maximum value of 0.3 ng per oocyte was attained by about 14 days of growth in oocytes 59 micrometers in diameter, well below the maximum diameter of 77 micrometers for full-grown oocytes. The stability of proteins synthesized in mid-growth phase oocytes was measured by labeling for 5 h with 35S-methionine and then following the decline of incorporated label during a 48h chase; 40% of the label decayed with a half-life of 11 h. and 60% was apparently stable. The two-dimensional electrophoretic patterns of labeled proteins synthesized by growing and full-grown oocytes were compared. The principal change was the appearance or great increase in intensity of several spots in full-grown oocytes as compared to growing oocytes. Egg proteins separated on a two-dimensional gel were visualized by silver staining. The cytoskeletal proteins actin, tubulin, and putative intermediate filament protein, as well as putative lactate dehydrogenase, were synthesized in growing and full-grown oocytes, and accumulated to form a significant portion of bulk egg protein.

摘要

通过将RNA样品与过量的与rRNA互补的3H-DNA杂交,测量了小鼠原始卵母细胞、三个生长阶段的卵母细胞、完全成熟的卵母细胞和排卵后的卵子中的rRNA含量。由于先前的研究已知rRNA是稳定的,因此将结果与卵母细胞生长天数作图表明,rRNA在生长的前9天以恒定速率合成,在最后5天合成速度快约1.5倍。直径59微米的卵母细胞在生长约14天时达到每个卵母细胞0.3纳克的最大值,远低于完全成熟卵母细胞77微米的最大直径。通过用35S-甲硫氨酸标记5小时,然后在48小时的追踪过程中跟踪掺入标记的下降情况,来测量生长中期卵母细胞中合成的蛋白质的稳定性;40%的标记以11小时的半衰期衰变,60%显然是稳定的。比较了生长中的和完全成熟的卵母细胞合成的标记蛋白质的二维电泳图谱。与生长中的卵母细胞相比,主要变化是完全成熟的卵母细胞中几个斑点的出现或强度大幅增加。二维凝胶上分离的卵蛋白通过银染可视化。细胞骨架蛋白肌动蛋白、微管蛋白和假定的中间丝蛋白,以及假定的乳酸脱氢酶,在生长中的和完全成熟的卵母细胞中合成,并积累形成大部分卵蛋白的重要组成部分。

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