Depolarization regulates adrenal preproenkephalin mRNA.
Depolarization regulates adrenal preproenkephalin mRNA.
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去极化调节肾上腺前脑啡肽原 mRNA。
DOI：
10.1073/pnas.82.23.8252
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发表时间：
1985
影响因子：
11.1
通讯作者：
Black,IB
中科院分区：
文献类型：
--
作者：
LaGamma,EF;White,JD;Adler,JE;Krause,JE;McKelvy,JF;Black,IB
用大鼠肾上腺髓质组织块培养法研究了神经肽基因表达的调控。培养3天后，（现在去神经）外植体表现出10倍的亮氨酸脑啡肽（leu-EK）含量增加。用放线菌酮抑制蛋白质合成完全阻断了上升，而用放线菌素D或α-鹅膏蕈碱抑制RNA合成抑制了50%的增加。用1-β-D-阿拉伯呋喃糖基胞嘧啶（阿糖胞苷）抑制DNA合成没有明显效果。为了确定leu-EK的增加是否与编码阿片肽前体的特定mRNA的增加有关，进行了印迹杂交分析。在不同的培养时间后检测到单一种类的前脑啡肽原mRNA。mRNA的量在培养2天后增加34倍，4天后增加74倍。因此，mRNA水平的上升先于leu-EK量的增加。肾上腺髓质的去极化，无论是升高钾或去甲肾上腺素，这可以防止亮氨酸-EK积累，抑制前脑啡肽原mRNA的量的增加。此外，藜芦定的作用被河豚毒素阻断，表明跨膜钠离子内流影响了信息量的增加。我们的研究表明，海拔的leu-EK在deleted（失神经支配）髓质与增加量的mRNA编码的肽前体，这些过程可以调节去极化。
The regulation of neuropeptide gene expression has been investigated by using rat adrenal medullae grown in explant culture. After 3 days in culture the (now denervated) explants exhibited a 10-fold increase in leucine-enkephalin (leu-EK) content. Inhibition of protein synthesis with cycloheximide completely blocked the rise, whereas inhibition of RNA synthesis with actinomycin D or alpha-amanitin inhibited the increase by 50%. Inhibition of DNA synthesis with 1-beta-D-arabinofuranosylcytosine (cytosine arabinoside) had no discernible effect. To determine whether the rise in leu-EK was associated with an increase in specific mRNA coding for the opiate peptide precursor, blot hybridization analysis was performed. A single species of preproenkephalin mRNA was detected after various culture periods. The amount of mRNA increased 34-fold after 2 days in culture and 74-fold after 4 days. Consequently, the rise in mRNA levels preceded the increase in the amount of leu-EK. Depolarization of the adrenal medullae with either elevated potassium or veratridine, which prevents leu-EK accumulation, inhibited the increase in the amount of preproenkephalin mRNA. Moreover, the effect of veratridine was blocked by tetrodotoxin, suggesting that transmembrane sodium ion influx affects the increase in the amount of message. Our studies suggest that elevation of leu-EK in explanted (denervated) medullae is associated with increased amounts of mRNA coding for the peptide precursor and that these processes can be regulated by depolarization.
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