Application of Recapitulation Theory to Secretion of Atrial Natriuretic Peptide

重演理论在心房钠尿肽分泌中的应用

基本信息

批准号：
02640584
负责人：
TAKEI Yoshio
金额：
$ 1.15万
依托单位：
Kitasato University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02640584/
关键词：
trial natriuretic peptide Eel Recapitulation theory Secretion Tissue culture Pulse-chase experiment Northern blotting Protein A-gold immunohistochemistry 組織培養法 cDNA

项目摘要

1. We isolated atrial natriuretic peptide(ANP)and ventricular natriuretic peptide(VNP)from the eel atrium and ventricle, respectively and determined their amino acid seguences. We also determined cDNA sequence of eel VNP.2. We established specific radioimm unoassay systems for eel ANP and VNP, and measured the levels in eel plasma, atria, ventricles and brains. The heart contains 3-fold greater level of ANP than that of VNP, but the plasma level of VNP was 3-foldgreater than that of ANP.3. Protein A-goldimmunocytochemistry revealed many secretory granules which contain ANP, but VNP-containing. . granules in the ventricle was scarce.4. Eel atrial myocytes in culture secrete a small amount of ANP into the medium but the amount stored in the cell is large. Eelventricular myocytes in culture secrete a large amount of VNP but store little VNP.5. Based on these results, we hypothesized that ANP is principally synthesized in the atrium, stored in the granules, and secreted upon exogenous stimulus(regulatory pathway), whereas most of the VNP synthesezed in the ventricle is secreted soon without storage in the granules (constitutive pathway).6. In order to substantiate this hypotheses, we are now conducting a pulse-chase experiment using labeled ^<32>S-Cysteine to examine the rate of secreion of ANP and VNP from eel atrium and ventricle in tissue culture. We also examine the expression of mRNA of ANP and VNP in the eel atrium and ventricle using a Northern blot analysis.7. In the fetal rat, ANP is secreted from the atrium and ventricle viaregulatory pathway and constitutive pathway, respectively, although adult ventricles do not produce ANP in mammals. Thus, the data from eels and fetal rats illustrate that the Biogenic law of Haeckel, i. e., "Ontogeny is a recapitulation of phylogeny" appears to beapplicable to the mode of hormone secretion.

1.我们分别从鳗鱼心房和心室中分离到心房钠尿肽（ANP）和心室钠尿肽（VNP），并测定了它们的氨基酸序列。我们还测定了鳗鱼VNP.2的cDNA序列。我们建立了鳗鱼ANP和VNP的特异性放射免疫分析系统，并测量了鳗鱼血浆、心房、心室和大脑中的水平。心脏中ANP的含量比VNP高3倍，但VNP的血浆水平比ANP高3倍。3。蛋白A-金免疫细胞化学显示许多分泌颗粒含有ANP，但也含有VNP。。心室颗粒稀少。4．培养中的鳗鱼心房肌细胞向培养基中分泌少量ANP，但细胞中储存的量很大。培养的脑室肌细胞分泌大量VNP，但储存少量VNP。5。基于这些结果，我们假设ANP主要在心房合成，储存在颗粒中，并在外源刺激下分泌（调节途径），而大部分在心室合成的VNP很快就会分泌，而不储存在颗粒中（组成性途径）。途径）6。为了证实这一假设，我们现在正在进行脉冲追踪实验，使用标记的^ 32 S-半胱氨酸来检查组织培养中鳗鱼心房和心室的ANP和VNP的分泌率。我们还使用Northern印迹分析检测了鳗鱼心房和心室中ANP和VNP mRNA的表达。7．在胎鼠中，ANP 分别通过调节途径和组成途径从心房和心室分泌，但在哺乳动物中成年心室不产生 ANP。因此，来自鳗鱼和胎鼠的数据说明了海克尔的生物起源定律，即。例如，“个体发育是系统发育的再现”似乎适用于激素分泌的模式。