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。我们分别从鳗中心房和心室分离出心房纳地尿肽（ANP）和心室亚催化肽（VNP），并确定其氨基酸seguence。我们还确定了EEL VNP.2的cDNA序列。我们为EEL ANP和VNP建立了特定的放射膜系统，并测量了EEL血浆，心房，心室和大脑的水平。心脏的ANP水平比VNP高3倍，但是VNP的血浆水平比ANP.3的血浆水平高3倍。蛋白质A-核细胞化学揭示了许多分泌颗粒，其中包含ANP但含有VNP。。心室中的颗粒很少。4。培养中的鳗鱼心肌细胞将少量的ANP分泌到培养基中，但存储在细胞中的量很大。培养中的脑室性心肌细胞分泌大量VNP，但储存小vnp.5。基于这些结果，我们假设ANP主要是在中庭中合成的，存储在颗粒中，并在外源刺激（调节途径）上分泌，而在心室中，大多数VNP合成的VNP很快就在没有存储的情况下分泌在颗粒（原始途径）。6.6 .6。为了证实这一假设，我们现在正在使用标记为 ^<32> S-半胱氨酸的标记的脉冲练习实验，以检查组织培养中鳗鱼中庭和心室中ANP和VNP的分隔率。我们还使用Northern印迹分析研究了ANP和VNP在鳗鱼中庭和心室中的mRNA的表达。7。在胎儿大鼠中，ANP分别从中庭和心室通过调节途径和组成途径分泌，尽管成年心室在哺乳动物中不产生ANP。因此，来自鳗鱼和胎儿大鼠的数据表明，海克尔的生物原则，即。例如，“个体发育是系统发育的概括”，似乎可以接受激素分泌的模式。