Molecular anatomy of functional structural expression of gap junctions.

间隙连接功能结构表达的分子解剖学。

• 批准号: 07407001
• 负责人: SHIBATA Yosaburo
• 金额: $ 20.42万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07407001/
• 关键词: gap junction; connexin; Cx43; Cx45; Cx40; Cx37; phosphorylation; intestinal pacemaker; 小腸ペースメーカー; 血管内皮; 小腸; コネキシン43; コネキシン45; コネキシン40; エナメル形成; 卵胞

Functional structures of gap junctions were analyzed by immunohistochemical, biochemical and molecular biological methods to examine the molecular diversity and distribution patterns of connexin protein family in various tissue and cell types as well as in different functional states.1) During ameloblast development of the rat upper incisor, connexin 43 in undifferentiated cells once disappears just before enamel formation and reappears after maturation of secretion activity.2) In granulosa cells of developing rat ovary, gap junctions are composed of Cx43 and Cx45 in various phosphorylation states. While Cx45 remains constant, Cx43 increases during maturations but disappears soon after ovulation. In addition, Cx40 detected in RNA blot in ovary is localized only in blood vessel wall.3) In rat mammary glands, Cx26 and Cx32 are colocalized between secretory acinar cells and Cx43 is localized between myoepithels. They gradually increase during pregnancy. Cx32 and Cx43 are induced dramatica … More lly just after delivery, and reach a maximum at the early lactation period. There seems to bea gap between mRNA and protein expression levels as well as the additional shift from P1 to P2 phosphorylation state of Cx43.4) In rat endocardium and blood vessels, endothelial cells can be classified as Cx40 dominant or Cx37 dominant type according to their connexin expression patterns.5) In rat trachea Cx37 is expressed in smooth muscle cells after 3weeks after birth and increases thereafter and colocalized with Cx43, but not with Cx40.6) In guinea pig, Cx43 is localized between fibroblast like cells in the outer layr of circular smooth muscles and deep muscular plexus of the small intestine and between submuscular plexus in the large intestine. These Cx43 containing cells are also c-Kit positive and interconnect the interstitial cells with intestinal pacemaker activity.This research project has successfully elucidated the diverse aspects of gap junction functions in the light of composite protein connexin family with specific physiological properties ; Gap junction protein expressions are characteristically regulated during developmental stages, in different functional states and between different tissues. They are controlled by various mechanisms such as expression levels or phosphorylation states. Finally, specific connexin molecules are identified in specialized functional cells such as pacemaker cells in heart and intestinal movement. Less

采用免疫组织化学、生物化学和分子生物学方法分析缝隙连接的功能结构，研究缝隙连接蛋白家族在不同组织、细胞类型和不同功能状态下的分子多样性和分布规律。1）大鼠上切牙成釉细胞发育过程中，未分化细胞中的连接蛋白43在釉质形成前消失，并在分泌活性成熟后重新出现。2）在发育中的大鼠卵巢的颗粒细胞中，缝隙连接由处于各种磷酸化状态的Cx43和Cx45组成。虽然Cx45保持不变，但Cx43在成熟期间增加，但在排卵后不久消失。在大鼠乳腺中，Cx26和Cx32共定位于腺泡细胞之间，Cx43定位于肌上皮细胞之间。在怀孕期间逐渐增加。Cx32和Cx43被戏剧性地诱导 ...更多信息 分娩后不久，泌乳早期达到最大值。在mRNA和蛋白质表达水平之间似乎存在差距，以及Cx43从P1磷酸化状态到P2磷酸化状态的额外转变。4）在大鼠内皮细胞和血管中，根据其连接蛋白表达模式，内皮细胞可分为Cx40显性或Cx37显性类型。在大鼠气管中，Cx37在出生后3周后在平滑肌细胞中表达，此后增加，并与Cx43共定位，但不与Cx40共定位。6）在豚鼠中，Cx43定位于小肠环状平滑肌外层和深肌丛的成纤维样细胞之间，大肠肌下丛之间。这些含Cx43的细胞也是c-Kit阳性的，并且将间质细胞与肠起搏器活动连接起来。本研究项目成功地阐明了缝隙连接蛋白家族具有特定生理特性的复合蛋白功能的多个方面;缝隙连接蛋白在发育阶段、不同功能状态和不同组织之间的表达具有特征性调控。它们受多种机制控制，如表达水平或磷酸化状态。最后，在专门的功能细胞如心脏和肠道运动中的起搏细胞中鉴定出特定的连接蛋白分子。少

项目成果

A,Kuraoka, T.Inai, K.Nishii, I.Yamanaka: "S.Y.Characterization of isolated guinea pig liver gap junctions. (Eds. Y Kanno, Shimadzu, Y Shibata), Elsevier, Amsterdam" 4. 111-114 (1995)

A、Kuraoka、T.Inai、K.Nishii、I.Yamanaka：“S.Y. 分离豚鼠肝脏间隙连接的表征。（Eds. Y Kanno、Shimadzu、Y Shibata），Elsevier，阿姆斯特丹”4. 111-114 (1995)

K.Manabe, H.Ito, H.Matsuda, A.Noma, Y.Shibata.: "Classification of ion channels in the luminal and abluminal membranes of guinea-pig endocardial endothelial cells" J.Physiol. 484. 41-52 (1995)

K.Manabe、H.Ito、H.Matsuda、A.Noma、Y.Shibata.：“豚鼠心内膜内皮细胞腔内和近腔膜中离子通道的分类”J.Physiol。

K.Nakamura et al.: "Specific localization of gap junction protein,connexin45,in the deep mus-cular plexus of dog and rat small intestine." Cell Tissue Res.(in press). (1998)

K.Nakamura 等人：“间隙连接蛋白 connexin45 在狗和大鼠小肠深肌丛中的特异性定位。”

A.Kuraoka et al.: "Characterization of isolated guinea pig liver gap junctions." Prog.Cell Res. 4. 111-114 (1995)

A.Kuraoka 等人：“离体豚鼠肝脏间隙连接的表征。”

K.Manabe et al.: "Classification of ion channels in the luminal and abluminal membranes of guinea-pig endocardial endothelial cells." J.Physiol.484. 41-52 (1995)

K.Manabe 等人：“豚鼠心内膜内皮细胞腔内膜和近腔膜中离子通道的分类。”