Expression pattern and signification of Cx43, beta-catenin and Smo in the second heart field

doi:10.12307/2024.143

Abstract

Abstract: BACKGROUND: The second heart field is crucial for the development of the embryonic heart. Abnormal development of the second heart field can result in multiple cardiac malformations. After Cx43 gene knockout, reduced formation and proliferation of cells of the second heart field can be observed, but the specific reason remains unclear.
OBJECTIVE: (1) To determine whether β-catenin, Smo and Cx43 were co-expressed in the second heart field and the endoderm, we observed the expression patterns of these proteins. (2) To explore whether Cx43 interacts with the Wnt/β-catenin pathway or the Shh pathway in the development of the second heart field.
METHODS: Serial paraffin sections of the mouse embryos at embryonic days 10-12 were selected for immunohistochemical staining, hematoxylin-eosin staining and immunofluorescence staining. The primitive gut of mouse embryos at embryonic day 11 was separated for western blot assay and co-immunoprecipitation.
RESULTS AND CONCLUSION: (1) Cx43 and Isl1 were co-expressed in some mesenchymal cells on the ventral side of the foregut and dorsal wall of the pericardial cavity of mouse embryos at embryonic days 10-12; Isl1 positive cells increased while Cx43 positive cells increased. (2) Cx43 and β-catenin were co-expressed in the ventral part of the endoderm at embryonic days 10-12. (3) Cx43 and Smo were co-expressed in the endoderm at embryonic days 10-12. (4) The co-immunoprecipitation results confirmed that there was an interaction between Cx43 and β-catenin, which suggested that Cx43 interacted with β-catenin to participate in the development of the second heart field.

Key words: mouse embryo, second heart field, outflow tract, Wnt/β-catenin pathway, Shh pathway, Cx43, co-immunoprecipitation

CLC Number:

Ding Zeyuan, Yan Yunan, Xie Jianshan, Shi Liang, Jing Ya, Yang Yanping. Expression pattern and signification of Cx43, beta-catenin and Smo in the second heart field[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(19): 3042-3048.

Figures/Tables 6

2.1 Cx43、Wnt/β-catenin通路与Shh通路组分的表达胚龄10 d，心管已成襻，从动脉端至静脉端，依次可见动脉囊、流出道、心室、心房与静脉窦。心管背侧的前肠壁由内胚层细胞构成。前肠的头端即原始咽横断面呈扁椭圆形，其背侧壁细胞呈单层扁平状，而腹侧的细胞为单层立方或柱状，尤其在腹侧正中，细胞形态由单层变为多层，呈Isl1阳性，局部基膜消失，内胚层腹侧可观察到少量散在分布的间充质细胞，见图1A-C，部分细胞呈Isl1阳性，见图1D。向心脏静脉端移行，前肠横断面呈管状，其腹侧部内胚层细胞表达Isl1，内胚层的腹侧Isl1阳性间充质细胞数量增加，见图1E。此外，Isl1表达也见于鳃弓核心间充质、心包腔背侧壁，见图1F，说明此期第二生心区由心包腔背侧壁脏壁中胚层、鳃弓核心间充质、前肠腹侧间充质细胞构成。β-catenin在前肠内胚层广泛表达，见图1G，H，其下游信号Lef1在前肠内胚层细胞染色呈阴性。Shh信号通路的受体Smo在咽内胚层的表达局限于两侧，见图1I。随着向前肠尾端移行，在前肠腹侧Isl1阳性间充质细胞丰富的区域，相邻的内胚层呈现明显的Smo表达，见图1J。Cx43在前肠内胚层的表达模式与Smo相似，在前肠头端的内胚层表达较弱，见图1K，至前肠尾端，Cx43在内胚层的表达变得明显，见图1L。在心管动脉端，心包腔背侧壁脏壁中胚层的Isl1表达延续至与之相连的流出道远端壁，MHC在Isl1阳性的流出道远端壁呈弱阳性表达，见图1C，M。在心管静脉端，Isl1阳性间充质细胞延续至心背系膜及与之相连的心房壁，见图1E，MHC在心房壁的Isl1阳性区尚未表达，见图1E，N。说明Isl1阳性细胞向心管动脉端和静脉端添加，并分化为心肌细胞。"

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