Tokunaga Kosuke, Akimoto Shota, Aiba Machiko, Nakagomi Mutsuki, Suzuki Takahiro, Tatsumi Ryuichi, Nakamura Mako
Muscle and Meat Science Laboratory, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University.
School of Interdisciplinary Science and Innovation, Kyushu University.
J Poult Sci. 2022 Oct 25;59(4):338-347. doi: 10.2141/jpsa.0220003.
Smooth muscle cells are widely distributed in the digestive organs of chickens. They exist as single cells, but adhere to each other to function synchronously. In this study, the expression of the gap junction protein connexin 43 (Cx43) in chicken gizzards was investigated at embryonic days (E) 10, E15, and E18. Gizzards have an abundance of smooth muscle cells because of their thick muscle layers, which enable easy analysis of the cells. Morphological observations and expression patterns of smooth muscle markers were confirmed. Next, we observed where the markers were localized in the gizzard tissue at E10, E15, and E18. Finally, the expression pattern of Cx43 in primary cultured smooth muscle cells from E15 gizzards was investigated. The analysis revealed the expression and localization of Cx43 and calponin 1 in the smooth muscle layers, and 3D analysis revealed dynamic changes in the localization pattern of Cx43 from E10 to E15. Cultured smooth muscle cells confirmed that Cx43 was expressed in the cell membrane and cytosol. In conclusion, Cx43 expression was identified in chicken gizzards at E10, E15, and E18, which was localized differently during development. The expression was broad at E10, and became restricted at E15 and E18. Primary culture of smooth muscle cells showed that Cx43 was present in the cell membrane and cytosol. This suggests that Cx43 is actively translated into the cytosol at E10, forming a hexamer, and shuttling the cell membrane to function as a gap junction.
平滑肌细胞广泛分布于鸡的消化器官中。它们以单个细胞形式存在，但彼此黏附以同步发挥功能。在本研究中，对鸡砂囊在胚胎第10天（E10）、第15天（E15）和第18天（E18）时缝隙连接蛋白连接蛋白43（Cx43）的表达进行了研究。砂囊因其肌肉层厚而含有大量平滑肌细胞，这便于对细胞进行分析。证实了平滑肌标志物的形态学观察和表达模式。接下来，我们观察了这些标志物在E10、E15和E18时在砂囊组织中的定位。最后，研究了来自E15砂囊的原代培养平滑肌细胞中Cx43的表达模式。分析揭示了Cx43和钙调蛋白1在平滑肌层中的表达和定位，三维分析揭示了从E10到E15时Cx43定位模式的动态变化。培养的平滑肌细胞证实Cx43在细胞膜和细胞质中表达。总之，在E10、E15和E18的鸡砂囊中均鉴定出Cx43的表达，其在发育过程中的定位有所不同。E10时表达广泛，而在E15和E18时受到限制。平滑肌细胞的原代培养表明Cx43存在于细胞膜和细胞质中。这表明Cx43在E10时被主动翻译到细胞质中，形成六聚体，并穿梭到细胞膜以发挥缝隙连接的功能。
