【摘要】：目的探討小鼠胚胎心流出道分隔過程中,前腸呼吸內胚層與咽前第二生心區(qū)細胞發(fā)育的形態(tài)學關系及機制。方法胚齡9~13d小鼠胚胎標本各6例,連續(xù)石蠟切片,用抗轉錄因子叉頭框蛋白A2(Foxa2)、抗胰島因子1(ISL-1)、抗patched1(Ptc1)、抗patched 2(Ptc2)、抗α-平滑肌肌動蛋白(α-SMA)及抗心肌肌球蛋白重鏈(MHC)抗體進行免疫組織化學及免疫熒光染色。結果胚齡9~9.5d,前腸腹側壁ISL-1陽性內胚層局部增厚,呼吸內胚層開始發(fā)育,ISL-1陽性間充質細胞緊隨其后開始出現在呼吸內胚層周圍的基質中。胚齡10~11.5d,呼吸內胚層向動脈囊方向生長延伸向喉-氣管溝演變,ISL-1陽性咽前間充質細胞圍繞呼吸內胚層呈對稱的特征性錐體形結構分布,錐體頂端突入動脈囊腔向主-肺動脈隔發(fā)育。在喉-氣管溝發(fā)育過程中,總能觀察到1條實心內胚層細胞索位于其腹側頂端,Ptc1和Ptc2主要局限于發(fā)育中的喉-氣管溝及實心細胞索表達,喉-氣管溝及實心細胞索的內胚層則位于錐體結構的中心。胚齡12~13d,在流出道水平前腸分隔形成氣管,內胚層細胞索逐漸消失,氣管上皮逐漸失去Ptc1和Ptc2表達,氣管腹側的ISL-1陽性間充質細胞密度明顯減低,并逐漸停止向流出道添加,動脈囊分隔完成。結論呼吸內胚層的分化發(fā)育與咽前ISL-1陽性第二生心區(qū)細胞的發(fā)育聚集密切耦聯。音猬因子(SHH)信號系統(tǒng)在呼吸內胚層發(fā)育過程中活躍程度較高,發(fā)育中的呼吸內胚層可能作為組織中心,通過SHH信號通路誘導ISL-1陽性細胞的聚集,并通過內胚層生長延伸造成的機械牽拉力驅動ISL-1陽性細胞遷移,參與流出道正常形態(tài)發(fā)生。
[Abstract]:Aim to investigate the morphological relationship and mechanism between the endoderm of pregut respiration and the cell development of prepharyngeal second heart during the separation of cardiac outflow tract in mouse embryos. Methods six embryonic specimens of 9-day-old 13-day-old mice were randomly divided into two groups: paraffin section, anti-Foxa2, anti-islet factor-1 (ISL-1), anti-patched1 (Ptc1) and anti-patched-2 (Ptc2). Anti-偽-smooth muscle actin (偽-SMA) and anti-myosin heavy chain (MHC) antibodies were stained by immunohistochemistry and immunofluorescence. Results ISL-1-positive endoderm was thickened in ventral wall of anterior intestine at 9-9. 5 days, respiratory endoderm began to develop, and ISL-1-positive mesenchymal cells began to appear in the matrix around respiratory endoderm. At the age of 10 to 11.5 days, the respiratory endoderm grew and extended toward the larynx-trachea sulcus, and the ISL-1-positive prepharyngeal mesenchymal cells showed symmetrical, characteristic cone-shaped distribution around the respiratory endoderm. The capsular cavity of the artery protruding into the apex of the pyramidal developed into the aortic pulmonary septum. During the development of larynx-trachea sulcus, a solid endoderm cell cord was always observed at its ventral tip. The expression of Ptc1 and Ptc2 was mainly confined to the developing laryngeal-tracheal sulcus and the solid cell cord during the development of larynx-tracheobronchial sulcus. The endoderm of larynx-trachea sulcus and solid cell cord is located in the center of pyramidal structure. At the age of 12 ~ 13 days, trachea was separated from the anterior intestine at the outflow tract level, the endoderm cell cord disappeared gradually, the expression of Ptc1 and Ptc2 was gradually lost in the trachea epithelium, and the density of ISL-1 positive mesenchymal cells in the ventral trachea was significantly decreased. And gradually stop adding to the outflow tract, arterial sac separation is complete. Conclusion the differentiation and development of respiratory endoderm are closely coupled with the development and aggregation of prepharyngeal ISL-1-positive secondary heart cells. The phonic hedgehog factor (SHH) signaling system is highly active in the development of respiratory endoderm. The developing respiratory endoderm may act as a tissue center and induce the aggregation of ISL-1 positive cells through SHH signaling pathway. The migration of ISL-1-positive cells was driven by mechanical pulling force caused by endoderm growth and extension, which was involved in the normal morphogenesis of outflow tract.
【作者單位】： 山西醫(yī)科大學組織學與胚胎學教研室;長春醫(yī)學高等專科學校解剖學教研室;山西醫(yī)學科學院神經內科;
【基金】：國家自然科學基金(30771141,31200899) 山西省自然科學基金(2015021189) 山西省優(yōu)勢重點學科建設經費(轉化醫(yī)學) 山西醫(yī)科大學博士啟動基金(03201410);山西醫(yī)科大學331基礎醫(yī)學科技培植基金(201423)
【分類號】：R321.5

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