Xác định các gen nhạy cảm với cơ học trong quá trình phát triển xương: sự thay đổi của các gen liên quan đến việc tái cấu trúc cytoskeleton và các con đường tín hiệu tế bào

Rebecca A. Rolfe1, Niamh C. Nowlan1, Elaine Kenny2, Paul Cormican2, Derek W. Morris2, Patrick J. Prendergast3, Daniel J. Kelly3, Paula Murphy3
1Department of Zoology, School of Natural Sciences, Trinity College Dublin, Dublin, Ireland
2TrinSeq, Institute of Molecular Medicine and Department of Psychiatry, Trinity College Dublin, Dublin, Ireland
3Trinity Centre for Bioengineering, School of Engineering, Trinity College Dublin, Dublin, Ireland

Tóm tắt

Tóm tắtĐặt vấn đềKích thích cơ học là cần thiết để điều chỉnh sự hình thành chính xác của bộ xương. Bài nghiên cứu này kiểm tra giả thuyết rằng kích thích cơ học của hệ thống xương phôi ảnh hưởng đến mức độ biểu hiện của các gen liên quan đến các con đường tín hiệu quan trọng trong sự phát triển theo cách tiếp cận toàn bộ bộ gen. Chúng tôi sử dụng mô hình chuột đột biến với kích thích cơ học thay đổi do sự thiếu vắng cơ bắp ở chi (Splotch-delayed), nơi mà các phôi không có cơ bắp cho thấy các khuyết tật cụ thể trong các yếu tố xương bao gồm sự vôi hóa chậm, sự thay đổi về kích thước và hình dạng của các mầm sụn và sự hợp nhất khớp. Chúng tôi đã sử dụng công cụ phân tích Microarray và RNA sequencing để xác định các gen biểu hiện khác biệt giữa các mô cánh tay phôi không có cơ bắp và đối chứng (TS23).Kết quảChúng tôi đã phát hiện rằng 680 gen độc lập giảm biểu hiện và 452 gen tăng biểu hiện trong cánh tay của các phôi Spd không có cơ so với các đối chứng cùng lứa (ít nhất gấp 2 lần; giá trị p điều chỉnh ≤0,05). Chúng tôi đã phân tích các bộ gen biểu hiện khác biệt này bằng cách sử dụng các chú thích Gene Ontology để xác định sự phong phú có ý nghĩa của các gen liên quan đến các quá trình sinh học cụ thể, cho thấy rằng việc loại bỏ các kích thích cơ học từ các co cơ ảnh hưởng đến các gen liên quan đến sự phát triển và phân hóa, kiến trúc cytoskeleton và tín hiệu tế bào. Trong số các con đường tín hiệu tế bào, con đường bị ảnh hưởng nhiều nhất là tín hiệu Wnt, với 34 gen, bao gồm 19 gen mục tiêu của con đường bị ảnh hưởng. Phân tích biểu hiện gen không gian cho thấy rằng cả gen mã hóa ligand Wnt (Wnt4) và một đối kháng của con đường (Sfrp2) đều được tăng cường cụ thể tại đường nối khớp đang phát triển, trong khi biểu hiện của một gen mục tiêu Wnt, Cd44, không còn có thể phát hiện được ở các phôi không có cơ bắp. Việc xác định 84 gen liên quan đến cytoskeleton giảm biểu hiện trong sự thiếu vắng của cơ cho thấy một số gen ứng cử viên có thể nhạy cảm với cơ học và có thể tham gia vào chuyển đổi cơ học, biến đổi một kích thích cơ học thành phản ứng phiên mã.Kết luậnCông trình này xác định các gen điều chỉnh phát triển chính bị ảnh hưởng bởi kích thích cơ học thay đổi, làm sáng tỏ các cơ chế phân tử giải thích kích thích cơ học trong suốt quá trình phát triển xương và cung cấp các nguồn lực quý giá để nghiên cứu thêm về cơ sở cơ chế của cơ chế điều chỉnh cơ học. Đặc biệt, nó làm nổi bật con đường tín hiệu Wnt như một điểm tích hợp tiềm năng giữa tín hiệu cơ học và phân tử, và các thành phần cytoskeleton như là các trung gian của phản ứng.

Từ khóa


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