研究報告
| 學年 | 97 |
|---|---|
| 學期 | 1 |
| 出版（發表）日期 | 2009-01-01 |
| 作品名稱 | 轉錄因子NF-Y, Capsulin與Musculin參與顏面肌肉軟骨發育之分子機制探討 |
| 作品名稱（其他語言） | Molecular Mechanisms of NF-Y, Capsulin and Musculin during Craniofacial Development |
| 著者 | 陳曜鴻 |
| 單位 | 淡江大學生命科學研究所 |
| 描述 | 計畫編號：NSC97-2313-B032-001-MY3 研究期間：200908~201007 研究經費：1,200,000 |
| 委託單位 | 行政院國家科學委員會 |
| 摘要 | 頭部肌肉與軟骨的發育起源於神經脊細胞，頭部的神經脊細胞來自於後 腦，然後經移動而進入咽弧，最後再與來自中胚層的間葉細胞以及周圍的上皮細 胞間進行交互作用，再經由某些未知基因的作用之下而形成頭部肌肉與骨骼的前 驅細胞。頭部的肌肉前驅細胞接著會受到MRF 基因群的調控進而形成肌肉；頭 部的骨骼前驅細胞接著會受到sox9a、runx2 與cbfa 等基因群的調控進而形成骨 骼。本期計畫欲研究之capsulin、musculin 與nf-y 等基因群，據推測，應該在神 經脊細胞與頭部肌肉與骨骼的前驅細胞之間，扮演重要的角色(Fig. 9)。這些研究 項目十分繁重，故擬分三年來完成下列目標。 第一年計畫內容涵蓋：(1)複殖出capsulin、musculin 與nf-y 等基因之cDNA 全長；(2)進行全個體雜交實驗，以便偵測capsulin、musculin 與nf-y 等基因在胚 胎發育早期的表現位置；(3)切片技術與組織染色；(4) capsulin、musculin 與nf-y 等基因結構之確定；(5)抗體免疫螢光染色實驗的建立。 第二年計畫內容涵蓋：(1)胚胎發育抑制劑的注射；(2)切片技術與組織染色； (3)抗體免疫螢光染色實驗的建立；(4)軟骨染色；(5)細胞凋亡實驗之建立。 第三年計畫內容涵蓋：(1)胚胎發育抑制劑的注射；(2)全長capsulin、 musculin、nf-y、myf5、myod 之mRNA 合成與注射；(3)軟骨染色；(4)抗體免疫 螢光染色實驗的建立；(5)切片技術與組織染色；(6) capsulin、musculin 與nf-y 參 與頭部肌肉軟骨發育機制之探討。 Cranial neural crest (CNC) cells play an important role during cranial myogenesis and cartilage development. CNC cell originates from hindbrain, migrates into pharyngeal arch, and then differentiates into muscle and cartilage under the control of some unknown genes. We hypothesize that capsulin, musculin and nf-y genes are upstream regulators that converting CNC into muscle and cartilage cell fates (Fig. 9). To confirm this hypothesis, we propose this 3-year project. Our proposal will be including: The first year: (1). Cloning the full length cDNA of capsulin, musculin and nf-y; (2). Carrying out whole mount in situ hybridization experiments to detect the endogenous expression patterns of capsulin, musculin and nf-y during early embryogenesis; (3). Using cryosection and immunohistochemical staining to further confirm the expression domain of each gene transcript; (4). Determination the genomic organizations of capsulin, musculin and nf-y genes; (5). Using antibodies staining to study the endogenous protein levels. The second year: (1). Morpholino injection to knockdown capsulin, musculin and nf-y; (2). Using cryosection and immunohistochemical staining to further confirm the expression domain of each gene transcript; (3). Using antibodies staining to study the endogenous protein levels; (4). Using alician blue staining to visualize the cartilages; (5). Carrying out TUNEL assay to detect the apoptotic cells. The third year: (1). Morpholino injection to knockdown capsulin, musculin and nf-y; (2). Synthesis of full length capsulin, musculin and nf-y mRNA and co-injection them with morpholino into zebrafish embryos; (3). Using alician blue staining to visualize the cartilages; (4). Using antibodies staining to study the endogenous protein levels; (5). Using cryosection and immunohistochemical staining to further confirm the expression domain of each gene transcript; (6) Discovery the molecular mechanisms of how capsulin, musculin and nf-y are involved during craniofacial development. |
| 關鍵字 | 神經脊細胞; 頭部肌肉; 斑馬魚; 軟骨; neural crest cells; cranial muscle; zebrafish; cartilage |
| 語言 | |
| 相關連結 |
機構典藏連結 ( http://tkuir.lib.tku.edu.tw:8080/dspace/handle/987654321/46930 ) |