心臟心肌疾病之致病機轉複雜，多種基因參與在其中，利用心臟衰竭模式小鼠透過基因定位的方式定與生物資訊軟體與表現模式資料庫比對，篩選出位於Hrtfm4基因座上，在心臟骨骼肌高度表現且與心臟衰竭高度相關的Alpk2基因。Alpk2基因屬於alpha kinase家族一員，當中還有Alpk1和Alpk3基因，胺基酸序列比對發現Alpk2與Alpk3（45%）相對於Alpk1（21%）有相對較高的同源性，而在先前的研究已知alpk3基因缺陷會導致小鼠心臟病變，暗示Alpk2也許與Alpk3對於心臟功能擁有類似功能或調控機制。基於初步的預測結果與理由，利用斑馬魚此種擁有近似於哺乳類閉鎖循環系統之脊椎生物研究Alpk2和Alpk3在心臟之功能。使用原位雜交染色確認alpk2/3基因其表現位置皆表現在心臟與骨骼肌。利用morpholino技術對alpk2/3做基因抑制，觀察到斑馬魚胚胎心臟發育出現左右不對稱以及心包膜水腫之缺陷情況，其調控器官左右不對稱的nodal相關基因仍然保持正常。為確認專一性，進一步利用CRISPR/Cas9技術嘗試建立基因剔除魚。雖然alpk2與alpk3具有相似表現模式，但是二者並不會相互地調控。本實驗結果推測alpk2與alpk3在心臟發育過程中，有著相似的功能，具有調控心臟管腔發育方向的功能，且二者無上下游調控之關係。

Heart failure is the most common outcome of heart diseases and the pathogenesis of heart failure is complex and polygenic. There are many genes participate in the process of heart failure pathogenesis. By using quantitative trait locus mapping to identify several loci (Hrtfm) in a heart failure mouse model that influence heart function and overall survival rate. In Hrtfm4, Alpk2 (alpha kinase 2) is a strong candidate gene, that is highly expressed in heart and skeletal muscles, suggests that Alpk2 might play an important role in heart function. Alpk2 is a gene that belong to Alpk family. There are three major members in the family, Alpk1, Alpk2 and Alpk3. Among all three Alpk proteins, Alpk2 shares a higher amino acid homology with Alpk3 (45%) than that with Alpk1 (21%). Alpk3 deficient mice has been reported to feature with cardiomyopathy. Based on these predictions and reasons, by using zebrafish embryo as the experimental animal model, took advantage for its closed circulation system to study the role of alpk2 and alpk3 on heart function. Both of alpk2 and alpk3 expressed on the skeletal muscle and heart in zebrafish embryo. Knockdown of alpk2 and alpk3 by using morpholino caused the pericardial edema and heart looping asymmetry. In order to clarify the mechanism of how alpk2 or alpk3 control the orientation of heart looping, the expression of southpaw, which is the left–right asymmetric master regulation signals during zebrafish early embryonic development was tested. The expression of southpaw is normal after the knockdown of alpk2 or alpk3. To confirm the specificity of the loss-of-fuction study that based on morpholino, by using CRISPR/Cas9 mutagenesis technique to generate alpk2 and alpk3 mutant zebrafish. Although alpk2 and alpk3 share the same expression pattern, Alpk2 and Alpk3 do not regulae each other in the process of heart early development. In this study, the expression pattern of alpk2 and alpk3 has been confirmed. The loss-of-function based on morpholino suggest that alpk2 and alpk3 will regulale the direction of heart looping, but alpk2 and alpk3 do not have the epistasis relationship to each other.

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