Desma ve Desmb Knockout Zebra Balığı Modellerinde Desmin İfadesinin İncelenmesi

Abstract

Desmin, a basic intermediate filament protein specific to muscle tissue, facilitates the transmission of force during muscle contraction and relaxation by forming a large network structure that allows the myofibrils to connect to the nucleus, sarcolemma and intracellular organelles. In animal models used to understand the underlying pathophysiology of DES-related diseases, it is necessary to analyze tissues in which orthologous genes are expressed. In the zebrafish, which has many advantages in modeling muscle diseases, there are two DES orthologous gene (desma and desmb). In our preliminary studies conducted by our group with in situ hybridization experiments in wild-type AB zebrafish, it has been shown that the desma and desmb transcripts have similar skeletal muscle expression pattern until 70th hour after fertilization; but after 70 hours, it was observed that the desma was expressed predominantly in skeletal muscle, while desmb was expressed highly in the smooth muscle. In the thesis study, it was aimed to investigate mRNA and protein expressions of desma and desmb in wild type AB, desma KO (-/-) and desmb KO (-/-) zebrafish embriyos. In the quantitative real-time PCR study, 99% of desma mRNA expression in desma KO (-/-) fishes and 98% of desmb mRNA expression in desmb KO (-/-) fishes were found to decrease relative to wild type AB fish. Whole-mount in situ hybridization and whole-mount immunofluorescence staining studies have shown that desma in desmb KO (-/-) zebrafish is also expressed in the gut, additional to skeletal and cardiac muscle. However, it was observed that in the desma KO (-/-) fish, the desmb was mostly expressed in smooth muscle from the 70th hour after fertilization and that no expression occurs in skeletal muscle in the absence of desma.