The zebrafish erythropoietin: Functional identification and biochemical characterization

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• 作者：Cheng-Ying Chu ; Chia-Hsiung Cheng ; Gen-Der Chen ; Yi-Chung Chen ; Chin-Chun Hung ; Kai-Yun Huang ; Chang-Jen Huang
• 关键词：Erythropoietin ; Zebrafish ; Morpholino oligonucleotide ; Glycoprotein ; Secretion ; Globin staining
• 刊名：FEBS Letters
• 出版年：2007
• 出版时间：4 September 2007
• 年：2007
• 卷：581
• 期：22
• 页码：4265-4271
• 全文大小：1090 K

文摘

In the present study, the zebrafish epo cDNA was cloned. The encoded protein displays 90 % , 55 % and 32 % identity to the Epo from carp, fugu and human, respectively. Through RT-PCR, the expression of zepo mRNA was mainly in the heart and liver. In the COS-1 cell transfection experiments, the recombinant zEpo-HA protein was efficiently secreted into the culture medium as a glycoprotein and the carbohydrate moiety can be cleaved by the treatment of peptide-N-glycosidase F (PNGase F). Using the morpholino approach, we showed that zepo morphants displayed severe anemia leading to high mortality during development. Such an effect can be significantly rescued by zepo RNA. Furthermore, in the absence of functional zEpo, the expression of specific markers for adult globin genes, such as αA1- and βA1-globin, but not the embryonic βe1-globin, was affected.