The image shows the severe defects in the forebrain tissue and eyes of a gibel carp’s embryo due to developmental inhibition caused by an overdose of the injected wile-type CagMdkb RNAs.
A research team at the CAS Wuhan Institute for Hydrobiology has made new progress in the genetic expression of the carp's embryo development. The work was reported as a cover story in a recent issue of the
International Journal of Developmental Biology.
The research paper, under the title of "Development Expression of CagMdkb during Gibel Carp Embryogenesis," is completed by YIN Jun and his colleagues under the tutelage of Prof. GUI Jianfang with Institute.
Midkine (Mdk) is a kind of secretive protein, playing a critical role in an animal's neurogenesis and found to take part in the formation of human tumor. In different vertebrates, however, it has been found its gene has varied patterns in genetic expression. In this study, the gibel carp (Carassius auratus gibelio) version of this protein, known as CagMdkb, has been cloned and characterized from an SMART cDNA library of the gibel carp's 10 somite-stage embryos. Its full length cDNA has 1,091 base pairs and encodes a sequence of 147 amino-acids, of which 97.3% are identical to those of zebrafish Mdkb at the amino acid level. In the course of the gibel carp's embryonic development, the analysis reveals that CagMdkb is first transcribed in gastrula-stage embryos and maintains a relatively stable expression level during the subsequent embryo-genesis. The Western blot analysis indicates a 19kDa maternal CagMdkb protein band and the zygotic CagMdkb protein are expressed from the gastrula stage. At around 10 somite stage, the 19kDa CagMdkb is processed to another protein band of 17kDa, probably a secreted form with the 21-residue signal peptide removed, becoming a matured protein. The maternal CagMdkb protein is found to be localized in each blastamere cell of early embryos. As soon as the embryo develops into its 18-somite stage, a new signal of synthesized protein appears in a pair of large neurons and in the later stages, CagMdkb protein is also extended to numerous small neutons in the forebrain, midbrain and hindbrain, as well as to nerve fibers in the spinal cord. A positioning analysis of co-localization with 3A10 antibody reveals the pair of large neurons are Mauthner neurons in which the CagMdkb immuno-reactivity is located. In addition, extopic expression ofCagMdkb in early embryos of gibel carp and zebrafish suppresses their head formation and CagMdkb function is found to depend on the secretory activity.
All of these findings indicate CagMdkb plays an important role in neural development of gibel carp embryo-genesis and there is functional conservation of Mdkb in the fish head formation.
The research has been jointly supported by the National Program for Basic Research (the "973 Program") and the National Natural Science Foundation of China.