Cloning, expression pattern and functional characterization of fused, an important kinase of the Hedgehog signalling pathway from Locusta migratoria (Orthoptera: Acridoidea)

Abstract

Locusta migratoria is one of the global destructive migratory agricultural pests. The species serves as a model insect for the study of morphology, physiology, developmental biology, and neurosciences and so on. In recent years, there has been growing interest in the research of the cell and molecular biology of L. migratoria, and a lot of genes of signalling pathways have been cloned. The Hedgehog (Hh) signalling pathway is essential for cellular proliferation and differentiation during embryonic development. Fused (Fu) is a conserved component of the Hh signalling pathway, and it plays a central role in embryonic development. The Fu gene was cloned from the grasshopper L. migratoria. The gene encodes a 717 amino acid protein. Then, quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expression pattern of the Lm-Fu gene at different development stages and tissues. The result showed that the gene was upregulated at the embryonic period and second instar, and the expression level was the highest in the head at the different tissues. Western blotting was used to assess the abundance of the Lm-Fu protein, which was expressed at different development stages. The embryogenesis of L. migratoria was examined by immunohistochemical staining, and the Lm-Fu protein was detected in the head, prothorax and legs. The results showed that the Lm-Fu gene plays an important role during embryonic development.

Keywords

• locusta migratoria
• lm-fu gene
• cloning
• expression
• western blot
• immunohistochemistry