| Lipid rafts is a microstructure domain riched in cholesterol,sphingomyelin as well as the specific proteins on the plasma membrane.Caveolae as a special type of Lipid rafts,is a plasma membrane invagination.Caveolae related membrane protein family of mammals includes three subfamilies,such as Caveolin,Flotillin and epidermal surface antigen(ESA).It has been reported that Flotillin-1 was related to axonal growth,learning-memory and tumor formation.Our previous study showed that Caveolin-1 as a molecular signature of caveolae,participated in proliferation,transformation and apoptosis of a variety of cells.It has been found that Flotillin-1 and Caveolin-1 both expressed in various types of invertebrates and vertebrates and they are highly conserved in evolution.Drosophila melanogaster is a complete metamorphosis insect,as a kind of important model organism,it contains at least 65% homologous genes of the human disease related genes.Therefore,studying the expression and distribution of these genes in different developmental stages of Drosophila is of great significance to further clarify the mechanism of human pathology.But the biological functions and the relationship between Flotillin-1 and Caveolin-1 in the development of Drosophila are not clear so far.In this paper,Drosophila melanogaster was used for the study.The physicochemical properties of Flotillin-1 protein of Drosophila melanogaster were analyzed using bioinformatics method,the expression levels and distributions of Flotillin-1 and Caveolin-1 in different developmental stages of it were detected utilizing Western blot and immunofluorescence,to explore the role and interaction of the two proteins in the development of Drosophila,which would provide experimental evidence for the further study of the molecular mechanism of the role of the two Caveolae related proteins.The results were as follows:(1)The Flotillin-1 of Drosophila melanogaster was highly conserved in evolution.The molecular weight of Flotillin-1 was 47.14 Kda,which isoelectric point was 5.56.It was a stable hydrophilic membrane protein and had several phosphorylation sites,but didn't have signal peptide and transmembrane region.(2)The expression levels and distributions of Flotillin-1 varied in different developmental stages of Drosophila melanogaster: the vivo distribution of Flotillin-1 was very little only in the fat bodies in larval stage,a little in the alimentary canal and fat bodies in pupa stage,mainly in the brain,ommateum,alimentary canal,genital gland and a little in the muscles in adult stage.While,the expression levels of Flotillin-1 in adults were significantly higher than those of the larvae and pupae,and the expression of the protein in the brain was extremely significantly higher than that in the thorax and abdomen of adult.(3)The expression levels and distributions of Caveolin-1 varied in different developmental stages of Drosophila melanogaster: the vivo distribution of Caveolin-1 was a little in the fat bodies in larval stage,a little in the brain and fat bodies in pupa stage,mainly in the brain,ommateum,muscles,alimentary canal and a little in the fat bodies in adult stage.The Caveolin-1 was mainly expressed in the adults of Drosophila melanogaster and the expression level of the female was significantly higher than that of the male.In addition,Flotillin-1 and Caveolin-1 both co-expressed in the brain,ommateum,alimentary canal and muscles in the adult.Conclusion: The expression levels and distributions of Flotillin-1 and Caveolin-1 varied in different developmental stages of Drosophila melanogaster.The Flotillin-1 and Caveolin-1 both are mainly expressed in the adults,especially in the brain and co-localized in the brain,ommateum,alimentary canal and muscles. |
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