Research On The Functions Of Anchor And Jumu In Drosophila Wing Development

Drosophila is an important model system for studying the underlying molecular mechanisms of morphogenesis and patterning at both the cellular and tissue levels.In particular,much has been learned about the genetic basis of the development of the wing.The wing is the largest organ in Drosophila tissue.Drosophila is a holometabolous insect whose wings are developed and differentiated by wing imaginal discs.The formation of insect wings is important to the migration,reproduction and transfer of insects,and expand the area of their lives.Therefore,the study of insect wing development not only helps us understand the process of insect metamorphosis,but also provides us with a broad theoretical basis for further exploration of the molecular mechanisms of organ morphogenesis and biological evolution.Drosophila Anchor(CG7510),which we have called anchor,is an integral membrane protein that is the ortholog of vertebrate GPR155 and shares more than 37%sequence identity with mammalian GPR155.Studies have shown that GPR155 is associated with the regulation of Huntington disease and autism spectrum disorders(ASD).In addition,our studies have shown that the knock-down of anchor induced abnormal phenotypes in Drosophila wing,but there is no report on the regulation of anchor in wing development.Jumeau(Jumu)is a member of the Forkhead(Fkh)transcription factor family,which is involved in the development of the cardiac progenitors of Drosophila,the establishment of the hematopoietic system,and the maintenance the homeostasis of the immune system.Previous studies have shown that the low expression of jumu results in abnormal development of the compound eyes,wings and bristles of Drosophila.Based on this,we further explored the regulation of Jumu in the development of Drosophila wings,providing a better understanding the functions of Fox family members in organ development.In this study,anchor and jumu mutant lines were used as research materials to analyze the abnormal phenotype of Drosophila adult wings.In addition to analyzing the localization of Anchor and Jumu genes in wing discs or the pupal wing tissue.And the process of the cell proliferation and apoptosis of the wing discs in the larval stage were detected by using immunohistochemistiy.The formation of intervein cells and wing hairs were also included in the study.The main results obtained are as follows:1.Knocking down anchor induces abnormal wing phenotypes in adult flies.2.The expression pattern of anchor through in situ hybridization expressed in a generalized manner in imaginal discs and pupal wings.3.Knocking down anchor resulted in an increased the number of the cell proliferation in imaginal discs and promoting the Wg levels.4.Knocking down anchor increased the levels of various target genes,such as p-Mad Sal and Omb in the wing discs and the expression levels of Dad and Brk were not changed;the expression levels of p-Mad and Dad were increased,but the expression level of Brk was decreased in the pupal wings,which indicates that the effects of anchor on BMP signalling was more significant in the pupal stages.5.Epistasis experiments revealed that the application of BMP signaling factors rescued the wing size and ectopic venation observed in the anchor-knockdown flies.6.jumu mutants showed small wings,the loss of the ACV,defective wing margins and multiple wing hairs.7.Jumu was expressed in the wing blade and wing margin during larval stages or expressed in intervein cells and localized to the nucleus during the pupal stages.8.Reductions in jumu expression in larval wing discs caused ectopic cell proliferation,increased Wg levels and a large number of apoptotic cells;the expression levels of p-JNK and puc were increased to cause ectopic activation of JNK signaling;the adult wing margin deletion was not related to apoptosis but resulted in a loss of Wnt signaling responses by reducing Cut levels.9.Jumu mutants performed the increasing levels of F-actin and reducing the levels of DE-cad and Rho1,which were resulted in multiple wing hairs.The above results indicated that both anchor and jumu genes are involved in the development of Drosophila wing,which performed a key role in the morphogenesis and cell differentiation of wings.They provided new clues for further investigation of the function of GPCRs in organ development,moreover gained a theoretical basis for studying the functions of Fox family members in organ development.