Charactertisation of the Drosophile Malvolio

Folwell, James Leigh (2001) Charactertisation of the Drosophile Malvolio. University of Southampton, Doctoral Thesis.

Abstract

The Drosophila malvolio gene was identified in a P-element mutagenesis screen for flies with taste defects, and was found to have high sequence identity with the mouse Nramp1 gene (Rodriguez et al., 1995), as well as expression in haemocytes. Nramp1 is involved in the innate immune response against certain bacterial pathogens, and belongs to a family of divalent metal cation transporters. Using the mvl-lacZ reporter line, I have confirmed and extended a number of previous observations made by Rodriguez et al., (1995). Expression was seen in cells of the amnioserosa, and cells associated with the antenno-maxillary complex stage 16 embryos. In larvae, expression was observed in the alimentary canal, proximal malpighian tubules, the chemosensory apparatus of the head, and the testis. In pupae, expression was confirmed in haemocytes. In the adult fly, expression was seen in the alimentary canal as well as the following chemosensory structures; the proboscis, the third antennal segment, and the labial palps. Antibodies were raised against the N-terminal 76 amino acids of the Malvolio protein, and the antiserum was affinity purified. The AP antiserum was then tested to confirm reactivity towards Malvolio alone. Immunohistochemistry was then performed using the AP anti-Malvolio antiserum. Expression of Malvolio was observed in macrophage-like cells, malpighian tubules, testis, brain, the amnioserosa of embryos, and the ventriculus of the larval and adult alimentary canal. In most tissues, the anti-Mv1 staining is punctate and subcellular in appearance. Sections of ventriculus, stained with anti-Mv1 antisera were analysed using transmission electron microscopy. This revealed anti-Mv1 staining associated with mitochondria, in the muscles which surround the ventriculus. The function of the malvolio gene is yet to be elucidated.

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