USE OF MICROARRAY HYBRIDIZATION TO IDENTIFY BRUGIA GENES INVOLVED IN MOSQUITO INFECTIVITY

Nematoda Genetic aspects;Filarial worms Molecular genetics;Brugia pahangi;Brugia malayi;Mosquitoes Pathogens;Mosquitoes as carriers of diseases

Griffiths, Kathryn GreerMichalski, Michelle ;

University of Wisconsin

关键词: Nematoda Genetic aspects;    Filarial worms Molecular genetics;    Brugia pahangi;    Brugia malayi;    Mosquitoes Pathogens;    Mosquitoes as carriers of diseases;   

Others  :  https://minds.wisconsin.edu/bitstream/handle/1793/46744/K%20Griffiths%20thesis.pdf?sequence=1&isAllowed=y

瑞士|英语

来源: University of Wisconsin

【 摘 要 】

Filarial nematodes are a family of insect-borne parasitic worms that cause anumber of diseases in man and animals. The most widespread filarial disease of humansis lymphatic filariasis, caused by worms in the genera Wuchereria and Brugia. Lymphaticfilariasis is an economic and social burden in endemic countries and affectsapproximately 119 million people worldwide. Brugia malayi and B. pahangimicrofilariae (mf) require a maturation period of at least five days in the mammalian hostbefore they can infect mosquito vectors. This maturation process correlates with changesin the surface composition of mf that likely are associated with changes in geneexpression. To test this hypothesis, we verified the differential infectivity of immature(<3 day) and mature (>30 day) Brugia mf for black-eyed Liverpool strain of Aedesaegypti (LVP), and then assessed transcriptome changes associated with microfilarialmaturation by competitively hybridizing microfilarial cDNAs to the B. malayioligonucleotide microarray. We identified transcripts that were more abundant inimmature (94 in B. pahangi and 29 in B. malayi) and mature (64 in B. pahangi and 14 inB. malayi) mf. In each case, >40% of Brugia transcripts shared no similarity to knowngenes, or were similar to genes with unknown function; the remaining transcripts werecategorized by putative function based on sequence similarity to known genes/proteins.Microfilarial maturation was not associated with demonstrable changes in the abundanceof transmembrane or secreted proteins; however, differences in transcript abundancewere observed for many that have predicted functions. For example, immature mf wereenriched for transcripts putatively associated with immune modulation,neurotransmission, transcription and cellular cytoskeleton elements. In mature mf, therewas an increase in transcripts potentially encoding hypodermal/muscle and surfacemolecules, i.e., cuticular collagens and sheath components. These finding lend support tothe underlying hypothesis that changes in microfilarial gene expression drive surfacemodifications that influence the parasite to begin development in compatible vectors.Brugia malayi genes corresponding to differentially abundant transcripts were identified,and transcript abundance validated by quantitative polymerase chain reaction. Thesestudies serve as a starting point towards gaining a better understanding of the parasiteside of the intricate parasite/mosquito relationship.