Identification of differentially expressed proteins between free-living and activated third-stage larvae of...

The disease caused by Haemonchus contortus, a blood-feeding nematode of small ruminants, is of major economic importance worldwide. The infective third-stage larva (L3) of this nematode is enclosed in a second cuticle. Once the L3 is ingested by the host, the outer cuticle undergoes an exsheathment process that marks the transition from the free-living stage to the parasitic stage. This study explored the changes in protein expression relative to this transition. Proteins extracted from free living L3 and exsheathed L3 (xL3) were analyzed by two dimensional differential gel electrophoresis (2D-DIGE). More than 2200 protein spots were recognized, and 124 of them was found to be differentially expressed (average ratio of xL3/L3>1.5 or xL3/L3<-1.5, p<0.05). Of these, 83 spots were up-regulated and 41 spots were down-regulated in xL3 when compared with L3. These differentially expressed spots were analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) or MALDI-TOF-MS/MS and 40 proteins were identified. To predict the functions of these identified proteins, they were assigned for gene ontology (GO) annotation. Results showed that the proteins may be involved in biological processes of reproduction, cellular organization or biogenesis, multi-cellular organismal processes, single-organism processes, metabolic processes, signaling, biological regulation, response to stimulus, cellular processes, biological adhesion, growth, locomotion, localization, developmental processes and multi-organism processes. Kyoto Encyclopedia of Genes and Genomes (KEGG) annotations were also performed, which was useful for exploring the process of metabolism and signal transduction pathways. This study indicated that some key alterations taking place, during the transition from L3 to xL3 may be interesting antiparasite targets, and some of the proteins involved in this process might be candidate antigens for vaccine development.