Identification and characterisation of the Mpeg1 homologue in the South African abalone, Haliotis midae

Abstract

Although Haliotis midae is the most economically important cultured abalone species in South Africa, infectious diseases have the potential to severely limit the production of this shellfish. Consequently, it is becoming increasingly important to characterise the abalone immune system in order to better understand their ability to combat infection. This study reports the identification and characterisation of a perforin-like protein, designated hmMpeg1, which is believed to be involved in the H. midae immune system. hmMpeg1 encodes for a 78 kDa protein that has significant sequence similarity to Mpeg proteins from other abalone species and includes the conserved cytolytic membrane attack complex/perforin (MACPF) domain of perforin. Real-time quantitative PCR (qPCR) analysis demonstrated expression of hmMpeg1 mRNA in haemocytes and epipodia samples from H. midae exposed to a heat-killed, Gram-negative bacterial pathogen, Vibrio anguillarum 5676. hmMpeg1 mRNA in haemocytes increased significantly 48 h post-infection (h.p.i) (8.2 fold; P < 0.05), coinciding with a decrease in the total number of circulating haemocytes, and reached a maximum at 96 h.p.i (17.2 fold; P < 0.05). Similarly, a significant increase in the level of hmMpeg1 mRNA occurred at 24 h.p.i in epipodia samples (3.8 fold; P < 0.05), reaching a maximum at 48 h.p.i (4.5 fold; P < 0.05). In addition, western blot analysis detected a significant increase in hmMpeg1 between 24 h.p.i (4.2 fold; P < 0.05) and 48 h.p.i (3.1 fold; P < 0.05) in the epipodia, and between 48 h.p.i (1.7 fold; P < 0.05) and 96 h.p.i (1.9 fold; P < 0.05) in haemocytes, sampled from abalone exposed to the abalone pathogen V. anguillarum 5676. The importance of hmMpeg1, in terms of its function and importance in the H. midae immune response, is discussed.