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Responses of Host Hemocytes During the Initiation of the Squid-Vibrio Symbiosis

Pacific Biomedical Research Center, Kewalo Marine Laboratory, University of Hawaii

^* To whom correspondence should be addressed, at Department of Medical Microbiology and Immunology, University of Wisconsin, 1300 University Avenue, Madison, WI 53706. E-mail: mjmcfallngai{at}wisc.edu

Within hours after colonization of the light organ of the squid Euprymna scolopes by its bacterial symbiont Vibrio fischeri, the symbiont triggers morphogenesis of the light organ. This process involves the induction of apoptosis in the cells of two superficial ciliated epithelial fields and the gradual regression of these surface structures over a 96-h period. In this study, microscopic examination of various squid tissues revealed that host hemocytes specifically migrate into the epithelial fields on the surface of the light organ, a process that begins before any other indication of symbiont-induced morphogenesis. Experimental manipulations of symbiont-signal delivery revealed that hemocyte infiltration alone is not sufficient to induce regression, and high numbers of hemocytes are not necessary for the induction of apoptosis or the initiation of regression. However, studies with mutant strains of V. fischeri that show a defect in the induction of hemocyte infiltration provided evidence that high numbers of hemocytes facilitate the regression of the epithelial fields. In addition, a change in hemocyte gene expression, as indicated by the up-regulation of the C8 subunit of the proteasome, correlates with the induction of light organ morphogenesis, suggesting that bacteria-induced molecular changes in the hemocytes are required for the participation of these host cells in the regression process.

Abbreviations: CAM, chloramphenicol • FSW, filter-sterilized seawater • PGN, peptidoglycan • WT, wild type