Extrusion of earthworm coelomocytes: comparison of the cell populations recovered from the species Lumbricus terrestris, Eisenia fetida and Octolasion tyrtaeum

Abstract

SummaryCoelomocytes were extruded from three earthworm species: Lumbricus terrestris, Eisenia fetida and Octolasion tyrtaeum. Featuring a simple low-vacuum holding device, the proposed methodology allows the recovery of cells with minimum risk of contamination by faecal material. The viability of O. tyrtaeum coelomocytes was highly reproducible (average 93%), with an average yield of 0.92 x 10 6 viable cells per earthworm. Cell viability for 1. terrestris and E. fetida averaged -68% but the cell yields were higher (respectively 1.67 x 10 6 and 1.28 x 10 6 ). Large inter-individual differences in cell yields were observed with 1. terrestris. Flow cytometric analyses indicated species to species differences in cell populations.Coelomocytes from E. fetida were the smallest with -57% of the total viable cells recovered being monitored between 2 and 10 tim. Large granulated cells I~20 tim) were detected in fairly large proportions in 1. terrestris and O. tyrtaeum [-52 and -96%, respectively I while they were less abundant in E. fetida (-9%). Using the vital dye neutral red to assess functional integrity, average cellular uptakes were significantly higher for 1. terrestris and O. tyrtaeum than for E. fetida (2.94, 2.66 and 0.64 tig/2 x 10 5 cells, respectively). In summary, the extrusion methodology herein described is applicable for the recovery of coelomocytes from a wide range of earthworm sizes and species. Moreover, this study strengthens the fact that extruded coelomocytes could be used for the evaluation of cell dysfunction and/or cell death following an in vitro and/or in vivo treatment. Keywords

Phagocytic Response of Terrestrial and Aquatic Invertebrates Following in Vitro Exposure to Trace Elements

“…Chateaureynaud-Duprat and Lestage (1980) also showed interesting pictures of L. terrestris leukocyte cells. Dioge`ne et al (1997) showed the flux cytometry patterns obtained for L. terrestris, E. fetida, and Octolasion tyrtaeum. They have reported the size distribution of extruded celomocyte cells, showing that L. terrestris had well-distributed size groups across the 5-to 430-mm range, whereas E. fetida cells were distributed mainly between the 2-to 15-mm particle diameters.…”

Biomarkers in Earthworms

“…Immunological responses are known to depend on many factors, but this has been scarcely studied in earthworms (Weeks et al 1992). The immunological response appears to be species dependent Diogene et al 1997), tissue dependent, and dependent on time of sampling (Goven et al 1994), but there appears to be no or only little seasonal dependency (Eyambe et al 1991;Goven et al 1993bGoven et al , 1994. For example, in copper-exposed L terrestris the immune response was more severely depressed in the coelomic fluid than in the coelomocytes (Goven et al 1994).…”

Evidence for the presence of thyroid stimulating hormone, thyroglobulin and their receptors in Eisenia fetida: a multilevel hormonal interface between the nervous system and the peripheral tissues

“…The previously described collecting solution was slightly modified. The extrusion buffer contained 71.2 mM NaCl, 50.4 mM guaicol-glyceryl-ether, 5 mM EGTA pH 7.3 (Eyambe et al 1991;Diogéne et al 1997). Animals were kept in this mixture for 5 min.…”