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A flow cytometric approach to the study of crustacean cellular immunity

Journal of Invertebrate Pathology

By:
, , and
DOI: 10.1006/jipa.2000.4960

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Abstract

Responses of hemocytes from the crayfish Procambarus zonangulus to stimulation by fungal cell walls (Zymosan A) were measured by flow cytometry. Changes in hemocyte physical characteristics were assessed flow cytometrically using forward- and sidescatter light parameters, and viability was measured by two-color fluorescent staining with calcein-AM and ethidium homodimer 1. The main effects of zymosan A on crayfish hemocytes were reduction in cell size and viability compared to control mixtures (hemocytes in buffer only). Adding diethyldithiocarbamic acid, an inhibitor of phenoloxidase, to hemocyte to zymosan mixtures delayed the time course of cell size reduction and cell death compared to zymosan-positive controls. The inclusion of trypsin inhibitor in reaction mixtures further delayed the reduction in hemocyte size and cell death, thereby indicating that a proteolytic cascade, along with prophenoloxidase activation, played a key role in generating signal molecules which mediate these cellular responses. In addition to traditional methods such as microscopy and protein chemistry, flow cytometry can provide a simple, reproducible, and sensitve method for evaluating invertebrate hemocyte responses to immunological stimuli.

Additional Publication Details

Publication type:
Article
Publication Subtype:
Journal Article
Title:
A flow cytometric approach to the study of crustacean cellular immunity
Series title:
Journal of Invertebrate Pathology
DOI:
10.1006/jipa.2000.4960
Volume
76
Issue:
2
Year Published:
2000
Language:
English
Contributing office(s):
National Wetlands Research Center
Description:
p. 112-119
Larger Work Type:
Article
Larger Work Subtype:
Journal Article
Larger Work Title:
Journal of Invertebrate Pathology
First page:
112
Last page:
119