Flow cytometric measurement of neutral red accumulation in earthworm coelomocytes: Novel assay for studies on heavy metal exposure

被引：26

作者：

Plytycz, Barbara ^{[1
]}

Klimek, Malgorzata ^{[1
]}

Homa, Joanna ^{[1
]}

Tylko, Grzegorz ^{[2
]}

Kolaczkowska, Elzbieta ^{[1
]}

机构：

[1] Jagiellonian Univ, Inst Zool, Dept Evolut Immunobiol, Krakow, Poland

[2] Jagiellonian Univ, Inst Zool, Dept Cytol & Histol, Krakow, Poland

来源：

EUROPEAN JOURNAL OF SOIL BIOLOGY | 2007年 / 43卷

关键词：

coelomocytes; amoebocytes; eleocytes; neutral red; riboflavin; flow cytometry; confocal microscopy;

D O I：

10.1016/j.ejsobi.2007.08.050

中图分类号：

Q14 [生态学（生物生态学）];

学科分类号：

071012 ; 0713 ;

摘要：

Neutral red (NR) is a vital stain accumulating in lysosomes of the cells, exemplified here by the coelomocytes non-invasively extruded from the earthworm coelomic cavity. Flow cytometry allows to distinguish two main populations of earthworm coelomocytes, namely hyaline and granular amoebocytes (A), and eleocytes/chloragocytes (E), the latter exhibiting riboflavin-derived auto-fluorescence. Coelomocytes of Dendrobaena veneta, Eisenia fetida, Allolobophora chlorotica, and Lumbricus sp. were incubated in vitro with copper chloride at 0 (controls) to 88 mu g/ml. Both amoebocytes and eleocytes accumulated neutral red, as evidenced by light and confocal microscopy and by flow cytometry. Flow cytometry allowed quantitative analysis of the adverse effects of an in vitro copper exposure on the intracellular neutral red accumulation in coelomocytes, phenomenon being completely prevented in the presence of metal-chelating agent, ethylenediaminetetraacetic acid (EDTA). In conclusion, flow cytometry may be used as an objective and fast tool for the measurement of NR accumulation in invertebrate cells, and the neutral red retention in the cells subjected to various stressors, including environmental pollution. (C) 2007 Published by Elsevier Masson SAS.

引用

页码：S116 / S120

页数：5

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