Metallothionein expression and Neutral Red uptake as biomarkers of metal exposure and effect in Eisenia fetida and Lumbricus terrestris exposed to Cd

2007 ◽  
Vol 43 ◽  
pp. S233-S238 ◽  
Author(s):  
Vega Asensio ◽  
Peter Kille ◽  
Andrew John Morgan ◽  
Manu Soto ◽  
Ionan Marigomez
Keyword(s):  
Eisenia Fetida ◽  
Lumbricus Terrestris ◽  
Neutral Red ◽  
Metal Exposure ◽  
Neutral Red Uptake

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

1997 ◽  
Vol 31 (4) ◽  
pp. 326-336 ◽  
Author(s):  
J. Diogéne ◽  
M. Dufour ◽  
G. G. Poirier ◽  
D. Nadeau
Keyword(s):  
Eisenia Fetida ◽  
Lumbricus Terrestris ◽  
Neutral Red ◽  
Cell Populations ◽  
Viable Cells ◽  
Wide Range ◽  
Holding Device ◽  
Octolasion Tyrtaeum

Coelomocytes 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 × 106 viable cells per earthworm. Cell viability for L. terrestris and E. fetida averaged ~68% but the cell yields were higher (respectively 1.67 × 106 and 1.28 × 106). Large inter-individual differences in cell yields were observed with L. 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 µm. Large granulated cells (≥20 µm) were detected in fairly large proportions in L. terrestris and O. tyrtaeum [~52 and ~96%, respectively) 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 L. terrestris and O. tyrtaeum than for E. fetida (2.94, 2.66 and 0.64 µg/2 × 105 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.

The EYTEX™ System and the Neutral Red Uptake Inhibition Assay in Cultured Hep G2 Cells as Alternative Methods for In Vivo Eye Irritation Following the EEC Protocol

1990 ◽  
Vol 17 (4) ◽  
pp. 325-333
Author(s):  
Paul J. Dierickx ◽  
Virginia C. Gordon
Keyword(s):  
Neutral Red ◽  
Alternative Methods ◽  
Inhibition Assay ◽  
Hep G2 ◽  
Hep G2 Cells ◽  
Eye Irritation ◽  
Uptake Inhibition ◽  
In Vitro Methods ◽  
Neutral Red Uptake

The neutral red uptake inhibition assay and the EYTEX™ system were investigated as alternative methods for the assessment of eye irritation, determined according to the EEC protocol. The 17 test chemicals used were mainly organic solvents. The xenobiotics were applied to Hep G2 cells for 24 hours at different concentrations. Neutral red uptake inhibition was then measured. The results are expressed as the NI50 value, which is the concentration of test compound required to induce a 50% reduction in neutral red uptake. The same chemicals were also tested as coded samples by the EYTEX™ test according to the manufacturer's directions. A nearly identical quantitative correlation was found for both in vitro methods with corneal opacity scores: r = 0.84 for EYTEX™ scores and r = 0.83 for log NI50, expressed in μg/ml. Whilst these correlations are certainly not perfect, it is clear that both in vitro methods can be used as valuable prescreening methods.

scholarly journals Biological Innocuousness of a Device developed to select the Optimum Spermatozoa to be used in the Treatment of Infertility

2021 ◽  
Author(s):  
Marisa A Cubilla ◽  
Hector A Guidobaldi ◽  
Laura C Giojalas
Keyword(s):  
Colony Formation ◽  
Physiological State ◽  
Neutral Red ◽  
Sperm Selection ◽  
Neutral Red Uptake

The sperm selection assay (SSA) is a method based on chemotaxis to obtain spermatozoa at the optimum physiological state to successfully fertilize the egg. It consists of a device made of acrylic and an attractant solution which includes progesterone. We evaluate potential cytotoxicity interactions by means of Neutral Red uptake, the MTT and colony formation assays, according to ISO normative. Here we showed that even stressing the conditions of the assays, the SSA device alone or together with the progesterone solutions employed, showed to be innocuous for the cells. Suggesting that SSA could be incorporated into the ART procedures.

The effect of hexaaza-and hexathia–macrocyclic ligands on transition metal cytotoxicity in human hepatoma-derived cultured cells

2002 ◽  
Vol 21 (8) ◽  
pp. 421-427 ◽  
Author(s):  
P W Smet ◽  
T F Pauwels ◽  
P J Dierickx
Keyword(s):  
Transition Metal ◽  
Cultured Cells ◽  
Copper Ions ◽  
Transition Metal Ions ◽  
Neutral Red ◽  
Macrocyclic Ligands ◽  
Human Hepatoma ◽  
Uptake Inhibition ◽  
Copper Zinc ◽  
Neutral Red Uptake

The effect of macrocyclic ligands on cytotoxic concentrations of the transition metal ions of copper, zinc, and cadmium was investigated. For this purpose, a hexaaza-[3,6,9,17,20,23-hexaazatricyclo[23.3.1.111,15] triaconta–1(29),11(30),12,14,25,27–hexaene (L2)] and hexathia-chelating ligand [1,4,7,10,13,16-hexathiacyclooctadecane (L3)] were used in the human hepatoma-derived HepG2 cell line. The cytotoxicity was measured by the neutral red uptake inhibition assay. First, the NI50 of the ligands, i.e., the concentration of the ligand inducing a 50% inhibition in neutral red uptake compared to control cells, was determined. In several metal/ligand combination experiments, the effects for L2 were difficult to interpret, whereas for L3 in combination with copper ions, a severe increase–and for zinc ions, a significant decrease of cell toxicity–relative to the metal control was observed. To further examine the different effects observed with L3 in combination with, respectively, Cu2+ and Zn2+, the glutathione (GSH) content was measured. The relative GSH content decreased as the concentration of L3 increased. It was proposed that the increased toxicity of the combination Cu2+ /L3 could be caused by the depletion of GSH and a subsequent inability to scavenge the produced reactive oxygen species (ROS). This hypothesis was supported by experiments during which vitamin E or C was added to the Cu2+ / L3 system.

scholarly journals Metallothionein Induction in the Coelomic Fluid of the EarthwormLumbricus terrestrisfollowing Heavy Metal Exposure: A Short Report

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
A. Calisi ◽  
M. G. Lionetto ◽  
E. De Lorenzis ◽  
A. Leomanni ◽  
T. Schettino
Keyword(s):  
Heavy Metal ◽  
Tissue Distribution ◽  
Physiological Condition ◽  
Lumbricus Terrestris ◽  
Polluted Soil ◽  
Metal Exposure ◽  
Coelomic Fluid ◽  
Short Report ◽  
Soil Monitoring ◽  
Heavy Metal Exposure

Earthworms are useful bioindicator organisms for soil biomonitoring. Recently the use of pollution biomarkers in earthworms has been increasingly investigated for soil monitoring and assessment. Earthworm coelomic fluid is particularly interesting from a toxicological perspective, because it is responsible for pollutant disposition and tissue distribution to the whole organism. The aim of the present work was to study the effect of heavy metal exposure on metallothionein (Mt) induction in the coelomic fluid ofLumbricus terrestrisin view of future use as sensitive biomarker suitable for application to metal polluted soil monitoring and assessment.L. terrestriscoelomic fluid showed a detectable Mt concentration of about4.0±0.6 μg/mL (mean ± SEM,n=10) in basal physiological condition. When the animals were exposed to CuSO4or CdCl2or to a mixture of the two metals in OECD soils for 72 h, the Mt specific concentration significantly (P<0.001) increased. The Mt response in the coelomic fluid perfectly reflected the commonly used Mt response in the whole organism when the two responses were compared on the same specimens. These findings indicate the suitability of Mt determination inL. terrestriscoelomic fluid as a sensitive biomarker for application to metal polluted soil monitoring and assessment.

Sign in / Sign up

Export Citation Format

Share Document