Dear all,
I am planning to investigate the effects of pollutants, substrate type and food quality on the physiology of farmland woodlice, and would like to include measures of oxidative stress. However, I am new to these model species, having primarily worked with birds and mammals, and I do not know what kind of markers could be measured, what are the particular constraints for this type of organisms and whether there are already standard protocols in these species?
If you had some foundational references on this topics, that would also be very appreciated!
Thanks a lot!
There are a variety of reactive oxygen species during oxidative stress in mammalian cells, such as, superoxide radicals, peroxynitrite, OCl- and so on. There are a bunch of fluorescent dyes that can detect these ROS. The links are here:
http://www.lifetechnologies.com/us/en/home/references/molecular-probes-the-handbook/probes-for-reactive-oxygen-species-including-nitric-oxide/generating-and-detecting-reactive-oxygen-species.html
Good luck!
Hi Jiansheng,
Thanks for the quick answer. However, I was rather enquiring about particular protocols and technical constraints when studying OS in isopods (a particular group of arthropods). I am quite familiar with the standard methods in birds and mammals.
First, you should decide on what tissue do your analysis. (The hemolymph of isopods could be a solution). The other issue is the method. About this, you must decide whether to search the element that generates oxidative stress or the effects of stress. In the first case would be good the DCFDA and an analysis of a flow cytometer, in the second you try to find oxidative damage to DNA. These are just some examples.
@Andrea Setini
The few papers I have read so far studied extracts or individual cells from the digestive glands, which would be interesting as our treatment would be applied on the feeding substrate of the woodlice.
On the other hand, the hemolymph might give us a better udnerstanding of what is going on at the whole organism level (or not, as the measures of OS on body fluids are always difficult to interpret). What kind of cells could be used for cytometry in the hemolymph? Are there any cells with functional cellular respiration in the hemolymph that we should be aware of when interpreting our results?
Haemocytes (sensu lato) are metabolically active cells and then can be used as biomarkers. I do not know the specific case of isopods but the idea is that you should make a brief preliminary study on subpopulations present (by flow cytometry) and then select only the portion that best responds to the ad hoc exposition.
one constraint in measuring oxidative stress in invertebrates is that particular organs give better measure of stress than other organs. Hence i suggest oxidative stress in hepatopancrease will give a good measure of stress in terrestrial isopods due to pollution and food quality. The hepatopancrease in invertebrates is analogous to the vertebrate liver and is critical for metabolism and detoxification.
We have worked on Biomarkers of oxidative stress but in the marine and terrestrial invertebrates (mussels, Annelida, crustaceans...) but not in mammals or birds!
however, There is several markers that could be developed in this aim, they are so simple and efficient: malinedialdehyde MDA, glutathione contents, some enzymatic activities: SOD superoxide dismutse, GSTs activities, GPX glutathione peroxydase... And other. In my opinion, these makers have been tested and validated in some mammls and birds.
Hi Charlotte Récapet,
I spent my Master and most of my PhD measuring terrestrial isopods biomarkers such as GST, GPx, CAT, LPO that give you information about oxidative stress in these organisms. Most of the works are already publish and you can download them from my Researchgate page. I hope his help.
Best Regards
- Badges
- Science topic
- Similar topics
- Biological Science
- Zoology
- Invertebrate Zoology
- Arthropods