Hello,
For a behavioral paradigm, if the main measure does not show significant difference, but other measure related to the paradigm shows significant difference, can the other measure be correlated with deficiency?
To be specific, in Light/Dark Box assay, mice did not show significant changes in amount of time spent in light zone, but did show significant differences in number of transitions between the two zones. Can I use this measure to prove my hypothesis?
Thanks,
Goutham Kodakandla
Hi,
Kindly refer the following:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3792196/
Number of transitions could be confounded by increased locomotor activity so you should also determine that your drug/treatment is not having this effect. Typically amount of time spent in the light versus dark side is the primary measure examined for indications that a drug treatment may increase/reduce anxiety-like behavior. You should be deciding on the measure you are interested in a priori and then running your experiment in order to either accept or reject the null hypothesis.
It could also be worthwhile to look at a video recording if such a thing exists for your experiments. Perhaps there are some artifacts in the behavior in either of the groups that you can spot there that could explain all the transitions.
In addition, your hypothesis should be stated in such a way that by running the experiment you get a clear answer as to whether you should reject or accept the null hypothesis.
Number of transitions is mainly linked to locomotor activity, but can be linked with anxiety too (kind of "risk taking"). If you want to be sure your mice are more or less anxious than control, maybe you can use other additionnal tests (omaze, or elevated plus maze).
Good luck with your mice !
Hi
Although you may have got your answer. My view is that you should repeat your study for main paradigm and do video recording this time. If you are getting same results again with fresh group of mice then you may decide for your hypothesis based on these results. Although, as specified by you the other tests have given significant differences then you may link the increased frequency to anti-anxiety effect rather than enhanced kinetic effect as it can be linked to reduced anxiety in a light/dark box. And you should go for some biochemical marker to rule out any animal to animal behavioral variability due to pharmacokinetic/dynamic differences.
Hope it made some sense to your problem.
Good Luck.....
Hi,
I have been using this test for the last 6 years with good results. I normally use 3 parameters, latency to first exit (i start with the animal on the less anxiogenic, black compartment), number of transitions and time spent in the lit compartment. you can additionally score time spent by the walls vs time in the central area of the DLB if using any software that allows you to do so.
Yes, you can use the said parameter, but as mentioned by others, you need to distinguish the anxiogenic and locomotor activity. One way of doing so is running an open-field test or, an easier and reliable alternative, a novel cage exploration test, which runs as described below
Animals are put on a cage similar to normal type II home cage, with little sawdust and illuminated by red (not thermal) light ($8/bulb, on normal lamps, for example - as mice do not see red and allow you to see, record and score) for the length of 5 min. you count the number of rearings, i.e. the number of times the animal stands on hindpaws and explores the surroundings, which also include those by the wall with this supporting animal standing up. a new rearing is counted when all fours are back on the floor and risen again.
Please note that when using the dark-light box paradigm it works best (more sensitive, without induction of locomotory side-effects) under low lighting, between 5-25 lux.
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