The weaknesses of this study involve the mortality rate of their test subjects. I was shocked to hear in class that somewhere around 25% of the mice died during testing from seizures. This makes sense considering they are just activating a whole bunch of neurons with a simple injection. It makes me wonder whether this could be effecting the behavior of the mice that did survive the experiment. For example, could their fear response with and without CNO be a natural anxiety developed from brain damage from the experiment? Secondly, the results from Fig 1D are somewhat concerning as it takes about an hour for the mice to respond to CNO which could mess with results. Additionally, the results rarely ever revealed over a 50% freezing which are nor very strong results. Despite that, because of their control experiments, and logical experimental outline, the results are still valid, just not as strong as we could hope for.
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