problem in the injection of the rotenone to induce PD. I inject it 1.5mg/kg 1:1 PEG and DMSO, which results in a high mortality rate.
Hi Hani Hafez,
You need to provide more information in the question. Was it i.p. or i.m. or s.c. or p.o. route? Which animal? How many times did you give this injection and how often? What was the age and weight of the animal? What are the concentrations of DMSO and PEG? PEG grade? The very simple answer is not to use both DMSO and PEG, but only one of thse. DMSO could be upto 30% (maximum, but with 1.5 mg doses per Kg, you can restrict it within 12%). You may use i.p. route in rats. Make the stock solution in 100% DMSO and dilute in saline or water to get the injection volume/concentration containing this amount of DMSO. Rotenone will have cardiotoxicity, and there will be fatalities with multiple injections, but not too high.
We use intracranial infsuion which allows you to use extremely small concentrations of rotenone to be delivered into median forebrain bundle (or intranigral or intrastriatal), with no cardiac toxicity but with specific nigrostriatal lesions. If infused unilaterally this will give a you a model with advantages so that yoiu could monitor drug actions in terms of ipasilateral and contralkateral rotation behaviors when administered with amphetamine or apomorphine, respectively. You may also use 'head swing test', or 'forelimb assymmetry test' or 'column test', methods which are non-drug induced behaviours for testing unilateral bias, as indication of unilateral nigrostriatal lesions. We have extensively used / published these methods for drug discovery programs.Good luck!
Thanks Prof MohanaKomur
I used 1:1 DMSO: PEG 30% and the injection was IP three times per week.
but I used DMSO 30%.
Further, should you send me the details and techniques for intracranial infusion.
thanks Hani
Dear Hani,
These are some of the citations of our methods paper. Please see the following three for intracranial infusion studies: Saravanan KS et al., Brain Res. 1049, 147-155, 2005; Sindhu KM et al., Brain Res. 1051, 25-34, 2005; Sindhu KM et al., Pharmacol. Biochem. Behav. 84; 321-329, 2006. The follwoing paper may be consulted for certain behaviour withour drugs: Tripathy D et al., PLoS ONE, 8(9): e72501, 2013.
The last one is available freely. In case you require the others' copies please let me know at [email protected], so that I can send these personally to you.
With best wishes,
Mohan
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