Hi Anna,

A couple of questions and a few comments for consideration:

-Diana

Hi Anna

This ref could be useful

Thiele SL, Warre R, Nash JE. Development of a unilaterally-lesioned 6-OHDA mouse model of Parkinson’s disease. J Vis Exp 2012. doi:10.3791/3234.

If injecting manually with a stereotaxic frame, I would recommend to increase the inception volume to 2 ul. Inject just above the target (MFB). Wait 5 min before gradually pulling needle out. Unilateral injection should deplete 95% DA (I recommend mastering the technique by assaying stratal DA using HPLC before going for locomotor behavior analysis.

Diana L Price Thank you for your answer 1. It is unilateral and the injection site is MFB 2. no 3. I have no amphetamine so I use apomorphine and rota-rod test 4. Thank you I have stored it so I will prepare it fresh next time

Navneet Ammal Kaidery Thank you for your reply, I'm trying to follow this protocol most likely it's a matter of practice I have just started and I am a first person in our laboratory who makes stereotactic delivery of 6-OHDA. To evaluate DA neurons depletion I'm using TH staining of paraffin embedded brain sections (SNpc and VTA region), rota-rod and apomorphine test.

Hi Anna,

I would like to add one more thing. There should be significant loss of dopamine in the striatum of the 6-OHDA injected site, so that proper contralateral rotation can be seen following subcutaneous apomorphine treatment. Sometimes, if the ear bars of the mice are not properly fixed in the stereotaxy apparatus, and the coordinate of MFB is altered, then the injected toxin might reach some other part of the brain, and consequently expected dopamine depletion can't be found. Secondly, like apomorphine, 6-OHDA should also be made fresh and dissolved in anti-oxidant solution like .01% ascorbic acid, otherwise 6-OHDA will be readily oxidized.