The MPTP intoxication protocol, the chronic L-DOPA treatment, the clinical assessments, the terminal procedure and the characterization of the extent of nigrostriatal denervation were conducted as previously published [3], [11], [12], [13], [14].
As a terminal procedure, the animal was deeply anesthetized and then injected i.v. with saline containing 10% sodium sulfide (200 mg/kg).
Injections were made as a terminal procedure, after optical imaging and electrophysiological recording, and targeted to patches physiologically identified as object-selective.
In addition, blood sampling to measure metabolite or hormone levels is generally a terminal procedure so repeated sampling is not possible.
100 μL caspase3/7-GLO reagent was directly added to each well of a 96-well plate as a terminal procedure (neurons are lysed) and incubated for 1 h.
In separate experiments, 100 μL of CellTiter-GLO assay reagent (Promega) was added directly to each well of a 96-well plate as a terminal procedure (neurons are lysed).
As terminal procedure, the knee cavity was washed with PBS (2×5 μl), the total number of leucocytes in the knee lavage was determined by counting the cells in a Neubauer chamber after staining with Turk's solution.
Blood for all studies was collected from the retroorbital venous plexus under anesthesia using isoflurane as previously described, with the exception of blood for thrombin antithrombin (TAT) evaluation, which was obtained under anesthesia from an open puncture of the inferior vena cava, performed as a terminal procedure.
As arterial catheterization is a terminal procedure, separate mice were anesthetized in the same way and a number 2 French micromanometer catheter (Mikro-tip SPR-838, Millar Instruments Inc., Houston, TX, USA) was advanced via the carotid artery into the ascending aorta to measure blood pressure.
