Does the combination of nicotine and ethanol facilitate or block dopaminergic neuron damage in Parkinson's disease models?

2019 Fiscal Year Research-status Report

• Project/Area Number: 19K10687
• Research Institution: Kagawa University
• Principal Investigator: モストファ ジャーマル 香川大学, 医学部, 助教 (50418802)
• Co-Investigator(Kenkyū-buntansha): 塚本 郁子 香川大学, 医学部, 寄付講座教員 (10183477)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: MPTP / Mortality / Monoamines / MPP / cell culture / cell viability

Outline of Annual Research Achievements

1. Optimization of MPTP dose in mice:
The mortality at a 20 mg/kg dose was substantial, from 90-100%. At 15 mg/kg, the mortality rate is reduced to 50%. The result showed that dopamine, DOPAC and HVA were reduced to 60-70% in the striatum using HPLC-ECD.
2. Cell viability assay in PC12 cells:
Incubation with various concentrations of MPP+ (0, 200, 500, and 1000 uM) at 24 and 48 h did not significantly affect viable cell count and the LDH activity, indicating that MPP+ does not cause PC12 cell toxicity.

Current Status of Research Progress

4: Progress in research has been delayed.

Reason

MPTP is a highly neurotoxic compound. MPTP preparation and its administration to animal should be conducted in a fume hood. From April 2020-March 2021, the draft chamber is not available due to the construction of the animal building.

Strategy for Future Research Activity

April 2020-March 2021: a. HPLC-ECD analysis for dopamine release in MPP treated PC12 cells, b. Immunoblot for TH phosphorylation at Ser31 and Ser40, PARP-1 and caspase in MPP treated PC12 cells.

April 2021-March 2022: a. HPLC-ECD analysis of dopamine, serotonin and their metabolites in MPTP treated mice, b.Immunoblot for TH phosphorylation at Ser31 and Ser40, PARP-1 and caspase in MPTP treated mice

Causes of Carryover

To peruse cell culture, need to buy the following things
1. PC12 cells, 2. Antibodies for WB, 3. IBend Flex western kit