Cell lines

differentiated SH-SY5Y neuroblastoma cells

Preparation method

The solubility of this compound in DMSO is ≥77.6mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reacting condition

50 nM; 21 days

Applications

In differentiated SH-SY5Y neuroblastoma cells, Rotenone (50 nM) produced a biphasic survival curve with ~40% loss by 6 days and a second decline to ~60% loss between 18 and 21 days. Mitochondrial movement velocities were reduced at 8 days.

Animal models

20–25-weekold female BALB/c mice

Dosage form

(rotenone: 0.35 mg/kg; DMSO: 9.86 mg/kg) or vehicle (DMSO: 9.86 mg/kg); injected into the right-side nasal cavity of mice; once a day for 2 weeks

Application

In mice, rotenone attenuated the olfactory function and retarded the inhibitory input into the mitral cells, which are output neurons in the olfactory bulb (OB). Rotenone also caused neurite degeneration of DA neurons in the substantia nigra.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

Rotenone is an inhibitor of the mitochondrial complex I electron transport chain with IC50 value of 1.7 - 2.2 μM.

Electron transport chain (ETC) is a series of compounds that transfer electrons from electron donors to electron acceptors and transfer protons (H+ ions) across a membrane. The proton gradient drives ATP synthesis. Complex I is one of the main sites of production of superoxide.

Rotenone is an inhibitor of the mitochondrial complex I electron transport chain. In the transformed cell line HEK 293 and cancer cell lines U87, rotenone (50 μM) induced cell death by 30% and 40% respectively in a dose dependent way, which was mediated by reactive oxygen species (ROS). Also, rotenone significantly induced autophagy formation [1]. In SH-SY5Y cells, rotenone induced cell apoptosis in a caspase-dependent way. Also, rotenone induced phosphorylation of p38 MAP kinase, c-Jun and JNK, which indicated activation of the p38 and JNK pathways [2]. In differentiated SH-SY5Y neuroblastoma cells, rotenone (50 nM) induced cell death by 60% and slowed mitochondrial movement. While rotenone didn’t induce the formation of resembling Lewy bodies [3].

References:
[1].  Chen Y, McMillan-Ward E, Kong J, et al. Mitochondrial electron-transport-chain inhibitors of complexes I and II induce autophagic cell death mediated by reactive oxygen species. J Cell Sci, 2007, 120(Pt 23): 4155-4166.
[2].  Newhouse K, Hsuan SL, Chang SH, et al. Rotenone-induced apoptosis is mediated by p38 and JNK MAP kinases in human dopaminergic SH-SY5Y cells. Toxicol Sci, 2004, 79(1): 137-146.
[3].  Borland MK, Trimmer PA, Rubinstein JD, et al. Chronic, low-dose rotenone reproduces Lewy neurites found in early stages of Parkinson's disease, reduces mitochondrial movement and slowly kills differentiated SH-SY5Y neural cells. Mol Neurodegener, 2008, 3: 21.