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Quantitative analysis of intraneuronal transport in human iPS neurons.

Journal of pharmacological sciences (2015-08-04)
Haruko Nakamura, Naoya Yamashita, Yuri Kanamaru, Takahiko Tachibana, Yuko Sekino, Sandy Chen, Toshiyuki Gotoh, Fumiaki Tanaka, Yoshio Goshima
ABSTRACT

Induced pluripotent stem (iPS) cells are promising tools to investigate disease mechanism and develop new drugs. Intraneuronal transport, which is fundamental for neuronal survival and function, is vulnerable to various pharmacological and chemical agents and is disrupted in some neurodegenerative disorders. We applied a quantification method for axonal transport by counting CM-DiI-labeled particles traveling along the neurite, which allowed us to monitor and quantitate, for the first time, intraneuronal transport in human neurons differentiated from iPS cells (iCell neurons). We evaluated the acute effects of several anti-neoplastic agents that have been previously shown to affect intraneuronal transport. Vincristine, paclitaxel and oxaliplatin decreased the number of moving particle along neurites. Cisplatin, however, produced no effect on intraneuronal transport, which is in contrast to our previous report indicating that it inhibits transport in chick dorsal root ganglion neurons. Our system may be a useful method for assessing intraneuronal transport and neurotoxicity in human iPS neurons.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
CDI, reagent grade
Sigma-Aldrich
cis-Diamineplatinum(II) dichloride, ≥99.9% trace metals basis
Sigma-Aldrich
CDI, ≥97.0% (T)
Sigma-Aldrich
5-Fluorouracil, ≥99% (HPLC), powder
Sigma-Aldrich
Fluorouracil, meets USP testing specifications
Sigma-Aldrich
5-Fluorouracil, Vetec, reagent grade, ≥99%
Sigma-Aldrich
Methotrexate, meets USP testing specifications
Sigma-Aldrich
Oxaliplatin, powder
Sigma-Aldrich
cis-Diammineplatinum(II) dichloride, crystalline
Sigma-Aldrich
trans-Platinum(II)diammine dichloride
SAFC
Methotrexate