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Axonal/Dendritic Growth and Circuit Formation

開催日 2014/9/11
時間 16:00 - 17:00
会場 Poster / Exhibition(Event Hall B)

Slow axonal growth in human iPSCs-derived neurons

  • P1-092
  • 六本木 麗子 / Reiko T. Roppongi:1 大原 由貴 / Yuki Ohara:1 小金澤 紀子 / Noriko Koganezawa:1 山崎 博幸 / Hiroyuki Yamazaki:1 大津 真生 / Mao Ootsu:1 佐藤 薫 / Kaoru Sato:2 関野 祐子 / Yuko Sekino:2 白尾 智明 / Tomoaki Shirao:1 
  • 1:群馬大学大学院 / Dept Neurobiol and Behav, Gunma Univ Grad Sch Med, Gunma, Japan 2:国立医薬品食品衛生研薬理 / Div Pharmacol, NIHS, Tokyo, Japan 

Recent advances in human induced pluripotent stem cells (hiPSCs) offer new possibilities for biomedical research and clinical applications. In this study, we analyzed neuronal development of hiPSCs-derived neurons particularly focusing on their early development stages. We cultured iCell Neuron (CDI) and compared their development with that of the primary cultured neurons derived from rat hippocampus. In 2 days in vitro (DIV), we observed three different stages which were stages 1, 2 and 3 in developmental classification proposed by Dotti. Most developed stage-3 neurons had several short neurites with one long neurite, which is destined for an axon. In the DIV-2 iCell Neuron, we observed all three stages of neurons similar to rat neurons, while the morphologies of iCell Neuron and rat neurons were different. To reveal the difference in the morphological development between them, neurite length and numbers, branching points, and axon length were measured sequentially. There were significant decreases in all parameters in iCell Neurons. It is suggested that iCell Neurons develop at a slower speed. However, there were quite a few neurons that have a neurite immunostained with anti-phosphorylated neurofilament antibody. This indicates that although the growth of the neurites of the iCell Neuron is slower, the polarization occurs normally compare to the rat neurons. We further double-labeled these cells for drebrin and F-actin. F-actin is the main structural component of filopodia and lamellipodia. Drebrin is known to play an important role in the neurite formation. We found that the localization patterns of F-actin and drebrin in growth cones of iCell Neuron were similar to those of rat neurons. To test whether there is a difference in the effect on F-actin severing and depolymerization drug Cytochalasin D on the growth cone. Cytochalasin D caused drebrin and F-actin to shift from the transitional zone to the distal edge of growth cone in rat neurons and iCell Neurons. These data suggest that although the axonal growth speed of iCell Neurons is slower, the axonal polarity formation and the cytoskeleton in the growth cone are normal.

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