TY - JOUR
T1 - Assay of Rab17 and its guanine nucleotide exchange factor Rabex-5 in the dendrites of hippocampal neurons
AU - Mori, Yasunori
AU - Fukuda, Mitsunori
N1 - Publisher Copyright:
© Springer Science+Business Media New York 2015.
PY - 2015
Y1 - 2015
N2 - Neurons are functionally and morphologically compartmentalized into axons and dendrites, and the localization of specifi c proteins within these compartments is critical to the proper formation of neuronal networks, which includes neurite morphogenesis and synapse formation. The small GTPase Rab17 is specifi cally localized in dendrites and is not found in axons, and it regulates the dendrite morphogenesis and postsynaptic development of mouse hippocampal neurons. However, the spatiotemporal regulation of Rab17 is poorly understood. We recently identifi ed Rabex-5, originally described as a Rab5-guanine nucleotide exchange factor (GEF), as a physiological Rab17-GEF that promotes translocation of Rab17 from the cell body to the dendrites of developing hippocampal neurons. Knockdown of Rab17 in mouse hippocampal neurons resulted in reductions in dendrite growth, branch numbers, fi lopodium density, and active synapse numbers. Knockdown of Rab17-GEF Rabex-5 in hippocampal neurons resulted in decreased targeting of Rab17 to the dendrites, which led to a reduction in dendrite growth. In this chapter we describe the assay procedures for analyzing Rab17 and Rabex-5 in cultured mouse hippocampal neurons, and we particularly focus on the measurement of total dendrite (or axon) length and total dendrite (or axon) branch numbers, fi lopodium density, number of active synapses, and dendritic Rab17 signals.
AB - Neurons are functionally and morphologically compartmentalized into axons and dendrites, and the localization of specifi c proteins within these compartments is critical to the proper formation of neuronal networks, which includes neurite morphogenesis and synapse formation. The small GTPase Rab17 is specifi cally localized in dendrites and is not found in axons, and it regulates the dendrite morphogenesis and postsynaptic development of mouse hippocampal neurons. However, the spatiotemporal regulation of Rab17 is poorly understood. We recently identifi ed Rabex-5, originally described as a Rab5-guanine nucleotide exchange factor (GEF), as a physiological Rab17-GEF that promotes translocation of Rab17 from the cell body to the dendrites of developing hippocampal neurons. Knockdown of Rab17 in mouse hippocampal neurons resulted in reductions in dendrite growth, branch numbers, fi lopodium density, and active synapse numbers. Knockdown of Rab17-GEF Rabex-5 in hippocampal neurons resulted in decreased targeting of Rab17 to the dendrites, which led to a reduction in dendrite growth. In this chapter we describe the assay procedures for analyzing Rab17 and Rabex-5 in cultured mouse hippocampal neurons, and we particularly focus on the measurement of total dendrite (or axon) length and total dendrite (or axon) branch numbers, fi lopodium density, number of active synapses, and dendritic Rab17 signals.
KW - Dendrite
KW - Hippocampal neuron
KW - Postsynapse
KW - Rab17
KW - Rabex-5
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U2 - 10.1007/978-1-4939-2569-8_20
DO - 10.1007/978-1-4939-2569-8_20
M3 - Article
C2 - 25800847
AN - SCOPUS:84925949460
VL - 1298
SP - 233
EP - 243
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
ER -