Small GTPase Rab17 Regulates Dendritic Morphogenesis and Postsynaptic Development of Hippocampal Neurons

Mori, Yasunori; Matsui, Takahide; Furutani, Yutaka; Yoshihara, Yoshihiro; Fukuda, Mitsunori

doi:10.1074/jbc.m111.314385

Cited by 39 publications

(50 citation statements)

References 44 publications

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“…Anti-Rab17 rabbit polyclonal antibody and anti-GFP guinea pig polyclonal antibody were prepared as described previously (21). Anti-Rabex-5 rabbit polyclonal antibody was raised against glutathione S-transferase (GST)-Rabex5-C (amino acid residues (aa) 208 -491) and affinity purified by exposure to antigen-bound Affi-Gel 10 beads (Bio-Rad) as described previously (24).…”

Section: Methodsmentioning

confidence: 99%

“…The cDNA of each of the Rabex-5 (or ALS2) constructs was subcloned into the pGAD-C1 vector, pEF-T7 vector (25), pEF-Myc vector (21), pEGFP-C1 vector (Clontech-Takara Bio Inc.), and/or pGEX-4T-3 vector (GE Healthcare). The pSilencer-EGFP vector (21) or pSilencer 2.1-U6 neo (Ambion, Austin, TX) encoding a mouse Rabex-5-shRNA (short hairpin RNA) (19-base target site 5Ј-GTTCAAGACATTGTTGAGA-3Ј), Rab5A-shRNA (19-base target site 5Ј-GCACAGTCCTATG-CAGATG-3Ј), Rab5B/C-shRNA (19-base target site 5Ј-GTTT-GAGATCTGGGACACA-3Ј), Rab21-shRNA (19-base target site 5Ј-TTTACTACCGAGATTCGAA-3Ј), and ALS2-shRNA (21-base target site 5Ј-GAACTCTTGCAAGATTTGTCA-3Ј) was constructed as described previously (21).…”

Section: Methodsmentioning

confidence: 99%

“…pEGFP-C1-Rabex-5 SR (an shRNA-resistant Rabex-5 mutant) and pEGFP-C1-Rabex-5 SR -D313A were produced by using the same method as described previously (21) and the following mutagenic oligonucleotides (substituted nucleotides are shown in italics): Rabex-5-SR-5Ј primer, 5Ј-CGGGATTG-CCAAGGAGGTACAGGATATAGTAGAAAAATACCCAC-TGGAG-3Ј, and Rabex-5-SR-3Ј primer, 5Ј-CTCCAGTGG-GTATTTTTCTACTATATCCTGTACCTCCTTGGCAATC-CCG-3Ј. pEF-FLAG-Rab17, pEF-Myc-Rab17, pEF-MycRab17-Q77L, pEGFP-C1-Rab17, pEGFP-C1-Rab17-Q77L, pmCherry-C1-Rab17, and pSilencer-EGFP-Rab17 were prepared as described previously (21).…”

Section: Methodsmentioning

confidence: 99%

“…pEF-FLAG-Rab17, pEF-Myc-Rab17, pEF-MycRab17-Q77L, pEGFP-C1-Rab17, pEGFP-C1-Rab17-Q77L, pmCherry-C1-Rab17, and pSilencer-EGFP-Rab17 were prepared as described previously (21). pEF-FLAG-Rab5A, pEF-FLAG-Rab5B, pEF-FLAG-Rab5C, and pEF-FLAG-Rab21 were also prepared as described previously (29,30).…”

Section: Methodsmentioning

confidence: 99%

“…Rab17 was originally described as an epithelial cell-specific Rab that regulates polarized trafficking (19,20), but the results of our previous study indicated that Rab17 is also expressed in mouse brain and that it regulates dendrite morphogenesis and postsynaptic development of hippocampal neurons (21). Because Rab17 is the only dendrite-specific Rab protein, unraveling the activation mechanism of Rab17 is crucial to a better understanding the molecular mechanism of dendrite outgrowth and branching.…”

mentioning

confidence: 99%

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Rabex-5 Protein Regulates Dendritic Localization of Small GTPase Rab17 and Neurite Morphogenesis in Hippocampal Neurons

Mori

Matsui²,

Fukuda

2013

Journal of Biological Chemistry

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Background: Small GTPase Rab17 regulates dendritic morphogenesis of hippocampal neurons, but its activation mechanism is completely unknown. Results: Expression of Rabex-5 in mouse hippocampal neurons promoted dendritic localization of Rab17, and Rabex-5 knockdown resulted in inhibition of neurite morphogenesis. Conclusion: Rabex-5 regulates neurite morphogenesis by activating Rab5 and Rab17. Significance: Our findings revealed a crucial role of Rabex-5 and its targets in neuronal development.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Rabex-5 Protein Regulates Dendritic Localization of Small GTPase Rab17 and Neurite Morphogenesis in Hippocampal Neurons

Mori

Matsui²,

Fukuda

2013

Journal of Biological Chemistry

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Cellular and molecular mechanisms regulating neuronal growth by brain‐derived neurotrophic factor

et al. 2016

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Brain-derived neurotrophic factor (BDNF) and its receptors TrkB and p75 regulate dendritic and axonal growth during development and maintenance of the mature nervous system; however, the cellular and molecular mechanisms underlying this process are not fully understood. In recent years, several advances have shed new light on the processes behind the regulation of BDNF-mediated structural plasticity including control of neuronal transcription, local translation of proteins, and regulation of cytoskeleton and membrane dynamics. In this review, we summarize recent advances in the field of BDNF signaling in neurons to induce neuronal growth. V C 2016 Wiley Periodicals, Inc.Key Words: BDNF; dendritic growth; signaling; cytoskeleton dynamics; membrane trafficking Introduction T he functionality of the nervous system (NS) depends on the extent of connectivity achieved by neurons. Although there is wide range of neuronal morphologies, neurons are specialized to form part of a given neuronal circuit and thus their morphology must adjust to this task [Gao, 2007]. In this regard, neurons are highly polarized cells composed of two main domains: the somato-dendritic compartment where neurons receive and integrate information from several axonal inputs and axons that are responsible for transmitting the action potential and conveying the information to the next relay in the network. A close relationship between the appropriate development of dendrites and axons and the functionality of the NS has been described. For example, alterations in dendrites, axonal inputs and dendritic spines are linked to neurodevelopmental and neuropathological conditions. Thus, the study of the mechanisms implicated in the regulation of neuronal morphology during development and maintenance of the adult NS is a focus of intense research [Armstrong et al., 1998;Wood et al., 2004;Bronfman et al., 2007;Dickstein et al., 2010;Cabeza et al., 2012;Eiland and McEwen, 2012;Kulkarni and Firestein, 2012].Neuronal morphology is regulated by both intrinsic and extrinsic factors, whose actions are overlapping. The intrinsic factors are described as the action of the genetic program of neurons that leads to a basic pattern of branching. In addition to this, there is an extensive list of molecules grouped as extrinsic factors that shape axonal and dendritic morphology throughout development and in response to sensory experience. These include bone morphogenetic family proteins (BMPs), semaphorins, Reelin, neurotrophins, and neuronal activity, among others [Jan and Jan, 2010]. The mechanisms underlying the effects of external cues to induce dendritic and axonal branching have only begun to be understood, and several steps including stimulation of intracellular signaling pathways to activate specific transcription factors, local synthesis of proteins and cellular membrane addition and turnover are involved. An interesting point of convergence is the effect of these external signals on the stability and dynamics of the cytoskeleton and the activity of molecular mo...

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Assay of Rab17 and Its Guanine Nucleotide Exchange Factor Rabex-5 in the Dendrites of Hippocampal Neurons

Mori

Fukuda

2015

Methods in Molecular Biology

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Neurons are functionally and morphologically compartmentalized into axons and dendrites, and the localization of specific 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 specifically 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 identified 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, filopodium 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, filopodium density, number of active synapses, and dendritic Rab17 signals.

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Small GTPase Rab17 Regulates Dendritic Morphogenesis and Postsynaptic Development of Hippocampal Neurons

Cited by 39 publications

References 44 publications

Rabex-5 Protein Regulates Dendritic Localization of Small GTPase Rab17 and Neurite Morphogenesis in Hippocampal Neurons

Rabex-5 Protein Regulates Dendritic Localization of Small GTPase Rab17 and Neurite Morphogenesis in Hippocampal Neurons

Cellular and molecular mechanisms regulating neuronal growth by brain‐derived neurotrophic factor

Assay of Rab17 and Its Guanine Nucleotide Exchange Factor Rabex-5 in the Dendrites of Hippocampal Neurons

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