Nerve Growth Factor Activates Persistent Rap1 Signaling in Endosomes

Wu, Chengbiao; Lai, Chun-Fai; Mobley, William C.

doi:10.1523/jneurosci.21-15-05406.2001

Cited by 220 publications

(185 citation statements)

References 52 publications

(59 reference statements)

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“…PC12 cells and were maintained as described (38). Embryonic DRG neurons were isolated from Sprague-Dawley fetal rats [embryonic day (E) 15 and 16], and cultured in DMEM containing 10% FCS and 50 ng/ml NGF as described by Chan et al (43).…”

Section: Methodsmentioning

confidence: 99%

“…To show that QD-NGF containing endosomes carry the NGF signal, we examined the distribution of these endosomes with respect to that of signaling proteins that were shown to mediate NGF signaling and associate with retrogradely transported endosomes under physiological conditions (11,38). For this study, we adopted a modified version of microfluidic nerve cell chamber (39), which enhances the ability to image and track QD signals (Fig.…”

Section: Most Qd-ngf Is Transported In Axons In Small 50-to 150-nm Vementioning

confidence: 99%

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One at a time, live tracking of NGF axonal transport using quantum dots

Cui¹,

Chen

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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338

336

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Retrograde axonal transport of nerve growth factor (NGF) signals is critical for the survival, differentiation, and maintenance of peripheral sympathetic and sensory neurons and basal forebrain cholinergic neurons. However, the mechanisms by which the NGF signal is propagated from the axon terminal to the cell body are yet to be fully elucidated. To gain insight into the mechanisms, we used quantum dot-labeled NGF (QD-NGF) to track the movement of NGF in real time in compartmentalized culture of rat dorsal root ganglion (DRG) neurons. Our studies showed that active transport of NGF within the axons was characterized by rapid, unidirectional movements interrupted by frequent pauses. Almost all movements were retrograde, but short-distance anterograde movements were occasionally observed. Surprisingly, quantitative analysis at the single molecule level demonstrated that the majority of NGFcontaining endosomes contained only a single NGF dimer. Electron microscopic analysis of axonal vesicles carrying QD-NGF confirmed this finding. The majority of QD-NGF was found to localize in vesicles 50 -150 nm in diameter with a single lumen and no visible intralumenal membranous components. Our findings point to the possibility that a single NGF dimer is sufficient to sustain signaling during retrograde axonal transport to the cell body.live imaging ͉ nerve growth factor ͉ single molecule imaging ͉ NGF signaling ͉ retrograde transport N erve growth factor (NGF) is produced and released by target tissues to activate specific receptors at the axon terminals of innervating neurons. In order for NGF to regulate gene expression and the survival of target neurons, a signal must be moved a considerable distance, in some cases Ͼ1,000-fold the diameter of the neuron cell body. The elucidation of the mechanism(s) used to transmit NGF signal from the terminals of axons to cell bodies of neurons is yet to be fully defined. In that retrograde NGF signaling is critical for the survival and maintenance of neurons of both the peripheral and central nervous systems, and the underlying mechanisms are likely to be shared by related neurotrophic factors, the issue remains one of the most significant and intriguing questions in neurobiology (1-16).In the past, radio-labeled NGF ( 125 I-NGF) has been used to study the binding, internalization, and axonal transport of NGF. These studies facilitated the measurement of transport rate and provided insights into the endocytic pathways used for NGF transport (8,(16)(17)(18)(19). Fluorescent labels such as rhodamine (20), Texas red (21), and Cy3 (22) were also used to track NGF movement in neurons. However, because these previous methods have limited spatial and temporal resolution, they provide only a coarse look at what is expected to be a very dynamic process. Moreover, few studies have examined NGF-containing endosomes during axonal transit. Some have suggested that NGF is transported principally within early endosomes (4, 5, 7-14) whereas others suggest that NGF is transported in organelles with com...

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Section: Methodsmentioning

confidence: 99%

Section: Most Qd-ngf Is Transported In Axons In Small 50-to 150-nm Vementioning

confidence: 99%

One at a time, live tracking of NGF axonal transport using quantum dots

Cui¹,

Chen

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

338

336

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“…Because Rap1 is required for NGF action in PC12 cells (Wu et al, 2001), the role of Rap1 activity in TIMP-2-mediated neuronal differentiation was examined using a dominant-negative mutant of Rap1, RapN17 (Vossler et al, 1997). PC12 cells were transfected with empty vectors or TIMP-2 alone or together with pDsRed-RapN17, and the extent of neurite outgrowth, as a hallmark of neuronal differentiation, was analyzed by fluorescence microscopy 72 h after transfection.…”

Section: Timp-2 Induces Growth Arrest Via Activation Of Integrin and mentioning

confidence: 99%

Tissue Inhibitor of Metalloproteinase-2 Promotes Neuronal Differentiation by Acting as an Anti-Mitogenic Signal

Pérez-Martı́nez

Jaworski²

2005

J. Neurosci.

110

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“…Interestingly, Rap1 appears to have a function in neurite outgrowth and neuronal polarization by mediating sustained MAPK pathway activation in addition to regulating the cytoskeleton (York et al, 1998;Kao et al, 2001;Wu et al, 2001;Schwamborn and Puschel, 2004). Further, Rap1 is also involved in the regulation of cell proliferation.…”

Section: Introductionmentioning

confidence: 99%

Anaplastic lymphoma kinase activates the small GTPase Rap1 via the Rap1-specific GEF C3G in both neuroblastoma and PC12 cells

et al. 2010

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Many different types of cancer originate from aberrant signaling from the anaplastic lymphoma kinase (ALK) receptor tyrosine kinase (RTK), arising through different translocation events and overexpression. Further, activating point mutations in the ALK domain have been recently reported in neuroblastoma. To characterize signaling in the context of the full-length receptor, we have examined whether ALK is able to activate Rap1 and contribute to differentiation/proliferation processes. We show that ALK activates Rap1 via the Rap1-specific guaninenucleotide exchange factor C3G, which binds in a constitutive complex with CrkL to activated ALK. The activation of the C3G/Rap1 pathway results in neurite outgrowth of PC12 cells, which is inhibited by either overexpression of Rap1GAP or siRNA-mediated knockdown of Rap1 itself or the guanine nucleotide exchange factor C3G. Significantly, this pathway also appears to function in the regulation of proliferation of neuroblastoma cells such as SK-N-SH and SH-SY5Y, because abrogation of Rap1 activity by Rap1-specific siRNA or overexpression of Rap1GAP reduces cellular growth. These results suggest that ALK activation of Rap1 may contribute to cell proliferation and oncogenesis of neuroblastoma driven by gain-of-function mutant ALK receptors.

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Nerve Growth Factor Activates Persistent Rap1 Signaling in Endosomes

Cited by 220 publications

References 52 publications

One at a time, live tracking of NGF axonal transport using quantum dots

One at a time, live tracking of NGF axonal transport using quantum dots

Tissue Inhibitor of Metalloproteinase-2 Promotes Neuronal Differentiation by Acting as an Anti-Mitogenic Signal

Anaplastic lymphoma kinase activates the small GTPase Rap1 via the Rap1-specific GEF C3G in both neuroblastoma and PC12 cells

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