One at a time, live tracking of NGF axonal transport using quantum dots.
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 NGF-containing 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.
Cui
Bianxiao
B
Department of Physics, Stanford University, Stanford, CA 94305, USA.
Wu
Chengbiao
C
Chen
Liang
L
Ramirez
Alfredo
A
Bearer
Elaine L
EL
Li
Wei-Ping
WP
Mobley
William C
WC
Chu
Steven
S
D000818:Animals; D001370:Axonal Transport / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D011992:Endosomes / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D008854:Microscopy, Electron; D020932:Nerve Growth Factor / Q000378:metabolism* / Q000494:pharmacology*; D045663:Quantum Dots; D051381:Rats; D017207:Rats, Sprague-Dawley; D020917:Receptor, trkA / Q000378:metabolism; D013997:Time Factors; D020696:rab5 GTP-Binding Proteins / Q000378:metabolism
D020932:Nerve Growth Factor; D020917:Receptor, trkA; D048049:Extracellular Signal-Regulated MAP Kinases; D020696:rab5 GTP-Binding Proteins
Science. 2002 Feb 22;295(5559):1536-9
11799202
J Cell Biol. 2002 Jan 21;156(2):233-9
11807088
J Cell Biol. 2002 May 13;157(4):679-91
12011113
Curr Opin Neurobiol. 2002 Jun;12(3):268-74
12049932
Hum Mol Genet. 2003 Apr 15;12(8):813-22
12668605
J Neurosci. 2003 Apr 15;23(8):3209-20
12716928
Neuron. 2003 Jul 3;39(1):57-68
12848932
Neuron. 2003 Jul 3;39(1):69-84
12848933
J Neurobiol. 2004 Feb 5;58(2):207-16
14704953
Nat Biotechnol. 2004 Feb;22(2):198-203
14704683
J Am Chem Soc. 2004 Feb 11;126(5):1324-5
14759174
Cell. 2004 Jul 23;118(2):243-55
15260993
Proc Natl Acad Sci U S A. 1977 Oct;74(10):4516-9
270699
J Neurosci. 1982 Apr;2(4):431-42
7069465
Neurosci Lett. 1983 Aug 19;39(1):1-4
6195560
J Neurochem. 1986 Feb;46(2):641-8
3001230
Brain Res. 1986 Jul;387(1):53-62
3742234
J Neurosci. 1997 Feb 15;17(4):1282-90
9006972
J Cell Biol. 1997 Jul 28;138(2):411-21
9230082
Cold Spring Harb Symp Quant Biol. 1996;61:389-406
9246468
Proc Natl Acad Sci U S A. 1997 Sep 2;94(18):9909-14
9275225
J Neurosci. 1997 Sep 15;17(18):7007-16
9278536
Brain Res. 1998 Jul 6;798(1-2):67-74
9666080
Brain Res Brain Res Protoc. 1999 Jan;3(3):308-12
9974146
J Neurosci. 1999 Sep 15;19(18):7889-900
10479691
J Neurosci. 2005 Mar 2;25(9):2181-91
15745944
Nano Lett. 2005 Apr;5(4):603-7
15826094
J Neurosci. 2005 May 25;25(21):5236-47
15917464
Nat Rev Neurosci. 2005 Aug;6(8):615-25
16062170
Nat Methods. 2005 Aug;2(8):599-605
16094385
Nat Methods. 2005 Oct;2(10):743-9
16179920
IEEE Trans Image Process. 2005 Sep;14(9):1384-95
16190473
Curr Opin Neurobiol. 2005 Oct;15(5):568-75
16150591
Biomaterials. 2006 Mar;27(9):1679-87
16318871
Hum Mol Genet. 2006 May 1;15(9):1451-63
16565160
Neuron. 2006 Jul 6;51(1):29-42
16815330
Science. 2000 Jun 16;288(5473):2048-51
10856219
Mol Biol Cell. 2000 Jul;11(7):2283-95
10888668
Methods Mol Biol. 2001;169:195-216
11142011
Cytometry. 2001 Mar 1;43(3):182-8
11170104
J Neurosci. 2001 Aug 1;21(15):5406-16
11466412
Annu Rev Neurosci. 2001;24:1217-81
11520933
Science. 2002 Feb 22;295(5559):1471-3
11859179