Ca2+-dependent regulation of rho GTPases triggers turning of nerve growth cones.
Cytoplasmic Ca2+ elevation and changes in Rho GTPase activity are both known to mediate axon guidance by extracellular factors, but the causal relationship between these two events has been unclear. Here we show that direct elevation of cytoplasmic Ca2+ by extracellular application of a low concentration of ryanodine, which activated Ca2+ release from intracellular stores, upregulated Cdc42/Rac, but downregulated RhoA, in cultured cerebellar granule cells and human embryonic kidney 293T cells. Chemoattractive turning of the growth cone triggered by a gradient of ryanodine was blocked by overexpression of mutant forms of Cdc42 but not of RhoA in Xenopus spinal cord neurons. Furthermore, Ca2+-induced GTPase activity correlated with activation of protein kinase C and required a basal activity of Ca2+/calmodulin-dependent protein kinase II. Thus, Rho GTPases may mediate axon guidance by linking upstream Ca2+ signals triggered by guidance factors to downstream cytoskeletal rearrangements.
Jin
Ming
M
Institute of Neuroscience, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, and Graduate School of the Chinese Academy of Sciences, Shanghai 200031, China.
Guan
Chen-bing
CB
Jiang
Yun-ai
YA
Chen
Gang
G
Zhao
Chun-tao
CT
Cui
Kai
K
Song
Yuan-quan
YQ
Wu
Chien-ping
CP
Poo
Mu-ming
MM
Yuan
Xiao-bing
XB
D000818:Animals; D001426:Bacterial Proteins / Q000494:pharmacology; D001427:Bacterial Toxins / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D002614:Chelating Agents / Q000494:pharmacology; D003952:Diagnostic Imaging / Q000379:methods; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D004625:Embryo, Nonmammalian; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000494:pharmacology; D020439:Growth Cones / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D009414:Nerve Growth Factors / Q000494:pharmacology; D000075388:Netrin-1; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D012433:Ryanodine / Q000494:pharmacology; D013116:Spinal Cord / Q000166:cytology; D019284:Thapsigargin / Q000494:pharmacology; D013997:Time Factors; D014162:Transfection / Q000379:methods; D025521:Tumor Suppressor Proteins / Q000494:pharmacology; D014981:Xenopus; D020764:cdc42 GTP-Binding Protein / Q000494:pharmacology; D020741:rho GTP-Binding Proteins / Q000378:metabolism*
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D019208:Brain-Derived Neurotrophic Factor; D002614:Chelating Agents; D004791:Enzyme Inhibitors; D009414:Nerve Growth Factors; D025521:Tumor Suppressor Proteins; C031815:toxB protein, Clostridium difficile; D049452:Green Fluorescent Proteins; D012433:Ryanodine; D000075388:Netrin-1; D004533:Egtazic Acid; D019284:Thapsigargin; D020764:cdc42 GTP-Binding Protein; D020741:rho GTP-Binding Proteins; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D002118:Calcium
Curr Opin Neurobiol. 1999 Jun;9(3):355-63
10395576
J Biol Chem. 2001 Jan 19;276(3):1677-80
11058583
Neuron. 2001 Oct 11;32(1):1-3
11604131
Cell. 2001 Oct 19;107(2):209-21
11672528
J Biol Chem. 2000 Jun 23;275(25):18732-8
10749865
J Cell Biol. 2001 Dec 10;155(6):1029-42
11733543
Neuroreport. 1997 May 27;8(8):2005-9
9223093
J Neurosci. 1998 Jul 1;18(13):4985-92
9634564
Prog Brain Res. 1994;102:49-60
7800832
Pharmacol Rev. 1997 Mar;49(1):1-51
9085308
J Biol Chem. 2002 Nov 8;277(45):43115-20
12183458
Nature. 2002 Mar 28;416(6879):442-7
11919635
J Neurosci. 1997 Jun 1;17(11):4094-100
9151726
J Neurosci. 2002 Jan 1;22(1):239-47
11756507
Nat Cell Biol. 2003 Jan;5(1):38-45
12510192
J Biol Chem. 2003 Oct 10;278(41):39413-21
12888567
Nat Neurosci. 2002 Dec;5(12):1302-8
12426574
Glia. 2001 Mar 15;33(4):288-97
11246227
Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):3035-40
10077632
Biophys J. 2002 Jan;82(1 Pt 1):50-63
11751295
Development. 2000 Mar;127(5):969-80
10662636
J Cell Biol. 2000 Apr 17;149(2):263-70
10769020
Nature. 2002 Dec 12;420(6916):629-35
12478284
Trends Cell Biol. 2004 Jun;14(6):320-30
15183189
Neuron. 2002 Jul 3;35(1):51-63
12123608
Cell Motil Cytoskeleton. 2002 Dec;53(4):293-316
12378540
Nature. 2000 Jan 6;403(6765):93-8
10638760
J Biol Chem. 1998 Aug 14;273(33):20685-8
9694808
Neuron. 2004 Sep 16;43(6):835-46
15363394
Science. 2001 Mar 9;291(5510):1983-7
11239161
J Biol Chem. 1999 Apr 30;274(18):12753-8
10212259
Cell. 1998 Mar 20;92(6):785-95
9529254
Curr Opin Neurobiol. 2001 Feb;11(1):103-10
11179879
Nat Cell Biol. 2001 Mar;3(3):E81-8
11231595
J Biol Chem. 2002 Apr 26;277(17):15207-14
11844789
Nature. 1994 Mar 10;368(6467):140-4
8139655
J Neurosci. 1996 Feb 1;16(3):1140-9
8558243
FEBS Lett. 1998 Jun 16;429(3):229-33
9662423
Neuron. 2001 Feb;29(2):441-52
11239434
Science. 2000 Feb 11;287(5455):1037-40
10669415
Science. 2002 Dec 6;298(5600):1959-64
12471249
Nature. 2000 Jan 6;403(6765):89-93
10638759
EMBO J. 1999 Feb 1;18(3):578-85
9927417
Nature. 2002 Nov 7;420(6911):74-8
12422217
J Neurosci. 1992 Apr;12(4):1253-61
1372932
Proc Natl Acad Sci U S A. 2002 Sep 17;99(19):12085-90
12196628
J Biol Chem. 1999 Jun 18;274(25):18055-9
10364257
Eur J Neurosci. 2002 Jun;15(12):1891-8
12099895
J Neurosci. 1995 Jan;15(1 Pt 2):511-9
7823159