Top

Published in:

Open Access 01-12-2016 | Research

Facilitation of axon outgrowth via a Wnt5a-CaMKK-CaMKIα pathway during neuronal polarization

Authors: Shin-ichiro Horigane, Natsumi Ageta-Ishihara, Satoshi Kamijo, Hajime Fujii, Michiko Okamura, Makoto Kinoshita, Sayaka Takemoto-Kimura, Haruhiko Bito

Published in: Molecular Brain | Issue 1/2016

Abstract

Background

Wnt5a, originally identified as a guidance cue for commissural axons, activates a non-canonical pathway critical for cortical axonal morphogenesis. The molecular signaling cascade underlying this event remains obscure.

Results

Through Ca²⁺ imaging in acute embryonic cortical slices, we tested if radially migrating cortical excitatory neurons that already bore primitive axons were sensitive to Wnt5a. While Wnt5a only evoked brief Ca²⁺ transients in immature neurons present in the intermediate zone (IZ), Wnt5a-induced Ca²⁺ oscillations were sustained in neurons that migrated out to the cortical plate (CP). We wondered whether this early Wnt5a-Ca²⁺ signaling during neuronal polarization has a morphogenetic consequence. During transition from round to polarized shape, Wnt5a administration to immature cultured cortical neurons specifically promoted axonal, but not dendritic, outgrowth. Pharmacological and genetic inhibition of the CaMKK-CaMKIα pathway abolished Wnt5a-induced axonal elongation, and rescue of CaMKIα in CaMKIα-knockdown neurons restored Wnt5a-mediated axon outgrowth.

Conclusions

This study suggests that Wnt5a activates Ca²⁺ signaling during a neuronal morphogenetic time window when axon outgrowth is critically facilitated. Furthermore, the CaMKK-CaMKIα cascade is required for the axonal growth effect of Wnt5a during neuronal polarization.

Available only for authorised users

Barnes AP, Polleux F. Establishment of axon-dendrite polarity in developing neurons. Annu Rev Neurosci. 2009;32:347–81.PubMedPubMedCentralCrossRef

Gordon MD, Nusse R. Wnt signaling: multiple pathways, multiple receptors, and multiple transcription factors. J Biol Chem. 2006;281:22429–33.PubMedCrossRef

Kikuchi A, Yamamoto H, Sato A. Selective activation mechanisms of Wnt signaling pathways. Trends Cell Biol. 2009;19:119–29.PubMedCrossRef

Freese JL, Pino D, Pleasure SJ. Wnt signaling in development and disease. Neurobiol Dis. 2010;38:148–53.PubMedPubMedCentralCrossRef

Salinas PC. Wnt signaling in the vertebrate central nervous system: from axon guidance to synaptic function. Cold Spring Harb Perspect Biol. 2012;4:a008003.PubMedPubMedCentralCrossRef

Kikuchi A, Yamamoto H, Sato A, Matsumoto S. Wnt5a: its signalling, functions and implication in diseases. Acta Physiol (Oxf). 2012;204:17–33.CrossRef

Yoshikawa S, McKinnon RD, Kokel M, Thomas JB. Wnt-mediated axon guidance via the Drosophila Derailed receptor. Nature. 2003;422:583–8.PubMedCrossRef

Keeble TR. The Wnt receptor Ryk is required for Wnt5a-mediated axon guidance on the contralateral side of the corpus callosum. J Neurosci. 2006;26:5840–8.PubMedCrossRef

Li L, Hutchins BI, Kalil K. Wnt5a induces simultaneous cortical axon outgrowth and repulsive axon guidance through distinct signaling mechanisms. J Neurosci. 2009;29:5873–83.PubMedPubMedCentralCrossRef

10.

Liu Y, Shi J, Lu C-C, Wang Z-B, Lyuksyutova AI, Song X, et al. Ryk-mediated Wnt repulsion regulates posterior-directed growth of corticospinal tract. Nat Neurosci. 2005;8:1151–9.PubMedCrossRef

11.

Hutchins BI, Li L, Kalil K. Wnt/calcium signaling mediates axon growth and guidance in the developing corpus callosum. Dev Neurobiol. 2011;71:269–83.PubMedPubMedCentralCrossRef

12.

Li L, Fothergill T, Hutchins BI, Dent EW, Kalil K. Wnt5a evokes cortical axon outgrowth and repulsive guidance by tau mediated reorganization of dynamic microtubules. Dev Neurobiol. 2014;74:797–817.PubMedPubMedCentralCrossRef

13.

Bodmer D, Levine-Wilkinson S, Richmond A, Hirsh S, Kuruvilla R. Wnt5a mediates nerve growth factor-dependent axonal branching and growth in developing sympathetic neurons. J Neurosci. 2009;29:7569–81.PubMedPubMedCentralCrossRef

14.

Nairn AC, Greengard P. Purification and characterization of Ca²⁺/calmodulin-dependent protein kinase I from bovine brain. J Biol Chem. 1987;262:7273–81.PubMed

15.

Yokokura H, Terada O, Naito Y, Hidaka H. Isolation and comparison of rat cDNAs encoding Ca²⁺/calmodulin-dependent protein kinase I isoforms. Biochim Biophys Acta. 1997;1338:8–12.PubMedCrossRef

16.

Takemoto-Kimura S, Terai H, Takamoto M, Ohmae S, Kikumura S, Segi E, et al. Molecular cloning and characterization of CLICK-III/CaMKIgamma, a novel membrane-anchored neuronal Ca²⁺/calmodulin-dependent protein kinase (CaMK). J Biol Chem. 2003;278:18597–605.PubMedCrossRef

17.

Ishikawa Y, Tokumitsu H, Inuzuka H, Murata-Hori M, Hosoya H, Kobayashi R. Identification and characterization of novel components of a Ca²⁺/calmodulin-dependent protein kinase cascade in HeLa cells. FEBS Lett. 2003;550:57–63.PubMedCrossRef

18.

Soderling TR. The Ca-calmodulin-dependent protein kinase cascade. Trends Biochem Sci. 1999;24:232–6.PubMedCrossRef

19.

Bito H, Takemoto-Kimura S. Ca²⁺/CREB/CBP-dependent gene regulation: a shared mechanism critical in long-term synaptic plasticity and neuronal survival. Cell Calcium. 2003;34:425–30.PubMedCrossRef

20.

Takemoto-Kimura S, Suzuki K, Kamijo S, Ageta-Ishihara N, Fujii H, Okuno H, et al. Differential roles for CaM kinases in mediating excitation-morphogenesis coupling during formation and maturation of neuronal circuits. Eur J Neurosci. 2010;32:224–30.PubMedCrossRef

21.

Ageta-Ishihara N, Takemoto-Kimura S, Nonaka M, Adachi-Morishima A, Suzuki K, Kamijo S, et al. Control of cortical axon elongation by a GABA-driven Ca²⁺/calmodulin-dependent protein kinase cascade. J Neurosci. 2009;29:13720–9.PubMedPubMedCentralCrossRef

22.

Takemoto-Kimura S, Ageta-Ishihara N, Nonaka M, Adachi-Morishima A, Mano T, Okamura M, et al. Regulation of dendritogenesis via a lipid-raft-associated Ca²⁺/calmodulin-dependent protein kinase CLICK-III/CaMKIgamma. Neuron. 2007;54:755–70.PubMedCrossRef

23.

Hatanaka Y, Namikawa T, Yamauchi K, Kawaguchi Y. Cortical divergent projections in mice originate from two sequentially generated, distinct populations of excitatory cortical neurons with different initial axonal outgrowth characteristics. Cereb Cortex. 2015; in press.

24.

Tabata H, Nakajima K. Efficient in utero gene transfer system to the developing mouse brain using electroporation: visualization of neuronal migration in the developing cortex. Neuroscience. 2001;103:865–72.PubMedCrossRef

25.

Saito T, Nakatsuji N. Efficient gene transfer into the embryonic mouse brain using in vivo electroporation. Dev Biol. 2001;240:237–46.PubMedCrossRef

26.

Wayman GA, Kaech S, Grant WF, Davare M, Impey S, Tokumitsu H, et al. Regulation of axonal extension and growth cone motility by calmodulin-dependent protein kinase I. J Neurosci. 2004;24:3786–94.PubMedCrossRef

27.

Schmitt JM, Wayman GA, Nozaki N, Soderling TR. Calcium activation of ERK mediated by calmodulin kinase I. J Biol Chem. 2004;279:24064–72.PubMedCrossRef

28.

Uboha NV, Flajolet M, Nairn AC, Picciotto MR. A calcium- and calmodulin-dependent kinase Ialpha/microtubule affinity regulating kinase 2 signaling cascade mediates calcium-dependent neurite outgrowth. J Neurosci. 2007;27:4413–23.PubMedCrossRef

29.

Nakamuta S, Funahashi Y, Namba T, Arimura N, Picciotto MR, Tokumitsu H, et al. Local application of neurotrophins specifies axons through inositol 1,4,5-trisphosphate, calcium, and Ca²⁺/calmodulin-dependent protein kinases. Sci Signal. 2011;4:ra76.PubMedCrossRef

30.

Wayman GA, Impey S, Marks D, Saneyoshi T, Grant WF, Derkach V, et al. Activity-dependent dendritic arborization mediated by CaM-kinase I activation and enhanced CREB-dependent transcription of Wnt-2. Neuron. 2006;50:897–909.PubMedCrossRef

31.

Saneyoshi T, Wayman G, Fortin D, Davare M, Hoshi N, Nozaki N, et al. Activity-dependent synaptogenesis: regulation by a CaM-kinase kinase/CaM-kinase I/betaPIX signaling complex. Neuron. 2008;57:94–107.PubMedPubMedCentralCrossRef

32.

Tokumitsu H, Chijiwa T, Hagiwara M, Mizutani A, Terasawa M, Hidaka H. KN-62, 1-[N, O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazi ne, a specific inhibitor of Ca2+/calmodulin-dependent protein kinase II. J Biol Chem. 1990;265:4315–20.PubMed

33.

Sumi M, Kiuchi K, Ishikawa T, Ishii A, Hagiwara M, Nagatsu T, et al. The newly synthesized selective Ca²⁺/calmodulin dependent protein kinase II inhibitor KN-93 reduces dopamine contents in PC12h cells. Biochem Biophys Res Commun. 1991;181:968–75.PubMedCrossRef

34.

Mochizuki H, Ito T, Hidaka H. Purification and characterization of Ca²⁺/calmodulin-dependent protein kinase V from rat cerebrum. J Biol Chem. 1993;268:9143–7.PubMed

35.

Enslen H, Sun P, Brickey D, Soderling S, Klamo E, Soderling T. Characterization of Ca²⁺/calmodulin-dependent protein kinase IV. Role in transcriptional regulation. J Biol Chem. 1994;269:15520–7.PubMed

36.

Tokumitsu H, Inuzuka H, Ishikawa Y, Ikeda M, Saji I, Kobayashi. STO-609, a specific inhibitor of the Ca²⁺/calmodulin-dependent protein kinase kinase. J Biol Chem. 2002;277:15813–8.PubMedCrossRef

37.

Kamata A, Sakagami H, Tokumitsu H, Sanda M, Owada Y, Fukunaga K, et al. Distinct developmental expression of two isoforms of Ca²⁺/calmodulin-dependent protein kinase kinases and their involvement in hippocampal dendritic formation. Neurosci Lett. 2007;423:143–8.PubMedCrossRef

38.

Tokumitsu H, Inuzuka H, Ishikawa Y, Kobayashi R. A single amino acid difference between alpha and beta Ca²⁺/calmodulin-dependent protein kinase kinase dictates sensitivity to the specific inhibitor, STO-609. J Biol Chem. 2003;278:10908–13.PubMedCrossRef

39.

Fujiwara Y, Hiraoka Y, Fujimoto T, Kanayama N, Magari M, Tokumitsu H. Analysis of distinct roles of CaMKK isoforms using STO-609-resistant mutants in living cells. Biochemistry. 2015;54:3969–77.PubMedCrossRef

40.

Boitard M, Bocchi R, Egervari K, Petrenko V, Viale B, Gremaud S, et al. Wnt signaling regulates Multipolar-to-Bipolar transition of migrating neurons in the cerebral cortex. Cell Rep. 2015;10:1349–61.PubMedCrossRef

41.

Witze ES, Connacher MK, Houel S, Schwartz MP, Morphew MK, Reid L, et al. Wnt5a directs polarized calcium gradients by recruiting cortical endoplasmic reticulum to the cell trailing edge. Dev Cell. 2013;26:645–57.PubMedCrossRef

42.

Cuitino L, Godoy JA, Farías GG, Couve A, Bonansco C, Fuenzalida M, et al. Wnt-5a modulates recycling of functional GABAA receptors on hippocampal neurons. J Neurosci. 2010;30:8411–20.PubMedCrossRef

43.

Wayman GA, Lee Y-S, Tokumitsu H, Silva AJ, Silva A, Soderling TR. Calmodulin-kinases: modulators of neuronal development and plasticity. Neuron. 2008;59:914–31.PubMedPubMedCentralCrossRef

44.

Namiki S, Sasaki T, Matsuki N, Ikegaya Y. Regional difference in stainability with calcium-sensitive acetoxymethyl-ester probes in mouse brain slices. Int J Neurosci. 2009;119:214–26.PubMedCrossRef

45.

Namiki S, Ikegaya Y. Current application and technology of functional multineuron calcium imaging. Biol Pharm Bull. 2009;32:1–9.PubMedCrossRef

46.

Ohkura M, Sasaki T, Sadakari J, Gengyo-Ando K, Kagawa-Nagamura Y, Kobayashi C, et al. Genetically encoded green fluorescent Ca²⁺ indicators with improved detectability for neuronal Ca²⁺ signals. PLoS One. 2012;7, e51286.PubMedPubMedCentralCrossRef

47.

Chen Y, Stevens B, Chang J, Milbrandt J, Barres BA, Hell JW. NS21: re-defined and modified supplement B27 for neuronal cultures. J Neurosci Methods. 2008;171:239–47.PubMedPubMedCentralCrossRef

48.

Blaeser F, Sanders MJ, Truong N, Ko S, Wu LJ, Wozniak DF, et al. Long-term memory deficits in Pavlovian fear conditioning in Ca²⁺/calmodulin kinase kinase alpha-deficient mice. Mol Cell Biol. 2006;26:9105–15.PubMedPubMedCentralCrossRef

49.

Peng I-C, Chen Z, Sun W, Li Y-S, Marin TL, Hsu P-H, et al. Glucagon regulates ACC activity in adipocytes through the CAMKKβ/AMPK pathway. Am J Physiol Endocrinol Metab. 2012;302:E1560–8.PubMedPubMedCentralCrossRef

Title: Facilitation of axon outgrowth via a Wnt5a-CaMKK-CaMKIα pathway during neuronal polarization
Authors: Shin-ichiro Horigane
Natsumi Ageta-Ishihara
Satoshi Kamijo
Hajime Fujii
Michiko Okamura
Makoto Kinoshita
Sayaka Takemoto-Kimura
Haruhiko Bito
Publication date: 01-12-2016
Publisher: BioMed Central
Published in: Molecular Brain / Issue 1/2016
Electronic ISSN: 1756-6606
DOI: https://doi.org/10.1186/s13041-016-0189-3

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Facilitation of axon outgrowth via a Wnt5a-CaMKK-CaMKIα pathway during neuronal polarization

Abstract

Background

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2016

Metabonomic profiles delineate potential role of glutamate-glutamine cycle in db/db mice with diabetes-associated cognitive decline

Glycine confers neuroprotection through microRNA-301a/PTEN signaling

Reviewer acknowledgement

Pathological classification of human iPSC-derived neural stem/progenitor cells towards safety assessment of transplantation therapy for CNS diseases

Dental noise exposed mice display depressive-like phenotypes

Positive geotactic behaviors induced by geomagnetic field in Drosophila