CXCL12/CXCR4 signaling pathway regulates cochlear development in neonatal mice

Zhang,Wen; Sun,Ji‑Zhou; Han,Yu; Chen,Jun; Liu,Hui; Wang,Ye; Yue,Bo; Chen,Yang

doi:10.3892/mmr.2016.5085

CXCL12/CXCR4 signaling pathway regulates cochlear development in neonatal mice

Authors:
- Wen Zhang
- Ji‑Zhou Sun
- Yu Han
- Jun Chen
- Hui Liu
- Ye Wang
- Bo Yue
- Yang Chen
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Affiliations: Department of Otolaryngology, Shaanxi Provincial People's Hospital, Xi'an, Shaanxi 710068, P.R. China, Department of Otolaryngology, Xi'an XD Group Hospital, Xi'an, Shaanxi 710077, P.R. China, Department of Otolaryngology, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China
Published online on: April 4, 2016 https://doi.org/10.3892/mmr.2016.5085
Pages: 4357-4364

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Abstract

Chemotactic cytokines (chemokines) are a highly conserved class of secreted signaling molecules that are important in various cellular processes. CXC chemokine ligand 12 (CXCL12) and its receptor, CXC chemokine receptor 4 (CXCR4) have been previously reported to be crucial for the establishment of neural networks in different neuronal systems. However, it is unclear whether the CXCL12/CXCR4 signaling pathway regulates the development of the cochlea. The current study investigated the effects of the CXCL12/CXCR4 signaling pathway on cochlear development in neonatal mice. The expression levels of CXCL12 and CXCR4 were detected using immunofluorescence, reverse transcription‑quantitative polymerase chain reaction and western blot analysis demonstrating that CXCL12 and CXCR4 expression were significantly increased during cochlear development in neonatal mice. Treatment of spiral ganglion neurons with CXCL12 significantly decreased the protein expression levels of caspase‑3 and cleaved caspase‑3, indicating that CXCL12/CXCR4 signaling increased cell survival of spiral ganglion neurons. Furthermore, CXCL12 treatment significantly increased the number and length of neurites extending from spiral ganglion neurons. By contrast, the in vitro effects of CXCL12 were significantly abrogated by AMD100, a CXCR4 antagonist. Additionally, inhibiting CXCL12/CXCR4 signaling in neonatal mice significantly reduced the cell number and altered the morphology of spiral ganglion neurons in vivo. Thus, the present study indicates that the CXCL12/CXCR4 signaling pathway is important during the development of cochleae in neonatal mice.

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1	Bok J, Zenczak C, Hwang CH and Wu DK: Auditory ganglion source of Sonic hedgehog regulates timing of cell cycle exit and differentiation of mammalian cochlear hair cells. Proc Natl Acad Sci USA. 110:13869–13874. 2013. View Article : Google Scholar : PubMed/NCBI
2	Tateya T, Imayoshi I, Tateya I, Hamaguchi K, Torii H, Ito J and Kageyama R: Hedgehog signaling regulates prosensory cell properties during the basal-to-apical wave of hair cell differentiation in the mammalian cochlea. Development. 140:3848–3857. 2013. View Article : Google Scholar : PubMed/NCBI
3	Barclay M, Ryan AF and Housley GD: Type I vs type II spiral ganglion neurons exhibit differential survival and neuritogenesis during cochlear development. Neural Dev. 6:332011. View Article : Google Scholar : PubMed/NCBI
4	Leake PA, Hradek GT, Hetherington AM and Stakhovskaya O: Brain-derived neurotrophic factor promotes cochlear spiral ganglion cell survival and function in deafened, developing cats. J Comp Neurol. 519:1526–1545. 2011. View Article : Google Scholar : PubMed/NCBI
5	Dabdoub A, Puligilla C, Jones JM, Fritzsch B, Cheah KS, Pevny LH and Kelley MW: Sox2 signaling in prosensory domain specification and subsequent hair cell differentiation in the developing cochlea. Proc Natl Acad Sci USA. 105:18396–18401. 2008. View Article : Google Scholar : PubMed/NCBI
6	Yang S, Edman LC, Sánchez-Alcañiz JA, Fritz N, Bonilla S, Hecht J, Uhlén P, Pleasure SJ, Villaescusa JC, Marín O and Arenas E: Cxcl12/Cxcr4 signaling controls the migration and process orientation of A9-A10 dopaminergic neurons. Development. 140:4554–4564. 2013. View Article : Google Scholar : PubMed/NCBI
7	Zou YR, Kottmann AH, Kuroda M, Taniuchi I and Littman DR: Function of the chemokine receptor CXCR4 in haematopoiesis and in cerebellar development. Nature. 393:595–599. 1998. View Article : Google Scholar : PubMed/NCBI
8	Nagasawa T: CXC chemokine ligand 12 (CXCL12) and its receptor CXCR4. J Mol Med Berl. 92:433–439. 2014. View Article : Google Scholar : PubMed/NCBI
9	Ma Q, Jones D, Borghesani PR, Segal RA, Nagasawa T, Kishimoto T, Bronson RT and Springer TA: Impaired B-lymphopoiesis, myelopoiesis, and derailed cerebellar neuron migration in CXCR4- and SDF-1-deficient mice. Proc Natl Acad Sci USA. 95:9448–9453. 1998. View Article : Google Scholar : PubMed/NCBI
10	Zhu Y, Matsumoto T, Mikami S, Nagasawa T and Murakami F: SDF1/CXCR4 signalling regulates two distinct processes of precerebellar neuronal migration and its depletion leads to abnormal pontine nuclei formation. Development. 136:1919–1928. 2009. View Article : Google Scholar : PubMed/NCBI
11	Stumm RK, Zhou C, Ara T, Lazarini F, Dubois-Dalcq M, Nagasawa T, Höllt V and Schulz S: CXCR4 regulates interneuron migration in the developing neocortex. J Neurosci. 23:5123–5130. 2003.PubMed/NCBI
12	Lieberam I, Agalliu D, Nagasawa T, Ericson J and Jessell TM: A Cxcl12-CXCR4 chemokine signaling pathway defines the initial trajectory of mammalian motor axons. Neuron. 47:667–679. 2005. View Article : Google Scholar : PubMed/NCBI
13	García-Hernández S, Potashner SJ and Morest DK: Role of fibroblast growth factor 8 in neurite outgrowth from spiral ganglion neurons in vitro. Brain Res. 1529:39–45. 2013. View Article : Google Scholar : PubMed/NCBI
14	Livak and Schmittgen: Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods. 25:402–408. 2001. View Article : Google Scholar
15	Xu N, Engbers J, Khaja S, Xu L, Clark JJ and Hansen MR: Influence of cAMP and protein kinase A on neurite length from spiral ganglion neurons. Hear Res. 283:33–44. 2012. View Article : Google Scholar :
16	Wang F, Gao X, Chen J, Liu SL, Wang FY, Hei RY, Chen Y and Qiu JH: Effect of early postnatal air-conduction auditory deprivation on the development and function of the rat spiral ganglion. J Laryngol Otol. 125:917–923. 2011. View Article : Google Scholar : PubMed/NCBI
17	Peng H, Kolb R, Kennedy JE and Zheng J: Differential expression of CXCL12 and CXCR4 during human fetal neural progenitor cell differentiation. J Neuroimmune Pharmacol. 2:251–258. 2007. View Article : Google Scholar : PubMed/NCBI
18	Schols D, Struyf S, Van Damme J, Esté JA, Henson G and De Clercq E: Inhibition of T-tropic HIV strains by selective antagonization of the chemokine receptor CXCR4. J Exp Med. 186:1383–1388. 1997. View Article : Google Scholar : PubMed/NCBI
19	Hartnick CJ, Staecker H, Malgrange B, Lefebvre PP, Liu W, Moonen G and Van de Water TR: Neurotrophic effects of BDNF and CNTF, alone and in combination, on postnatal day 5 rat acoustic ganglion neurons. J Neurobiol. 30:246–254. 1996. View Article : Google Scholar : PubMed/NCBI
20	Mullen LM, Pak KK, Chavez E, Kondo K, Brand Y and Ryan AF: Ras/p38 and PI3K/Akt but not Mek/Erk signaling mediate BDNF-induced neurite formation on neonatal cochlear spiral ganglion explants. Brain Res. 1430:25–34. 2012. View Article : Google Scholar
21	Aletsee C, Beros A, Mullen L, Palacios S, Pak K, Dazert S and Ryan AF: Ras/MEK but not p38 signaling mediates NT-3-induced neurite extension from spiral ganglion neurons. J Assoc Res Otolaryngol. 2:377–387. 2001. View Article : Google Scholar
22	Li M and Ransohoff RM: Multiple roles of chemokine CXCL12 in the central nervous system: A migration from immunology to neurobiology. Prog Neurobiol. 84:116–131. 2008. View Article : Google Scholar : PubMed/NCBI
23	Banisadr G, Fontanges P, Haour F, Kitabgi P, Rostène W and Mélik Parsadaniantz S: Neuroanatomical distribution of CXCR4 in adult rat brain and its localization in cholinergic and dopaminergic neurons. Eur J Neurosci. 16:1661–1671. 2002. View Article : Google Scholar : PubMed/NCBI
24	Krumbholz M, Theil D, Cepok S, Hemmer B, Kivisäkk P, Ransohoff RM, Hofbauer M, Farina C, Derfuss T, Hartle C, et al: Chemokines in multiple sclerosis: CXCL12 and CXCL13 up-regulation is differentially linked to CNS immune cell recruitment. Brain. 129:200–211. 2006. View Article : Google Scholar
25	Stumm RK, Rummel J, Junker V, Culmsee C, Pfeiffer M, Krieglstein J, Höllt V and Schulz S: A dual role for the SDF-1/CXCR4 chemokine receptor system in adult brain: Isoform-selective regulation of SDF-1 expression modulates CXCR4-dependent neuronal plasticity and cerebral leukocyte recruitment after focal ischemia. J Neurosci. 22:5865–5878. 2002.PubMed/NCBI
26	Oh SB, Tran PB, Gillard SE, Hurley RW, Hammond DL and Miller RJ: Chemokines and glycoprotein120 produce pain hypersensitivity by directly exciting primary nociceptive neurons. J Neurosci. 21:5027–5035. 2001.PubMed/NCBI
27	Limatola C, Giovannelli A, Maggi L, Ragozzino D, Castellani L, Ciotti MT, Vacca F, Mercanti D, Santoni A and Eusebi F: SDF-1α-mediated modulation of synaptic transmission in rat cerebellum. Eur J Neurosci. 12:2497–2504. 2000. View Article : Google Scholar : PubMed/NCBI
28	Gillard SE, Lu M, Mastracci RM and Miller RJ: Expression of functional chemokine receptors by rat cerebellar neurons. J Neuroimmunol. 124:16–28. 2002. View Article : Google Scholar : PubMed/NCBI
29	Ragozzino D, Renzi M, Giovannelli A and Eusebi F: Stimulation of chemokine CXC receptor 4 induces synaptic depression of evoked parallel fibers inputs onto Purkinje neurons in mouse cerebellum. J Neuroimmunol. 127:30–36. 2002. View Article : Google Scholar : PubMed/NCBI
30	Kilpatrick LA, Zhu J, Lee FS and Lang H: Role of stromal cell-derived factor-1 expression in the injured mouse auditory nerve. Otolaryngol Head Neck Surg. 145:1007–1015. 2011. View Article : Google Scholar : PubMed/NCBI
31	Zhang PZ, He Y, Jiang XW, Chen FQ, Chen Y, Xue T, Zhou K, Li X, Wang Y, Wu YX, et al: Up-regulation of stromal cell-derived factor-1 enhances migration of transplanted neural stem cells to injury region following degeneration of spiral ganglion neurons in the adult rat inner ear. Neurosci Lett. 534:101–106. 2013. View Article : Google Scholar