Skip to main content

Advertisement

SpringerLink
Log in

Obligatory Activation of SRC and JNK by GDNF for Survival and Axonal Outgrowth of Postnatal Intestinal Neurons

  • Original Research
  • Published:
Cellular and Molecular Neurobiology Aims and scope Submit manuscript

Abstract

The neurotrophin GDNF acts through its co-receptor RET to direct embryonic development of the intestinal nervous system. Since this continues in the post-natal intestine, co-cultures of rat enteric neurons and intestinal smooth muscle cells were used to examine how receptor activation mediates neuronal survival or axonal extension. GDNF-mediated activation of SRC was essential for neuronal survival and axon outgrowth and activated the major downstream signaling pathways. Selective inhibition of individual pathways had little effect on survival but JNK activation was required for axonal maintenance, extension or regeneration. This was localized to axonal endings and retrograde transport was needed for central JUN activation and subsequent axon extension. Collectively, GDNF signaling supports neuronal survival via SRC activation with multiple downstream events, with JNK signaling mediating structural plasticity. These pathways may limit neuron death and drive subsequent regeneration during challenges in vivo such as intestinal inflammation, where supportive strategies could preserve intestinal function.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Abbreviations

CNS:

Central nervous system

DMEM:

Dulbecco’s minimal essential medium

ENS:

Enteric nervous system

GDNF:

Glial cell line derived neurotrophic factor

ISMC:

Intestinal smooth muscle cell

JNK:

C-Jun N-terminal kinase

LY:

LY294002

PD:

PD98059

RET:

“Re-arranged on transfection”

SB:

SB203580

SP:

SP600125

TNBS:

Trinitrobenzene sulphonic acid

References

  • Airaksinen MS, Saarma M (2002) The GDNF family: signalling, biological functions and therapeutic value. Nat Rev Neurosci 3:383–394

    Article  CAS  PubMed  Google Scholar 

  • Ammori JB, Zhang WZ, Li JY, Chai BX, Mulholland MW (2008) Effects of ghrelin on neuronal survival in cells derived from dorsal motor nucleus of the vagus. Surgery 144:159–167

    Article  PubMed  Google Scholar 

  • Anitha M, Gondha C, Sutliff R, Parsadanian A, Mwangi S, Sitaraman SV, Srinivasan S (2006) GDNF rescues hyperglycemia-induced diabetic enteric neuropathy through activation of the PI3K/Akt pathway. J Clin Invest 116:344–356

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Aszmann OC, Winkler T, Korak K, Lassmann H, Frey M (2004) The influence of GDNF on the timecourse and extent of motoneuron loss in the cervical spinal cord after brachial plexus injury in the neonate. Neurol Res 26:211–217

    Article  CAS  PubMed  Google Scholar 

  • Bain J, Plater L, Elliott M, Shpiro N, Hastie CJ, McLauchlan H, Klevernic I, Arthur JS, Alessi DR, Cohen P (2007) The selectivity of protein kinase inhibitors: a further update. Biochem J 408:297–315

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Baloh RH, Enomoto H, Johnson EM Jr, Milbrandt J (2000) The GDNF family ligands and receptors - implications for neural development. Curr Opin Neurobiol 10:103–110

    Article  CAS  PubMed  Google Scholar 

  • Barnat M, Enslen H, Propst F, Davis RJ, Soares S, Nothias F (2010) Distinct roles of c-Jun N-terminal kinase isoforms in neurite initiation and elongation during axonal regeneration. J Neurosci 30:7804–7816

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Blennerhassett MG, Vignjevic P, Vermillion DL, Collins SM (1992) Inflammation causes hyperplasia and hypertrophy in smooth muscle of rat small intestine. Am J Physiol 262:G1041–G1046

    CAS  PubMed  Google Scholar 

  • Blennerhassett MG, Lourenssen SR, Parlow LRG, Ghasemlou N, Winterborn AN (2017) Analgesia and mouse strain influence neuromuscular plasticity in inflamed intestine. Neurogastroenterol Motil 29:1–12

    Article  CAS  PubMed  Google Scholar 

  • Bonafiglia QA, Lourenssen SR, Hurlbut DJ, Blennerhassett MG (2018) Epigenetic modification of intestinal smooth muscle cell phenotype during proliferation. Am J Physiol Cell Physiol 315:C722–C733

    Article  CAS  PubMed  Google Scholar 

  • Bonanomi D, Chivatakarn O, Bai G, Abdesselem H, Lettieri K, Marquardt T, Pierchala BA, Pfaff SL (2012) Ret is a multifunctional coreceptor that integrates diffusible- and contact-axon guidance signals. Cell 148:568–582

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Brat DJ, Brimijoin S (1992) A paradigm for examining toxicant effects on viability, structure, and axonal transport of neurons in culture. Mol Neurobiol 6:125–135

    Article  CAS  PubMed  Google Scholar 

  • Danzi MC, Mehta ST, Dulla K, Zunino G, Cooper DJ, Bixby JL, Lemmon VP (2018) The effect of Jun dimerization on neurite outgrowth and motif binding. Mol Cell Neurosci 92:114–127

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dinsmore CJ, Soriano P (2018) MAPK and PI3K signaling: At the crossroads of neural crest development. Dev Biol 10.

  • Durbec P, Marcos-Gutierrez CV, Kilkenny C, Grigoriou M, Wartiowaara K, Suvanto P, Smith D, Ponder B, Costantini F, Saarma M (1996) GDNF signalling through the Ret receptor tyrosine kinase. Nature 381:789–793

    Article  CAS  PubMed  Google Scholar 

  • de Santa Barbara P, van den Brink GR, Roberts DJ (2002) Molecular etiology of gut malformations and diseases. Am J Med Genet 115:221–230

    Article  PubMed  Google Scholar 

  • Encinas M, Tansey MG, Tsui-Pierchala BA, Comella JX, Milbrandt J, Johnson EM Jr (2001) c-Src is required for glial cell line-derived neurotrophic factor (GDNF) family ligand-mediated neuronal survival via a phosphatidylinositol-3 kinase (PI-3K)-dependent pathway. J Neurosci 21:1464–1472

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gabella G (2002) Development of visceral smooth muscle. Results Probl Cell Differ 38:1–37

    Article  PubMed  Google Scholar 

  • Gougeon PY, Lourenssen S, Han TY, Nair DG, Ropeleski MJ, Blennerhassett MG (2013) The pro-inflammatory cytokines IL-1beta and TNFalpha are neurotrophic for enteric neurons. J Neurosci 33:3339–3351

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Han TY, Lourenssen S, Miller KG, Blennerhassett MG (2015) Intestinal smooth muscle phenotype determines enteric neuronal survival via GDNF expression. Neuroscience 290:357–368

    Article  CAS  PubMed  Google Scholar 

  • Hao MM, Bergner AJ, Nguyen HTH, Dissanayake P, Burnett LE, Hopkins CD, Zeng K, Young HM, Stamp LA (2019) Role of JNK, MEK and adenylyl cyclase signalling in speed and directionality of enteric neural crest-derived cells. Dev Biol 455:362–368

    Article  CAS  PubMed  Google Scholar 

  • Hossain MI, Kamaruddin MA, Cheng HC (2012) Aberrant regulation and function of Src family tyrosine kinases: their potential contributions to glutamate-induced neurotoxicity. Clin Exp Pharmacol Physiol 39:684–691

    Article  CAS  PubMed  Google Scholar 

  • Hossain MI, Hoque A, Lessene G, Aizuddin KM, Chu PW, Ng IH, Irtegun S, Ng DC, Bogoyevitch MA, Burgess AW, Hill AF, Cheng HC (2015) Dual role of Src kinase in governing neuronal survival. Brain Res 1594:1–14

    Article  CAS  Google Scholar 

  • Hou XY, Liu Y, Zhang GY (2007) PP2, a potent inhibitor of Src family kinases, protects against hippocampal CA1 pyramidal cell death after transient global brain ischemia. Neurosci Lett 420:235–239

    Article  CAS  PubMed  Google Scholar 

  • Ibanez CF, Andressoo JO (2017) Biology of GDNF and its receptors - Relevance for disorders of the central nervous system. Neurobiol Dis 97:80–89

    Article  CAS  PubMed  Google Scholar 

  • Jo H, Mondal S, Tan D, Nagata E, Takizawa S, Sharma AK, Hou Q, Shanmugasundaram K, Prasad A, Tung JK, Tejeda AO, Man H, Rigby AC, Luo HR (2012) Small molecule-induced cytosolic activation of protein kinase Akt rescues ischemia-elicited neuronal death. Proc Natl Acad Sci U S A 109:10581–10586

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lake JI, Heuckeroth RO (2013) Enteric nervous system development: migration, differentiation, and disease. Am J Physiol Gastrointest Liver Physiol 305:G1-24

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lin A, Lourenssen S, Stanzel RD, Blennerhassett MG (2005a) Nerve growth factor sensitivity is broadly distributed among myenteric neurons of the rat colon. J Comp Neurol 490:194–206

    Article  CAS  PubMed  Google Scholar 

  • Lin A, Lourenssen S, Stanzel RD, Blennerhassett MG (2005b) Selective loss of NGF-sensitive neurons following experimental colitis. Exp Neurol 191:337–343

    Article  CAS  PubMed  Google Scholar 

  • Lourenssen SR, Blennerhassett MG (2020) M2 macrophages and phenotypic modulation of intestinal smooth muscle cells characterize inflammatory stricture formation in rats. Am J Pathol 190:1843–1858

    Article  CAS  PubMed  Google Scholar 

  • Lourenssen S, Wells RW, Blennerhassett MG (2005) Differential responses of intrinsic and extrinsic innervation of smooth muscle cells in rat colitis. Exp Neurol 195:497–507

    Article  CAS  PubMed  Google Scholar 

  • Lourenssen S, Miller KG, Blennerhassett MG (2009) Discrete responses of myenteric neurons to structural and functional damage by neurotoxins in vitro. Am J Physiol Gastrointest Liver Physiol 297:G228–G239

    Article  CAS  PubMed  Google Scholar 

  • Mahar M, Cavalli V (2018) Intrinsic mechanisms of neuronal axon regeneration. Nat Rev Neurosci 19:323–337

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Marlow SL, Blennerhassett MG (2006) Deficient innervation characterizes intestinal strictures in a rat model of colitis. Exp Mol Pathol 80:54–66

    Article  CAS  PubMed  Google Scholar 

  • Meka DP, Muller-Rischart AK, Nidadavolu P, Mohammadi B, Motori E, Ponna SK, Aboutalebi H, Bassal M, Annamneedi A, Finckh B, Miesbauer M, Rotermund N, Lohr C, Tatzelt J, Winklhofer KF, Kramer ER (2015) Parkin cooperates with GDNF/RET signaling to prevent dopaminergic neuron degeneration. J Clin Invest 125:1873–1885

    Article  PubMed  PubMed Central  Google Scholar 

  • Moore MW, Klein RD, Farinas I, Sauer H, Armanini M, Phillips H, Reichardt LF, Ryan AM, Carver-Moore K, Rosenthal A (1996) Renal and neuronal abnormalities in mice lacking GDNF. Nature 382:76–79

    Article  CAS  PubMed  Google Scholar 

  • Rodrigues DM, Li AY, Nair DG, Blennerhassett MG (2011) Glial cell line-derived neurotrophic factor is a key neurotrophin in the postnatal enteric nervous system. Neurogastroenterol Motil 23:44–56

    Article  Google Scholar 

  • Runeberg-Roos P, Saarma M (2007) Neurotrophic factor receptor RET: structure, cell biology, and inherited diseases. Ann Med 39:572–580

    Article  CAS  PubMed  Google Scholar 

  • Sanchez MP, Silos-Santiago I, Frisen J, He B, Lira SA, Barbacid M (1996) Renal agenesis and the absence of enteric neurons in mice lacking GDNF. Nature 382:70–73

    Article  CAS  PubMed  Google Scholar 

  • Sanovic S, Lamb DP, Blennerhassett MG (1999) Damage to the enteric nervous system in experimental colitis. Am J Pathol 155:1051–1057

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sariola H, Saarma M (2003) Novel functions and signalling pathways for GDNF. J Cell Sci 116:3855–3862

    Article  CAS  PubMed  Google Scholar 

  • Srinivasan S, Anitha M, Mwangi S, Heuckeroth RO (2005) Enteric neuroblasts require the phosphatidylinositol 3-kinase/Akt/Forkhead pathway for GDNF-stimulated survival. Mol Cell Neurosci 29:107–119

    Article  CAS  PubMed  Google Scholar 

  • Stanzel RD, Lourenssen S, Nair DG, Blennerhassett MG (2010) Mitogenic factors promoting intestinal smooth muscle cell proliferation. Am J Physiol Cell Physiol 299:C805–C817

    Article  CAS  PubMed  Google Scholar 

  • Suvanto P, Hiltunen JO, Arumae U, Moshnyakov M, Sariola H, Sainio K, Saarma M (1996) Localization of glial cell line-derived neurotrophic factor (GDNF) mRNA in embryonic rat by in situ hybridization. Eur J Neurosci 8:816–822

    Article  CAS  PubMed  Google Scholar 

  • Takadera T, Fujibayashi M, Koriyama Y, Kato S (2012) Apoptosis induced by SRC-family tyrosine kinase inhibitors in cultured rat cortical cells. Neurotox Res 21:309–316

    Article  CAS  PubMed  Google Scholar 

  • Tansey MG, Baloh RH, Milbrandt J, Johnson EM Jr (2000) GFRalpha-mediated localization of RET to lipid rafts is required for effective downstream signaling, differentiation, and neuronal survival. Neuron 25:611–623

    Article  CAS  PubMed  Google Scholar 

  • Wang P, Chen FX, Du C, Li CQ, Yu YB, Zuo XL, Li YQ (2015) Increased production of BDNF in colonic epithelial cells induced by fecal supernatants from diarrheic IBS patients. Sci Rep 5:10121. https://doi.org/10.1038/srep10121.:10121

    Article  PubMed  PubMed Central  Google Scholar 

  • Wang P, Du C, Chen FX, Li CQ, Yu YB, Han T, Akhtar S, Zuo XL, Tan XD, Li YQ (2016) BDNF contributes to IBS-like colonic hypersensitivity via activating the enteroglia-nerve unit. Sci Rep 6:20320. https://doi.org/10.1038/srep20320.:20320

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Young HM, Hearn CJ, Farlie PG, Canty AJ, Thomas PQ, Newgreen DF (2001) GDNF is a chemoattractant for enteric neural cells. Dev Biol 229:503–516

    Article  CAS  PubMed  Google Scholar 

  • Yu YB, Zuo XL, Zhao QJ, Chen FX, Yang J, Dong YY, Wang P, Li YQ (2012) Brain-derived neurotrophic factor contributes to abdominal pain in irritable bowel syndrome. Gut 61:685–694

    Article  CAS  PubMed  Google Scholar 

  • Zoumboulakis D, Cirella KR, Gougeon PY, Lourenssen SR, Blennerhassett MG (2020) MMP-9 processing of intestinal smooth muscle-derived GDNF is required for neurotrophic action on enteric neurons. Neuroscience 443:8–18

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the Natural Sciences and Engineering Research Council (NSERC) of Canada.

Author information

Authors and Affiliations

  1. Gastrointestinal Diseases Research Unit and Queen’s University, Kingston, ON, K7L 2V7, Canada

    M. G. Blennerhassett & S. R. Lourenssen

  2. Department of Medicine, GIDRU Wing, Kingston General Hospital, Queen’s University, Kingston, ON, K7L2V7, Canada

    M. G. Blennerhassett

Authors
  1. M. G. Blennerhassett
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S. R. Lourenssen
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Both authors contributed to the planning, experimentation and writing of the manuscript.

Corresponding author

Correspondence to M. G. Blennerhassett.

Ethics declarations

Conflict of interest

The authors declare no conflicts of interest or competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Blennerhassett, M.G., Lourenssen, S.R. Obligatory Activation of SRC and JNK by GDNF for Survival and Axonal Outgrowth of Postnatal Intestinal Neurons. Cell Mol Neurobiol 42, 1569–1583 (2022). https://doi.org/10.1007/s10571-021-01048-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10571-021-01048-9

Keywords

Search

Navigation