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, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Central nervous system
Dulbecco’s minimal essential medium
Enteric nervous system
Glial cell line derived neurotrophic factor
Intestinal smooth muscle cell
C-Jun N-terminal kinase
“Re-arranged on transfection”
Trinitrobenzene sulphonic acid
Airaksinen MS, Saarma M (2002) The GDNF family: signalling, biological functions and therapeutic value. Nat Rev Neurosci 3:383–394
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
Gabella G (2002) Development of visceral smooth muscle. Results Probl Cell Differ 38:1–37
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
Han TY, Lourenssen S, Miller KG, Blennerhassett MG (2015) Intestinal smooth muscle phenotype determines enteric neuronal survival via GDNF expression. Neuroscience 290:357–368
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
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
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
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
Ibanez CF, Andressoo JO (2017) Biology of GDNF and its receptors - Relevance for disorders of the central nervous system. Neurobiol Dis 97:80–89
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
Lake JI, Heuckeroth RO (2013) Enteric nervous system development: migration, differentiation, and disease. Am J Physiol Gastrointest Liver Physiol 305:G1-24
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
Lin A, Lourenssen S, Stanzel RD, Blennerhassett MG (2005b) Selective loss of NGF-sensitive neurons following experimental colitis. Exp Neurol 191:337–343
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
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
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
Mahar M, Cavalli V (2018) Intrinsic mechanisms of neuronal axon regeneration. Nat Rev Neurosci 19:323–337
Marlow SL, Blennerhassett MG (2006) Deficient innervation characterizes intestinal strictures in a rat model of colitis. Exp Mol Pathol 80:54–66
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
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
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
Runeberg-Roos P, Saarma M (2007) Neurotrophic factor receptor RET: structure, cell biology, and inherited diseases. Ann Med 39:572–580
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
Sanovic S, Lamb DP, Blennerhassett MG (1999) Damage to the enteric nervous system in experimental colitis. Am J Pathol 155:1051–1057
Sariola H, Saarma M (2003) Novel functions and signalling pathways for GDNF. J Cell Sci 116:3855–3862
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
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
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
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
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
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
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
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
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
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
This work was supported by the Natural Sciences and Engineering Research Council (NSERC) of Canada.
Conflict of interest
The authors declare no conflicts of interest or competing interests.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
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 (2021). https://doi.org/10.1007/s10571-021-01048-9
- Retrograde transport
- Receptor tyrosine kinase
- Enteric nervous system