{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T22:46:40Z","timestamp":1784242000142,"version":"3.55.0"},"reference-count":50,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2021,9,18]],"date-time":"2021-09-18T00:00:00Z","timestamp":1631923200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["DFG, Ri-817\/12-1 and 817\/15-1; HLR; KR 3807\/7-1; SMK"],"award-info":[{"award-number":["DFG, Ri-817\/12-1 and 817\/15-1; HLR; KR 3807\/7-1; SMK"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Else\u2013Kr\u00f6ner\u2013Fresenius Foundation","award":["2017"],"award-info":[{"award-number":["2017"]}]},{"name":"Graduate School of Life Sciences (GSLS), University of Wuerzburg","award":["2018, 2019"],"award-info":[{"award-number":["2018, 2019"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The blood\u2013nerve barrier and myelin barrier normally shield peripheral nerves from potentially harmful insults. They are broken down during nerve injury, which contributes to neuronal damage. Netrin-1 is a neuronal guidance protein with various established functions in the peripheral and central nervous systems; however, its role in regulating barrier integrity and pain processing after nerve injury is poorly understood. Here, we show that chronic constriction injury (CCI) in Wistar rats reduced netrin-1 protein and the netrin-1 receptor neogenin-1 (Neo1) in the sciatic nerve. Replacement of netrin-1 via systemic or local administration of the recombinant protein rescued injury-induced nociceptive hypersensitivity. This was prevented by siRNA-mediated knockdown of Neo1 in the sciatic nerve. Mechanistically, netrin-1 restored endothelial and myelin, but not perineural, barrier function as measured by fluorescent dye or fibrinogen penetration. Netrin-1 also reversed the decline in the tight junction proteins claudin-5 and claudin-19 in the sciatic nerve caused by CCI. Our findings emphasize the role of the endothelial and myelin barriers in pain processing after nerve damage and reveal that exogenous netrin-1 restores their function to mitigate CCI-induced hypersensitivity via Neo1. The netrin-1-neogenin-1 signaling pathway may thus represent a multi-target barrier protector for the treatment of neuropathic pain.<\/jats:p>","DOI":"10.3390\/ijms221810090","type":"journal-article","created":{"date-parts":[[2021,9,21]],"date-time":"2021-09-21T08:04:23Z","timestamp":1632211463000},"page":"10090","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Netrin-1 as a Multitarget Barrier Stabilizer in the Peripheral Nerve after Injury"],"prefix":"10.3390","volume":"22","author":[{"given":"Jeremy Tsung-Chieh","family":"Chen","sequence":"first","affiliation":[{"name":"Center for Interdisciplinary Pain Medicine, Department of Anesthesiology, Intensive Care, Emergency Medicine and Pain Therapy, University Hospital of W\u00fcrzburg, 97080 W\u00fcrzburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lea","family":"Schmidt","sequence":"additional","affiliation":[{"name":"Center for Interdisciplinary Pain Medicine, Department of Anesthesiology, Intensive Care, Emergency Medicine and Pain Therapy, University Hospital of W\u00fcrzburg, 97080 W\u00fcrzburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christina","family":"Sch\u00fcrger","sequence":"additional","affiliation":[{"name":"Center for Interdisciplinary Pain Medicine, Department of Anesthesiology, Intensive Care, Emergency Medicine and Pain Therapy, University Hospital of W\u00fcrzburg, 97080 W\u00fcrzburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammed K.","family":"Hankir","sequence":"additional","affiliation":[{"name":"Department of Experimental Surgery, University Hospital of W\u00fcrzburg, 97080 W\u00fcrzburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1293-2484","authenticated-orcid":false,"given":"Susanne M.","family":"Krug","sequence":"additional","affiliation":[{"name":"Clinical Physiology\/Nutritional Medicine, Campus Benjamin Franklin, Charit\u00e9-Universit\u00e4tsmedizin Berlin, 12203 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4867-0188","authenticated-orcid":false,"given":"Heike L.","family":"Rittner","sequence":"additional","affiliation":[{"name":"Center for Interdisciplinary Pain Medicine, Department of Anesthesiology, Intensive Care, Emergency Medicine and Pain Therapy, University Hospital of W\u00fcrzburg, 97080 W\u00fcrzburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,9,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1253","DOI":"10.4161\/cc.9.7.11072","article-title":"Netrin-1, a missing link between chronic inflammation and tumor progression","volume":"9","author":"Paradisi","year":"2010","journal-title":"Cell Cycle"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"544","DOI":"10.1016\/j.molmed.2012.07.001","article-title":"Netrin-1 in the developing enteric nervous system and colorectal cancer","volume":"18","author":"Ko","year":"2012","journal-title":"Trends Mol. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1598","DOI":"10.1093\/brain\/awv092","article-title":"Netrin 1 regulates blood-brain barrier function and neuroinflammation","volume":"138","author":"Podjaski","year":"2015","journal-title":"Brain"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3006","DOI":"10.1172\/JCI84767","article-title":"Truncated netrin-1 contributes to pathological vascular permeability in diabetic retinopathy","volume":"126","author":"Miloudi","year":"2016","journal-title":"J. Clin. Investig."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e24956","DOI":"10.4161\/tisb.24956","article-title":"Barriers of the peripheral nerve","volume":"1","author":"Peltonen","year":"2013","journal-title":"Tissue Barriers"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1007\/s00424-016-1920-8","article-title":"Barrier function in the peripheral and central nervous system\u2014A review","volume":"469","author":"Reinhold","year":"2017","journal-title":"Pflugers Arch."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"113244","DOI":"10.1016\/j.expneurol.2020.113244","article-title":"Characteristics of the nerve barrier and the blood dorsal root ganglion barrier in health and disease","volume":"327","author":"Reinhold","year":"2020","journal-title":"Exp. Neurol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1369\/jhc.2009.951681","article-title":"Tight junction proteins in human Schwann cell autotypic junctions","volume":"57","author":"Alanne","year":"2009","journal-title":"J. Histochem. Cytochem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"954","DOI":"10.1016\/j.pain.2014.01.026","article-title":"Blood-nerve barrier dysfunction contributes to the generation of neuropathic pain and allows targeting of injured nerves for pain relief","volume":"155","author":"Lim","year":"2014","journal-title":"Pain"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1177\/0022034516679395","article-title":"Hedgehog Pathway-Mediated Vascular Alterations Following Trigeminal Nerve Injury","volume":"96","author":"Moreau","year":"2017","journal-title":"J. Dent. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1097\/j.pain.0000000000000444","article-title":"Early alterations of Hedgehog signaling pathway in vascular endothelial cells after peripheral nerve injury elicit blood-nerve barrier disruption, nerve inflammation, and neuropathic pain development","volume":"157","author":"Moreau","year":"2016","journal-title":"Pain"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"936","DOI":"10.3389\/fnins.2018.00936","article-title":"Quantitative and Microstructural Changes of the Blood-Nerve Barrier in Peripheral Neuropathy","volume":"12","author":"Reinhold","year":"2018","journal-title":"Front. Neurosci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1519","DOI":"10.3389\/fphys.2018.01519","article-title":"Peripheral Nerve Injury Induces Dynamic Changes of Tight Junction Components","volume":"9","author":"Wang","year":"2018","journal-title":"Front. Physiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1038","DOI":"10.3389\/fnins.2018.01038","article-title":"Functional and Structural Changes of the Blood-Nerve-Barrier in Diabetic Neuropathy","volume":"12","author":"Richner","year":"2018","journal-title":"Front. Neurosci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"890","DOI":"10.4103\/1673-5374.184452","article-title":"Role of myelin auto-antigens in pain: A female connection","volume":"11","author":"Shubayev","year":"2016","journal-title":"Neural Regen. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"a020487","DOI":"10.1101\/cshperspect.a020487","article-title":"Schwann Cells: Development and Role in Nerve Repair","volume":"7","author":"Jessen","year":"2015","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"553","DOI":"10.5966\/sctm.2013-0011","article-title":"Plastic fantastic: Schwann cells and repair of the peripheral nervous system","volume":"2","author":"Kim","year":"2013","journal-title":"Stem Cells Transl. Med."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Dun, X.P., and Parkinson, D.B. (2017). Role of Netrin-1 Signaling in Nerve Regeneration. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18030491"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1057","DOI":"10.1016\/j.bbrc.2007.08.143","article-title":"Netrin-1 induces proliferation of Schwann cells through Unc5b receptor","volume":"362","author":"Lee","year":"2007","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1016\/j.bbrc.2015.06.140","article-title":"Netrin-1 induces the migration of Schwann cells via p38 MAPK and PI3K-Akt signaling pathway mediated by the UNC5B receptor","volume":"464","author":"Lv","year":"2015","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"380","DOI":"10.7555\/JBR.29.20140076","article-title":"Netrin-1 overexpression in bone marrow mesenchymal stem cells promotes functional recovery in a rat model of peripheral nerve injury","volume":"29","author":"Ke","year":"2015","journal-title":"J. Biomed. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1160","DOI":"10.1016\/j.bbadis.2019.01.008","article-title":"Tissue plasminogen activator and neuropathy open the blood-nerve barrier with upregulation of microRNA-155-5p in male rats","volume":"1865","author":"Reinhold","year":"2019","journal-title":"Biochim. Biophys. Acta Mol. Basis Dis."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"E2018","DOI":"10.1073\/pnas.1120800109","article-title":"Transient opening of the perineurial barrier for analgesic drug delivery","volume":"109","author":"Hackel","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1111\/nyas.13436","article-title":"Blood-spinal cord barrier breakdown and pericyte deficiency in peripheral neuropathy","volume":"1405","author":"Sauer","year":"2017","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1083\/jcb.200302070","article-title":"Size-selective loosening of the blood-brain barrier in claudin-5-deficient mice","volume":"161","author":"Nitta","year":"2003","journal-title":"J. Cell Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1237","DOI":"10.1007\/s00109-021-02091-1","article-title":"Selective blood-nerve barrier leakiness with claudin-1 and vessel-associated macrophage loss in diabetic polyneuropathy","volume":"99","author":"Sauer","year":"2021","journal-title":"J. Mol. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1083\/jcb.200501154","article-title":"Tight junctions in Schwann cells of peripheral myelinated axons: A lesson from claudin-19-deficient mice","volume":"169","author":"Miyamoto","year":"2005","journal-title":"J. Cell Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1002\/ana.24086","article-title":"Abnormal junctions and permeability of myelin in PMP22-deficient nerves","volume":"75","author":"Guo","year":"2014","journal-title":"Ann. Neurol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"17002","DOI":"10.1038\/nrdp.2017.2","article-title":"Neuropathic pain","volume":"3","author":"Colloca","year":"2017","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1038\/nrn.2018.2","article-title":"Microglia in neuropathic pain: Cellular and molecular mechanisms and therapeutic potential","volume":"19","author":"Inoue","year":"2018","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.ejphar.2009.04.028","article-title":"B vitamins alleviate indices of neuropathic pain in diabetic rats","volume":"612","author":"Jolivalt","year":"2009","journal-title":"Eur. J. Pharmacol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1002\/1096-9861(20000904)424:4<563::AID-CNE1>3.0.CO;2-U","article-title":"Damage to the cytoskeleton of large diameter sensory neurons and myelinated axons in vincristine-induced painful peripheral neuropathy in the rat","volume":"424","author":"Topp","year":"2000","journal-title":"J. Comp. Neurol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"700","DOI":"10.3389\/fnins.2017.00700","article-title":"Netrin-1 Ameliorates Blood-Brain Barrier Impairment Secondary to Ischemic Stroke via the Activation of PI3K Pathway","volume":"11","author":"Yu","year":"2017","journal-title":"Front. Neurosci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"6490","DOI":"10.1172\/JCI132372","article-title":"Astrocytic neogenin\/netrin-1 pathway promotes blood vessel homeostasis and function in mouse cortex","volume":"130","author":"Yao","year":"2020","journal-title":"J. Clin. Investig."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e005198","DOI":"10.1161\/JAHA.116.005198","article-title":"Netrin-1 Preserves Blood-Brain Barrier Integrity Through Deleted in Colorectal Cancer\/Focal Adhesion Kinase\/RhoA Signaling Pathway Following Subarachnoid Hemorrhage in Rats","volume":"6","author":"Xie","year":"2017","journal-title":"J. Am. Heart Assoc."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1136\/gutjnl-2011-300012","article-title":"Neuronal guidance molecule netrin-1 attenuates inflammatory cell trafficking during acute experimental colitis","volume":"61","author":"Aherne","year":"2012","journal-title":"Gut"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1006\/exnr.1999.7292","article-title":"Netrin-1 and peripheral nerve regeneration in the adult rat","volume":"161","author":"Madison","year":"2000","journal-title":"Exp. Neurol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1503","DOI":"10.1002\/glia.21194","article-title":"Schwann cells direct peripheral nerve regeneration through the Netrin-1 receptors, DCC and Unc5H2","volume":"59","author":"Webber","year":"2011","journal-title":"Glia"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.ijpharm.2012.01.023","article-title":"Changes in protein and mRNA expression levels of claudin family after mucosal lesion by intestinal ischemia\/reperfusion","volume":"426","author":"Takizawa","year":"2012","journal-title":"Int. J. Pharm."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1262","DOI":"10.1097\/j.pain.0000000000002122","article-title":"Differential impact of keratinocytes and fibroblasts on nociceptor degeneration and sensitization in small fiber neuropathy","volume":"162","author":"Kress","year":"2021","journal-title":"Pain"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"E19","DOI":"10.1038\/nature22293","article-title":"Can we predict protein from mRNA levels?","volume":"547","author":"Fortelny","year":"2017","journal-title":"Nature"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1038\/nrg3185","article-title":"Insights into the regulation of protein abundance from proteomic and transcriptomic analyses","volume":"13","author":"Vogel","year":"2012","journal-title":"Nat. Rev. Genet."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1186\/s12974-018-1123-7","article-title":"Acute- and late-phase matrix metalloproteinase (MMP)-9 activity is comparable in female and male rats after peripheral nerve injury","volume":"15","author":"Remacle","year":"2018","journal-title":"J. Neuroinflamm."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3489","DOI":"10.1074\/jbc.RA119.007389","article-title":"The microRNAs let-7 and miR-9 down-regulate the axon-guidance genes Ntn1 and Dcc during peripheral nerve regeneration","volume":"294","author":"Wang","year":"2019","journal-title":"J. Biol. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1038\/ni.1683","article-title":"Hypoxia-inducible factor-dependent induction of netrin-1 dampens inflammation caused by hypoxia","volume":"10","author":"Rosenberger","year":"2009","journal-title":"Nat. Immunol."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Toque, H.A., Fernandez-Flores, A., Mohamed, R., Caldwell, R.B., Ramesh, G., and Caldwell, R.W. (2017). Netrin-1 is a novel regulator of vascular endothelial function in diabetes. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0186734"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"355","DOI":"10.2215\/CJN.02870310","article-title":"Renal, ocular, and neuromuscular involvements in patients with CLDN19 mutations","volume":"6","author":"Faguer","year":"2011","journal-title":"Clin. J. Am. Soc. Nephrol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1386","DOI":"10.1053\/j.gastro.2004.07.022","article-title":"Claudin-1 gene mutations in neonatal sclerosing cholangitis associated with ichthyosis: A tight junction disease","volume":"127","author":"Baala","year":"2004","journal-title":"Gastroenterology"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2812","DOI":"10.1007\/s00415-019-09496-6","article-title":"Reduced gene expression of netrin family members in skin and sural nerve specimens of patients with painful peripheral neuropathies","volume":"266","author":"Schubert","year":"2019","journal-title":"J. Neurol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/0165-0270(94)90144-9","article-title":"Quantitative assessment of tactile allodynia in the rat paw","volume":"53","author":"Chaplan","year":"1994","journal-title":"J. Neurosci. Methods"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/18\/10090\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:01:45Z","timestamp":1760166105000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/18\/10090"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,9,18]]},"references-count":50,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2021,9]]}},"alternative-id":["ijms221810090"],"URL":"https:\/\/doi.org\/10.3390\/ijms221810090","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,9,18]]}}}