https://doi.org/10.4081/ejh.2023.3846
Expression and role of cocaine-amphetamine regulated transcript (CART) in the proliferation of biliary epithelium
Submitted: 4 August 2023
Accepted: 25 September 2023
Published: 19 October 2023
Accepted: 25 September 2023
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All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
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Cholangiocytes, the epithelial cells that line the biliary tree, can proliferate under the stimulation of several factors through both autocrine and paracrine pathways. The cocaine-amphetamine-regulated-transcript (CART) peptide has several physiological functions, and it is widely expressed in several organs. CART increases the survival of hippocampal neurons by upregulating brain-derived neurotrophic factor (BDNF), whose expression has been correlated to the proliferation rate of cholangiocytes. In the present study, we aimed to evaluate the expression of CART and its role in modulating cholangiocyte proliferation in healthy and bile duct ligated (BDL) rats in vivo, as well as in cultured normal rat cholangiocytes (NRC) in vitro. Liver samples from both healthy and BDL (1 week) rats, were analyzed by immunohistochemistry and immunofluorescence for CART, CK19, TrkB and p75NTR BDNF receptors. PCNA staining was used to evaluate the proliferation of the cholangiocytes, whereas TUNEL assay was used to evaluate biliary apoptosis. NRC treated or not with CART were used to confirm the role of CART on cholangiocytes proliferation and the secretion of BDNF. Cholangiocytes proliferation, apoptosis, CART and TrkB expression were increased in BDL rats, compared to control rats. We found a higher expression of TrkB and p75NTR, which could be correlated with the proliferation rate of biliary tree during BDL. The in vitro study demonstrated increased BDNF secretion by NRC after treatment with CART compared with control cells. As previously reported, proliferating cholangiocytes acquire a neuroendocrine phenotype, modulated by several factors, including neurotrophins. Accordingly, CART may play a key role in the remodeling of biliary epithelium during cholestasis by modulating the secretion of BDNF.
Alpini G, Prall RT, LaRusso NF. The pathobiology of biliary epithelia. In: Arias IM, Boyer JL, Chisari FV, Fausto N, Jakoby W, Schachter D, Editors. The liver: biology and pathobiology, 4th edition. Philadelphia, Lippincott Williams & Wilkins. 2001. p. 421-35.
Casini A, Leone S, Vaccaro R, Vivacqua G, Ceci L, Pannarale L, et al. The emerging role of ferroptosis in liver cancers. Life (Basel) 2022;12:2128.
DOI: https://doi.org/10.3390/life12122128
Alpini G, Lenzi R, Sarkozi L, Tavoloni N. Biliary physiology in rats with bile ductular cell hyperplasia. Evidence for a secretory function of proliferated bile ductules. J Clin Invest 1988;81:569-78.
DOI: https://doi.org/10.1172/JCI113355
Munshi MK, Priester S, Gaudio E, Yang F, Alpini G, Mancinelli R, et al. Regulation of biliary proliferation by neuroendocrine factors: implications for the pathogenesis of cholestatic liver diseases. Am J Pathol 2011;178:472-84.
DOI: https://doi.org/10.1016/j.ajpath.2010.09.043
Mancinelli R, Glaser S, Francis H, Carpino G, Franchitto A, Vetuschi A, et al. Ischemia reperfusion of the hepatic artery induces the functional damage of large bile ducts by changes in the expression of angiogenic factors. Am J Physiol Gastrointest Liver Physiol 2015;309:G865-73.
DOI: https://doi.org/10.1152/ajpgi.00015.2015
Alvaro D, Mancino MG, Glaser S, Gaudio E, Marzioni M, Francis H, et al. Proliferating cholangiocytes: a neuroendocrine compartment in the diseased liver. Gastroenterology 2007;132:415-31.
DOI: https://doi.org/10.1053/j.gastro.2006.07.023
Francis H, Onori P, Gaudio E, Franchitto A, DeMorrow S, Venter J, et al. H3 histamine receptor-mediated activation of protein kinase Calpha inhibits the growth of cholangiocarcinoma in vitro and in vivo. Mol Cancer Res 2009;7:1704-13.
DOI: https://doi.org/10.1158/1541-7786.MCR-09-0261
Glaser S, Meng F, Han Y, Onori P, Chow BK, Francis H, et al. Secretin stimulates biliary cell proliferation by regulating expression of microRNA 125b and microRNA let7a in mice. Gastroenterology 2014;146:1795-808.e12.
DOI: https://doi.org/10.1053/j.gastro.2014.02.030
Gaudio E, Barbaro B, Alvaro D, Glaser S, Francis H, Ueno Y, et al. Vascular endothelial growth factor stimulates rat cholangiocyte proliferation via an autocrine mechanism. Gastroenterology 2006;130:1270-82.
DOI: https://doi.org/10.1053/j.gastro.2005.12.034
Elmquist JK, Coppari R, Balthasar N, Ichinose M, Lowell BB. Identifying hypothalamic pathways controlling food intake, body weight, and glucose homeostasis. J Comp Neurol 2005;493:63-71.
DOI: https://doi.org/10.1002/cne.20786
Dudek M, Ziarniak K and Sliwowska JH. Kisspeptin and metabolism: the brain and beyond. Front Endocrinol (Lausanne) 2018;9:145.
DOI: https://doi.org/10.3389/fendo.2018.00145
Vivacqua G, Renzi A, Carpino G, Franchitto A, Gaudio E. Expression of brain derivated neurotrophic factor and of its receptors: TrKB and p75NT in normal and bile duct ligated rat liver. Ital J Anat Embryol 2014;119:111-29.
DeMorrow S, Meng F, Venter J, Leyva-Illades D, Francis H, Frampton G, et al. Neuropeptide Y inhibits biliary hyperplasia of cholestatic rats by paracrine and autocrine mechanisms. Am J Physiol Gastrointest Liver Physiol 2013;305:G250-7.
DOI: https://doi.org/10.1152/ajpgi.00140.2013
Casini A, Vaccaro R, D'Este L, Sakaue Y, Bellier JP, Kimura H, et al. Immunolocalization of choline acetyltransferase of common type in the central brain mass of Octopus vulgaris. Eur J Histochem 2012;56:e34.
DOI: https://doi.org/10.4081/ejh.2012.e34
Douglass J, McKinzie AA, Couceyro P. PCR differential display identifies a rat brain mRNA that is transcriptionally regulated by cocaine and amphetamine. J Neurosci 1995;15:2471-81.
DOI: https://doi.org/10.1523/JNEUROSCI.15-03-02471.1995
Kuhar MJ, Adams LD, Hunter RG, Vechia SD, Smith Y. CART peptides. Regul Pept 2000;89:1-6.
DOI: https://doi.org/10.1016/S0167-0115(00)00096-3
Dominguez G. The CART gene: structure and regulation. Peptides 2006;27:1913-8.
DOI: https://doi.org/10.1016/j.peptides.2006.01.025
Dylag T, Kotlinska J, Rafalski P, Pachuta A, Silberring J. The activity of CART peptide fragments. Peptides 2006;27:1926-33.
DOI: https://doi.org/10.1016/j.peptides.2005.10.025
Murphy KG, Abbott CR, Mahmoudi M, Hunter R, Gardiner JV, Rossi M, et al. Quantification and synthesis of cocaine- and amphetamine-regulated transcript peptide (79-102)-like immunoreactivity and mRNA in rat tissues. J Endocrinol 2000;166:659-68.
DOI: https://doi.org/10.1677/joe.0.1660659
Wierup N, Gunnarsdottir A, Ekblad E, Sundler F. Characterisation of CART-containing neurons and cells in the porcine pancreas, gastro-intestinal tract, adrenal and thyroid glands. BMC Neurosci 2007;8:51.
DOI: https://doi.org/10.1186/1471-2202-8-51
Kasacka I and Piotrowska Z. Evaluation of density and distribution of CART-immunoreactive structures in gastrointestinal tract of hypertensive rats. Biofactors 2012;38:407-15.
DOI: https://doi.org/10.1002/biof.1037
Ekblad E, Kuhar M, Wierup N, Sundler F. Cocaine- and amphetamine-regulated transcript: distribution and function in rat gastrointestinal tract. Neurogastroenterol Motil 2003;15:545-57.
DOI: https://doi.org/10.1046/j.1365-2982.2003.00437.x
Vaccaro R, Severi C, Serra G, Carabotti M, Casini A, Chirletti P, et al. Endocrine cells distribution in human proximal small intestine: an immunohistochemical and morphometrical study. Ital J Anat Embryol 2016;121:112-21.
Casini A, Mancinelli R, Mammola CL, Pannarale L, Chirletti P, Onori P, et al. Distribution of alpha-synuclein in normal human jejunum and its relations with the chemosensory and neuroendocrine system. Eur J Histochem 2021;65:3310.
DOI: https://doi.org/10.4081/ejh.2021.3310
Jia J, Chen X, Zhu W, Luo Y, Hua Z, Xu Y. CART protects brain from damage through ERK activation in ischemic stroke. Neuropeptides 2008;42:653-61.
DOI: https://doi.org/10.1016/j.npep.2008.05.006
Lakatos A, Prinster S, Vicentic A, Hall RA, Kuhar MJ. Cocaine- and amphetamine-regulated transcript (CART) peptide activates the extracellular signal-regulated kinase (ERK) pathway in AtT20 cells via putative G-protein coupled receptors. Neurosci Lett 2005;384:198-202.
DOI: https://doi.org/10.1016/j.neulet.2005.04.072
Wu B, Hu S, Yang M, Pan H, Zhu S. CART peptide promotes the survival of hippocampal neurons by upregulating brain-derived neurotrophic factor. Biochem Biophys Res Commun 2006;347:656-61.
DOI: https://doi.org/10.1016/j.bbrc.2006.06.117
Hashemi P, Ahmadi S. Alpha-pinene moderates memory impairment induced by kainic acid via improving the BDNF/TrkB/CREB signaling pathway in rat hippocampus. Front Mol Neurosci 2023;16:1202232.
DOI: https://doi.org/10.3389/fnmol.2023.1202232
Moya-Alvarado G, Tiburcio-Felix R, Ibanez MR, Aguirre-Soto AA, Guerra MV, Wu C, et al. BDNF/TrkB signaling endosomes in axons coordinate CREB/mTOR activation and protein synthesis in the cell body to induce dendritic growth in cortical neurons. Elife 2023;12:e77455.
DOI: https://doi.org/10.7554/eLife.77455
Abels M, Riva M, Shcherbina L, Fischer AT, Banke E, Degerman E, et al. Overexpressed beta cell CART increases insulin secretion in mouse models of insulin resistance and diabetes. Peptides 2022;151:170747.
DOI: https://doi.org/10.1016/j.peptides.2022.170747
Sathanoori R, Olde B, Erlinge D, Goransson O and Wierup N. Cocaine- and amphetamine-regulated transcript (CART) protects beta cells against glucotoxicity and increases cell proliferation. J Biol Chem 2013;288:3208-18.
DOI: https://doi.org/10.1074/jbc.M112.437145
Dhillo WS, Small CJ, Stanley SA, Jethwa PH, Seal LJ, Murphy KG, et al. Hypothalamic interactions between neuropeptide Y, agouti-related protein, cocaine- and amphetamine-regulated transcript and alpha-melanocyte-stimulating hormone in vitro in male rats. J Neuroendocrinol 2002;14:725-30.
DOI: https://doi.org/10.1046/j.1365-2826.2002.00832.x
Francis H, Glaser S, Demorrow S, Gaudio E, Ueno Y, Venter J, et al. Small mouse cholangiocytes proliferate in response to H1 histamine receptor stimulation by activation of the IP3/CaMK I/CREB pathway. Am J Physiol Cell Physiol 2008;295:C499-513.
DOI: https://doi.org/10.1152/ajpcell.00369.2007
Mancinelli R, Franchitto A, Glaser S, Meng F, Onori P, Demorrow S, et al. GABA induces the differentiation of small into large cholangiocytes by activation of Ca(2+) /CaMK I-dependent adenylyl cyclase 8. Hepatology 2013;58:251-63.
DOI: https://doi.org/10.1002/hep.26308
Glaser S, Gaudio E, Renzi A, Mancinelli R, Ueno Y, Venter J, et al. Knockout of the neurokinin-1 receptor reduces cholangiocyte proliferation in bile duct-ligated mice. Am J Physiol Gastrointest Liver Physiol 2011;301:G297-305.
DOI: https://doi.org/10.1152/ajpgi.00418.2010
Zhou T, Wu N, Meng F, Venter J, Giang TK, Francis H, et al. Knockout of secretin receptor reduces biliary damage and liver fibrosis in Mdr2(-/-) mice by diminishing senescence of cholangiocytes. Lab Invest 2018;98:1449-64.
DOI: https://doi.org/10.1038/s41374-018-0093-9
Jin W. Regulation of BDNF-TrkB Signaling and potential therapeutic strategies for Parkinson's disease. J Clin Med 2020;9:257.
DOI: https://doi.org/10.3390/jcm9010257
Colucci-D'Amato L, Speranza L and Volpicelli F. Neurotrophic factor BDNF, physiological functions and therapeutic potential in depression, neurodegeneration and brain cancer. Int J Mol Sci 2020;21:7777.
DOI: https://doi.org/10.3390/ijms21207777
Fayard B, Loeffler S, Weis J, Vogelin E, Kruttgen A. The secreted brain-derived neurotrophic factor precursor pro-BDNF binds to TrkB and p75NTR but not to TrkA or TrkC. J Neurosci Res 2005;80:18-28.
DOI: https://doi.org/10.1002/jnr.20432
Sen A, Bettegowda A, Jimenez-Krassel F, Ireland JJ, Smith GW. Cocaine- and amphetamine-regulated transcript regulation of follicle-stimulating hormone signal transduction in bovine granulosa cells. Endocrinology 2007;148:4400-10.
DOI: https://doi.org/10.1210/en.2007-0332
Lin L, Sun D, Chang J, Ma M, Zhou X, Zhao M, et al. Cocaine and amphetamineregulated transcript (CART) is associated with dopamine and is protective against ischemic stroke. Mol Med Rep 2018;18:3298-304.
DOI: https://doi.org/10.3892/mmr.2018.9296
Xu Y, Zhang W, Klaus J, Young J, Koerner I, Sheldahl LC, et al. Role of cocaine- and amphetamine-regulated transcript in estradiol-mediated neuroprotection. Proc Natl Acad Sci USA 2006;103:14489-94.
DOI: https://doi.org/10.1073/pnas.0602932103
Liu Z, Huang D, Zhang M, Chen Z, Jin J, Huang S, et al. Cocaine- and amphetamine-regulated transcript promotes the differentiation of mouse bone marrow-derived mesenchymal stem cells into neural cells. BMC Neurosci 2011;12:67.
DOI: https://doi.org/10.1186/1471-2202-12-67
Jiang H, Niu F, Zheng Y and Xu Y. CART mitigates oxidative stress and DNA damage in memory deficits of APP/PS1 mice via upregulating betaamyloid metabolismassociated enzymes. Mol Med Rep 2021;23:280.
DOI: https://doi.org/10.3892/mmr.2021.11919
Wierup N, Kuhar M, Nilsson BO, Mulder H, Ekblad E, Sundler F. Cocaine- and amphetamine-regulated transcript (CART) is expressed in several islet cell types during rat development. J Histochem Cytochem 2004;52:169-77.
DOI: https://doi.org/10.1177/002215540405200204
Landerholm K, Shcherbina L, Falkmer SE, Jarhult J and Wierup N. Expression of cocaine- and amphetamine-regulated transcript is associated with worse survival in small bowel carcinoid tumors. Clin Cancer Res 2012;18:3668-76.
DOI: https://doi.org/10.1158/1078-0432.CCR-11-2513
Landerholm K, Falkmer SE, Jarhult J, Sundler F, Wierup N. Cocaine- and amphetamine-regulated transcript in neuroendocrine tumors. Neuroendocrinology 2011;94:228-36.
DOI: https://doi.org/10.1159/000329044
Wierup N, Abels M, Shcherbina L, Lindqvist A. The role of CART in islet biology. Peptides 2022;149:170708.
DOI: https://doi.org/10.1016/j.peptides.2021.170708
Fan L, Zhou L. AG490 protects cerebral ischemia/reperfusion injury via inhibiting the JAK2/3 signaling pathway. Brain Behav 2021;11:e01911.
DOI: https://doi.org/10.1002/brb3.1911
Prakash Y, Thompson MA, Meuchel L, Pabelick CM, Mantilla CB, Zaidi S, et al. Neurotrophins in lung health and disease. Expert Rev Respir Med 2010;4:395-411.
DOI: https://doi.org/10.1586/ers.10.29
Cefis M, Chaney R, Quirie A, Santini C, Marie C, Garnier P, et al. Endothelial cells are an important source of BDNF in rat skeletal muscle. Sci Rep 2022;12:311.
DOI: https://doi.org/10.1038/s41598-021-03740-8
Irfan M, Kim JH, Druzinsky RE, Ravindran S and Chung S. Complement C5aR/LPS-induced BDNF and NGF modulation in human dental pulp stem cells. Sci Rep 2022;12:2042.
DOI: https://doi.org/10.1038/s41598-022-06110-0
D'Este L, Casini A, Kimura S, Bellier JP, Ito E, Kimura H, et al. Immunohistochemical demonstration of cholinergic structures in central ganglia of the slug (Limax maximus, Limax valentianus). Neurochem Int 2011;58:605-11.
DOI: https://doi.org/10.1016/j.neuint.2011.02.002
Patel TD, Jackman A, Rice FL, Kucera J, Snider WD. Development of sensory neurons in the absence of NGF/TrkA signaling in vivo. Neuron 2000;25:345-57.
DOI: https://doi.org/10.1016/S0896-6273(00)80899-5
Marchetti L, Bonsignore F, Gobbo F, Amodeo R, Calvello M, Jacob A, et al. Fast-diffusing p75(NTR) monomers support apoptosis and growth cone collapse by neurotrophin ligands. Proc Natl Acad Sci USA 2019;116:21563-72.
DOI: https://doi.org/10.1073/pnas.1902790116
Wang Z, Wu J, Hu Z, Luo C, Wang P, Zhang Y, et al. Dexmedetomidine Alleviates lipopolysaccharide-induced acute kidney injury by inhibiting p75NTR-mediated oxidative stress and apoptosis. Oxid Med Cell Longev 2020;2020:5454210.
DOI: https://doi.org/10.1155/2020/5454210
Reichardt LF. Neurotrophin-regulated signalling pathways. Philos Trans R Soc Lond B Biol Sci 2006;361:1545-64.
DOI: https://doi.org/10.1098/rstb.2006.1894
Yuan JH, Pan F, Chen J, Chen CE, Xie DP, Jiang XZ, et al. Neuroprotection by plumbagin involves BDNF-TrkB-PI3K/Akt and ERK1/2/JNK pathways in isoflurane-induced neonatal rats. J Pharm Pharmacol 2017;69:896-906.
DOI: https://doi.org/10.1111/jphp.12681
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All animal experiments were performed following the regulations of the protocols approved by Baylor Scott & White Institutional Animal Care and Use CommitteeSupporting Agencies
Sapienza University of Rome, ItalyHow to Cite
Casini, A., Vivacqua, G., Vaccaro, R., Renzi, A., Leone, S., Pannarale, L., Franchitto, A., Onori, P., Mancinelli, R., & Gaudio, E. (2023). Expression and role of cocaine-amphetamine regulated transcript (CART) in the proliferation of biliary epithelium. European Journal of Histochemistry, 67(4). https://doi.org/10.4081/ejh.2023.3846
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