Identification of intrinsic primary afferent neurons in mouse jejunum
Carina Guimarães de Souza Melo
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorEvan N. Nicolai
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorConstanza Alcaino
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorTiffany J. Cassmann
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorSara T. Whiteman
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorAlec M. Wright
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorKatie E. Miller
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorSimon J. Gibbons
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorArthur Beyder
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorCorresponding Author
David R. Linden
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Correspondence
David R. Linden, Department of Physiology and Biomedical Engineering, Mayo Clinic College of Medicine, 200 First Street SW, Rochester, MN 55905, USA.
Email: [email protected]
Search for more papers by this authorCarina Guimarães de Souza Melo
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorEvan N. Nicolai
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorConstanza Alcaino
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorTiffany J. Cassmann
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorSara T. Whiteman
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorAlec M. Wright
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorKatie E. Miller
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorSimon J. Gibbons
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorArthur Beyder
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Search for more papers by this authorCorresponding Author
David R. Linden
Department of Physiology and Biomedical Engineering and Enteric NeuroScience Program, Mayo Clinic College of Medicine, Rochester, MN, USA
Correspondence
David R. Linden, Department of Physiology and Biomedical Engineering, Mayo Clinic College of Medicine, 200 First Street SW, Rochester, MN 55905, USA.
Email: [email protected]
Search for more papers by this authorFunding information
National Institutes of Health R01DK106011 R03DK119683 K08DK106456Optical Microscopy Core of the Mayo Clinic Center for Cell Signaling in Gastroenterology P30DK084567Department of Defense W81XWH-18-1-0218Brazilian Research Foundation CNPq 200879/2015-4.
Abstract
Background
The gut is the only organ system with intrinsic neural reflexes. Intrinsic primary afferent neurons (IPANs) of the enteric nervous system initiate intrinsic reflexes, form gut-brain connections, and undergo considerable neuroplasticity to cause digestive diseases. They remain inaccessible to study in mice in the absence of a selective marker. Advillin is used as a marker for primary afferent neurons in dorsal root ganglia. The aim of this study was to test the hypothesis that advillin is expressed in IPANs of the mouse jejunum.
Methods
Advillin expression was assessed with immunohistochemistry and using transgenic mice expressing an inducible Cre recombinase under the advillin promoter were used to drive tdTomato and the genetically encoded calcium indicator GCaMP5. These mice were used to characterize the morphology and physiology of advillin-expressing enteric neurons using confocal microscopy, calcium imaging, and whole-cell patch-clamp electrophysiology.
Key Results
Advillin is expressed in about 25% of myenteric neurons of the mouse jejunum, and these neurons demonstrate the requisite properties of IPANs. Functionally, they demonstrate calcium responses following mechanical stimuli of the mucosa and during antidromic action potentials. They have Dogiel type II morphology with neural processes that mostly remain within the myenteric plexus, but also project to the mucosa and express NeuN and calcitonin gene-related peptide (CGRP), but not nNOS.
Conclusions and Inferences
Advillin marks jejunal IPANs providing accessibility to this important neuronal population to study and model digestive disease.
DISCLOSURE
No competing interests declared.
Supporting Information
| Filename | Description |
|---|---|
| nmo13989-sup-0001-FigureS1.tifTIFF image, 18.9 MB | Figure S1 |
| nmo13989-sup-0002-FigureS2.tifTIFF image, 11.5 MB | Figure S2 |
| nmo13989-sup-0003-FigureS3.tifTIFF image, 14.6 MB | Figure S3 |
| nmo13989-sup-0004-VideoS1.mp4MPEG-4 video, 14.9 MB | Video S1 |
| nmo13989-sup-0005-VideoS2.mp4MPEG-4 video, 11.9 MB | Video S2 |
| nmo13989-sup-0006-VideoS3.mp4MPEG-4 video, 10 MB | Video S3 |
| nmo13989-sup-0007-VideoS4.mp4MPEG-4 video, 9.9 MB | Video S4 |
| nmo13989-sup-0008-supcap.docxWord document, 14.8 KB | Supplementary Material |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
References
- 1Furness JB, Jones C, Nurgali K, Clerc N. Intrinsic primary afferent neurons and nerve circuits within the intestine. Prog Neurogibol. 2004; 72: 143-164.
- 2Bertrand PP, Kunze WA, Bornstein JC, Furness JB. Electrical mapping of the projections of intrinsic primary afferent neurones to the mucosa of the guinea-pig small intestine. Neurogastroenterol Motil. 1998; 10: 533-541.
- 3Kunze WA, Clerc N, Furness JB, Gola M. The soma and neurites of primary afferent neurons in the guinea-pig intestine respond differentially to deformation. J Physiol. 2000; 526(Pt 2): 375-385.
- 4Kunze WA, Furness JB, Bertrand PP, Bornstein JC. Intracellular recording from myenteric neurons of the guinea-pig ileum that respond to stretch. J Physiol. 1998; 506(Pt 3): 827-842.
- 5Katayama Y, North RA, Williams JT. The action of substance P on neurons of the myenteric plexus of the guinea-pig small intestine. Proc R Soc Lond B Biol Sci. 1979; 206: 191-208.
- 6Johnson SM, Katayama Y, North RA. Multiple actions of 5-hydroxytryptamine on myenteric neurones of the guinea-pig ileum. J Physiol. 1980; 304: 459-470.
- 7Bertrand PP, Bornstein JC. ATP as a putative sensory mediator: activation of intrinsic sensory neurons of the myenteric plexus via P2X receptors. J Neurosci. 2002; 22: 4767-4775.
- 8Baldassano S, Wang GD, Mule F, Wood JD. Glucagon-like peptide-1 modulates neurally evoked mucosal chloride secretion in guinea pig small intestine in vitro. Am J Physiol Gastrointest Liver Physiol. 2012; 302: G352-G358.
- 9Mao YK, Kasper DL, Wang B, Forsythe P, Bienenstock J, Kunze WA. Bacteroides fragilis polysaccharide a is necessary and sufficient for acute activation of intestinal sensory neurons. Nat Commun. 2013; 4: 1465.
- 10Xia Y, Hu HZ, Liu S, Ren J, Zafirov DH, Wood JD. IL-1beta and IL-6 excite neurons and suppress nicotinic and noradrenergic neurotransmission in guinea pig enteric nervous system. J Clin Invest. 1999; 103: 1309-1316.
- 11Manning BP, Sharkey KA, Mawe GM. Effects of PGE2 in guinea pig colonic myenteric ganglia. Am J Physiol Gastrointest Liver Physiol. 2002; 283: G1388-G1397.
- 12Hu HZ, Gao N, Liu S, et al. Action of bradykinin in the submucosal plexus of guinea pig small intestine. J Pharmacol Exp Ther. 2004; 309: 320-327.
- 13Perez-Burgos A, Mao YK, Bienenstock J, Kunze WA. The gut-brain axis rewired: Adding a functional vagal nicotinic "Sensory Synapse". FASEB J. 2014; 28: 3064-3074.
- 14Brierley SM, Linden DR. Neuroplasticity and dysfunction after gastrointestinal inflammation. Nat Rev Gastroenterol Hepatol. 2014; 11: 611-627.
- 15Qu ZD, Thacker M, Castelucci P, Bagyanszki M, Epstein ML, Furness JB. Immunohistochemical analysis of neuron types in the mouse small intestine. Cell Tissue Res. 2008; 334: 147-161.
- 16Hillsley K, Kenyon JL, Smith TK. Ryanodine-sensitive stores regulate the excitability of ah neurons in the myenteric plexus of guinea-pig ileum. J Neurophysiol. 2000; 84: 2777-2785.
- 17Hirst GD, Holman ME, Spence I. Two types of neurones in the myenteric plexus of duodenum in the guinea-pig. J Physiol. 1974; 236: 303-326.
- 18Linden DR, Sharkey KA, Mawe GM. Enhanced excitability of myenteric AH neurones in the inflamed guinea-pig distal colon. J Physiol. 2003; 547: 589-601.
- 19Nishi S, North RA. Intracellular recording from the myenteric plexus of the guinea-pig ileum. J Physiol. 1973; 231: 471-491.
- 20Mao Y, Wang B, Kunze W. Characterization of myenteric sensory neurons in the mouse small intestine. J Neurophysiol. 2006; 96: 998-1010.
- 21Ren J, Bian X, Devries M, et al. P2X2 subunits contribute to fast synaptic excitation in myenteric neurons of the mouse small intestine. J Physiol. 2003; 552: 809-821.
- 22Rugiero F, Mistry M, Sage D, et al. Selective expression of a persistent tetrodotoxin-resistant Na+ current and Nav1.9 subunit in myenteric sensory neurons. J Neurosci. 2003; 23: 2715-2725.
- 23Marks PW, Arai M, Bandura JL, Kwiatkowski DJ. Advillin (P92): a new member of the gelsolin/villin family of actin regulatory proteins. J Cell Sci. 1998; 111(Pt 15): 2129-2136.
- 24Hasegawa H, Abbott S, Han BX, Qi Y, Wang F. Analyzing somatosensory axon projections with the sensory neuron-specific advillin gene. J Neurosci. 2007; 27: 14404-14414.
- 25Lau J, Minett MS, Zhao J, et al. Temporal control of gene deletion in sensory ganglia using a tamoxifen-inducible Advillin-Cre-ERT2 recombinase mouse. Mol Pain. 2011; 7: 100.
- 26Hunter DV, Smaila BD, Lopes DM, Takatoh J, Denk F, Ramer MS. Advillin is expressed in all adult neural crest-derived neurons. Eneuro. 2018; 5:ENEURO.0077-18.2018.
- 27Gee JM, Smith NA, Fernandez FR, et al. Imaging activity in neurons and glia with a Polr2a-based and Cre-dependent Gcamp5g-IRES-Tdtomato reporter mouse. Neuron. 2014; 83: 1058-1072.
- 28Taniguchi H, He M, Wu P, et al. A resource of Cre driver lines for genetic targeting of gabaergic neurons in cerebral cortex. Neuron. 2011; 71: 995-1013.
- 29Van Nassauw L, Wu M, De Jonge F, Adriaensen D, Timmermans JP. Cytoplasmic, but not nuclear, expression of the neuronal nuclei (NeuN) antibody is an exclusive feature of Dogiel type II neurons in the guinea-pig gastrointestinal tract. Histochem Cell Biol. 2005; 124: 369-377.
- 30Furness JB, Robbins HL, Xiao J, Stebbing MJ, Nurgali K. Projections and chemistry of Dogiel type II neurons in the mouse colon. Cell Tissue Res. 2004; 317: 1-12.
- 31Brierley SM, Jones RC 3rd, Gebhart GF, Blackshaw LA. Splanchnic and pelvic mechanosensory afferents signal different qualities of colonic stimuli In mice. Gastroenterology. 2004; 127: 166-178.
- 32Smith TK, Kang SH, Vanden BP. Calcium channels in enteric neurons. Curr Opin Pharmacol. 2003; 3: 588-593.
- 33Rugiero F, Gola M, Kunze WA, Reynaud JC, Furness JB, Clerc N. Analysis of whole-cell currents by patch clamp of guinea-pig myenteric neurones in intact ganglia. J Physiol. 2002; 538: 447-463.
- 34Song ZM, Brookes SJ, Costa M. Identification of myenteric neurons which project to the mucosa of the guinea-pig small intestine. Neurosci Lett. 1991; 129: 294-298.
- 35Costa M, Brody K, Brookes SJH. A new marker for enteric primary afferent neurones. Neurogastroenterol Motil. 2001; 13: 383.
- 36Sang Q, Young HM. Chemical coding of neurons in the myenteric plexus and external muscle of the small and large intestine of the mouse. Cell Tissue Res. 1996; 284: 39-53.
- 37Moghimzadeh E, Ekman R, Hakanson R, Yanaihara N, Sundler F. Neuronal gastrin-releasing peptide in the mammalian gut and pancreas. Neuroscience. 1983; 10: 553-563.
- 38Hendriks R, Bornstein JC, Furness JB. An electrophysiological study of the projections of putative sensory neurons within the myenteric plexus of the guinea pig ileum. Neurosci Lett. 1990; 110: 286-290.
- 39Zappia KJ, O'Hara CL, Moehring F, Kwan KY, Stucky CL. Sensory neuron-specific deletion of TRPA1 results in mechanical cutaneous sensory deficits. Eneuro. 2017; 4:ENEURO.0069-16.2017.
Citing Literature
