Itch-associated peptides: RNA-Seq and bioinformatic analysis of natriuretic precursor peptide B and gastrin releasing peptide in dorsal root and trigeminal ganglia, and the spinal cord

Molecular Pain

Michael J Iadarola²  Andrew J Mannes²  Mark A Hoon¹  Santosh Mishra¹  Jean Thierry-Mieg³  Danielle Thierry-Mieg³  Samridhi C Goswami² 

[1] Molecular Genetics Unit, Laboratory of Sensory Biology, National Institute of Dental and Craniofacial Research, NIH, Bethesda, MD, USA;Department of Perioperative Medicine, Building 10, Room 2C401, MSC 1510, Clinical Center, NIH, 10 Center Drive, Bethesda, MD 20892, USA;National Center for Biomedical Information, NIH, Bethesda, MD, USA

关键词: Cross-reaction;    Immunocytochemistry;    Peptidylglycine alpha-amidating monooxygenase;    RNA-Seq unified mapper;    PolyA⁺ mRNA;    RNA-Seq;    Npr3;    Npr2;    Npr1;    Nppc;    Nppb;    Nppa;    Tac1;    NMBR;    NMB;    GRPR;    GRP;    Ranatensin;    Bombesin;    Pruritis;   

Others  :  1161773
DOI  :  10.1186/1744-8069-10-44

 received in 2014-05-21, accepted in 2014-06-25,  发布年份 2014

【 摘 要 】

Background

Three neuropeptides, gastrin releasing peptide (GRP), natriuritic precursor peptide B (NPPB), and neuromedin B (NMB) have been proposed to play roles in itch sensation. However, the tissues in which these peptides are expressed and their positions in the itch circuit has recently become the subject of debate. Here we used next-gen RNA-Seq to examine the expression of transcripts coding for GRP, NPPB, NMB, and other peptides in DRG, trigeminal ganglion, and the spinal cord as well as expression levels for their cognate receptors in these tissues.

Results

RNA-Seq demonstrates that GRP is not transcribed in mouse, rat, or human sensory ganglia. NPPB, which activates natriuretic peptide receptor 1 (NPR1), is well expressed in mouse DRG and less so in rat and human, whereas NPPA, which also acts on the NPR1 receptor, is expressed in all three species. Analysis of transcripts expressed in the spinal cord of mouse, rat, and human reveals no expression of Nppb, but unambiguously detects expression of Grp and the GRP-receptor (Grpr). The transcripts coding for NMB and tachykinin peptides are among the most highly expressed in DRG. Bioinformatics comparisons using the sequence of the peptides used to produce GRP-antibodies with proteome databases revealed that the C-terminal primary sequence of NMB and Substance P can potentially account for results from previous studies which showed GRP-immunostaining in the DRG.

Conclusions

RNA-Seq corroborates a primary itch afferent role for NPPB in mouse and potentially NPPB and NPPA in rats and humans, but does not support GRP as a primary itch neurotransmitter in mouse, rat, or humans. As such, our results are at odds with the initial proposal of Sun and Chen (2007) that GRP is expressed in DRG. By contrast, our data strongly support an itch pathway where the itch-inducing actions of GRP are exerted through its release from spinal cord neurons.

【 授权许可】

   
2014 Goswami et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

【 参考文献 】

• [1]Sun YG, Chen ZF: A gastrin-releasing peptide receptor mediates the itch sensation in the spinal cord. Nature 2007, 448(7154):700-703.
• [2]Sun YG, Zhao ZQ, Meng XL, Yin J, Liu XY, Chen ZF: Cellular basis of itch sensation. Science 2009, 325(5947):1531-1534.
• [3]Fleming MS, Ramos D, Han SB, Zhao J, Son YJ, Luo W: The majority of dorsal spinal cord gastrin releasing peptide is synthesized locally whereas neuromedin B is highly expressed in pain- and itch-sensing somatosensory neurons. Mol Pain 2012, 8:52.
• [4]Mishra SK, Hoon MA: The cells and circuitry for itch responses in mice. Science 2013, 340(6135):968-971.
• [5]Su PY, Ko MC: The role of central gastrin-releasing peptide and neuromedin B receptors in the modulation of scratching behavior in rats. J Pharmacol Exp Ther 2011, 337(3):822-829.
• [6]Mishra SK, Tisel SM, Orestes P, Bhangoo SK, Hoon MA: TRPV1-lineage neurons are required for thermal sensation. EMBO J 2011, 30(3):582-593.
• [7]Mishra SK, Holzman S, Hoon MA: A nociceptive signaling role of neuromedin B. J Neurosci 2012, 32(25):8686-8695.
• [8]Hammer P, Banck MS, Amberg R, Wang C, Petznick G, Luo S, Khrebtukova I, Schroth GP, Beyerlein P, Beutler AS: mRNA-seq with agnostic splice site discovery for nervous system transcriptomics tested in chronic pain. Genome Res 2010, 20(6):847-860.
• [9]Grant GR, Farkas MH, Pizarro AD, Lahens NF, Schug J, Brunk BP, Stoeckert CJ, Hogenesch JB, Pierce EA: Comparative analysis of RNA-Seq alignment algorithms and the RNA-Seq unified mapper (RUM). Bioinformatics 2011, 27(18):2518-2528.
• [10]Bradbury AF, Finnie MD, Smyth DG: Mechanism of C-terminal amide formation by pituitary enzymes. Nature 1982, 298(5875):686-688.
• [11]Eipper BA, Milgram SL, Husten EJ, Yun HY, Mains RE: Peptidylglycine alpha-amidating monooxygenase: a multifunctional protein with catalytic, processing, and routing domains. Protein Sci 1993, 2(4):489-497.
• [12]Steinhoff MS, von Mentzer B, Geppetti P, Pothoulakis C, Bunnett NW: Tachykinins and their receptors: contributions to physiological control and the mechanisms of disease. Physiol Rev 2014, 94(1):265-301.
• [13]Misono KS, Philo JS, Arakawa T, Ogata CM, Qui Y, Ogawa H, Young HS: Structure, signaling mechanism and regulation of the natriuretic peptide receptor guanylate cyclase. FEBS J 2011, 278(11):1818-1829.
• [14]Potter LR, Hosch SA, Dickey DM: Natriuretic peptides, their receptors, and cyclic guanosine monophosphate-dependent signaling functions. Endocr Rev 2006, 27:10.
• [15]Erspamer V: Discovery, isolation, and characterization of bombesin-like peptides. Ann N Y Acad Sci 1988, 547:3-9.
• [16]Liu XY, Wan L, Huo FQ, Barry DM, Li H, Zhao ZQ, Chen ZF: B-type natriuretic peptide is neither itch-specific nor functions upstream of the GRP-GRPR signaling pathway. Mol Pain 2014, 10(1):4.
• [17]Wada E, Way J, Lebacq-Verheyden AM, Battey JF: Neuromedin B and gastrin-releasing peptide mRNAs are differentially distributed in the rat nervous system. J Neurosci 1990, 10(9):2917-2930.
• [18]Mitchell K, Iadarola MJ: RT-PCR analysis of pain genes: use of gel-based RT-PCR for studying induced and tissue-enriched gene expression. Methods Mol Biol 2010, 617:279-295.
• [19]Loo L, Shepherd AJ, Mickle AD, Lorca RA, Shutov LP, Usachev YM, Mohapatra DP: The C-type natriuretic peptide induces thermal hyperalgesia through a noncanonical Gβγ-dependent modulation of TRPV1 channel. J Neurosci 2012, 32(35):11942-11955.
• [20]Akiyama T, Cartsens E: Neural processing of itch. Neuroscience 2013, 10(250):697-714.
• [21]Bautista DM, Wilson SR, Hoon MA: Why we scratch an itch: the molecules, cells and circuits of itch. Nat Neurosci 2014, 17(2):175-182.
• [22]Jensen RT, Battey JF, Spindel ER, Benya RV: International Union of Pharmacology. LXVIII. Mammalian bombesin receptors: nomenclature, distribution, pharmacology, signaling, and functions in normal and disease states. Pharmacol Rev 2008, 60(1):1-42.
• [23]Sukhtankar DD, Ko MC: Physiological function of gastrin-releasing peptide and neuromedin B receptors in regulating itch scratching behavior in the spinal cord of mice. PLoS One 2013, 8(6):e67422.
• [24]Noguchi K, Ruda MA: Gene regulation in an ascending nociceptive pathway: inflammation-induced increase in preprotachykinin mRNA in rat lamina I spinal projection neurons. J Neurosci 1992, 12(7):2563-2572.
• [25]Benya RV, Wada E, Battey JF, Fathi Z, Wang LH, Mantey SA, Coy DH, Jensen RT: Neuromedin B receptors retain functional expression when transfected into BALB 3T3 fibroblasts: analysis of binding, kinetics, stoichiometry, modulation by guanine nucleotide-binding proteins, and signal transduction and comparison with natively expressed receptors. Mol Pharmacol 1992, 42(6):1058-1068.
• [26]Moser HR, Giesler GJ Jr: Characterization of pruriceptive trigeminothalamic tract neurons in rats. J Neurophysiol 2014, 111(8):1574-1589.
• [27]Panula P, Yang HY, Costa E: Neuronal location of the bombesin-like immunoreactivity in the central nervous system of the rat. Regul Pept 1982, 4(5):275-283.
• [28]Timmes TR, Rothbaum R, Kirti , Silva CY, Bhawan J, Cummins DL, Wolpowitz D: Gastrin-releasing peptide-expressing nerves comprise subsets of human cutaneous Aδ and C fibers that may sense pruritus. J Invest Dermatol 2013, 133(11):2645-2647.
• [29]Baroni A, Perfetto B, Canozo N, Braca A, Farina E, Melito A, De Maria S, Carteni M: Bombesin: a possible role in wound repair. Peptides 2008, 29(7):1157-1166.
• [30]Yamaguchi Y, Hosokawa K, Nakatani Y, Sano S, Yoshikawa K, Itami S: Gastrin-releasing peptide, a bombesin-like neuropeptide, promotes cutaneous wound healing. Dermatol Surg 2002, 28(4):314-319.