Abstract
Parasitic nematode infections are treated using anthelmintic drugs, some of which target nicotinic acetylcholine receptors (nAChRs) located in different parasite tissues. The limited arsenal of anthelmintic agents and the prevalence of drug resistance imply that future defense against parasitic infections will depend on the discovery of novel targets and therapeutics. Previous studies have suggested that Ascaris suum ACR-16 nAChRs are a suitable target for the development of antinematodal drugs. In this study, we characterized the pharmacology of the Ancylostoma caninum ACR-16 receptor using two-electrode voltage-clamp electrophysiology. This technique allowed us to study the effects of cholinergic agonists and antagonists on the nematode nAChRs expressed in Xenopus laevis oocytes. Aca-ACR-16 was not sensitive to many of the existing cholinomimetic anthelmintics (levamisole, oxantel, pyrantel, and tribendimidine). 3-Bromocytisine was the most potent agonist (> 130% of the control acetylcholine current) on the Aca-ACR-16 nAChR but, unlike Asu-ACR-16, oxantel did not activate the receptor. The mean time constants of desensitization for agonists on Aca-ACR-16 were longer than the rates observed in Asu-ACR-16. In contrast to Asu-ACR-16, the A. caninum receptor was completely inhibited by DHβE and moderately inhibited by α-BTX. In conclusion, we have successfully reconstituted a fully functional homomeric nAChR, ACR-16, from A. caninum, a model for human hookworm infections. The pharmacology of the receptor is distinct from levamisole-sensitive nematode receptors. The ACR-16 homologue also displayed some pharmacological differences from Asu-ACR-16. Hence, A. caninum ACR-16 may be a valid target site for the development of anthelmintics against hookworm infections.
Similar content being viewed by others
References
Abongwa M, Buxton SK, Courtot E, Charvet CL, Neveu C, McCoy CJ et al (2016) Pharmacological profile of Ascaris suum ACR-16, a new homomeric nicotinic acetylcholine receptor widely distributed in Ascaris tissues. Br J Pharmacol 173(16):2463–2477. https://doi.org/10.1111/bph.13524
Abongwa M, Martin RJ, Robertson AP (2017) A brief review on the mode of action of antinematodal drugs. Acta Vet (Beogr) 67(2):137–152. https://doi.org/10.1515/acve-2017-0013
Aceves J, Erlij D, Martínez-Marañón R (1970) The mechanism of the paralysing action of tetramisole on Ascaris somatic muscle. Br J Pharmacol 38(3):602–607
Albonico M, Smith PG, Ercole E, Hall A, Chwaya HM, Alawi KS et al (1995) Rate of reinfection with intestinal nematodes after treatment of children with mebendazole or albendazole in a highly endemic area. Trans R Soc Trop Med Hyg 89(5):538–541
Albonico M, Bickle Q, Ramsan M, Montresor A, Savioli L, Taylor M (2003) Efficacy of mebendazole and levamisole alone or in combination against intestinal nematode infections after repeated targeted mebendazole treatment in Zanzibar. Bull World Health Organ 81(5):343–352
Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W et al (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25(17):3389–3402
Aubry ML, Cowell P, Davey MJ, Shevde S (1970) Aspects of the pharmacology of a new anthelmintic: pyrantel. Br J Pharmacol 38(2):332–344
Baermann G (1917) A simple method for the detection of Ankylostomum (nematode) larvae in soil tests. Javasche Boekhandel & Drukkerij, Batavia
Ballivet M, Alliod C, Bertrand S, Bertrand D (1996) Nicotinic acetylcholine receptors in the NematodeCaenorhabditis elegans. J Mol Biol 258(2):261–269. https://doi.org/10.1006/jmbi.1996.0248
Bartsch SM, Hotez PJ, Asti L, Zapf KM, Bottazzi ME, Diemert DJ et al (2016) The global economic and health burden of human hookworm infection. PLoS Negl Trop Dis 10(9):e0004922. https://doi.org/10.1371/journal.pntd.0004922
Bennett A, Guyatt H (2000) Reducing intestinal nematode infection: efficacy of albendazole and mebendazole. Parasitol Today 16(2):71–74
Bethony J, Brooker S, Albonico M, Geiger SM, Loukas A, Diemert D et al (2006) Soil-transmitted helminth infections: ascariasis, trichuriasis, and hookworm. Lancet 367(9521):1521–1532. https://doi.org/10.1016/S0140-6736(06)68653-4
Blaxter M (2000) Genes and genomes of Necator americanus and related hookworms. Int J Parasitol 30(4):347–355. https://doi.org/10.1016/S0020-7519(99)00198-8
Boulin T, Gielen M, Richmond JE, Williams DC, Paoletti P, Bessereau JL (2008) Eight genes are required for functional reconstitution of the Caenorhabditis elegans levamisole-sensitive acetylcholine receptor. Proc Natl Acad Sci USA 105(47):18590–18595. https://doi.org/10.1073/pnas.0806933105
Bowman DD, Montgomery SP, Zajac AM, Eberhard ML, Kazacos KR (2010) Hookworms of dogs and cats as agents of cutaneous larva migrans. Trends Parasitol 26(4):162–167. https://doi.org/10.1016/j.pt.2010.01.005
Brooker S, Hotez PJ, Bundy DA (2008) Hookworm-related anaemia among pregnant women: a systematic review. PLoS Negl Trop Dis 2(9):e291. https://doi.org/10.1371/journal.pntd.0000291
Campbell SJ, Biritwum NK, Woods G, Velleman Y, Fleming F, Stothard JR (2018) Tailoring water, sanitation, and hygiene (WASH) targets for soil-transmitted helminthiasis and schistosomiasis control. Trends Parasitol 34(1):53–63. https://doi.org/10.1016/j.pt.2017.09.004
Charvet CL, Guégnard F, Courtot E, Cortet J, Neveu C (2018) Nicotine-sensitive acetylcholine receptors are relevant pharmacological targets for the control of multidrug resistant parasitic nematodes. Int J Parasitol Drugs Drug Resist 8(3):540–549. https://doi.org/10.1016/j.ijpddr.2018.11.003
Choudhary S, Marjianović DS, Wong CR, Zhang X, Abongwa M, Coats JR et al (2019) Menthol acts as a positive allosteric modulator on nematode levamisole sensitive nicotinic acetylcholine receptors. Int J Parasitol Drugs Drug Resist 9:44–53. https://doi.org/10.1016/j.ijpddr.2018.12.005
Conder GA, Campbell WC (1995) Chemotherapy of nematode infections of veterinary importance, with special reference to drug resistance. Adv Parasitol 35:1–84
Corringer PJ, Le Novere N, Changeux JP (2000) Nicotinic receptors at the amino acid level. Annu Rev Pharmacol Toxicol 40:431–458. https://doi.org/10.1146/annurev.pharmtox.40.1.431
De Clercq D, Sacko M, Behnke J, Gilbert F, Dorny P, Vercruysse J (1997a) Failure of mebendazole in treatment of human hookworm infections in the Southern Region of Mali. Am J Trop Med Hyg 57(1):25–30. https://doi.org/10.4269/ajtmh.1997.57.25
De Clercq D, Sacko M, Behnke J, Gilbert F, Dorny P, Vercruysse J (1997b) Failure of mebendazole in treatment of human hookworm infections in the southern region of Mali. Am J Trop Med Hyg 57(1):25–30
de Silva NR, Brooker S, Hotez PJ, Montresor A, Engels D, Savioli L (2003) Soil-transmitted helminth infections: updating the global picture. Trends Parasitol 19(12):547–551
Dias SRC, Cunha DES, da Silva SM, dos Santos HA, Fujiwara RT, Rabelo ÉML (2013) Evaluation of parasitological and immunological aspects of acute infection by Ancylostoma caninum and Ancylostoma braziliense in mixed-breed dogs. Parasitol Res 112(6):2151–2157. https://doi.org/10.1007/s00436-013-3370-y
Diemert DJ, Bethony JM, Hotez PJ (2008) Hookworm vaccines. Clin Infect Dis 46(2):282–288. https://doi.org/10.1086/524070
Epe C (2009) Intestinal nematodes: biology and control. Vet Clin North Am Small Anim Pract 39(6):1091–1107. https://doi.org/10.1016/j.cvsm.2009.07.002
Flohr C, Tuyen LN, Lewis S, Minh TT, Campbell J, Britton J et al (2007) Low efficacy of mebendazole against hookworm in Vietnam: two randomized controlled trials. Am J Trop Med Hyg 76(4):732–736
Francis MM, Evans SP, Jensen M, Madsen DM, Mancuso J, Norman KR et al (2005) The Ror receptor tyrosine kinase CAM-1 is required for ACR-16-mediated synaptic transmission at the C. elegans neuromuscular junction. Neuron 46(4):581–594. https://doi.org/10.1016/j.neuron.2005.04.010
Geerts S, Gryseels B (2000) Drug resistance in human helminths: current situation and lessons from livestock. Clin Microbiol Rev 13(2):207–222
Gibson DG, Young L, Chuang RY, Venter JC, Hutchison CA 3rd, Smith HO (2009) Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods 6(5):343–345. https://doi.org/10.1038/nmeth.1318
Giniatullin R, Nistri A, Yakel JL (2005) Desensitization of nicotinic ACh receptors: shaping cholinergic signaling. Trends Neurosci 28(7):371–378. https://doi.org/10.1016/j.tins.2005.04.009
Global Burden of Disease Study, C (2015) Global, regional, and national incidence, prevalence, and years lived with disability for 301 acute and chronic diseases and injuries in 188 countries, 1990–2013: a systematic analysis for the Global Burden of Disease Study 2013. Lancet 386(9995):743–800. https://doi.org/10.1016/S0140-6736(15)60692-4
Guyatt HL, Brooker S, Peshu N, Shulman CE (2000) Hookworm and anaemia prevalence. Lancet 356(9247):2101
Harrow ID, Gration KAF (1985) Mode of action of the anthelmintics morantel, pyrantel and levamisole on muscle cell membrane of the nematode Ascaris suum. Pestic Sci 16(6):662–672. https://doi.org/10.1002/ps.2780160612
Hewitson JP, Maizels RM (2014) Vaccination against helminth parasite infections. Expert Rev Vaccines 13(4):473–487. https://doi.org/10.1586/14760584.2014.893195
Hofmann K, Stoffel W (1993) TMbase—a database of membrane spanning proteins segments. Biol. Chem. Hoppe-Seyler 374:166
Hotez P (2008) Hookworm and poverty. Ann N Y Acad Sci 1136:38–44. https://doi.org/10.1196/annals.1425.000
Hotez PJ, Pritchard DI (1995) Hookworm infection. Sci Am 272(6):68. https://doi.org/10.1038/scientificamerican0695-68
Hotez PJ, Alvarado M, Basanez MG, Bolliger I, Bourne R, Boussinesq M et al (2014) The global burden of disease study 2010: interpretation and implications for the neglected tropical diseases. PLoS Negl Trop Dis 8(7):e2865. https://doi.org/10.1371/journal.pntd.0002865
Jones AK, Davis P, Hodgkin J, Sattelle DB (2007) The nicotinic acetylcholine receptor gene family of the nematode Caenorhabditis elegans: an update on nomenclature. Invertebr Neurosci 7(2):129–131. https://doi.org/10.1007/s10158-007-0049-z
Kaminsky R, Ducray P, Jung M, Clover R, Rufener L, Bouvier J et al (2008) A new class of anthelmintics effective against drug-resistant nematodes. Nature 452(7184):176–180. https://doi.org/10.1038/nature06722
Kaplan RM (2004) Drug resistance in nematodes of veterinary importance: a status report. Trends Parasitol 20(10):477–481. https://doi.org/10.1016/j.pt.2004.08.001
Keiser J, Utzinger J (2008) Efficacy of current drugs against soil-transmitted helminth infections: systematic review and meta-analysis. JAMA 299(16):1937–1948. https://doi.org/10.1001/jama.299.16.1937
Knopp S, Steinmann P, Keiser J, Utzinger J (2012) Nematode infections: soil-transmitted helminths and trichinella. Infect Dis Clin North Am 26(2):341–358. https://doi.org/10.1016/j.idc.2012.02.006
Krücken J, Fraundorfer K, Mugisha JC, Ramünke S, Sifft KC, Geus D et al (2017) Reduced efficacy of albendazole against Ascaris lumbricoides in Rwandan schoolchildren. Int J Parasitol Drugs Drug Resist 7(3):262–271. https://doi.org/10.1016/j.ijpddr.2017.06.001
Landmann JK, Prociv P (2003) Experimental human infection with the dog hookworm, Ancylostoma caninum. Med J Aust 178(2):69–71
Levin ED (2002) Nicotinic receptor subtypes and cognitive function. J Neurobiol 53(4):633–640. https://doi.org/10.1002/neu.10151
Li Z, Liu J, Zheng M, Xu XZS (2014) Encoding of both analog- and digital-like behavioral outputs by one C. elegans interneuron. Cell 159(4):751–765. https://doi.org/10.1016/j.cell.2014.09.056
Loukas A, Hotez PJ, Diemert D, Yazdanbakhsh M, McCarthy JS, Correa-Oliveira R et al (2016) Hookworm infection. Nat Rev Dis Primers 2:16088. https://doi.org/10.1038/nrdp.2016.88
Martin RJ, Verma S, Levandoski M, Clark CL, Qian H, Stewart M et al (2005) Drug resistance and neurotransmitter receptors of nematodes: recent studies on the mode of action of levamisole. Parasitology 131(Suppl):S71–S84. https://doi.org/10.1017/s0031182005008668
Martin RJ, Robertson AP, Buxton SK, Beech RN, Charvet CL, Neveu C (2012) Levamisole receptors: a second awakening. Trends Parasitol 28(7):289–296. https://doi.org/10.1016/j.pt.2012.04.003
Menzies SK, Rodriguez A, Chico M, Sandoval C, Broncano N, Guadalupe I et al (2014) Risk factors for soil-transmitted helminth infections during the first 3 years of life in the tropics; findings from a birth cohort. PLoS Negl Trop Dis 8(2):e2718. https://doi.org/10.1371/journal.pntd.0002718
Nemzek JA, Lester PA, Wolfe AM, Dysko RC, Myers DD (2015) Chapter 12—Biology and diseases of dogs. In: Fox JG, Anderson LC, Otto GM, Pritchett-Corning KR, Whary MT (eds) Laboratory animal medicine, 3rd edn. Academic Press, Boston, pp 511–554
Petersen TN, Brunak S, von Heijne G, Nielsen H (2011) SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat Methods 8(10):785–786. https://doi.org/10.1038/nmeth.1701
Picciotto MR, Addy NA, Mineur YS, Brunzell DH (2008) It is not “either/or”: activation and desensitization of nicotinic acetylcholine receptors both contribute to behaviors related to nicotine addiction and mood. Prog Neurobiol 84(4):329–342. https://doi.org/10.1016/j.pneurobio.2007.12.005
Prociv P, Croese J (1996) Human enteric infection with Ancylostoma caninum: hookworms reappraised in the light of a “new” zoonosis. Acta Trop 62(1):23–44
Pullan RL, Brooker SJ (2012) The global limits and population at risk of soil-transmitted helminth infections in 2010. Parasit Vectors 5:81. https://doi.org/10.1186/1756-3305-5-81
Pullan RL, Smith JL, Jasrasaria R, Brooker SJ (2014) Global numbers of infection and disease burden of soil transmitted helminth infections in 2010. Parasit Vectors 7:37. https://doi.org/10.1186/1756-3305-7-37
Quick MW, Lester RA (2002) Desensitization of neuronal nicotinic receptors. J Neurobiol 53(4):457–478. https://doi.org/10.1002/neu.10109
Raymond V, Mongan NP, Sattelle DB (2000) Anthelmintic actions on homomer-forming nicotinic acetylcholine receptor subunits: chicken α7 and ACR-16 from the nematode Caenorhabditis elegans. Neuroscience 101(3):785–791. https://doi.org/10.1016/S0306-4522(00)00279-7
Reynoldson JA, Behnke JM, Pallant LJ, Macnish MG, Gilbert F, Giles S et al (1997) Failure of pyrantel in treatment of human hookworm infections (Ancylostoma duodenale) in the Kimberley region of North West Australia. Acta Trop 68(3):301–312. https://doi.org/10.1016/S0001-706X(97)00106-X
Richmond JE, Jorgensen EM (1999) One GABA and two acetylcholine receptors function at the C. elegans neuromuscular junction. Nat Neurosci 2(9):791–797. https://doi.org/10.1038/12160
Sievers F, Wilm A, Dineen D, Gibson TJ, Karplus K, Li W et al (2011) Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol Syst Biol 7:539. https://doi.org/10.1038/msb.2011.75
Thompson AJ, Lester HA, Lummis SC (2010) The structural basis of function in Cys-loop receptors. Q Rev Biophys 43(4):449–499. https://doi.org/10.1017/S0033583510000168
Touroutine D, Fox RM, Von Stetina SE, Burdina A, Miller DM, Richmond JE (2005) acr-16 encodes an essential subunit of the levamisole-resistant nicotinic receptor at the Caenorhabditis elegans neuromuscular junction. J Biol Chem 280(29):27013–27021
Traversa D (2012) Pet roundworms and hookworms: a continuing need for global worming. Parasit Vectors 5:91. https://doi.org/10.1186/1756-3305-5-91
Verma S, Kashyap SS, Robertson AP, Martin RJ (2017) Functional genomics in Brugia malayi reveal diverse muscle nAChRs and differences between cholinergic anthelmintics. Proc Natl Acad Sci USA 114(21):5539–5544. https://doi.org/10.1073/pnas.1619820114
WHO (2015) Investing to overcome the global impact of neglected tropical diseases: third WHO report on neglected diseases. World Health Organisation, Geneva
Wolstenholme AJ (2011) Ion channels and receptor as targets for the control of parasitic nematodes. Int J Parasitol Drugs Drug Resist 1(1):2–13. https://doi.org/10.1016/j.ijpddr.2011.09.003
Wolstenholme AJ, Fairweather I, Prichard R, von Samson-Himmelstjerna G, Sangster NC (2004) Drug resistance in veterinary helminths. Trends Parasitol 20(10):469–476. https://doi.org/10.1016/j.pt.2004.07.010
Zheng F, Robertson AP, Abongwa M, Yu EW, Martin RJ (2016) The Ascaris suum nicotinic receptor, ACR-16, as a drug target: four novel negative allosteric modulators from virtual screening. Int J Parasitol Drugs Drug Resist 6(1):60–73. https://doi.org/10.1016/j.ijpddr.2016.02.001
Acknowledgements
This research was funded by NIH R21AI092185 to APR, NIH RO1AI047194 and the E. A. Benbrook Foundation for Pathology and Parasitology to RJM. We would like to thank Dr. Claude L. Charvet, INRA, Université Tours, Nouzilly, France for the generous gift of pTB207 vector.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that this work has no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Choudhary, S., Tipton, J.G., Abongwa, M. et al. Pharmacological characterization of a homomeric nicotinic acetylcholine receptor formed by Ancylostoma caninum ACR-16. Invert Neurosci 19, 11 (2019). https://doi.org/10.1007/s10158-019-0231-0
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s10158-019-0231-0