Original Article

Main Pathways and Ion Channels Differentially Expressed in the Transcriptome of Male and Female Adult Angiostrongylus cantonensis using a Deep Sequencing Approach


Background: The adult stage is an important period in the life cycle of Angiostrongylus cantonensis, as it is at this stage that male and female worms produce thousands of fertilized eggs daily.

Methods: To explore the transcriptional details of adult male and female A. cantonensis, three groups of male and female adult worms were collected, and their transcriptome profiles were analyzed using an Illumina next-generation sequencing platform. A total of 283,910,174 clean reads were obtained, and 137,626 unigenes and 237,059 transcripts were then generated. Unigenes were successfully annotated by querying the Gene Ontology (GO), the Kyoto Encyclopedia of Genes and Genomes (KEGG), NCBI non-redundant protein sequences (NR), PFAM, STRING, and SWISS-PROT databases. Then, differentially expressed genes (DEGs) between the 2 genders were identified. The GO and KEGG databases were used for DEG annotation, and a number of DEG annotations were enriched.

Results: The results obtained from querying DEGs using the GO and KEGG databases revealed that male and female adult worms exhibited differences in metabolism and production. Protein phosphorylation, ion transport, and calcium transport were all significantly enriched according to GO annotation. A number of other pathways were also enriched according to KEGG enrichment annotation, including the pentose phosphate pathway, nitrogen metabolism, oocyte meiosis pathway, neuroactive ligand-receptor interaction, calcium signaling pathway, transforming growth factor β (TGF-β) signaling pathway etc.

Conclusion: We hypothesized that the nervous system of the worm plays a key role in the physiological regulation of adult A. cantonensis, and based on this, the function of the calcium-signaling pathway should be investigated.

1. Aekphachaisawat N, Sawanyawisuth K, Khamsai S, et al . An ecological study of eosinophilic meningitis caused by the nematode, Angiostrongylus cantonensis (chen, 1935) (nematoda: Metastrongylidae). Parasitol Int. 2019; 72:101944.
2. Barratt J, Chan D, Sandaradura I, et al. Angiostrongylus cantonensis: A review of its distribution, molecular biology and clinical significance as a human pathogen. Parasitology. 2016; 143(9):1087-118.
3. Sohal RJ, Gilotra TS, Lui F. Angiostrongylus cantonensis (angiostrongliasis). StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2021 Jan. 2021 Sep 25.
4. Prociv P, Turner M. Neuroangiostrongyliasis: The "subarachnoid phase" and its implications for anthelminthic therapy. Am J Trop Med Hyg. 2018; 98(2):353-359.
5. Yao L, Xi S, Zhong-Dao W. [advances in pathogenic mechanisms of Angiostrongylus cantonensis infection]. Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi. 2019; 31(1):98-104.
6. Jeremias WJ, Araujo FMG, Queiroz FR, et al. Comparative sequence analysis reveals regulation of genes in developing schistosomula of Schistosoma mansoni exposed to host portal serum. PLoS One. 2017; 12(6):e0178829.
7. Lucas ER, Miles A, Harding NJ, et al. Whole-genome sequencing reveals high complexity of copy number variation at insecticide resistance loci in malaria mosquitoes. Genome Res. 2019;29:1250-1261.
8. Anderson L, Amaral MS, Beckedorff F, et al. Schistosoma mansoni egg, adult male and female comparative gene expression analysis and identification of novel genes by rna-seq. PLoS Negl Trop Dis. 2015; 9(12):e0004334.
9. Cai P, Liu S, Piao X, et al. Comprehensive transcriptome analysis of sex-biased expressed genes reveals discrete biological and physiological features of male and female Schistosoma japonicum. PLoS Negl Trop Dis. 2016; 10(4):e0004684.
10. West SM, Mecenas D, Gutwein M, et al. Developmental dynamics of gene expression and alternative polyadenylation in the Caenorhabditis elegans germline. Genome Biol. 2018; 19(1):8.
11. Wang J, Yu Y, Shen H, et al. Dynamic transcriptomes identify biogenic amines and insect-like hormonal regulation for mediating reproduction in Schistosoma japonicum. Nat Commun. 2017;8:14693.
12. Waisberg M, Lobo FP, Cerqueira GC, et al. Microarray analysis of gene expression induced by sexual contact in Schistosoma mansoni. BMC genomics. 2007; 8:181.
13. Luck AN, Anderson KG, McClung CM, et al. Tissue-specific transcriptomics and proteomics of a filarial nematode and its wolbachia endosymbiont. BMC genomics. 2015; 16:920.
14. Fitzpatrick JM, Johansen MV, Johnston DA, et al. Gender-associated gene expression in two related strains of Schistosoma japonicum. Molecular and Biochemical Parasitology. 2004;136:191-209.
15. Fitzpatrick JM, Johnston DA, Williams GW, et al. An oligonucleotide microarray for transcriptome analysis of Schistosoma mansoni and its application/use to investigate gender-associated gene expression. Mol Biochem Parasitol. 2005; 141(1):1-13.
16. Moertel L, McManus DP, Piva TJ, et al. Oligonucleotide microarray analysis of strain- and gender-associated gene expression in the human blood fluke, Schistosoma japonicum. Mol Cell Probes. 2006; 20(5):280-9.
17. Piao X, Cai P, Liu S, et al. Global expression analysis revealed novel gender-specific gene expression features in the blood fluke parasite Schistosoma japonicum. PLoS One. 2011; 6(4):e18267.
18. Phuphisut O, Ajawatanawong P, Limpanont Y, et al. Transcriptomic analysis of male and female Schistosoma mekongi adult worms. Parasites & Vectors. 2018;11:018-3086.
19. McVeigh P, Kimber MJ, Novozhilova E, et al. Neuropeptide signalling systems in flatworms. Parasitology. 2005;131:S41-55.
20. McVeigh P, Mair GR, Atkinson L, et al. Discovery of multiple neuropeptide families in the phylum platyhelminthes. Int J Parasitol. 2009; 39(11):1243-52.
21. Doshi S, Price E, Landis J, et al. Neuropeptide signaling regulates the susceptibility of developing C. elegans to anoxia. Free Radic Biol Med. 2019;131:197-208.
22. Lu Z, Sessler F, Holroyd N, et al. Schistosome sex matters: A deep view into gonad-specific and pairing-dependent transcriptomes reveals a complex gender interplay. Sci Rep. 2016;6:31150.
23. Alvarez J, Alvarez-Illera P, Garcia-Casas P, et al. The role of ca(2+) signaling in aging and neurodegeneration: Insights from Caenorhabditis elegans models. Cells. 2020; 9(1): 204.
24. Williams PDE, Zahratka JA, Rodenbeck M, et al. Serotonin disinhibits a Caenorhabditis elegans sensory neuron by suppressing ca(2+)-dependent negative feedback. J Neurosci. 2018; 38(8):2069-2080.
25. Goodman MB, Sengupta P. How Caenorhabditis elegans senses mechanical stress, temperature, and other physical stimuli. Genetics. 2019; 212(1):25-51.
26. Takeishi A, Takagaki N, Kuhara A. Temperature signaling underlying thermotaxis and cold tolerance in Caenorhabditis elegans. J Neurogenet. 2020; 34(3-4):351-362.
27. Pereira Mda C, Morais S, Sequeiros J, et al. Large-scale functional rnai screen in Caenorhabditis elegans identifies tgf-beta and notch signaling pathways as modifiers of cacna1a. ASN Neuro. 2016; 8(2):1759091416637025.
28. Monsivais D, Matzuk MM, Pangas SA. The tgf-beta family in the reproductive tract. Cold Spring Harb Perspect Biol. 9(10): a022251.
29. Savage-Dunn C, Padgett RW. The tgf-beta family in Caenorhabditis elegans. Cold Spring Harb Perspect Biol. 2017; 9(6):a022178.
30. Magalhaes LG, Morais ER, Machado CB, et al. Uncovering notch pathway in the parasitic flatworm Schistosoma mansoni. Parasitol Res. 2016; 115(10):3951-61.
31. Wong C, Roy R. Ampk regulates developmental plasticity through an endogenous small RNA pathway in Caenorhabditis elegans. Int J Mol Sci. 2020; 21(6):2238.
32. Viollet B, Foretz M. Animal models to study ampk. Exp Suppl. 2016;107:441-469.
33. Lin XG, Ming M, Chen MR, et al . Unc-31/caps docks and primes dense core vesicles in Caenorhabditis elegans neurons. Biochem Biophys Res Commun. 2010; 397(3):526-31.
34. Buddell T, Friedman V, Drozd CJ, et al. An autism-causing calcium channel variant functions with selective autophagy to alter axon targeting and behavior. PLoS Genet. 2019; 15(12):e1008488.
IssueVol 16 No 4 (2021) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijpa.v16i4.7874
Angiostrongylus cantonensis Adult stage Transcriptome Calcium signaling pathway

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
Guo Y, Zhou HC, Dong HY, Yao YL, Xu BY, Zhao Y. Main Pathways and Ion Channels Differentially Expressed in the Transcriptome of Male and Female Adult Angiostrongylus cantonensis using a Deep Sequencing Approach. Iran J Parasitol. 2021;16(4):610-620.