Comparative Transcriptome Sequencing Analysis of Hirudo nipponia in Different Growth Periods

Hirudo nipponia is the only blood-sucking leech included in Chinese Pharmacopoeia having distinct features of anticoagulation, exorcizing blood stasis, and promoting menstruation. Despite such significant characteristics, very little is known about its molecular genetics and related physiological mechanisms. In this study, the transcriptomes of H. nipponia at three developmental stages (larvae, young, and adults), revealed a total of 1,348 differentially expressed genes (DEGs), 223 differentially expressed lncRNAs, and 88 novel mRNAs. A significant diverse gene expression patterns were observed at different developmental stages which were analyzed by differential gene expression trends, and the overall gene expression trends consist of three overall down-regulated trends, and two overall up-regulated trends. Furthermore, the GO and KEGG enrichment functional annotation analysis revealed that these DEGs were mainly associated with protein hydrolysis, signal transduction, energy metabolism, and lipid metabolism while growth, development, metabolism, and reproduction-related DEGs were also found. Additionally, real-time quantitative PCR results confirmed deep sequencing results based on the relative expression levels of nine randomly selected genes. This is the first transcriptome-based comprehensive study of H. irudo nipponia at different developmental stages which provided considerable deep understanding related to gene expression patterns and their relevant developmental pathways, neurodevelopmental and reproductive characteristics of the leech.

[1]  Qingyou Liu,et al.  Comprehensive Transcriptome Sequencing Analysis of Hirudinaria manillensis in Different Growth Periods , 2022, Frontiers in Physiology.

[2]  Jorge Verdín,et al.  An F-Actin Mega-Cable Is Associated With the Migration of the Sperm Nucleus During the Fertilization of the Polarity-Inverted Central Cell of Agave inaequidens , 2021, Frontiers in Plant Science.

[3]  Jichen Zhao,et al.  Growth trait gene analysis of kuruma shrimp (Marsupenaeus japonicus) by transcriptome study. , 2021, Comparative biochemistry and physiology. Part D, Genomics & proteomics.

[4]  M. Irving,et al.  Myosin-based regulation of twitch and tetanic contractions in mammalian skeletal muscle , 2021, eLife.

[5]  Qi Li,et al.  Characterization of paramyosin protein structure and gene expression during myogenesis in Pacific oyster (Crassostrea gigas). , 2021, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[6]  Hongjun Yang,et al.  Characterization and analysis of the transcriptome in Opisina arenosella from different developmental stages using single-molecule real-time transcript sequencing and RNA-seq. , 2020, International journal of biological macromolecules.

[7]  D. Kovar,et al.  F-Actin Cytoskeleton Network Self-Organization Through Competition and Cooperation. , 2020, Annual review of cell and developmental biology.

[8]  Yu-LanYeh,et al.  Leech extract: a candidate cardioprotective against hypertension-induced cardiac hypertrophy and fibrosis. , 2020, Journal of ethnopharmacology.

[9]  F. F. De-Miguel,et al.  Transcriptional profiling of identified neurons in leech , 2020, bioRxiv.

[10]  A. Filipek,et al.  Binding of S100A6 to actin and the actin–tropomyosin complex , 2020, Scientific Reports.

[11]  M. Ceylan Effects of maternal age on reproductive performance of the southern medicinal leech, Hirudo verbana Carena, 1820. , 2020, Animal Reproduction Science.

[12]  E. Korn,et al.  Muscle myosins form folded monomers, dimers, and tetramers during filament polymerization in vitro , 2020, Proceedings of the National Academy of Sciences.

[13]  Kyle R. Eberlin,et al.  Leech Therapy Following Digital Replantation and Revascularization. , 2020, The Journal of hand surgery.

[14]  M. Rynkiewicz,et al.  Protein-Protein Docking Reveals Dynamic Interactions of Tropomyosin on Actin Filaments. , 2020, Biophysical journal.

[15]  O. Shefi,et al.  Brief Electrical Stimulation Triggers an Effective Regeneration of Leech CNS , 2020, eNeuro.

[16]  D. Heeley,et al.  Demonstration of beta-tropomyosin (Tpm2) and duplication of the alpha-slow tropomyosin gene (TPM3) in Atlantic salmon Salmo salar. , 2020, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[17]  M. Pasalar,et al.  Medicinal leech therapy in venous congestion and various ulcer forms: Perspectives of Western, Persian and Indian medicine , 2019, Journal of traditional and complementary medicine.

[18]  G. Pertea,et al.  GFF Utilities: GffRead and GffCompare. , 2020, F1000Research.

[19]  N. Sağlam Internal and External Morphological Characteristics of the Medicinal Leech Species Hirudo sulukii and Hirudo verbana , 2019, Turkiye parazitolojii dergisi.

[20]  J. Kretzberg,et al.  Non-synaptic Plasticity in Leech Touch Cells , 2019, Front. Physiol..

[21]  J. Squire Special Issue: The Actin-Myosin Interaction in Muscle: Background and Overview , 2019, International journal of molecular sciences.

[22]  Soon Cheol Park,et al.  Spatiotemporal Expression of Anticoagulation Factor Antistasin in Freshwater Leeches , 2019, International journal of molecular sciences.

[23]  Hakan Murat Büyükçapar,et al.  Investigation of reproductive efficiency, growth performance and survival of the southern medicinal leech, Hirudo verbana Carena, 1820 fed with mammalian and poultry blood. , 2019, Animal reproduction science.

[24]  S. Kvist,et al.  The salivary transcriptome of Limnobdella mexicana (Annelida: Clitellata: Praobdellidae) and orthology determination of major leech anticoagulants , 2019, Parasitology.

[25]  Debin Wang,et al.  In-depth profiles of bioactive large molecules in saliva secretions of leeches determined by combining salivary gland proteome and transcriptome data. , 2019, Journal of proteomics.

[26]  J. Hammer,et al.  Origin, Organization, Dynamics, and Function of Actin and Actomyosin Networks at the T Cell Immunological Synapse. , 2019, Annual review of immunology.

[27]  S. Singh,et al.  Medical leech therapy in Ayurveda and biomedicine – A review , 2019, Journal of Ayurveda and integrative medicine.

[28]  Zeng-hui Lu,et al.  Transcriptomic analysis of the salivary gland of medicinal leech Hirudo nipponia , 2018, PloS one.

[29]  Lijiang Yang,et al.  Comparative transcriptomic analysis reveals the mechanism of leech environmental adaptation. , 2018, Gene.

[30]  Jia Gu,et al.  fastp: an ultra-fast all-in-one FASTQ preprocessor , 2018, bioRxiv.

[31]  J. Xiang,et al.  Actin genes and their expression in pacific white shrimp, Litopenaeus vannamei , 2018, Molecular Genetics and Genomics.

[32]  Aylin Uskudar Guclu,et al.  Medicinal leech therapy—an overall perspective , 2017, Integrative medicine research.

[33]  Shuji Takahashi,et al.  Comprehensive analyses of hox gene expression in Xenopus laevis embryos and adult tissues , 2017, Development, growth & differentiation.

[34]  M. Salzet,et al.  Neuro‐immune lessons from an annelid: The medicinal leech , 2017, Developmental and comparative immunology.

[35]  Jinyan Huang,et al.  Inhibition of the nuclear export of p65 and IQCG in leukemogenesis by NUP98-IQCG , 2016, Frontiers of Medicine.

[36]  T. Minokawa,et al.  Characterization of paramyosin and thin filaments in the smooth muscle of acorn worm, a member of hemichordates. , 2016, Journal of biochemistry.

[37]  Steven L Salzberg,et al.  HISAT: a fast spliced aligner with low memory requirements , 2015, Nature Methods.

[38]  S. Salzberg,et al.  StringTie enables improved reconstruction of a transcriptome from RNA-seq reads , 2015, Nature Biotechnology.

[39]  H. Taylor,et al.  The Role of Hox Genes in Female Reproductive Tract Development, Adult Function, and Fertility. , 2015, Cold Spring Harbor perspectives in medicine.

[40]  D. Robledo,et al.  Analysis of qPCR reference gene stability determination methods and a practical approach for efficiency calculation on a turbot (Scophthalmus maximus) gonad dataset , 2014, BMC Genomics.

[41]  J. Schimenti,et al.  IQ Motif-Containing G (Iqcg) Is Required for Mouse Spermiogenesis , 2013, G3: Genes, Genomes, Genetics.

[42]  Alexander Hillisch,et al.  Oral, direct thrombin and factor Xa inhibitors: the replacement for warfarin, leeches, and pig intestines? , 2011, Angewandte Chemie.

[43]  I. Fournier,et al.  Multiple Changes in Peptide and Lipid Expression Associated with Regeneration in the Nervous System of the Medicinal Leech , 2011, PloS one.

[44]  N. Bols,et al.  An evaluation of potential reference genes for stability of expression in two salmonid cell lines after infection with either Piscirickettsia salmonis or IPNV , 2010, BMC Research Notes.

[45]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[46]  S. de Mateo,et al.  Identification of proteomic differences in asthenozoospermic sperm samples. , 2008, Human reproduction.

[47]  A. Mújica,et al.  F‐actin involvement in guinea pig sperm motility , 2007, Molecular reproduction and development.

[48]  W. Bo Studies on growth and reproduction of Hirudinaria manillensis in Guangdong Province , 2002 .

[49]  Mestranol. , 2020, IARC monographs on the evaluation of the carcinogenic risk of chemicals to humans.