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Construction of a medicinal leech transcriptome database and its application to the identification of leech homologs of neural and innate immune genes
Journal article   Open access   Peer reviewed

Construction of a medicinal leech transcriptome database and its application to the identification of leech homologs of neural and innate immune genes

Eduardo R Macagno, Terry Gaasterland, Lee Edsall, Vineet Bafna, Marcelo B Soares, Todd Scheetz, Thomas Casavant, Corinne Da Silva, Patrick Wincker, Aurélie Tasiemski, …
BMC genomics, Vol.11(1), pp.407-407
06/25/2010
DOI: 10.1186/1471-2164-11-407
PMCID: PMC2996935
PMID: 20579359
url
https://doi.org/10.1186/1471-2164-11-407View
Published (Version of record) Open Access

Abstract

The medicinal leech, Hirudo medicinalis, is an important model system for the study of nervous system structure, function, development, regeneration and repair. It is also a unique species in being presently approved for use in medical procedures, such as clearing of pooled blood following certain surgical procedures. It is a current, and potentially also future, source of medically useful molecular factors, such as anticoagulants and antibacterial peptides, which may have evolved as a result of its parasitizing large mammals, including humans. Despite the broad focus of research on this system, little has been done at the genomic or transcriptomic levels and there is a paucity of openly available sequence data. To begin to address this problem, we constructed whole embryo and adult central nervous system (CNS) EST libraries and created a clustered sequence database of the Hirudo transcriptome that is available to the scientific community. A total of approximately 133,000 EST clones from two directionally-cloned cDNA libraries, one constructed from mRNA derived from whole embryos at several developmental stages and the other from adult CNS cords, were sequenced in one or both directions by three different groups: Genoscope (French National Sequencing Center), the University of Iowa Sequencing Facility and the DOE Joint Genome Institute. These were assembled using the phrap software package into 31,232 unique contigs and singletons, with an average length of 827 nt. The assembled transcripts were then translated in all six frames and compared to proteins in NCBI's non-redundant (NR) and to the Gene Ontology (GO) protein sequence databases, resulting in 15,565 matches to 11,236 proteins in NR and 13,935 matches to 8,073 proteins in GO. Searching the database for transcripts of genes homologous to those thought to be involved in the innate immune responses of vertebrates and other invertebrates yielded a set of nearly one hundred evolutionarily conserved sequences, representing all known pathways involved in these important functions. The sequences obtained for Hirudo transcripts represent the first major database of genes expressed in this important model system. Comparison of translated open reading frames (ORFs) with the other openly available leech datasets, the genome and transcriptome of Helobdella robusta, shows an average identity at the amino acid level of 58% in matched sequences. Interestingly, comparison with other available Lophotrochozoans shows similar high levels of amino acid identity, where sequences match, for example, 64% with Capitella capitata (a polychaete) and 56% with Aplysia californica (a mollusk), as well as 58% with Schistosoma mansoni (a platyhelminth). Phylogenetic comparisons of putative Hirudo innate immune response genes present within the Hirudo transcriptome database herein described show a strong resemblance to the corresponding mammalian genes, indicating that this important physiological response may have older origins than what has been previously proposed.
Central Nervous System - metabolism Species Specificity Hirudo medicinalis - genetics Humans RNA, Messenger - genetics Immunity, Innate - genetics Databases, Genetic Gene Expression Profiling Hirudo medicinalis - immunology Antigens, CD - genetics Hirudo medicinalis - embryology Sequence Homology, Nucleic Acid Central Nervous System - immunology Adaptive Immunity - genetics Central Nervous System - physiology Animals Regeneration - genetics Toll-Like Receptors - genetics Receptors, Pattern Recognition - genetics Expressed Sequence Tags - metabolism Antimicrobial Cationic Peptides - genetics Cytokines - genetics Databases, Nucleic Acid

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