title: First transcriptome of the testis-vas deferens-male accessory gland and proteome of the spermatophore from Dermacentor variabilis (Acari: Ixodidae).
abstract: Ticks are important vectors of numerous human diseases and animal diseases. Feeding stimulates spermatogenesis, mating and insemination of male factors that trigger female reproduction. The physiology of male reproduction and its regulation of female development are essentially a black box. Several transcriptomes have catalogued expression of tick genes in the salivary glands, synganglion and midgut but no comprehensive investigation has addressed male reproduction and mating. Consequently, a new global approach using transcriptomics, proteomics, and quantitative gene expression is needed to understand male reproduction and stimulation of female reproduction.This first transcriptome to the reproductive biology of fed male ticks, Dermacentor variabilis, was obtained by 454 pyrosequencing (563,093 reads, 12,804 contigs). Gene Ontology (Biological Processes level III) recognized 3,866 transcripts in 73 different categories; spermiogenesis; spermatogenesis; peptidases, lipases and hydrolases; oxidative and environmental stress; immune defense; and protein binding. Reproduction-associated genes included serine/threonine kinase, metalloendoproteinases, ferritins, serine proteases, trypsin, cysteine proteases, serpins, a cystatin, GPCR and others. qRT-PCR showed significant upregulation from unfed versus fed adult male reproductive organs of zinc metalloprotease, astacin metalloprotease and serine protease, enzymes important in spermiogenesis and mating activity in insects, as well as a GPCR with the greatest similarity to a SIFamide receptor known to be important in regulating courtship behavior in Drosophila. Proteomics on these organs and the spermatophore by tryptic digestion/Liquid chromatography/Mass spectrometry/Mass spectrometry (LC/MS/MS) demonstrated expression of many of the same messages found by 454 sequencing, supporting their identification, and revealed differences in protein distribution in the reproductive system versus the spermatophore. We found Efα but no EF β in the transcriptome and neither of these proteins in the spermatophore. Thus, the previously described model for male regulation of female reproduction may not apply to other ticks. A new paradigm is needed to explain male stimulation of female tick reproduction.
authors.affiliation: Department of Biological Sciences, Old Dominion University, Norfolk, Virginia, United States of America. dsonensh@odu.edu
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Sonenshine
authors.forename: Brooke W
authors.initials: BW
authors.lastname: Bissinger
authors.forename: Noble
authors.initials: N
authors.lastname: Egekwu
authors.forename: Kevin V
authors.initials: KV
authors.lastname: Donohue
authors.forename: Sayed M
authors.initials: SM
authors.lastname: Khalil
authors.forename: R Michael
authors.initials: RM
authors.lastname: Roe
mesh_terms: D000595:Amino Acid Sequence; D000825:Animal Structures / Q000378:metabolism*; D000818:Animals; D020451:Contig Mapping; D003870:Dermacentor / Q000235:genetics* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008297:Male; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D020543:Proteome / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D013093:Spermatogonia / Q000378:metabolism*; D013737:Testis / Q000378:metabolism*; D059467:Transcriptome; D014649:Vas Deferens / Q000378:metabolism*
chemical_list: D006360:Heat-Shock Proteins; D020543:Proteome