Identification of Male Gametogenesis Expressed Genes from the Scallop Nodipecten subnodosus by Suppressive Subtraction Hybridization and Pyrosequencing

Type Article
Date 2013-09
Language English
Author(s) Llera-Herrera Raul1, Garcia-Gasca Alejandra2, Abreu-Goodger Cei3, Huvet ArnaudORCID4, Ibarra Ana M.1
Affiliation(s) 1 : Ctr Invest Biol Noroeste, Aquaculture Genet & Breeding Lab, La Paz, Baja California, Mexico.
2 : Ctr Invest Alimentac & Desarrollo, Mazatlan, Sinaloa, Mexico.
3 : Inst Politecn Nacl CINVESTAV IPN, Ctr Invest & Estudios Avanzados, Lab Nacl Genom Biodiversidad Langebio, Guanajuato, Mexico.
4 : IFREMER, Ctr Bretagne, Lab Sci Environm Marin, Plouzane, France.
Source Plos One (1932-6203) (Public Library Science), 2013-09 , Vol. 8 , N. 9 , P. e73176
DOI 10.1371/journal.pone.0073176
WOS© Times Cited 19
Abstract Despite the great advances in sequencing technologies, genomic and transcriptomic information for marine non-model species with ecological, evolutionary, and economical interest is still scarce. In this work we aimed to identify genes expressed during spermatogenesis in the functional hermaphrodite scallop Nodipecten subnodosus (Mollusca: Bivalvia: Pectinidae), with the purpose of obtaining a panel of genes that would allow for the study of differentially transcribed genes between diploid and triploid scallops in the context of meiotic arrest and reproductive sterility. Because our aim was to isolate genes involved in meiosis and other testis maturation-related processes, we generated suppressive subtractive hybridization libraries of testis vs. inactive gonad. We obtained 352 and 177 ESTs by clone sequencing, and using pyrosequencing (454-Roche) we maximized the identified ESTs to 34,276 reads. A total of 1,153 genes from the testis library had a blastx hit and GO annotation, including genes specific for meiosis, spermatogenesis, sex-differentiation, and transposable elements. Some of the identified meiosis genes function in chromosome pairing (scp2, scp3), recombination and DNA repair (dmc1, rad51, ccnb1ip1/hei10), and meiotic checkpoints (rad1, hormad1, dtl/cdt2). Gene expression analyses in different gametogenic stages in both sexual regions of the gonad of meiosis genes confirmed that the expression was specific or increased towards the maturing testis. Spermatogenesis genes included known testis-specific ones (kelch-10, shippo1, adad1), with some of these known to be associated to sterility. Sex differentiation genes included one of the most conserved genes at the bottom of the sex-determination cascade (dmrt1). Transcript from transposable elements, reverse transcriptase, and transposases in this library evidenced that transposition is an active process during spermatogenesis in N. subnodosus. In relation to the inactive library, we identified 833 transcripts with functional annotation related to activation of the transcription and translation machinery, as well as to germline control and maintenance.
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