A targeted in situ hybridization screen identifies putative seminal fluid proteins in a simultaneously hermaphroditic flatworm
Abstract Background Along with sperm, in many taxa ejaculates also contain large numbers of seminal fluid proteins (SFPs). SFPs and sperm are transferred to the mating partner, where they are thought to play key roles in mediating post-mating sexual selection. They modulate the partner’s behavior an...
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| Format: | Article |
| Language: | English |
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BMC
2018-05-01
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| Series: | BMC Evolutionary Biology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12862-018-1187-0 |
| _version_ | 1818956124021325824 |
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| author | Michael Weber Julia Wunderer Birgit Lengerer Robert Pjeta Marcelo Rodrigues Lukas Schärer Peter Ladurner Steven A. Ramm |
| author_facet | Michael Weber Julia Wunderer Birgit Lengerer Robert Pjeta Marcelo Rodrigues Lukas Schärer Peter Ladurner Steven A. Ramm |
| author_sort | Michael Weber |
| collection | DOAJ |
| description | Abstract Background Along with sperm, in many taxa ejaculates also contain large numbers of seminal fluid proteins (SFPs). SFPs and sperm are transferred to the mating partner, where they are thought to play key roles in mediating post-mating sexual selection. They modulate the partner’s behavior and physiology in ways that influence the reproductive success of both partners, thus potentially leading to sexual conflict. Despite the presumed general functional and evolutionary significance of SFPs, their identification and characterization has to date focused on just a few animal groups, predominantly insects and mammals. Moreover, until now seminal fluid profiling has mainly focused on species with separate sexes. Here we report a comprehensive screen for putative SFPs in the simultaneously hermaphroditic flatworm Macrostomum lignano. Results Based on existing transcriptomic data, we selected 150 transcripts known to be (a) predominantly expressed in the tail region of the worms, where the seminal fluid-producing prostate gland cells are located, and (b) differentially expressed in social environments differing in sperm competition level, strongly implying that they represent a phenotypically plastic aspect of male reproductive allocation in this species. For these SFP candidates, we then performed whole-mount in situ hybridization (ISH) experiments to characterize tissue-specific expression. In total, we identified 98 transcripts that exhibited prostate-specific expression, 76 of which we found to be expressed exclusively in the prostate gland cells; additional sites of expression for the remaining 22 included the testis or other gland cells. Bioinformatics analyses of the prostate-limited candidates revealed that at least 64 are predicted to be secretory proteins, making these especially strong candidates to be SFPs that are transferred during copulation. Conclusions Our study represents a first comprehensive analysis using a combination of transcriptomic and ISH screen data to identify SFPs based on transcript expression in seminal fluid-producing tissues. We thereby extend the range of taxa for which seminal fluid has been characterized to a flatworm species with a sequenced genome and for which several methods such as antibody staining, transgenesis and RNA interference have been established. Our data provide a basis for testing the functional and evolutionary significance of SFPs. |
| first_indexed | 2024-12-20T10:48:58Z |
| format | Article |
| id | doaj.art-75100686641745bba77231f8508b2b53 |
| institution | Directory Open Access Journal |
| issn | 1471-2148 |
| language | English |
| last_indexed | 2024-12-20T10:48:58Z |
| publishDate | 2018-05-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Evolutionary Biology |
| spelling | doaj.art-75100686641745bba77231f8508b2b532022-12-21T19:43:19ZengBMCBMC Evolutionary Biology1471-21482018-05-0118111310.1186/s12862-018-1187-0A targeted in situ hybridization screen identifies putative seminal fluid proteins in a simultaneously hermaphroditic flatwormMichael Weber0Julia Wunderer1Birgit Lengerer2Robert Pjeta3Marcelo Rodrigues4Lukas Schärer5Peter Ladurner6Steven A. Ramm7Evolutionary Biology, Bielefeld UniversityInstitute of Zoology and Center of Molecular Biosciences Innsbruck, University of InnsbruckInstitute of Zoology and Center of Molecular Biosciences Innsbruck, University of InnsbruckInstitute of Zoology and Center of Molecular Biosciences Innsbruck, University of InnsbruckInstitute of Zoology and Center of Molecular Biosciences Innsbruck, University of InnsbruckEvolutionary Biology, Zoological Institute, University of BaselInstitute of Zoology and Center of Molecular Biosciences Innsbruck, University of InnsbruckEvolutionary Biology, Bielefeld UniversityAbstract Background Along with sperm, in many taxa ejaculates also contain large numbers of seminal fluid proteins (SFPs). SFPs and sperm are transferred to the mating partner, where they are thought to play key roles in mediating post-mating sexual selection. They modulate the partner’s behavior and physiology in ways that influence the reproductive success of both partners, thus potentially leading to sexual conflict. Despite the presumed general functional and evolutionary significance of SFPs, their identification and characterization has to date focused on just a few animal groups, predominantly insects and mammals. Moreover, until now seminal fluid profiling has mainly focused on species with separate sexes. Here we report a comprehensive screen for putative SFPs in the simultaneously hermaphroditic flatworm Macrostomum lignano. Results Based on existing transcriptomic data, we selected 150 transcripts known to be (a) predominantly expressed in the tail region of the worms, where the seminal fluid-producing prostate gland cells are located, and (b) differentially expressed in social environments differing in sperm competition level, strongly implying that they represent a phenotypically plastic aspect of male reproductive allocation in this species. For these SFP candidates, we then performed whole-mount in situ hybridization (ISH) experiments to characterize tissue-specific expression. In total, we identified 98 transcripts that exhibited prostate-specific expression, 76 of which we found to be expressed exclusively in the prostate gland cells; additional sites of expression for the remaining 22 included the testis or other gland cells. Bioinformatics analyses of the prostate-limited candidates revealed that at least 64 are predicted to be secretory proteins, making these especially strong candidates to be SFPs that are transferred during copulation. Conclusions Our study represents a first comprehensive analysis using a combination of transcriptomic and ISH screen data to identify SFPs based on transcript expression in seminal fluid-producing tissues. We thereby extend the range of taxa for which seminal fluid has been characterized to a flatworm species with a sequenced genome and for which several methods such as antibody staining, transgenesis and RNA interference have been established. Our data provide a basis for testing the functional and evolutionary significance of SFPs.http://link.springer.com/article/10.1186/s12862-018-1187-0Seminal fluidFlatwormIn situ hybridizationProstateSex allocationSexual selection |
| spellingShingle | Michael Weber Julia Wunderer Birgit Lengerer Robert Pjeta Marcelo Rodrigues Lukas Schärer Peter Ladurner Steven A. Ramm A targeted in situ hybridization screen identifies putative seminal fluid proteins in a simultaneously hermaphroditic flatworm BMC Evolutionary Biology Seminal fluid Flatworm In situ hybridization Prostate Sex allocation Sexual selection |
| title | A targeted in situ hybridization screen identifies putative seminal fluid proteins in a simultaneously hermaphroditic flatworm |
| title_full | A targeted in situ hybridization screen identifies putative seminal fluid proteins in a simultaneously hermaphroditic flatworm |
| title_fullStr | A targeted in situ hybridization screen identifies putative seminal fluid proteins in a simultaneously hermaphroditic flatworm |
| title_full_unstemmed | A targeted in situ hybridization screen identifies putative seminal fluid proteins in a simultaneously hermaphroditic flatworm |
| title_short | A targeted in situ hybridization screen identifies putative seminal fluid proteins in a simultaneously hermaphroditic flatworm |
| title_sort | targeted in situ hybridization screen identifies putative seminal fluid proteins in a simultaneously hermaphroditic flatworm |
| topic | Seminal fluid Flatworm In situ hybridization Prostate Sex allocation Sexual selection |
| url | http://link.springer.com/article/10.1186/s12862-018-1187-0 |
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