A targeted in situ hybridization screen identifies putative seminal fluid proteins in a simultaneously hermaphroditic flatworm.
A targeted in situ hybridization screen identifies putative seminal fluid proteins in a simultaneously hermaphroditic flatworm.
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DOI：
10.1186/s12862-018-1187-0
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发表时间：
2018-05-30
影响因子：
3.4
通讯作者：
Ramm SA
中科院分区：
文献类型：
--
作者：
Weber M;Wunderer J;Lengerer B;Pjeta R;Rodrigues M;Schärer L;Ladurner P;Ramm SA
在许多分类群中，除精子外，射精还含有大量的精液蛋白（SFPs）。SFPs和精子被转移到配偶身上，在那里它们被认为在调解交配后的性选择中起着关键作用。它们调节伴侣的行为和生理，从而影响双方的繁殖成功，从而潜在地导致性冲突。尽管SFPs被认为具有普遍的功能和进化意义，但迄今为止，它们的鉴定和表征只集中在少数动物群体上，主要是昆虫和哺乳动物。此外，到目前为止，精液分析主要集中在具有不同性别的物种上。在这里，我们报道了对同时雌雄同体的扁形虫（Macrostomum lignano）中推测的SFPs的全面筛选。基于现有的转录组学数据，我们选择了150个已知的转录本(a)主要表达于蠕虫尾部区域，即产生精液的前列腺细胞所在的位置，以及(b)在精子竞争水平不同的社会环境中差异表达，这强烈暗示它们代表了该物种雄性生殖分配的表型可塑性方面。对于这些SFP候选蛋白，我们进行了全载原位杂交（ISH）实验来表征组织特异性表达。总的来说，我们确定了98个表现出前列腺特异性表达的转录本，其中76个我们发现仅在前列腺细胞中表达；其余22个表达位点包括睾丸或其他腺体细胞。对前列腺受限候选蛋白的生物信息学分析显示，预计至少有64种是分泌蛋白，这使得这些候选蛋白特别有可能是在交配过程中转移的sftp。我们的研究首次综合分析了转录组学和ISH筛选数据，根据精液产生组织中的转录表达来鉴定SFPs。因此，我们将精液特征分类群的范围扩展到具有基因组测序的扁虫物种，并且已经建立了抗体染色，转基因和RNA干扰等几种方法。我们的数据为测试SFPs的功能和进化意义提供了基础。本文的在线版本（10.1186/s12862-018-1187-0）包含补充材料，授权用户可以使用。
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. 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. 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. The online version of this article (10.1186/s12862-018-1187-0) contains supplementary material, which is available to authorized users.
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