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Quantitative and phosphorylated proteomics data on the dynamic changes of ovarian developmental proteins in female haemaphysalis longicornis

ABSTRACT: Background: Haemaphysalis longicornis is an obligate blood-feeding ectoparasite that can spread a variety of pathogens, causing serious damage to humans, livestock, and wildlife. With extremely high reproductive capability, female ticks can ingest a large amount of blood from their hosts, causing extreme harm. The degree of ovarian development of females directly affects the number of eggs laid by female ticks. Studying the regulatory mechanism of tick ovary development is of great significance for effectively controlling tick populations and thus reducing tick damage. Methods: In this study, quantitative proteomics based on data independent acquisition (DIA) was used to systematically study dynamic changes in the expression levels of all proteins and in protein phosphorylation during the ovarian development of engorged females. Results: In ovary development, the synergistic action of many proteins is required to rapidly achieve ovarian development and egg formation. Leucine aminopeptidase, acting as a hydrolase, provides amino acids for ovarian development by hydrolysing proteins or peptides. When it was knocked down by RNAi, ovarian development was inhibited. Conclusions: The results of this study found many proteins that play important role during ovarian development, and also reveal the importance of protein phosphorylation in ovarian development, which is an important reproductive strategy in which ticks can rapidly and efficiently convert large amounts of blood in the ovaries into egg-producing raw materials and ultimately produce many eggs.

ORGANISM(S): Haemaphysalis Longicornis

SUBMITTER: Hui Wang

PROVIDER: PXD015267 | iProX | Sun Sep 01 00:00:00 BST 2019

REPOSITORIES: iProX

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Project description:Tick saliva contains many bioactive molecules that are involved in attachment to the host, blood feeding and transmission of pathogens. MicroRNAs (miRNAs) are a class of short non-coding RNAs with a length of 19-24 nucleotides. They act as regulators of gene expression by binding to their target mRNA at the post-transcriptional level and control a variety of cellular functions, including regulation of growth, metabolism, and development. The detection and characterizations of miRNAs from tick saliva may help explain the molecular mechanisms involved in the interaction between ticks, pathogens, and hosts. They may also contribute to the discovery of vaccines, which can control ticks and the pathogens they transmit. An RNA library was generated from the saliva of fed adult Haemaphysalis longicornis ticks, and it contained 17.4 million clean reads of 18-30 nucleotides. Overall, 319 known miRNAs and 1 novel miRNA were found. The 10 most abundantly expressed miRNAs present in tick saliva were miR-100_2, miR-315, miR-184_1, miR-100-5p_2, miR-5307, miR-184-3p_3, Let-7-5p_6, miR-71_5, miR-1-3p_6, and miR-10-5p_2. The miR-375, one of the abundantly expressed, was subjected to Quantitative Real-Time PCR analysis (qRT-PCR) in various tick developmental stages, as well as in different tissues isolated from adult ticks. The expression of miR-375 in different tick development stages was highest in unfed nymphs and lowest in the egg stage. In the tissues of adult ticks, miR-375 was most highly expressed in the salivary gland. To investigate the possible role of miR-375, Ant-375 was used to inhibit the miR-375. Treated group (Ant-375) had a reduced number of eggs (t(10)= 2.652, P=0.0242), eggs that were partially desiccated, and reduced egg hatchability (t(10)=2.272, P=0.044) compared to Ms-Ant and the non-injected control. This is the first study to investigate the miRNAs profile in tick saliva and the role of miR-375 in H. longicornis. The identification and characterization of miRNA in tick saliva may help to reveal the molecular mechanisms of interactions among ticks, pathogens, and hosts and suggest new vaccine strategies to control tick borne diseases.

Project description:Female Aedes aegypti mosquitoes impose a severe global public health burden as primary vectors of multiple viral and parasitic pathogens. Under optimal environmental conditions, Aedes aegypti females have access to human hosts that provide blood proteins for egg development, conspecific males that provide sperm for fertilization, and freshwater that serves as an egg-laying substrate suitable for offspring survival. As global temperatures rise, Aedes aegypti females are faced with climate challenges, like intense droughts and intermittent precipitation, which create unpredictable and suboptimal conditions for the egg-laying step of their reproductive cycle. Aedes aegypti mosquitoes nonetheless show remarkable reproductive resilience, but how they achieve this is unknown. Here we show that under drought-like conditions simulated in the laboratory, mated, blood-fed Aedes aegypti females carrying mature eggs retain them in their ovaries for extended periods, while maintaining the viability of these eggs until they can be deposited in freshwater. Using transcriptomic and proteomic profiling of Aedes aegypti ovaries, we identify two previously uncharacterized genes – here named tweedledee and tweedledum – that show ovary-enriched, temporally-restricted expression during egg retention. These genes are mosquito-specific, linked within a syntenic locus, and rapidly evolving under positive selection, raising the possibility that they serve an adaptive function. Using loss-of-function mutagenesis to disrupt both genes, we show that, tweedledee and tweedledum, which encode secreted proteins, are specifically required for extended retention of viable eggs, such as during intermittent precipitation or drought. These results highlight an elegant example of taxon-restricted genes at the heart of an important adaptation that equips Aedes aegypti females with “insurance” to, when contextually appropriate, flexibly extend their reproductive sequence without losing reproductive capacity, thus allowing this species to exploit diverse and unpredictable/chaotic/changing habitats.

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Quantitative and phosphorylated proteomics data on the dynamic changes of ovarian developmental proteins in female haemaphysalis longicornis

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