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Protein regulation mechanism of Haemaphysalis longicornis in response to high temperature

ABSTRACT: Ticks, as obligate blood-sucking ectoparasitic arthropods, are intricately linked to ecosystem stability. Global warming significantly impacts tick survival and reproduction, affecting individual behavior, population physiology, adaptive evolution, and altering the likelihood of tick-borne pathogen transmission, thereby posing a threat to public health. To explore the impact of elevated temperatures on ticks and the associated protein regulatory mechanisms, a quantitative proteomic analysis using the DIA technique was conducted on the ovary, salivary gland, and midgut of H. longicornis exposed to high temperatures. The findings revealed substantial changes in protein expression under high temperature stress, impacting various crucial biological processes. Specifically, Ubiquitin carboxyl-terminal hydrolase (UCH) exhibited upregulation across all tested high temperatures. Knocking down UCH through RNA interference resulted in decreased heat tolerance in H. longicornis, underscoring the essential role of UCH in maintaining normal physiological functions of ticks in high-temperature environments. This study sheds light on the physiological mechanisms employed by ticks to adapt to climate change and aids in the prevention and management of tick-borne diseases.

ORGANISM(S): Haemaphysalis Longicornis

SUBMITTER: Hui Wang

PROVIDER: PXD062795 | iProX | Thu Apr 10 00:00:00 BST 2025

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:Ticks are a kind of obligate blood-sucking in vitro parasitic arthropods, which can carry and transmit a variety of extremely harmful pathogens. They greatly threaten the health of humans and wildlife and seriously affect livestock production. In recent years, with global warming, the growth, reproduction, and behavior of ticks have been significantly affected. Therefore, it is particularly important to study how high temperature affects ticks and how ticks adopt effective behavioral strategies and physiological adjustment mechanisms to cope with high temperatures. Phosphorylation, the most common posttranslational modification of proteins, plays an important role in cell signal transduction, gene expression, and rapid regulation of the cell cycle. The regulation strategy of protein phosphorylation under high-temperature conditions is of great significance to organisms. In order to explore the effect of high temperature on ticks, this study systematically studied the phosphorylation changes of proteins in the salivary gland, midgut, ovary, and malpighian tubules of ticks at different temperatures of 26°C, 36°C and 45°C by quantitative proteomics. The continuously up-regulated and down-regulated phosphorylated proteins under the influence of high temperature were selected and subjected to GO analysis, pathway analysis, Motif analysis, and kinase prediction. The results showed that each tissue of ticks has its own unique way of coping with high-temperature stress. In the four tissues, a large number of phosphorylated proteins were enriched in terms and pathways related to splicing and regulation of RNA splicing. It may indicate that the protein diversity caused by RNA alternative splicing plays an important role in the response of ticks to high-temperature stress. In addition, RNAi results showed that PKC, GRK, and CDC37 can interact with each other and play an important role in the process of H. longicornis resisting high temperature. CDC37 can also improve the survival rate of H. longicornis in high temperatures. This study lays a foundation for revealing the molecular regulation mechanism of ticks in response to high-temperature stress and is of great significance for a comprehensive understanding of the molecular regulation strategies of ticks to adapt to global warming.

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Protein regulation mechanism of Haemaphysalis longicornis in response to high temperature

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