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Targeting TtVgR via siRNA Knockdown Elicits Ovarian Cell Death in the Tri-spine Horseshoe Crab

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Abstract

The vitellogenin present in the bloodstream undergoes internalization into developing oocytes through the vitellogenin receptor (VgR), a process mediated by receptor-mediated endocytosis. VgR plays a crucial role in facilitating the accumulation of vitellogenin and the maturation of oocytes. In this study, we characterized a Tachypleus tridentatus vitellogenin receptor (TtVgR) gene from the tri-spine horseshoe crab, revealing a length of 1956 bp and encoding 652 amino acid residues with 12 exons. TtVgR has a molecular weight of 64.26 kDa and an isoelectric point of 5.95. Predictions indicate 85 phosphorylation sites and 7 glycosylation sites within TtVgR. Transcriptional analysis demonstrated specific expression of TtVgR in the ovary and yellow connective tissue. TtVgR was identified and distributed in the plasma membrane of oocytes. The siRNA-mediated TtVgR knockdown significantly reduced the transcriptional activity of TtVgR. This depletion induced excessive ROS production, resulting in DNA damage in ovarian primary cells. TUNEL and flow cytometry analyses confirmed ovarian cell apoptosis following TtVgR knockdown, indicating DNA damage in ovarian primary cells. These findings underscore the importance of TtVgR in ovarian cell development, suggesting its potential involvement in vitellogenesis and oocyte maturation. This knowledge may inform innovative breeding strategies and contribute to the sustainable management and conservation of the tri-spine horseshoe crab.

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Funding

This research was financially supported by a research grant from the National Natural Science Foundation of China (42366006); High-level Talent Scientific Research Start-Up Fund Project of Beibu Gulf University (2022KYQD10); Key Laboratory of Tropical Marine Ecosystem and Bioresource, Ministry of Natural Resources (2021QN05); and Marine Science Program for Guangxi First-Class Discipline, Beibu Gulf University (DRA002, TRA001).

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Authors and Affiliations

  1. Guangxi Key Laboratory of Beibu Gulf Marine Biodiversity Conservation, College of Marine Sciences, Beibu Gulf University, Qinzhou City, 535011, Guangxi, China

    Kianann Tan, Boyu Su, Chen Zhan, Xin Yang & Kit Yue Kwan

  2. Key Laboratory of Tropical Marine Ecosystem and Bioresource, Ministry of Natural Resources, Fourth Institute of Oceanography, Beihai, 536000, Guangxi, China

    Xiaowan Ma

  3. Higher Institution Centre of Excellence (HICoE), Institute of Tropical Aquaculture and Fisheries, Universiti Malaysia Terengganu, 21030, Kuala Nerus City, Terengganu, Malaysia

    Khor Waiho

  4. Borneo Marine Research Institute, Universiti Malaysia Sabah, Kota Kinabalu City, Sabah, Malaysia

    Leong-Seng Lim

Authors
  1. Kianann Tan
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  2. Xiaowan Ma
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  3. Boyu Su
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  4. Chen Zhan
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  5. Xin Yang
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  7. Leong-Seng Lim
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  8. Kit Yue Kwan
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Contributions

Kianann Tan: Writing – original draft, Funding acquisition, Conceptualization, Resources, Supervision. Xiaowan Ma: Writing – original draft, Funding acquisition, Resources. Boyu Su: Formal analysis, Visualization. Chen Zhan: Formal analysis, Visualization. Xin Yang: Investigation, Methodology. Khor Waiho: Conceptualization. Leong-Seng Lim: Writing – review & editing. Kit Yue Kwan: Project administration, Supervision, Funding acquisition, Resources, Writing – review & editing.

Corresponding author

Correspondence to Kit Yue Kwan.

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All experiments in this study were conducted under a protocol approved by the Institutional Animal Care and Use Committee (IACUC, China).

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Tan, K., Ma, X., Su, B. et al. Targeting TtVgR via siRNA Knockdown Elicits Ovarian Cell Death in the Tri-spine Horseshoe Crab. Mar Biotechnol (2024). https://doi.org/10.1007/s10126-024-10319-7

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