Developmental and comparative immunology最新文献_第3页

Exploring the immunological functions of thioredoxin domain-containing protein 17 (TXNDC17) in chub mackerel (Scomber japonicus): Immune response and cellular redox homeostasis 探索鲭鱼含硫氧还蛋白结构域蛋白 17 (TXNDC17) 的免疫功能：免疫反应与细胞氧化还原平衡。

IF 2.7 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105303

Arthika Kalaichelvan , Jeongeun Kim , Gaeun Kim , Ji Hun Lee , H.M.V. Udayantha , Y.K. Kodagoda , W.A.D.L.R. Warnakula , G.A.N.P. Ganepola , Yuhwan Jo , U.P.E. Arachchi , B.P.M. Vileka Jayamali , Qiang Wan , Sumi Jung , Jehee Lee

All organisms have evolved sophisticated antioxidant networks and enzymes to counteract reactive radicals, among which thioredoxin (Trx) systems are especially noteworthy. Thioredoxin domain-containing protein 17 (TXNDC17) is a ubiquitously expressed enzyme with oxidoreductase activity belonging to the Trx protein family. This study successfully uncovered and analyzed the TXNDC17 gene in Scomber japonicus (SjTXNDC17). The gene consists of a 372-base-pair coding sequence that encodes a protein of 123 amino acids, with an estimated molecular weight of 14.1 kDa. Structural analysis revealed that SjTXNDC17 contains a TRX-related protein 14 domain with two redox-responsive cysteine residues in the ⁴²WCPDC⁴⁶ motif. Spatial expression analysis indicated that SjTXNDC17 had the highest constitutive expression in the brain. Stimulation with polyinosinic-polycytidylic acid (poly I:C), Vibrio harveyi, and Streptococcus iniae, significantly upregulated the mRNA levels of SjTXNDC17 in the head kidney. The antioxidant activity of the recombinant SjTXNDC17 protein was evidenced by 2,2-Diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radical scavenging, insulin reduction, and cupric ion-reducing antioxidant capacity assays. SjTXNDC17 overexpression in fathead minnow (FHM) cells significantly reduced reactive oxygen species (ROS) levels and decreased apoptosis. The anti-apoptotic effect was driven by the upregulation of the Bcl2 gene and the downregulation of the Bax gene, as well as the suppression of JNK signaling pathway genes. Moreover, overexpression of SjTXNDC17 facilitated M2 polarization and suppressed nitric oxide production in macrophages. Collectively, these results demonstrate that SjTXNDC17 plays a crucial role in both the immune response and cellular redox balance in Scomber japonicus.

所有生物都进化出了复杂的抗氧化网络和酶来对抗活性自由基，其中硫氧还蛋白（Trx）系统尤其值得注意。含硫氧还蛋白17（TXNDC17）是一种普遍表达的具有氧化还原酶活性的酶，属于Trx蛋白家族。本研究成功发现并分析了日本鲭的 TXNDC17 基因（SjTXNDC17）。该基因由 372 个碱基对的编码序列组成，可编码 123 个氨基酸的蛋白质，分子量约为 14.1 kDa。结构分析表明，SjTXNDC17 含有一个 TRX 相关蛋白 14 结构域，42WCPDC46 基序中有两个氧化还原反应半胱氨酸残基。空间表达分析表明，SjTXNDC17 在大脑中的组成表达量最高。在多聚肌苷酸（poly I:C）、哈维弧菌和猪链球菌的刺激下，头肾中 SjTXNDC17 的 mRNA 水平显著上调。重组 SjTXNDC17 蛋白的抗氧化活性通过 2,2-二苯基-1-吡啶-肼水合物（DPPH）自由基清除、胰岛素还原和铜离子还原抗氧化能力试验得到了证明。在黑头鲦鱼（FHM）细胞中过表达 SjTXNDC17 能显著降低活性氧（ROS）水平并减少细胞凋亡。这种抗凋亡效应是由 Bcl2 基因的上调和 Bax 基因的下调以及 JNK 信号通路基因的抑制所驱动的。此外，SjTXNDC17 的过表达促进了巨噬细胞的 M2 极化并抑制了一氧化氮的产生。总之，这些结果表明，SjTXNDC17 在日本鲭鱼的免疫反应和细胞氧化还原平衡中起着至关重要的作用。

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引用次数: 0

WTAP, a conserved m6A writer, can promote the antiviral immunity of Miichthys miiuy WTAP是一种保守的m6A转录因子，可促进米氏蝇的抗病毒免疫。

IF 2.7 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105310

Hanfu Gong , Wenxin Li , Shang Geng , Tianjun Xu , Yuena Sun

N6-methyladenosine (m⁶A) is one of the most prevalent modifications found in eukaryotic mRNA and has been implicated in the regulation of cell proliferation, development, invasion, apoptosis, and immunity. In this study, we first conducted a structural and evolutionary analysis of Wilms' tumour 1-associating protein (WTAP) in vertebrates, and the results showed that WTAP in vertebrates is conserved particularly in mammals and fish. We subsequently investigated the involvement of WTAP in the antiviral immune response of fish and discovered that the expression of Miichthys miiuy (mmiWTAP) decreased in response to stimulation with Siniperca chuatsi rhabdovirus (SCRV) and poly(I:C). Immunofluorescence assays revealed that mmiWTAP was distributed in both the nucleus and the cytoplasm. Furthermore, overexpression of mmiWTAP enhanced the mRNA expression of MAVS and antiviral genes, thereby inhibiting SCRV replication. The beneficial effects of WTAP on MAVS and antiviral factors were disrupted upon introduction of cycloleucine, a methylation inhibitor, suggesting that the positive regulatory role of mmiWTAP in the antiviral immune response is reliant on its methyltransferase activity. These findings provide new insights into the involvement of m⁶A regulatory networks in fish antiviral immunity.

n6 -甲基腺苷（m6A）是真核生物mRNA中最常见的修饰之一，与细胞增殖、发育、侵袭、凋亡和免疫的调控有关。在本研究中，我们首先对脊椎动物的Wilms' tumor 1- associated protein （WTAP）进行了结构和进化分析，结果表明，脊椎动物的WTAP在哺乳动物和鱼类中具有保守性。我们随后研究了WTAP在鱼类抗病毒免疫应答中的作用，发现在受到chuatsi横纹肌病毒（SCRV）和poly（I:C）的刺激后，Miichthys miuy （mimwtap）的表达降低。免疫荧光分析显示，mimwtap在细胞核和细胞质中均有分布。此外，过表达mmiWTAP可增强MAVS和抗病毒基因的mRNA表达，从而抑制SCRV的复制。引入甲基化抑制剂环亮氨酸后，WTAP对MAVS和抗病毒因子的有益作用被破坏，这表明mmiWTAP在抗病毒免疫应答中的积极调节作用依赖于其甲基转移酶活性。这些发现为m6A调控网络参与鱼类抗病毒免疫提供了新的见解。

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引用次数: 0

Transcriptomics reveals crowding stress inhibit the immune defense of the head kidney of the pearl gentian grouper juvenile through NF-κB signal pathway 转录组学揭示拥挤胁迫通过NF-κB信号通路抑制龙胆石斑鱼幼鱼头肾免疫防御。

IF 2.7 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105299

Wenyang Li , Ao Li , Xianhong Zhang , Fan Fei , Xiaoqiang Gao , Yingying Fang , Shuquan Cao , Hongjun Yang , Wensheng Li , Baoliang Liu

Crowding stress is a significant welfare factor affecting aquatic animals in recirculating aquaculture systems. Little is known regarding the influence of prolonged crowding stress on the immunity of juvenile pearl gentian groupers. However, research exploring the potential mechanisms through which crowding stress affects fish immune function is limited. Therefore, this study aims to investigate the effect of crowding stress on the immune stress of the pearl gentian grouper juvenile (♀Epinephelus fuscoguttatus × ♂ Epinephelus lanceolatus) under prolonged conditions. We focused on the pearl gentian grouper juvenile and selected low- and high-density groups as the experimental breeding densities. Research shows that crowding stress increases the activities of alkaline acid plum and acid phosphatase, reduces the activities of lysozyme and immunoglobulin M content. RNA sequencing and comparative transcriptomic analyses were employed to explore changes in the gene expression of juvenile pearl gentian groupers subjected to crowding stress. Differential gene expression analyses between the low- and high-density groups identified 5777 unigenes that were differentially expressed following crowding stress, with 3216 and 2561 upregulated and downregulated, respectively. In the GO and KEGG enrichment analyses, many of the enriched signaling pathways related to genes were associated with immunity and oxidative stress. In addition, the combined analyses of enzyme activity and transcriptomics indicated that crowding stress suppressed the immune function of juvenile pearl gentian groupers, reducing their immune ability. Overall, these findings offer new insights into the molecular mechanisms underlying crowding stress tolerance in juvenile pearl gentian grouper, suggesting that the NF-κB pathway plays a crucial role in the immune response of the head kidney of the pearl gentian grouper to long-term crowding stress.

拥挤胁迫是影响循环水养殖系统中水生动物福利的重要因素。长期的拥挤胁迫对龙胆石斑鱼幼鱼免疫力的影响尚不清楚。然而，关于拥挤胁迫影响鱼类免疫功能的潜在机制的研究是有限的。因此，本研究旨在研究拥挤胁迫对长时间条件下龙胆石斑鱼幼鱼（♀Epinephelus fuscoguttatus ×♂Epinephelus lanceolatus）免疫应激的影响。以龙胆石斑鱼幼鱼为研究对象，选择低密度和高密度两组作为实验养殖密度。研究表明，拥挤胁迫增加了碱性酸梅和酸性磷酸酶的活性，降低了溶菌酶活性和免疫球蛋白M含量。通过RNA测序和比较转录组学分析，探讨了拥挤胁迫下龙胆石斑鱼幼鱼基因表达的变化。低密度组和高密度组之间的差异基因表达分析发现，拥挤胁迫下差异表达的单一基因有5777个，分别有3216个和2561个上调和下调。在GO和KEGG富集分析中，许多与基因相关的富集信号通路与免疫和氧化应激有关。此外，酶活性和转录组学分析表明，拥挤胁迫抑制了龙胆石斑鱼幼鱼的免疫功能，降低了它们的免疫能力。总之，这些发现为龙胆石斑鱼幼鱼对拥挤胁迫耐受的分子机制提供了新的见解，表明NF-κB通路在龙胆石斑鱼头肾对长期拥挤胁迫的免疫应答中起着至关重要的作用。

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引用次数: 0

Ontogeny of the organized nasopharynx-associated lymphoid tissue in rainbow trout 虹鳟鱼有组织鼻咽相关淋巴组织的个体发生。

IF 2.7 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105298

Benjamin J. Garcia , Narmin Musayeva , Alexis Reyes , Chrysler Martinez , Yago Serra dos Santos , Irene Salinas

Understanding the ontogeny of teleost mucosa-associated lymphoid tissues (MALT) is critical for determining the earliest timepoint for effective mucosal vaccination of young fish. Here, we describe the developmental sequence that leads to the formation of an organized MALT structure in rainbow trout, the organized nasopharynx-associated lymphoid tissue (O-NALT). Control rainbow trout were sampled between 340 and 1860 degree days (DD) and routine histology and immunofluorescence staining were used to determine cellular changes in immune cells in the nasal cavity as well as O-NALT formation. We identified that O-NALT is first seeded by CD8α⁺ T cells at 700 DD followed by IgM⁺ B cells and cd4-2b⁺ cells at 1000 DD. Histomorphologically, trout O-NALT is fully formed at 1400 DD. Whole body gene expression analyses uncovered waves of igmh, cd4-2b, and cd8a expression that recapitulate the cellular seeding sequence of O-NALT by specific lymphocyte subsets. Our results indicate that 1) O-NALT formation results from a specific sequence of lymphocyte subset colonization pioneered by CD8α⁺ T cells and 2) the presence of the full O-NALT structure at 1400 DD may mark this timepoint as the earliest developmental stage at which mucosal vaccines can induce long lasting, specific immune responses.

了解硬骨鱼粘膜相关淋巴组织（MALT）的个体发生对于确定幼鱼有效粘膜疫苗接种的最早时间点至关重要。在这里，我们描述了导致虹鳟鱼形成有组织MALT结构的发育序列，即有组织的鼻咽相关淋巴组织（O-NALT）。对照虹鳟鱼在340 - 1860度（DD）之间取样，采用常规组织学和免疫荧光染色测定鼻腔免疫细胞的细胞变化以及O-NALT的形成。我们发现O-NALT首先由CD8α+ T细胞在700 DD时植入，然后是IgM+ B细胞和cd4-2b+细胞在1000 DD时植入。组织形态学上，鳟鱼O-NALT在1400 DD时完全形成。全身基因表达分析揭示了igmh、cd4-2b和cd8a的表达波，这些表达波通过特定淋巴细胞亚群总结了O-NALT的细胞植入序列。我们的研究结果表明，1)O-NALT的形成是由CD8α+ T细胞主导的淋巴细胞亚群定殖的特定序列引起的，2)在1400dd出现完整的O-NALT结构可能标志着这个时间点是黏膜疫苗可以诱导持久的特异性免疫反应的最早发育阶段。

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引用次数: 0

HSC70 functions as a negatively regulator in IFN signaling pathway via suppressing K63-linked ubiquitination of RIG-I in black carp HSC70通过抑制黑鱼体内与k63相关的RIG-I泛素化，在IFN信号通路中发挥负调控作用。

IF 2.7 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105300

Jiaxin Fu , Nianfeng chen , Tian Qin , Yixin Chen , Ji Liu , Hui Wu , Jun Yan , Jun Xiao , Jun Zou , Hao Feng

Heat shock cognate 70 (HSC70), a highly conserved molecular chaperone in the heat shock protein 70 (HSP70) family, plays an essential role in maintaining the homeostasis of the cellular environment. Furthermore, although previous studies have investigated potential function of HSC70 in innate antiviral immunity, further research is still required to fully elucidate its role. In this study, we cloned and characterized the HSC70 homolog gene from black carp (Mylopharyngodon piceus), which consists of 1950 nucleotides encoding 650 amino acids, migrates at approximately 71 kDa on SDS-PAGE, and is distributed in the cytoplasm. In response to different stimuli (SVCV, poly (I:C) and LPS), the transcription level of black carp HSC70 (bcHSC70) all increased to a certain extent. Luciferase reporter assay demonstrated that co-transfected bcHSC70 obviously reduced activity of interferon (IFN) promoters mediated by most factors in the RLRs pathway, and further qRT-PCR and plaque assay indicated that co-transfection of bcHSC70 with bcRIG-I decreased the bcRIG-I-mediated IFN transcription and antiviral ability resisting spring viremia of carp virus (SVCV), whereas knockdown of bcHSC70 improves the host cellular antiviral activity. Noteworthily, co-immunoprecipitation (co-IP) assay and immunofluorescence (IF) assay confirmed bcHSC70 interacts with bcRIG-I, and weaken K63-linked polyubiquitination of bcRIG-I. In summary, our study revealed that HSC70 negatively regulates IFN signaling pathway through impairing K63-linked ubiquitination of RIG-I in black carp, which provides an important basis for exploring innate immune regulatory mechanisms in teleost fish.

热休克同源蛋白70 （HSC70）是热休克蛋白70 （HSP70）家族中高度保守的分子伴侣，在维持细胞环境的稳态中起重要作用。此外，尽管先前的研究已经研究了HSC70在先天抗病毒免疫中的潜在功能，但仍需要进一步的研究来充分阐明其作用。本研究从黑鱼（Mylopharyngodon piceus）中克隆并鉴定了HSC70同源基因，该基因由1950个核苷酸组成，编码650个氨基酸，在SDS-PAGE上迁移约71 kDa，分布于细胞质中。在不同刺激（SVCV、poly （I:C）和LPS）下，黑鱼HSC70 （bcHSC70）的转录水平均有一定程度的升高。荧光素酶报告基因实验表明，共转染bcHSC70可明显降低RLRs通路中大部分因子介导的干扰素（IFN）启动子的活性，qRT-PCR和斑块实验进一步表明，bcHSC70与bcrig - 1共转染可降低bcrig - 1介导的IFN转录和抗鲤春病毒血症（SVCV）的抗病毒能力，而敲低bcHSC70可提高宿主细胞的抗病毒活性。值得注意的是，共免疫沉淀（co-IP）和免疫荧光（IF）实验证实了bcHSC70与bcrig - 1相互作用，并减弱了bcrig - 1的k63连锁多泛素化。综上所述，我们的研究揭示了HSC70通过破坏黑鱼体内k63相关的RIG-I泛素化而负调控IFN信号通路，为探索硬骨鱼的先天免疫调节机制提供了重要依据。

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引用次数: 0

First report of Peroxiredoxin-5 (PRDX5) in starry flounder (Platichthys stellatus): Molecular features and expression analysis 首次报道星鲽体内的过氧化物歧化酶 5 (PRDX5)：分子特征和表达分析。

IF 2.7 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105291

Min-Young Sohn , Kwang-Min Choi , Ji-Min Jeong , Gyoungsik Kang , Won-Sik Woo , Kyung-Ho Kim , Ha-Jeong Son , Hye-Seong Kim , Jun-ichi Hikima , Chan-Il Park

In this study, the Peroxiredoxin-5 (PRDX5) gene from starry flounder (Platichthys stellatus, PsPRDX5) was identified through next-generation sequencing (NGS) and verified via cloning and sequencing. The predicted amino acid sequence, derived from the cDNA, was confirmed by multiple sequence alignment and phylogenetic analysis, establishing its homology with related sequences. PsPRDX5 mRNA was expressed across all examined tissues in healthy starry flounder, with distinct tissue-specific expression profiles. Following artificial infection with viral hemorrhagic septicemia virus (VHSV) and Streptococcus parauberis PH0710, significant alterations in PsPRDX5 mRNA expression were observed. VHSV infection resulted in a pronounced downregulation of PsPRDX5 in most major tissues, except for a significant upregulation in the intestine at 7 days post-infection. Conversely, infection with S. parauberis PH0710 led to a significant upregulation of PsPRDX5 in nearly all tissues. These results suggest that the differential expression of PsPRDX5 in response to VHSV and S. parauberis PH0710 reflects pathogen-specific immune and antioxidant defense mechanisms, highlighting the role of PsPRDX5 in the immune response of starry flounder to both viral and bacterial infections.

本研究通过新一代测序（NGS）鉴定了星鲽的过氧化还原酶-5（Peroxiredoxin-5，PRDX5）基因，并通过克隆和测序进行了验证。通过多序列比对和系统进化分析，证实了从 cDNA 中推导出的氨基酸序列，并确定了其与相关序列的同源性。PsPRDX5 mRNA在健康星鲽的所有受检组织中均有表达，并具有明显的组织特异性表达特征。人工感染病毒性出血性败血症病毒（VHSV）和副猪链球菌 PH0710 后，PsPRDX5 mRNA 的表达发生了显著变化。VHSV 感染导致 PsPRDX5 在大多数主要组织中的表达明显下调，但在感染后 7 天，肠道中的表达明显上调。相反，感染副猪嗜血杆菌 PH0710 后，PsPRDX5 在几乎所有组织中都显著上调。这些结果表明，PsPRDX5对VHSV和S. parauberis PH0710的不同表达反映了病原体特异性免疫和抗氧化防御机制，突出了PsPRDX5在星鲽对病毒和细菌感染的免疫反应中的作用。

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引用次数: 0

Caspase-8 promotes innate immunity in the Chinese mitten crab by regulating the expression of antimicrobial peptides and apoptosis in hemocyte Caspase-8通过调控抗菌肽表达和血细胞凋亡促进中华绒螯蟹先天免疫。

IF 2.7 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105308

Guangyu Wang , Guoqing Shen , Chaohui Xu , Yanan Guo , Wen Zhang , Qun Wang , Youting Zhu

In mammals, caspase-8 primarily functions as an initiator caspase that regulates apoptosis, while in Drosophila, the caspase-8 ortholog DREDD not only induces apoptosis during development but also regulates antimicrobial peptides (AMPs) expression during Gram-negative bacterial infection-induced immune responses. However, the immune-related function of caspase-8 in the crustacean remains unknown. In the present study, the open reading frame of EsCaspase-8 was cloned from the Chinese mitten crab (Eriocheir sinensis). The deduced EsCaspase-8 protein sequence contained only one death effector domain (DED) and a cysteine aspartase cysteine structural domain. The EsCaspase-8 expression was significantly induced after 6 h of Vibrio parahaemolyticus infection and continued to 24 h in hemocyte. Knocking down EsCaspase-8 expression in hemocytes significantly inhibited Relish's nuclear translocation and suppressed the expression of AMPs, including Crustin 1, Crustin 2, Lysosome, and double WAP domain, after V. parahaemolyticus infection. Furthermore, the knockdown of EsCaspase-8 in vivo significantly inhibited hemocyte apoptosis post-bacterial infection. These results demonstrated that EsCaspase-8 can promote antibacterial activities by regulating the expression of AMPs through activation of Relish nuclear translocation in Chinese mitten crabs, thus acting as a critical positive regulator in innate immunity. In addition, EsCaspase-8 also has the function of inducing hemocyte apoptosis. These findings expand our understanding of the molecular mechanisms underlying crustacean immune responses and provide a foundation for future research to improve disease resistance.

在哺乳动物中，caspase-8主要作为一种启动caspase调节细胞凋亡，而在果蝇中，caspase-8同源基因DREDD不仅在发育过程中诱导细胞凋亡，而且在革兰氏阴性细菌感染诱导的免疫反应中调节抗菌肽（AMPs）的表达。然而，caspase-8在甲壳类动物中的免疫相关功能尚不清楚。本研究从中华绒螯蟹（Eriocheir sinensis）中克隆了EsCaspase-8的开放阅读框。推断的EsCaspase-8蛋白序列仅包含一个死亡效应域（DED）和一个半胱氨酸天冬氨酸半胱氨酸结构域。EsCaspase-8在感染副溶血性弧菌6 h后显著表达，并持续到24 h。在副溶血性弧菌感染后，敲低EsCaspase-8在血细胞中的表达，显著抑制了津津的核易位，抑制了包括Crustin 1、Crustin 2、溶酶体和双WAP结构域在内的amp的表达。此外，在体内敲低EsCaspase-8可显著抑制细菌感染后的血细胞凋亡。以上结果表明，EsCaspase-8通过激活中华绒螯蟹的津津核易位，调控AMPs的表达，从而促进了中华绒螯蟹的抗菌活性，在先天免疫中起着重要的正调控作用。此外，EsCaspase-8还具有诱导血细胞凋亡的功能。这些发现扩大了我们对甲壳类动物免疫反应的分子机制的理解，并为今后提高疾病抵抗力的研究提供了基础。

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引用次数: 0

Infectious myonecrosis virus (IMNV) induces upregulation of RNAi-related genes in white shrimp Penaeus vannamei 传染性肌坏死病毒（IMNV）诱导凡纳滨对虾rnai相关基因表达上调。

IF 2.7 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105296

Rubens Galdino Feijó , Jhonatas Teixeira Viana , Rodrigo Maggioni , Luis Fernando Marins

Infectious myonecrosis virus (IMNV) still causes significant economic and social losses in American and Asian shrimp farming. In this work, we investigated the transcription patterns of Sid-1, Dicer-2 and Argonaute-2 genes from the RNAi mechanism in Penaeus vannamei naturally infected with IMNV, and injected with inoculum containing 1.02 × 10⁵, 1.02 × 10⁴ or 1.02 × 10³ IMNV copies‧μL⁻¹. We observed that infection with increasing IMNV concentrations affected the transcription levels of these key genes. However, the viral load did not decrease during the experiment. We suggest that changes in Sid-1 mRNA expression could be used as marker of viral replication for evaluating sanitary status in P. vannamei farming.

传染性肌坏死病毒（IMNV）仍然给美国和亚洲的虾养殖业造成重大的经济和社会损失。本研究从RNAi机制研究了Sid-1、Dicer-2和Argonaute-2基因在自然感染了IMNV并注射含有1.02×105、1.02×104或1.02×103 IMNV拷贝·μL-1的接种剂的南美对虾（Penaeus vannamei）体内的转录模式。我们观察到，随着感染病毒浓度的增加，这些关键基因的转录水平受到影响。然而，在实验过程中，病毒载量并没有减少。我们认为Sid-1 mRNA表达的变化可以作为病毒复制的标记物，用于评价凡纳米养殖的卫生状况。

引用次数: 0

CgmiR307 involved in the regulation of Nrf2-dependent oxidative response in the Pacific oyster Crassostrea gigas under high-temperature stress CgmiR307参与高温胁迫下太平洋牡蛎长牡蛎nrf2依赖性氧化反应的调控。

IF 2.7 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105306

Keli Zhou , Lei Gao , Pingan Ge , Ling Wang , Lu Liu , Jiayu Ye , Hairu Xu , Lingling Wang , Linsheng Song

miRNA, a type of endogenous small non-coding RNA, is involved in the response to various environmental stresses through post-transcriptional regulation. In the present study, the role of CgmiR307 in the regulation of oxidative response under high-temperature stress by targeting CgNrf2 was investigated in the Pacific oyster Crassostrea gigas. The binding site of CgmiR307 were predicted at 1799–1818 bp in the 3′-UTR of CgNrf2, and the binding activity of CgmiR307 with the mRNA of CgNrf2 was further proved by the dual-luciferase reporter assay. The expression levels of CgmiR307 and CgNrf2 in gill were significantly higher than in other tissues, and exhibited significant fluctuations and variations after exposure to 28 °C. There was a significant reduction in the expression levels of CgSOD, and CgCAT in gill, as well as the activities of SOD, CAT, and T-AOC, while ROS and MDA contents significantly increased in CgNrf2-RNAi oysters. After CgmiR307 agomir injection and high-temperature stress, the expression levels of CgNrf2, CgSOD and CgCAT in gill, the activities of SOD and CAT and T-AOC decreased significantly, while ROS and MDA content significantly increased. After CgmiR307 antagomir injection and high-temperature stress, the changes in the parameters of oxidative response shown exactly the opposite trend. These results demonstrated that CgmiR307 was involved in the regulation of oxidative response by inhibiting the mRNA expression of CgNrf2 under high-temperature stress.

miRNA是一种内源性小非编码RNA，通过转录后调控参与对各种环境胁迫的反应。本研究在太平洋牡蛎长牡蛎中研究了CgmiR307靶向CgNrf2调控高温胁迫下氧化反应的作用。在CgNrf2的3'-UTR中预测了CgmiR307的结合位点在1799 - 1818 bp，并通过双荧光素酶报告基因实验进一步证实了CgmiR307与CgNrf2 mRNA的结合活性。CgmiR307和CgNrf2在鳃中的表达水平明显高于其他组织，并在28°C后呈现明显的波动和变化。CgNrf2-RNAi牡蛎鳃中CgSOD、CgCAT的表达水平显著降低，SOD、CAT和T-AOC活性显著降低，ROS和MDA含量显著升高。注射CgmiR307 agomir和高温胁迫后，鳃中CgNrf2、CgSOD和CgCAT的表达水平以及SOD、CAT和T-AOC的活性显著降低，ROS和MDA含量显著升高。注射CgmiR307和高温应激后，氧化反应参数的变化趋势完全相反。这些结果表明，在高温胁迫下，CgmiR307通过抑制CgNrf2 mRNA的表达参与了氧化反应的调控。

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引用次数: 0

Molecular dynamics and spatial response of proliferation and apoptosis in wound healing and early intestinal regeneration of sea cucumber Apostichopus japonicus 刺参创面愈合和早期肠道再生过程中增殖和凋亡的分子动力学及空间响应

IF 2.7 3区农林科学 Q1 FISHERIES

Developmental and comparative immunology

Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105297

Fang Su , Igor Yu. Dolmatov , Wei Cui , Hongsheng Yang , Lina Sun

The sea cucumber, Apostichopus japonicus, exhibits significant regenerative capabilities. To ensure survival and reduce metabolic costs under adverse conditions, A. japonicus can expel intestine, respiratory trees and other internal organs. It takes only 14 days to regenerate a fully connected, lumen-containing intestine. Despite numerous reports characterizing the cellular events in intestinal regeneration, limited investigation has been conducted on the molecular events that occur during wound healing and the initial stages of regeneration after evisceration. Here, we identified differentially expressed genes (DEGs) during wound healing (6 h post-evisceration, Aj6hpe) and early intestinal regeneration (Aj1dpe, Aj3dpe, Aj7dpe). Cell proliferation and apoptosis were detected by EdU and TUNEL assays, respectively. Results demonstrated that calcium ion and neuroactive ligand-receptor interaction were involved in the transmission of injury signals from evisceration to Aj1dpe. The main events occurring in the wound healing and early regeneration process were autophagy, apoptosis, dedifferentiation, migration and shutdown of feeding. Cell proliferation was primarily observed during the lumen formation stage. Maximal number of apoptotic cells were found during wound healing stage (6 hpe - 1 dpe). Consequently, the immune response is mainly mobilized by neural regulation after evisceration. Our findings bridge the gap between evisceration and regeneration, illuminating the molecular events that mediate damage response and initiate regeneration. This study significantly advances our understanding of the mechanisms underlying intestinal regeneration.

海参Apostichopus japonicus表现出显著的再生能力。为了在不利条件下保证生存和降低代谢成本，日本刺参可以排出肠道、呼吸树等内脏器官。只需要14天就能再生出一个完全连接的、含管腔的肠道。尽管有许多报道描述了肠道再生中的细胞事件，但对伤口愈合和内脏切除后再生初始阶段发生的分子事件进行了有限的调查。在这里，我们鉴定了伤口愈合（内脏取出后6小时，Aj6hpe）和早期肠道再生（Aj1dpe, Aj3dpe, Aj7dpe）期间的差异表达基因（DEGs）。EdU和TUNEL分别检测细胞增殖和凋亡。结果表明，钙离子和神经活性配体受体相互作用参与了损伤信号从内脏到Aj1dpe的传递。在创面愈合和早期再生过程中发生的主要事件是自噬、凋亡、去分化、迁移和停止进食。细胞增殖主要发生在管腔形成阶段。创面愈合期凋亡细胞数最多（6 hpe - 1 dpe）。因此，内脏切除后的免疫反应主要是通过神经调节来调动的。我们的发现弥合了内脏和再生之间的差距，阐明了介导损伤反应和启动再生的分子事件。这项研究极大地促进了我们对肠道再生机制的理解。

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