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A novel digestive protease chymotrypsin-like serine contributes to anti-BmNPV activity in silkworm (Bombyx mori) 一种新型消化蛋白酶糜蛋白酶样丝氨酸有助于提高家蚕(Bombyx Mori)的抗 BmNPV 活性。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105301
Wei-Wei Kong , Yu-Liang Yan , Cai-Ping Hou , Tao Hong , Yi-Sheng Wang , Xin Xu , Shi-Huo Liu , Jia-Ping Xu
Serine proteases (SPs) are important proteases in the digestive system of lepidopteran insects. They play important roles in protein digestion, coagulation, signal transduction, hormone activation, inflammation and development. Blood-borne pyosis caused by Bombyx mori nuclear polyhedrosis virus (BmNPV) has caused serious harm to sericulture. At present, the scientific problems of BmNPV infection and silkworm resistance to BmNPV infection have been the focus of many scientists, but the molecular mechanism needs further research and exploration. Based on the results of label-free quantitative protein proteomics of the midgut digestive juice of different resistant strains in our laboratory, we successfully screened a differentially expressed candidate protein (DEP), B. mori chymotrypsin-like serine protease (BmCLSP), and comprehensively analyzed the biological characteristics and anti-BmNPV function of BmCLSP. The open reading frame (ORF) of BmCLSP is 891 bp, encoding 296 amino acid residues. The analysis of the domain structure showed that there was a signal peptide and a trypsin-like serine protease domain, Tryp_SPC, in the BmCLSP protein. Semi-quantitative and real-time fluorescence quantitative PCR analysis showed that the BmCLSP gene was highly expressed in the fifth instar larvae of silkworm, and specifically expressed in the midgut. The expression level of BmCLSP in the BmNPV resistant strain A35 was higher than that in the sensitive strain P50. Virus amplification analysis showed that the relative expression level of VP39 was significantly lower than that of the control group after infection of silkworm larvae and BmN cells with BmNPV treated with recombinant BmCLSP at an appropriate concentration. Furthermore, our overexpression of BmCLSP in BmN cells significantly inhibited the expansion of BmNPV. In summary, the results of this study indicate that BmCLSP has anti-BmNPV activity in silkworm, and can significantly inhibit the proliferation of BmNPV in silkworm. It offers a promising avenue for silkworm anti-virus breeding.
丝氨酸蛋白酶(SPs)是鳞翅目昆虫消化系统中的重要蛋白酶。它们在蛋白质消化、凝血、信号转导、激素激活、炎症和发育中发挥重要作用。家蚕核型多角体病毒(Bombyx mori nuclear polyhedrosis virus, BmNPV)引起的血源性化脓病对养蚕业造成了严重危害。目前,关于BmNPV感染和家蚕对BmNPV感染的抗性的科学问题一直是众多科学家关注的焦点,但其分子机制还需要进一步的研究和探索。基于本实验室对不同耐药菌株中肠消化液进行无标记定量蛋白质组学分析,成功筛选了一种差异表达候选蛋白(DEP)——家蚕凝乳胰蛋白酶样丝氨酸蛋白酶(BmCLSP),并综合分析了BmCLSP的生物学特性和抗bmnpv功能。BmCLSP的开放阅读框(ORF)长度为891 bp,编码296个氨基酸残基。结构域结构分析表明,BmCLSP蛋白中存在一个信号肽和一个胰蛋白酶样丝氨酸蛋白酶结构域Tryp_SPC。半定量和实时荧光定量PCR分析表明,BmCLSP基因在家蚕5龄幼虫中高表达,并在中肠特异性表达。BmCLSP在BmNPV抗性菌株A35中的表达量高于敏感菌株P50。病毒扩增分析表明,用适当浓度的重组BmCLSP处理过的BmNPV感染家蚕幼虫和BmN细胞后,VP39的相对表达量显著低于对照组。此外,我们在BmN细胞中过表达BmCLSP可显著抑制BmNPV的扩增。综上所述,本研究结果表明,BmCLSP在家蚕体内具有抗BmNPV活性,并能显著抑制BmNPV在家蚕体内的增殖。这为蚕种抗病毒育种提供了一条很有前途的途径。
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引用次数: 0
Expression of FcμR by bovine mononuclear blood leukocytes 牛单核血白细胞中FcμR的表达。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105304
Florence B. Gilbert, Rodrigo P. Martins, Pascal Rainard
The receptor for IgM has been identified a few years ago, but its expression by bovine mononuclear cells has not yet been studied. We used rabbit antibodies against bovine FcμR to begin to fill this gap. Anti-FcμR antibodies bound to B lymphocytes and monocytes, although less than to neutrophils. Nonclassical and intermediate monocytes (CD172apos CD16pos) bound nonspecifically to rabbit antibodies, complicating analysis, but they bound more anti-FcμR antibodies than control antibodies, indicating that they also express the FcμR. They also express more C5a receptors than classical monocytes. Anti-FcμR antibodies did not bind to CD3pos αβT lymphocytes (both CD4pos and CD8pos) and γδT cells. At low temperature but not at physiological temperature, purified bovine IgM bound to all monocytes and strongly to all B cells, but hardly to CD3pos T cells. Monocytes and B cells bound human IgA, but IgA did not compete, whereas unlabeled bovine IgM competed for binding of labeled IgM. This supports the role of the FcμR, and not the FαμR, in IgM binding. Finally, we showed that monocytes were able to ingest bacteria opsonized with serum deprived of IgG, indicating their ability to perform IgM-dependent phagocytosis. In conclusion, surface expression of FcμR by unstimulated blood leukocytes was demonstrated on B cells and monocytes, but not on T cells.
IgM的受体在几年前已被发现,但其在牛单核细胞中的表达尚未被研究。我们使用兔抗牛FcμR抗体开始填补这一空白。抗fc μ r抗体与B淋巴细胞和单核细胞结合,但与中性粒细胞结合较少。非经典和中间单核细胞(CD172apos CD16pos)与兔抗体非特异性结合,使分析复杂化,但它们结合的抗FcμR抗体多于对照抗体,表明它们也表达FcμR。它们也比传统的单核细胞表达更多的C5a受体。抗fc μ r抗体不与CD3pos αβT淋巴细胞(CD4pos和CD8pos)和γδT细胞结合。在低温而非生理温度下,纯化的牛IgM与所有单核细胞结合,与所有B细胞结合强烈,但与CD3pos T细胞结合不强烈。单核细胞和B细胞结合人IgA,但IgA不竞争,而未标记的牛IgM则竞争与标记的IgM结合。这支持了FcμR在IgM结合中的作用,而不是FαμR。最后,我们发现单核细胞能够摄取被血清中IgG剥夺而调理的细菌,这表明它们有能力进行igm依赖性吞噬。综上所述,未受刺激的血白细胞在B细胞和单核细胞表面表达FcμR,而在T细胞表面不表达。
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引用次数: 0
Haemolymphatic tissues of captive boa constrictor (Boa constrictor): Morphological features in healthy individuals and with boid inclusion body disease 圈养大蟒蛇的血淋巴组织:健康个体和Boid包涵体病的形态学特征。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105302
E. Dervas , E. Michalopoulou , J. Hepojoki , T. Thiele , F. Baggio , U. Hetzel , A. Kipar
Knowledge on the structure and composition of the haematopoietic tissue (HT) is essential to understand the basic immune functions of the immune system in any species. For reptiles, it is extremely limited, hence we undertook an in-depth in situ investigation of the HT (bone marrow, thymus, spleen, lymphatic tissue of the alimentary tract) in the common boa (Boa constrictor). We also assessed age- and disease-related changes, with a special focus on Boid Inclusion Body Disease, a highly relevant reptarenavirus-associated disease in boid snakes. The HT was subjected to gross, histological and ultrastructural examination, including special stains to highlight collagen and reticulin fibers and iron, immunohistochemistry, in situ hybridization and morphometric analyses. In general, the HT was dominated by T cells and lacked a clear structural organization, such as follicle formation. BIBD was associated with significantly higher cellularity and a granulomatous response in the spleen, and the presence of virus-infected haematopoietic cells in the bone marrow, suggesting the latter as a persistent source of viremia.
了解造血组织(HT)的结构和组成对于了解任何物种免疫系统的基本免疫功能至关重要。对于爬行动物来说,这是非常有限的,因此我们对普通蟒蛇(蟒蛇)的HT(骨髓、胸腺、脾脏、消化道淋巴组织)进行了深入的原位调查。我们还评估了年龄和疾病相关的变化,特别关注类蛇包涵体疾病,这是一种与类蛇中reprepavirus相关的高度相关的疾病。对HT进行大体、组织学和超微结构检查,包括特殊染色以突出胶原、网状蛋白纤维和铁,免疫组织化学,原位杂交和形态计量学分析。一般情况下,HT以T细胞为主,缺乏清晰的结构组织,如卵泡形成。BIBD与脾脏中明显较高的细胞性和肉芽肿反应以及骨髓中存在病毒感染的造血细胞相关,表明后者是病毒血症的持续来源。
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引用次数: 0
Effects of Saprolegnia parasitica on pathological damage and metabolism of Epithelioma papulosum cyprini cell 寄生腐毛虫对鲤丘疹上皮瘤细胞病理损伤及代谢的影响。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105311
Yali Wang , Xiaoning Gao , Tianewi Wang , Yangyang Zhang , Kun Hu
<div><div>Saprolegniasis is a common fungal disease in aquaculture. It will form white flocculent hyphae on the skin of fish, and the hyphae may grow inward and penetrate into muscle tissue, which will reduce the immunity of the body and eventually lead to death. However, there are still some gaps in the mechanism of the fish body surface against the invasion of <em>Saprolegnia</em>. This study explored the defense mechanism of <em>Epithelioma papulosum cyprini</em> cell (EPC) in the process of <em>Saprolegnia parasitica</em> infection from the perspective of pathogenic bacteria and host cells, so as to provide a theoretical basis for further exploring the mechanism of host resistance to <em>S</em>. <em>parasitica</em> invasion. The EPC cell was used as the research object. The EPC cells were treated with 1 × 10<sup>6</sup> CFU/mL of <em>S</em>. <em>parasitica</em> for 0, 6, 12, 24, 48 and 72 h. Cell viability and cell membrane damage were detected, and the non-specific immune enzyme activity in the cells was detected. Based on the above research, the apoptosis genes and antioxidant genes in the cells were detected to analyze the effect of <em>S</em>. <em>parasitica</em> on the metabolism of the EPC cells. The results showed that with the prolongation of the co-culture time of <em>S</em>. <em>parasitica</em> and cells, the cell viability gradually decreased and the cell membrane integrity was destroyed, but at the same time, the activity of non-specific immune enzymes increased to resist the infection of <em>S</em>. <em>parasitica</em>. In addition, the detection of EPC apoptosis gene <em>casp3a</em> and <em>CTSD</em> showed that the relative content of <em>casp3a</em> gene increased significantly at 24 h and reached the maximum value of the culture time (<em>P</em> < 0.05). The content of <em>CTSD</em> gene increased significantly at 12 h and reached the maximum value (<em>P</em> < 0.05). The results of antioxidant immune genes <em>serpinh1a</em> and <em>gpx1a</em> were opposite to the structure of apoptotic genes. The content of <em>serpinh1a</em> and <em>gpx1a</em> genes decreased significantly at 12 h (<em>P</em> < 0.05), but with the prolongation of culture time, the content increased significantly at 24 h and 48 h (<em>P</em> < 0.05). After stimulation of EPC cells by <em>S</em>. <em>parasitica</em>, the differential metabolites were mainly concentrated in Lipids, Compounds with biological roles and Phytochemical compounds. The KEGG pathway mainly focused on ABC transporters, Glycerophospholipid metabolism, Cysteine and methionine metabolism, Glycine, serine and threonine metabolism, Purine metabolism. In general, <em>S</em>. <em>parasitica</em> can affect cell activity, destroy the cell membrane of EPC cells, and cause apoptosis. However, EPC cells can also resist the invasion of <em>S</em>. <em>parasitica</em> by regulating their own non-specific immunity and their own metabolites, thereby protecting the body from the infectio
腐殖菌病是水产养殖中一种常见的真菌病。它会在鱼的皮肤上形成白色絮状菌丝,菌丝可能向内生长并渗入肌肉组织,从而降低机体的免疫力,最终导致死亡。然而,对鱼体表抵御腐毛虫入侵的机制仍有一些空白。本研究从病原菌和寄主细胞的角度探讨了cyprini上皮瘤乳头状瘤细胞(epelioma papulosum cyprini cell, EPC)在寄生病霉感染过程中的防御机制,为进一步探索寄主抵抗寄生病霉侵袭的机制提供理论依据。以EPC细胞为研究对象。以1×106 CFU/mL的寄生弧菌处理EPC细胞0、6、12、24、48和72 h,检测细胞活力和细胞膜损伤情况,检测细胞非特异性免疫酶活性。在此基础上,检测细胞中凋亡基因和抗氧化基因,分析寄生葡萄球菌对EPC细胞代谢的影响。结果表明,随着寄生弧菌与细胞共培养时间的延长,细胞活力逐渐下降,细胞膜完整性被破坏,但同时非特异性免疫酶活性增强,抵抗寄生弧菌的感染。此外,对EPC细胞凋亡基因casp3a和CTSD的检测显示,casp3a基因的相对含量在24 h时显著增加,达到培养时间的最大值(P
{"title":"Effects of Saprolegnia parasitica on pathological damage and metabolism of Epithelioma papulosum cyprini cell","authors":"Yali Wang ,&nbsp;Xiaoning Gao ,&nbsp;Tianewi Wang ,&nbsp;Yangyang Zhang ,&nbsp;Kun Hu","doi":"10.1016/j.dci.2024.105311","DOIUrl":"10.1016/j.dci.2024.105311","url":null,"abstract":"&lt;div&gt;&lt;div&gt;Saprolegniasis is a common fungal disease in aquaculture. It will form white flocculent hyphae on the skin of fish, and the hyphae may grow inward and penetrate into muscle tissue, which will reduce the immunity of the body and eventually lead to death. However, there are still some gaps in the mechanism of the fish body surface against the invasion of &lt;em&gt;Saprolegnia&lt;/em&gt;. This study explored the defense mechanism of &lt;em&gt;Epithelioma papulosum cyprini&lt;/em&gt; cell (EPC) in the process of &lt;em&gt;Saprolegnia parasitica&lt;/em&gt; infection from the perspective of pathogenic bacteria and host cells, so as to provide a theoretical basis for further exploring the mechanism of host resistance to &lt;em&gt;S&lt;/em&gt;. &lt;em&gt;parasitica&lt;/em&gt; invasion. The EPC cell was used as the research object. The EPC cells were treated with 1 × 10&lt;sup&gt;6&lt;/sup&gt; CFU/mL of &lt;em&gt;S&lt;/em&gt;. &lt;em&gt;parasitica&lt;/em&gt; for 0, 6, 12, 24, 48 and 72 h. Cell viability and cell membrane damage were detected, and the non-specific immune enzyme activity in the cells was detected. Based on the above research, the apoptosis genes and antioxidant genes in the cells were detected to analyze the effect of &lt;em&gt;S&lt;/em&gt;. &lt;em&gt;parasitica&lt;/em&gt; on the metabolism of the EPC cells. The results showed that with the prolongation of the co-culture time of &lt;em&gt;S&lt;/em&gt;. &lt;em&gt;parasitica&lt;/em&gt; and cells, the cell viability gradually decreased and the cell membrane integrity was destroyed, but at the same time, the activity of non-specific immune enzymes increased to resist the infection of &lt;em&gt;S&lt;/em&gt;. &lt;em&gt;parasitica&lt;/em&gt;. In addition, the detection of EPC apoptosis gene &lt;em&gt;casp3a&lt;/em&gt; and &lt;em&gt;CTSD&lt;/em&gt; showed that the relative content of &lt;em&gt;casp3a&lt;/em&gt; gene increased significantly at 24 h and reached the maximum value of the culture time (&lt;em&gt;P&lt;/em&gt; &lt; 0.05). The content of &lt;em&gt;CTSD&lt;/em&gt; gene increased significantly at 12 h and reached the maximum value (&lt;em&gt;P&lt;/em&gt; &lt; 0.05). The results of antioxidant immune genes &lt;em&gt;serpinh1a&lt;/em&gt; and &lt;em&gt;gpx1a&lt;/em&gt; were opposite to the structure of apoptotic genes. The content of &lt;em&gt;serpinh1a&lt;/em&gt; and &lt;em&gt;gpx1a&lt;/em&gt; genes decreased significantly at 12 h (&lt;em&gt;P&lt;/em&gt; &lt; 0.05), but with the prolongation of culture time, the content increased significantly at 24 h and 48 h (&lt;em&gt;P&lt;/em&gt; &lt; 0.05). After stimulation of EPC cells by &lt;em&gt;S&lt;/em&gt;. &lt;em&gt;parasitica&lt;/em&gt;, the differential metabolites were mainly concentrated in Lipids, Compounds with biological roles and Phytochemical compounds. The KEGG pathway mainly focused on ABC transporters, Glycerophospholipid metabolism, Cysteine and methionine metabolism, Glycine, serine and threonine metabolism, Purine metabolism. In general, &lt;em&gt;S&lt;/em&gt;. &lt;em&gt;parasitica&lt;/em&gt; can affect cell activity, destroy the cell membrane of EPC cells, and cause apoptosis. However, EPC cells can also resist the invasion of &lt;em&gt;S&lt;/em&gt;. &lt;em&gt;parasitica&lt;/em&gt; by regulating their own non-specific immunity and their own metabolites, thereby protecting the body from the infectio","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"162 ","pages":"Article 105311"},"PeriodicalIF":2.7,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142902712","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Characterization and functional analysis of interleukin-6 and its receptor subunits (IL-6Rα and IL-6Rβ) in the yellow drum, Nibea alibiflora 白介素-6及其受体亚基(IL-6Rα和IL-6Rβ)的鉴定与功能分析。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105305
Jiaxin Liu , Mingming Wenren , Xu Zhou , Dongdong Xu , Changfeng Chi , Zhenming Lü , Huihui Liu
Interleukin 6 (IL-6) is one of the cytokines found to be multifunctional and biologically effective, regulating immune and inflammatory response by interacting with receptors to transmit signals. In this study, the full-length cDNAs of IL-6 (named as NaIL-6) and its receptors IL-6R and gp130 (named as NaIL-6Rα and NaIL-6Rβ) of Nibea albiflora were acquired and they possessed the typical symbolic motifs similar to its teleost orthologues in multiple sequence comparisons. The phylogenetic trees showed that NaIL-6 and its receptors clustered with their counterparts in bony fish, and had the closest affinity to Larimichthys crocea. Real-time PCR indicated that NaIL-6, NaIL-6Rα and NaIL-6Rβ were widely expressed in different tissues, among which NaIL-6 was highly expressed in the liver, NaIL-6Rα showed the highest expression in the kidney and NaIL-6Rβ was reflected in the liver. Following stimulation by Vibrio parahaemolyticus, Vibrio alginolyticus, or Polyinosinic-polycytidylic acid (Poly (I:C)) infection, the mRNA expression of all three genes were greatly up-regulated over time. The cell localization analysis showed that NaIL-6 distributed in cytoplasm and cell membrane, and NaIL-6Rα located on the cell membrane. After co-transfection of NaIL-6- mCherry and NaIL-6Rα-EGFP, they co-expressed as orange at the same position for their possibility of spatial interactions on the cellular membrane. By GST-Pull down with the purified target proteins, the association between NaIL-6-His and NaIL-6Rα-GST was confirmed, which may be utilized for further functional study. Taken together, the results indicated that the biological functions of NaIL-6 and its receptors NaIL-6Rα and NaIL-6Rβ involved in the immunologic mechanism in N.albiflora, which would provide a basis for the research of immune functions and signaling pathway of IL-6 and its receptors in fish.
白细胞介素6 (Interleukin 6, IL-6)是一种被发现具有多功能性和生物有效性的细胞因子,通过与受体相互作用传递信号来调节免疫和炎症反应。本研究获得了Nibea albiflora的IL-6(命名为nai -6)及其受体IL-6R和gp130(命名为nai - 6r α和nai - 6r β)的全长cdna,经多序列比较,它们具有与其骨鱼同源物相似的典型符号基序。系统进化树显示,在硬骨鱼中,NaIL-6及其受体与相应的受体聚集在一起,与Larimichthys crocea亲缘关系最密切。Real-time PCR结果显示,nai -6、nai - 6r α和nai - 6r β在不同组织中广泛表达,其中nai -6在肝脏中高表达,nai - 6r α在肾脏中表达最高,nai - 6r β在肝脏中也有表达。在副溶血性弧菌、溶藻弧菌或聚肌酸-多胞酸(Poly (I:C))感染刺激后,这三个基因的mRNA表达随着时间的推移而大幅上调。细胞定位分析显示,nai -6分布在细胞质和细胞膜上,而nai - 6r α位于细胞膜上。在共转染nai -6- mCherry和nai - 6r α- egfp后,由于它们在细胞膜上的空间相互作用的可能性,它们在同一位置共表达为橙色。通过纯化的靶蛋白进行GST-Pull down,证实了nai -6- his与nai - 6r α- gst之间的关联,可用于进一步的功能研究。综上所述,上述结果提示IL-6及其受体il - 6r α和il - 6r β的生物学功能参与了白颡鱼的免疫机制,为研究IL-6及其受体在鱼类体内的免疫功能和信号通路提供依据。
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引用次数: 0
Corrigendum to “Transcriptomics reveals crowding stress inhibit the immune defense of the head kidney of the pearl gentian grouper juvenile through NF-κB signal pathway” (162), January 2025, 105299 “转录组学揭示拥挤胁迫通过NF-κB信号通路抑制珍珠龙胆石斑鱼幼鱼头肾的免疫防御”(162),2025年1月,105299。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2025-01-01 DOI: 10.1016/j.dci.2024.105309
Wenyang Li , Ao Li , Xianhong Zhang , Fan Fei , Xiaoqiang Gao , Yingying Fang , Shuquan Cao , Hongjun Yang , Wensheng Li , Baoliang Liu
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引用次数: 0
Transcriptome-wide identification and characterization of Toll pathway genes in Riptortus pedestris (Hemiptera: Alydidae) 步蛇(半翅目:叶蝉科)Toll通路基因的全转录组鉴定与表征
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-11-28 DOI: 10.1016/j.dci.2024.105294
Yipeng Ren , Wenhao Dong , Juhong Chen , Wenjun Bu , Huaijun Xue
The Toll pathway was first identified in Drosophila and plays an essential role in defense against infection by various pathogens. To date, various noncoding RNAs (ncRNAs) have been demonstrated to maintain immune homeostasis by regulating several target genes in the insect Toll pathway. However, the characterization and function of Toll pathway genes involved in the response to environmental changes at the posttranscriptional level associated with gut bacterial changes in Riptortus pedestris, which is a significant pest of soybeans, remain unclear. In this study, we identified and classified six Toll genes into three subtypes with typical Toll domain arrangements, including a Toll/interleukin receptor (TIR) domain, a transmembrane domain, and multiple leucine-rich repeat (LRR) domains; in addition, only one positive selection site was found in hemipteran sPP-Tolls, and a total of five downstream members in the Toll signaling pathway were selected and characterized. The expression patterns revealed that all these genes were widely expressed at all developmental stages of R. pedestris, and they presented variable expression levels among the different feeding treatments in the R. pedestris gut. Our comprehensive prediction analysis revealed that there are sixty miRNA‒mRNA interaction pairs, including fifty-six miRNA and six Toll pathway genes (P‒Toll1, sP‒Toll, Myd88, Pelle, Tube, and Cactus), and a ceRNA network comprising two lncRNA‒miRNA‒Toll pairs was constructed in response to environmental changes. Finally, the expression of some above genes and ncRNAs from the ceRNA network exhibited positive or negative association with the most changes in gut bacterial genera via Pearson correlation analysis. These findings provide valuable insights into how the Toll pathway of R. pedestris is involved in environmental adaptation at the posttranscriptional level and identifies new avenues for developing more effective methods for pest control through integration with gut bacteria.
Toll通路最早是在果蝇中发现的,在防御各种病原体感染方面起着重要作用。迄今为止,各种非编码rna (ncRNAs)已被证明通过调节昆虫Toll通路中的几个靶基因来维持免疫稳态。然而,在大豆的重要害虫Riptortus pestris中,Toll通路基因的特征和功能仍不清楚,这些基因参与了与肠道细菌变化相关的转录后水平的环境变化反应。在这项研究中,我们鉴定并将6个Toll基因分为3个亚型,具有典型的Toll结构域排列,包括Toll/白细胞介素受体(TIR)结构域、跨膜结构域和多个富亮氨酸重复结构域;此外,在半翼体spp -Toll中只发现了一个阳性选择位点,总共选择并表征了Toll信号通路的五个下游成员。结果表明,上述基因均广泛表达于小檗发育的各个阶段,且在不同摄食处理下在小檗肠道内的表达水平存在差异。我们的综合预测分析显示,miRNA - mrna有60对相互作用对,包括56对miRNA和6个Toll通路基因(p - tol1、sP-Toll、Myd88、Pelle、Tube和Cactus),并构建了一个由两个lncRNA-miRNA-Toll对组成的ceRNA网络,以响应环境变化。最后,通过Pearson相关分析,上述部分基因和来自ceRNA网络的ncrna的表达与肠道细菌属的大多数变化呈正相关或负相关。这些发现为研究r.t histris Toll通路如何参与转录后水平的环境适应提供了有价值的见解,并为开发通过与肠道细菌整合更有效的害虫控制方法确定了新的途径。
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引用次数: 0
CgANT2 regulates mitophagy of oyster haemocyte response against bacterial stimulation CgANT2调节牡蛎血细胞对细菌刺激的自噬反应。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-11-28 DOI: 10.1016/j.dci.2024.105295
Wei Wu , Xiaoqian Lv , Jiejie Sun , Zihan Wang , Miren Dong , Lingling Wang , Linsheng Song
Adenine nucleotide translocator (ANT) is a major molecule in the inner membrane of mitochondria that plays an important role in regulating mitophagy. In the present study, a conserved ANT2 homologue (designated as CgANT2) was identified and functionally characterized in oyster Crassostrea gigas. There were three typical Mito_carr tandem repeats in CgANT2. The mRNA expression levels of CgANT2 in haemocytes increased significantly at 24 and 72 h after Vibrio splendidus stimulation. Its protein was abundantly expressed in granulocytes and was observed to be colocalized with mitochondria. When CgANT2 expression was suppressed by injection with its dsRNA, there was an increased mitochondrial reactive oxygen species (mtROS) production and mitochondrial permeability transition pore (mPTP) opening, while the mRNA expression levels of mitophagy-related genes (CgPINK1 and CgParkin) and the percentage of mitophagy in haemocytes all decreased significantly. These results indicated that CgANT2 regulated mtROS production and mPTP opening, thereby inducing mitophagy in the oyster haemocyte response against V. splendidus stimulation.
腺嘌呤核苷酸转运子(Adenine nucleotide translocator, ANT)是线粒体内膜上的一种重要分子,在线粒体自噬过程中起着重要的调节作用。在本研究中,我们在牡蛎长牡蛎中发现了一个保守的ANT2同源物(命名为CgANT2),并对其进行了功能表征。在CgANT2中存在3个典型的Mito_carr串联重复序列。脾弧菌刺激后24和72 h,细胞中CgANT2 mRNA表达水平显著升高。其蛋白在粒细胞中大量表达,并与线粒体共定位。注射CgANT2 dsRNA抑制其表达后,线粒体活性氧(mtROS)产生增加,线粒体通透性过渡孔(mPTP)开放,线粒体自噬相关基因(CgPINK1和CgParkin) mRNA表达水平和血细胞自噬百分比均显著降低。这些结果表明,CgANT2调节mtROS的产生和mPTP的开放,从而在牡蛎血细胞对脾弧菌刺激的反应中诱导线粒体自噬。
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引用次数: 0
Parkin is a critical factor in grouper immune response to virus infection 帕金蛋白是石斑鱼对病毒感染免疫反应的关键因素
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-11-26 DOI: 10.1016/j.dci.2024.105293
Xiaoxia Lei , Siting Wu , Zhuqing Xu , Qiongyue Xu , Helong Cao , Zhouling Zhan , Qiwei Qin , Jingguang Wei
Parkin is an E3 ubiquitinated ligase that mainly participates in mitophagy and plays an essential biological role in organisms. To investigate Parkin's function in fish, a Parkin homolog was cloned from Epinephelus coioides (EcParkin). The open reading frame (ORF) of EcParkin consists of 1461 nucleotides and encodes a protein of 486 amino acids, with a predicted molecular weight of 53.32 kDa. EcParkin was highly expressed in the heart, kidney, and head kidney of healthy groupers, especially in the heart. The expression levels of EcParkin were upregulated after Singapore grouper iridovirus (SGIV) and red-spotted grouper nervous necrosis virus (RGNNV) infection. Intracellular localization studies revealed that EcParkin is distributed in both the cytoplasm and nucleus of GS cells. Overexpression of EcParkin promoted SGIV and RGNNV replication in vitro, while knockdown of EcParkin inhibited SGIV and RGNNV replication. EcParkin suppressed the promoter activities of IFN-β, ISRE, and NF-κB, as well as the expression of interferon-related factors and inflammatory cytokines. EcParkin was found to colocalize and interact with EcMDA5, EcMAVS, EcTBK1, EcIRF3, and EcIRF7. Additionally, EcParkin enhanced LC3-II production in GS cells. These findings suggest that EcParkin may play a crucial role in the antiviral innate immunity and cellular autophagy of fish.
Parkin是一种E3泛素化连接酶,主要参与有丝分裂,在生物体中具有重要的生物学作用。为了研究Parkin在鱼类中的功能,从Epinephelus coioides (EcParkin)中克隆了一个Parkin同源基因。EcParkin的开放阅读框(ORF)由1461个核苷酸组成,编码486个氨基酸的蛋白,预测分子量为53.32 kDa。EcParkin在健康石斑鱼的心脏、肾脏和头肾中高度表达,尤其是在心脏中。新加坡石斑鱼虹膜病毒(SGIV)和红斑石斑鱼神经坏死病毒(RGNNV)感染后,EcParkin的表达水平上调。细胞内定位研究表明,EcParkin分布在GS细胞的细胞质和细胞核中。EcParkin的过表达促进了SGIV和RGNNV的体外复制,而EcParkin的低表达抑制了SGIV和RGNNV的体外复制。EcParkin抑制IFN-β、ISRE、NF-κB启动子活性,抑制干扰素相关因子和炎症因子的表达。EcParkin被发现与EcMDA5、EcMAVS、EcTBK1、EcIRF3和EcIRF7共定位并相互作用。此外,EcParkin还能增强GS细胞中LC3-II的生成。这些发现提示EcParkin可能在鱼类的抗病毒先天免疫和细胞自噬中起重要作用。
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引用次数: 0
Analysis of the potential long-lasting effects of probiotic Debaryomyces hansenii CBS 8339 on trained immunity in newborn goats 分析益生菌 Debaryomyces hansenii CBS 8339 对新生山羊训练有素的免疫力的潜在长期影响。
IF 2.7 3区 农林科学 Q1 FISHERIES Pub Date : 2024-11-20 DOI: 10.1016/j.dci.2024.105292
Miriam Angulo , Carlos Angulo
Trained immunity has been described as the memory capacity of the innate immune system. Several microbial components have been shown to induce trained immunity. Research on the potential of probiotics to trigger these effects has been limited to a few in vitro studies but remains completely unknown in vivo. Components from the probiotic Debaryomyces hansenii CBS 8339 (Dh) have been shown to induce innate immune memory in goat kids and calves. In the present study, stimulating innate immune cells from newborn goats with probiotic Dh increased respiratory burst activity and nitric oxide production, while cell phagocytosis was unaffected. Glucose uptake was enhanced in goat's cells stimulated with Dh, but lactate production was decreased. In newborn goats, after the training scheme (via oral probiotic administration), cell phagocytosis, nitric oxide production and glycolysis — through the upregulation of AKT and HIF1A gene expression, glucose consumption and lactate production— were enhanced. The expression of IL1B gene was similar between the D. hansenii and control groups. Moreover, the potential long-lasting effects were assessed 30 days after initiation of the training scheme. Cell phagocytosis, respiratory burst and myeloperoxidase activity were enhanced, while glycolytic parameters remained unaffected. Altogether, the results of the present study suggest that the immune training scheme may induce trained immunity by the probiotic D. hansenii in newborn goats. However, our findings were not conclusive regarding the long-lasting (one-month) effects of trained immunity by probiotics.
训练免疫被描述为先天免疫系统的记忆能力。一些微生物成分已被证明能诱导训练有素的免疫力。关于益生菌触发这些效应的潜力的研究仅限于一些体外研究,但对体内的研究仍完全未知。益生菌德巴氏酵母菌(Debaryomyces hansenii)CBS 8339(Dh)中的成分已被证明能诱导山羊幼崽和小牛的先天免疫记忆。在本研究中,用益生菌 Dh 刺激新生山羊的先天性免疫细胞可增加呼吸爆发活动和一氧化氮的产生,而细胞吞噬功能不受影响。受 Dh 刺激的山羊细胞对葡萄糖的吸收增强,但乳酸盐的产生减少。通过口服益生菌)训练计划后,新生山羊的细胞吞噬、一氧化氮产生和糖酵解(通过上调 AKT 和 HIF1A 基因表达、葡萄糖消耗和乳酸盐产生)均得到增强。D. hansenii组和对照组的IL1B基因表达相似。此外,在训练计划开始 30 天后,还对潜在的持久效果进行了评估。细胞吞噬功能、呼吸爆发和髓过氧化物酶活性得到了增强,而糖酵解参数未受影响。总之,本研究的结果表明,免疫训练计划可通过益生菌 D. hansenii 诱导新生山羊产生训练有素的免疫力。然而,我们的研究结果并不能确定益生菌训练免疫的效果是否持久(一个月)。
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引用次数: 0
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Developmental and comparative immunology
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