Developmental and comparative immunology最新文献

{"title":"Expression of FcμR by bovine mononuclear blood leukocytes","authors":"Florence B. Gilbert,&nbsp;Rodrigo P. Martins,&nbsp;Pascal Rainard","doi":"10.1016/j.dci.2024.105304","DOIUrl":"10.1016/j.dci.2024.105304","url":null,"abstract":"<div><div>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 (CD172a^pos CD16^pos) 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 CD3^pos αβT lymphocytes (both CD4^pos and CD8^pos) 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 CD3^pos 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.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"162 ","pages":"Article 105304"},"PeriodicalIF":2.7,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142823989","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Haemolymphatic tissues of captive boa constrictor (Boa constrictor): Morphological features in healthy individuals and with boid inclusion body disease","authors":"E. Dervas ,&nbsp;E. Michalopoulou ,&nbsp;J. Hepojoki ,&nbsp;T. Thiele ,&nbsp;F. Baggio ,&nbsp;U. Hetzel ,&nbsp;A. Kipar","doi":"10.1016/j.dci.2024.105302","DOIUrl":"10.1016/j.dci.2024.105302","url":null,"abstract":"<div><div>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 (<em>Boa constrictor</em>). 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.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"162 ","pages":"Article 105302"},"PeriodicalIF":2.7,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142853501","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"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}

{"title":"Characterization and functional analysis of interleukin-6 and its receptor subunits (IL-6Rα and IL-6Rβ) in the yellow drum, Nibea alibiflora","authors":"Jiaxin Liu ,&nbsp;Mingming Wenren ,&nbsp;Xu Zhou ,&nbsp;Dongdong Xu ,&nbsp;Changfeng Chi ,&nbsp;Zhenming Lü ,&nbsp;Huihui Liu","doi":"10.1016/j.dci.2024.105305","DOIUrl":"10.1016/j.dci.2024.105305","url":null,"abstract":"<div><div>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 <em>NaIL-6</em>) and its receptors IL-6R and gp130 (named as <em>NaIL-6Rα</em> and <em>NaIL-6Rβ</em>) of <em>Nibea albiflora</em> were acquired and they possessed the typical symbolic motifs similar to its teleost orthologues in multiple sequence comparisons. The phylogenetic trees showed that <em>NaIL-6</em> and its receptors clustered with their counterparts in bony fish, and had the closest affinity to <em>Larimichthys crocea</em>. Real-time PCR indicated that <em>NaIL-6</em>, <em>NaIL-6Rα</em> and <em>NaIL-6Rβ</em> were widely expressed in different tissues, among which <em>NaIL-6</em> was highly expressed in the liver, <em>NaIL-6Rα</em> showed the highest expression in the kidney and <em>NaIL-6Rβ</em> was reflected in the liver. Following stimulation by <em>Vibrio parahaemolyticus</em>, <em>Vibrio alginolyticus</em>, 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 <em>NaIL-6</em> and its receptors <em>NaIL-6Rα</em> and <em>NaIL-6Rβ</em> involved in the immunologic mechanism in <em>N.albiflora</em>, which would provide a basis for the research of immune functions and signaling pathway of IL-6 and its receptors in fish.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"162 ","pages":"Article 105305"},"PeriodicalIF":2.7,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142881529","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}

{"title":"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","authors":"Wenyang Li ,&nbsp;Ao Li ,&nbsp;Xianhong Zhang ,&nbsp;Fan Fei ,&nbsp;Xiaoqiang Gao ,&nbsp;Yingying Fang ,&nbsp;Shuquan Cao ,&nbsp;Hongjun Yang ,&nbsp;Wensheng Li ,&nbsp;Baoliang Liu","doi":"10.1016/j.dci.2024.105309","DOIUrl":"10.1016/j.dci.2024.105309","url":null,"abstract":"","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"162 ","pages":"Article 105309"},"PeriodicalIF":2.7,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142902709","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}

{"title":"Transcriptome-wide identification and characterization of Toll pathway genes in Riptortus pedestris (Hemiptera: Alydidae)","authors":"Yipeng Ren ,&nbsp;Wenhao Dong ,&nbsp;Juhong Chen ,&nbsp;Wenjun Bu ,&nbsp;Huaijun Xue","doi":"10.1016/j.dci.2024.105294","DOIUrl":"10.1016/j.dci.2024.105294","url":null,"abstract":"<div><div>The Toll pathway was first identified in <em>Drosophila</em> 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 <em>Riptortus pedestris</em>, which is a significant pest of soybeans, remain unclear. In this study, we identified and classified six <em>Toll</em> 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 <em>R. pedestris</em>, and they presented variable expression levels among the different feeding treatments in the <em>R. pedestris</em> gut. Our comprehensive prediction analysis revealed that there are sixty miRNA‒mRNA interaction pairs, including fifty-six miRNA and six Toll pathway genes (<em>P‒Toll1</em>, <em>sP‒Toll</em>, <em>Myd88</em>, <em>Pelle</em>, <em>Tube</em>, and <em>Cactus</em>), 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 <em>R. pedestris</em> 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.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"162 ","pages":"Article 105294"},"PeriodicalIF":2.7,"publicationDate":"2024-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142759412","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}

{"title":"CgANT2 regulates mitophagy of oyster haemocyte response against bacterial stimulation","authors":"Wei Wu ,&nbsp;Xiaoqian Lv ,&nbsp;Jiejie Sun ,&nbsp;Zihan Wang ,&nbsp;Miren Dong ,&nbsp;Lingling Wang ,&nbsp;Linsheng Song","doi":"10.1016/j.dci.2024.105295","DOIUrl":"10.1016/j.dci.2024.105295","url":null,"abstract":"<div><div>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 <em>Cg</em>ANT2) was identified and functionally characterized in oyster <em>Crassostrea gigas</em>. There were three typical Mito_carr tandem repeats in <em>Cg</em>ANT2. The mRNA expression levels of <em>Cg</em>ANT2 in haemocytes increased significantly at 24 and 72 h after <em>Vibrio splendidus</em> stimulation. Its protein was abundantly expressed in granulocytes and was observed to be colocalized with mitochondria. When <em>Cg</em>ANT2 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 (<em>Cg</em>PINK1 and <em>Cg</em>Parkin) and the percentage of mitophagy in haemocytes all decreased significantly. These results indicated that <em>Cg</em>ANT2 regulated mtROS production and mPTP opening, thereby inducing mitophagy in the oyster haemocyte response against <em>V. splendidus</em> stimulation.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"162 ","pages":"Article 105295"},"PeriodicalIF":2.7,"publicationDate":"2024-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142754770","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}

{"title":"Parkin is a critical factor in grouper immune response to virus infection","authors":"Xiaoxia Lei ,&nbsp;Siting Wu ,&nbsp;Zhuqing Xu ,&nbsp;Qiongyue Xu ,&nbsp;Helong Cao ,&nbsp;Zhouling Zhan ,&nbsp;Qiwei Qin ,&nbsp;Jingguang Wei","doi":"10.1016/j.dci.2024.105293","DOIUrl":"10.1016/j.dci.2024.105293","url":null,"abstract":"<div><div>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 <em>Epinephelus coioides</em> (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 <em>in vitro</em>, 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.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"162 ","pages":"Article 105293"},"PeriodicalIF":2.7,"publicationDate":"2024-11-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142746448","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}

{"title":"Analysis of the potential long-lasting effects of probiotic Debaryomyces hansenii CBS 8339 on trained immunity in newborn goats","authors":"Miriam Angulo ,&nbsp;Carlos Angulo","doi":"10.1016/j.dci.2024.105292","DOIUrl":"10.1016/j.dci.2024.105292","url":null,"abstract":"<div><div>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 <em>in vitro</em> studies but remains completely unknown <em>in vivo</em>. Components from the probiotic <em>Debaryomyces hansenii</em> 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 <em>AKT</em> and <em>HIF1A</em> gene expression, glucose consumption and lactate production— were enhanced. The expression of <em>IL1B</em> gene was similar between the <em>D. hansenii</em> 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 <em>D. hansenii</em> in newborn goats. However, our findings were not conclusive regarding the long-lasting (one-month) effects of trained immunity by probiotics.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"162 ","pages":"Article 105292"},"PeriodicalIF":2.7,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142692558","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}

{"title":"Establishment of a novel clonal GFP-expressing transgenic ginbuna crucian carp","authors":"Ren Uehara ,&nbsp;Shinji Takeda ,&nbsp;Daichi Oku ,&nbsp;Ryo Sasaki ,&nbsp;Masaru Murakami ,&nbsp;Hajime Shiba ,&nbsp;Fumihiko Katakura ,&nbsp;Tadaaki Moritomo","doi":"10.1016/j.dci.2024.105290","DOIUrl":"10.1016/j.dci.2024.105290","url":null,"abstract":"<div><div>The clonal triploid ginbuna crucian carp <em>Carassius auratus langsdorfii</em>, a naturally occurring gynogenetic fish, is suitable for cell transplantation studies to reveal the roles of stem cells and immune cells. To ensure long-term traceability of donor cells within recipient fish, we have established a transgenic ginbuna line that expresses green fluorescent protein (GFP). The <em>Xenopus laevis ef1a</em> promoter was introduced for regulating GFP expression. <em>Tol2</em> transposon-based transgenesis to ginbuna embryos resulted in producing a putative founder fish (F0) in a mosaic fluorescent fashion; the frequency of germline transmission was 14.9%. All embryos of GFP-positive offspring (F1)-derived F2 generation expressed GFP widely across the body. The result of Southern blot analysis showed that the transgene was present on a single DNA fragment of equivalent size among F1 and F2 individuals tested, indicating that the transgene was stably transmitted without translocation. Analysis of the fluorescence intensity of organs obtained from F1 and F2 juveniles using fluorescence microscope showed that eyes, brain, skeletal muscle, heart and gonad exhibited a strong GFP fluorescence while gill, spleen and intestine gave a weak signal; no fluorescence was observed in erythrocytes. Flow cytometric analyses of peripheral leukocytes from F1 and F2 adult fish revealed all cell populations expressed GFP. Scale grafts from the transgenic fish to the wild-type fish exhibited persistent engraftment. Together, our transgenic line can be a powerful tool for studying cellular dynamics by cell transplantation and provide a solid basis for further immunological research advances in teleost.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"162 ","pages":"Article 105290"},"PeriodicalIF":2.7,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142686405","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}