首页 > 最新文献

Biological Research最新文献

英文 中文
Combined transcriptomics and proteomics unveil the impact of vitamin C in modulating specific protein abundance in the mouse liver. 转录组学和蛋白质组学的结合揭示了维生素 C 对调节小鼠肝脏中特定蛋白质丰度的影响。
IF 6.7 2区 生物学 Q1 BIOLOGY Pub Date : 2024-05-12 DOI: 10.1186/s40659-024-00509-x
Lucie Aumailley, Antoine Bodein, Pauline Adjibade, Mickaël Leclercq, Sylvie Bourassa, Arnaud Droit, Rachid Mazroui, Michel Lebel

Background: Vitamin C (ascorbate) is a water-soluble antioxidant and an important cofactor for various biosynthetic and regulatory enzymes. Mice can synthesize vitamin C thanks to the key enzyme gulonolactone oxidase (Gulo) unlike humans. In the current investigation, we used Gulo-/- mice, which cannot synthesize their own ascorbate to determine the impact of this vitamin on both the transcriptomics and proteomics profiles in the whole liver. The study included Gulo-/- mouse groups treated with either sub-optimal or optimal ascorbate concentrations in drinking water. Liver tissues of females and males were collected at the age of four months and divided for transcriptomics and proteomics analysis. Immunoblotting, quantitative RT-PCR, and polysome profiling experiments were also conducted to complement our combined omics studies.

Results: Principal component analyses revealed distinctive differences in the mRNA and protein profiles as a function of sex between all the mouse cohorts. Despite such sexual dimorphism, Spearman analyses of transcriptomics data from females and males revealed correlations of hepatic ascorbate levels with transcripts encoding a wide array of biological processes involved in glucose and lipid metabolisms as well as in the acute-phase immune response. Moreover, integration of the proteomics data showed that ascorbate modulates the abundance of various enzymes involved in lipid, xenobiotic, organic acid, acetyl-CoA, and steroid metabolism mainly at the transcriptional level, especially in females. However, several proteins of the mitochondrial complex III significantly correlated with ascorbate concentrations in both males and females unlike their corresponding transcripts. Finally, poly(ribo)some profiling did not reveal significant enrichment difference for these mitochondrial complex III mRNAs between Gulo-/- mice treated with sub-optimal and optimal ascorbate levels.

Conclusions: Thus, the abundance of several subunits of the mitochondrial complex III are regulated by ascorbate at the post-transcriptional levels. Our extensive omics analyses provide a novel resource of altered gene expression patterns at the transcriptional and post-transcriptional levels under ascorbate deficiency.

背景:维生素 C(抗坏血酸)是一种水溶性抗氧化剂,也是各种生物合成和调节酶的重要辅助因子。与人类不同的是,小鼠可以通过关键酶古洛内酯氧化酶(Gulo)合成维生素 C。在目前的研究中,我们使用了不能自己合成抗坏血酸的 Gulo/- 小鼠,以确定这种维生素对全肝脏转录组学和蛋白质组学特征的影响。研究包括在饮用水中添加次优或最优抗坏血酸浓度的 Gulo-/- 小鼠组。在小鼠四个月大时收集雌性和雄性小鼠的肝脏组织,并进行转录组学和蛋白质组学分析。我们还进行了免疫印迹、定量 RT-PCR 和多聚体分析实验,以补充我们的综合全息研究:结果:主成分分析表明,所有小鼠组群的 mRNA 和蛋白质图谱在性别功能上存在明显差异。尽管存在这种性别二态性,但对雌性和雄性的转录组学数据进行斯皮尔曼分析后发现,肝脏抗坏血酸水平与编码葡萄糖和脂质代谢以及急性期免疫反应所涉及的一系列生物过程的转录本存在相关性。此外,蛋白质组学数据整合显示,抗坏血酸主要在转录水平上调节参与脂质、异生物、有机酸、乙酰-CoA 和类固醇代谢的各种酶的丰度,尤其是在雌性动物中。然而,线粒体复合体 III 的几种蛋白质与抗坏血酸浓度在雄性和雌性中都有显著的相关性,这与其相应的转录物不同。最后,聚核糖谱分析结果显示,在接受次优和最优抗坏血酸水平治疗的 Gulo-/- 小鼠中,这些线粒体复合体 III mRNA 的富集差异并不明显:因此,线粒体复合体 III 的几个亚基的丰度在转录后水平上受抗坏血酸的调控。我们广泛的全局分析为抗坏血酸缺乏时转录和转录后水平基因表达模式的改变提供了新的资源。
{"title":"Combined transcriptomics and proteomics unveil the impact of vitamin C in modulating specific protein abundance in the mouse liver.","authors":"Lucie Aumailley, Antoine Bodein, Pauline Adjibade, Mickaël Leclercq, Sylvie Bourassa, Arnaud Droit, Rachid Mazroui, Michel Lebel","doi":"10.1186/s40659-024-00509-x","DOIUrl":"10.1186/s40659-024-00509-x","url":null,"abstract":"<p><strong>Background: </strong>Vitamin C (ascorbate) is a water-soluble antioxidant and an important cofactor for various biosynthetic and regulatory enzymes. Mice can synthesize vitamin C thanks to the key enzyme gulonolactone oxidase (Gulo) unlike humans. In the current investigation, we used Gulo<sup>-/-</sup> mice, which cannot synthesize their own ascorbate to determine the impact of this vitamin on both the transcriptomics and proteomics profiles in the whole liver. The study included Gulo<sup>-/-</sup> mouse groups treated with either sub-optimal or optimal ascorbate concentrations in drinking water. Liver tissues of females and males were collected at the age of four months and divided for transcriptomics and proteomics analysis. Immunoblotting, quantitative RT-PCR, and polysome profiling experiments were also conducted to complement our combined omics studies.</p><p><strong>Results: </strong>Principal component analyses revealed distinctive differences in the mRNA and protein profiles as a function of sex between all the mouse cohorts. Despite such sexual dimorphism, Spearman analyses of transcriptomics data from females and males revealed correlations of hepatic ascorbate levels with transcripts encoding a wide array of biological processes involved in glucose and lipid metabolisms as well as in the acute-phase immune response. Moreover, integration of the proteomics data showed that ascorbate modulates the abundance of various enzymes involved in lipid, xenobiotic, organic acid, acetyl-CoA, and steroid metabolism mainly at the transcriptional level, especially in females. However, several proteins of the mitochondrial complex III significantly correlated with ascorbate concentrations in both males and females unlike their corresponding transcripts. Finally, poly(ribo)some profiling did not reveal significant enrichment difference for these mitochondrial complex III mRNAs between Gulo<sup>-/-</sup> mice treated with sub-optimal and optimal ascorbate levels.</p><p><strong>Conclusions: </strong>Thus, the abundance of several subunits of the mitochondrial complex III are regulated by ascorbate at the post-transcriptional levels. Our extensive omics analyses provide a novel resource of altered gene expression patterns at the transcriptional and post-transcriptional levels under ascorbate deficiency.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"57 1","pages":"26"},"PeriodicalIF":6.7,"publicationDate":"2024-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11088995/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140911169","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Novel role of LLGL2 silencing in autophagy: reversing epithelial-mesenchymal transition in prostate cancer. LLGL2沉默在自噬中的新作用:逆转前列腺癌的上皮-间质转化。
IF 6.7 2区 生物学 Q1 BIOLOGY Pub Date : 2024-05-08 DOI: 10.1186/s40659-024-00499-w
Geum-Lan Hong, Kyung-Hyun Kim, Yae-Ji Kim, Hui-Ju Lee, Sung-Pil Cho, Seung-Yun Han, Seung Woo Yang, Jong-Soo Lee, Shin-Kwang Kang, Jae-Sung Lim, Ju-Young Jung

Purpose: Prostate cancer (PCa) is a major urological disease that is associated with significant morbidity and mortality in men. LLGL2 is the mammalian homolog of Lgl. It acts as a tumor suppressor in breast and hepatic cancer. However, the role of LLGL2 and the underlying mechanisms in PCa have not yet been elucidated. Here, we investigate the role of LLGL2 in the regulation of epithelial-mesenchymal transition (EMT) in PCa through autophagy in vitro and in vivo.

Methods: PC3 cells were transfected with siLLGL2 or plasmid LLGL2 and autophagy was examined. Invasion, migration, and wound healing were assessed in PC3 cells under autophagy regulation. Tumor growth was evaluated using a shLLGL2 xenograft mouse model.

Results: In patients with PCa, LLGL2 levels were higher with defective autophagy and increased EMT. Our results showed that the knockdown of LLGL2 induced autophagy flux by upregulating Vps34 and ATG14L. LLGL2 knockdown inhibits EMT by upregulating E-cadherin and downregulating fibronectin and α-SMA. The pharmacological activation of autophagy by rapamycin suppressed EMT, and these effects were reversed by 3-methyladenine treatment. Interestingly, in a shLLGL2 xenograft mouse model, tumor size and EMT were decreased, which were improved by autophagy induction and worsened by autophagy inhibition.

Conclusion: Defective expression of LLGL2 leads to attenuation of EMT due to the upregulation of autophagy flux in PCa. Our results suggest that LLGL2 is a novel target for alleviating PCa via the regulation of autophagy.

目的:前列腺癌(PCa)是一种主要的泌尿系统疾病,与男性严重的发病率和死亡率有关。LLGL2 是哺乳动物 Lgl 的同源物。它是乳腺癌和肝癌的肿瘤抑制因子。然而,LLGL2在PCa中的作用及其内在机制尚未阐明。在此,我们研究了LLGL2在PCa中通过体外和体内自噬调节上皮-间质转化(EMT)的作用:方法:用 siLLGL2 或质粒 LLGL2 转染 PC3 细胞并检测自噬。在自噬调控下,对 PC3 细胞的侵袭、迁移和伤口愈合进行了评估。使用 shLLGL2 异种移植小鼠模型评估肿瘤生长情况:结果:在PCa患者中,LLGL2水平较高,自噬缺陷和EMT增加。我们的研究结果表明,敲除LLGL2可通过上调Vps34和ATG14L诱导自噬通量。LLGL2敲除可通过上调E-cadherin、下调纤连蛋白和α-SMA来抑制EMT。雷帕霉素对自噬的药理激活抑制了EMT,而3-甲基腺嘌呤处理则逆转了这些效应。有趣的是,在shLLGL2异种移植小鼠模型中,肿瘤大小和EMT均有所减小,自噬诱导可改善肿瘤大小和EMT,而自噬抑制则会恶化肿瘤大小和EMT:结论:LLGL2表达缺陷会导致自噬通量上调,从而减轻PCa的EMT。我们的研究结果表明,LLGL2是通过调节自噬缓解PCa的一个新靶点。
{"title":"Novel role of LLGL2 silencing in autophagy: reversing epithelial-mesenchymal transition in prostate cancer.","authors":"Geum-Lan Hong, Kyung-Hyun Kim, Yae-Ji Kim, Hui-Ju Lee, Sung-Pil Cho, Seung-Yun Han, Seung Woo Yang, Jong-Soo Lee, Shin-Kwang Kang, Jae-Sung Lim, Ju-Young Jung","doi":"10.1186/s40659-024-00499-w","DOIUrl":"10.1186/s40659-024-00499-w","url":null,"abstract":"<p><strong>Purpose: </strong>Prostate cancer (PCa) is a major urological disease that is associated with significant morbidity and mortality in men. LLGL2 is the mammalian homolog of Lgl. It acts as a tumor suppressor in breast and hepatic cancer. However, the role of LLGL2 and the underlying mechanisms in PCa have not yet been elucidated. Here, we investigate the role of LLGL2 in the regulation of epithelial-mesenchymal transition (EMT) in PCa through autophagy in vitro and in vivo.</p><p><strong>Methods: </strong>PC3 cells were transfected with siLLGL2 or plasmid LLGL2 and autophagy was examined. Invasion, migration, and wound healing were assessed in PC3 cells under autophagy regulation. Tumor growth was evaluated using a shLLGL2 xenograft mouse model.</p><p><strong>Results: </strong>In patients with PCa, LLGL2 levels were higher with defective autophagy and increased EMT. Our results showed that the knockdown of LLGL2 induced autophagy flux by upregulating Vps34 and ATG14L. LLGL2 knockdown inhibits EMT by upregulating E-cadherin and downregulating fibronectin and α-SMA. The pharmacological activation of autophagy by rapamycin suppressed EMT, and these effects were reversed by 3-methyladenine treatment. Interestingly, in a shLLGL2 xenograft mouse model, tumor size and EMT were decreased, which were improved by autophagy induction and worsened by autophagy inhibition.</p><p><strong>Conclusion: </strong>Defective expression of LLGL2 leads to attenuation of EMT due to the upregulation of autophagy flux in PCa. Our results suggest that LLGL2 is a novel target for alleviating PCa via the regulation of autophagy.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"57 1","pages":"25"},"PeriodicalIF":6.7,"publicationDate":"2024-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11077766/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140891243","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
High-fat diet, microbiome-gut-brain axis signaling, and anxiety-like behavior in male rats. 雄性大鼠的高脂饮食、微生物组-肠-脑轴信号传导和焦虑样行为。
IF 6.7 2区 生物学 Q1 BIOLOGY Pub Date : 2024-05-06 DOI: 10.1186/s40659-024-00505-1
Sylvana I S Rendeiro de Noronha, Lauro Angelo Gonçalves de Moraes, James E Hassell, Christopher E Stamper, Mathew R Arnold, Jared D Heinze, Christine L Foxx, Margaret M Lieb, Kristin E Cler, Bree L Karns, Sophia Jaekel, Kelsey M Loupy, Fernanda C S Silva, Deoclécio Alves Chianca-Jr, Christopher A Lowry, Rodrigo Cunha de Menezes

Obesity, associated with the intake of a high-fat diet (HFD), and anxiety are common among those living in modern urban societies. Recent studies suggest a role of microbiome-gut-brain axis signaling, including a role for brain serotonergic systems in the relationship between HFD and anxiety. Evidence suggests the gut microbiome and the serotonergic brain system together may play an important role in this response. Here we conducted a nine-week HFD protocol in male rats, followed by an analysis of the gut microbiome diversity and community composition, brainstem serotonergic gene expression (tph2, htr1a, and slc6a4), and anxiety-related defensive behavioral responses. We show that HFD intake decreased alpha diversity and altered the community composition of the gut microbiome in association with obesity, increased brainstem tph2, htr1a and slc6a4 mRNA expression, including in the caudal part of the dorsomedial dorsal raphe nucleus (cDRD), a subregion previously associated with stress- and anxiety-related behavioral responses, and, finally, increased anxiety-related defensive behavioral responses. The HFD increased the Firmicutes/Bacteroidetes ratio relative to control diet, as well as higher relative abundances of Blautia, and decreases in Prevotella. We found that tph2, htr1a and slc6a4 mRNA expression were increased in subregions of the dorsal raphe nucleus in the HFD, relative to control diet. Specific bacterial taxa were associated with increased serotonergic gene expression in the cDRD. Thus, we propose that HFD-induced obesity is associated with altered microbiome-gut-serotonergic brain axis signaling, leading to increased anxiety-related defensive behavioral responses in rats.

肥胖(与摄入高脂肪饮食(HFD)有关)和焦虑是生活在现代城市社会中的人的共同特征。最近的研究表明,微生物组-肠道-大脑轴信号传导,包括大脑血清素能系统在高脂饮食与焦虑之间的关系中发挥作用。有证据表明,肠道微生物组和大脑血清素能系统可能在这一反应中共同发挥重要作用。在此,我们对雄性大鼠进行了为期九周的高氟日粮方案,随后分析了肠道微生物组的多样性和群落组成、脑干血清素能基因表达(ph2、htr1a 和 slc6a4)以及与焦虑相关的防御性行为反应。我们的研究表明,摄入高氟日粮会降低肠道微生物组的α多样性并改变其群落组成,从而导致肥胖;增加脑干tph2、htr1a和slc6a4 mRNA的表达,包括在背内侧背侧剑突核(ctRD)尾部的表达,该亚区以前与压力和焦虑相关的行为反应有关;最后,增加焦虑相关的防御性行为反应。与对照组饮食相比,HFD 增加了固着菌/类杆菌的比例,并提高了 Blautia 的相对丰度,降低了 Prevotella 的丰度。我们发现,与对照组饮食相比,HFD 食物中背侧剑突核亚区域的 tph2、htr1a 和 slc6a4 mRNA 表达增加。特定细菌类群与 cDRD 中血清素能基因表达的增加有关。因此,我们认为高密度脂蛋白胆固醇诱导的肥胖与微生物组-肠道-血清素能脑轴信号的改变有关,从而导致大鼠焦虑相关防御性行为反应的增加。
{"title":"High-fat diet, microbiome-gut-brain axis signaling, and anxiety-like behavior in male rats.","authors":"Sylvana I S Rendeiro de Noronha, Lauro Angelo Gonçalves de Moraes, James E Hassell, Christopher E Stamper, Mathew R Arnold, Jared D Heinze, Christine L Foxx, Margaret M Lieb, Kristin E Cler, Bree L Karns, Sophia Jaekel, Kelsey M Loupy, Fernanda C S Silva, Deoclécio Alves Chianca-Jr, Christopher A Lowry, Rodrigo Cunha de Menezes","doi":"10.1186/s40659-024-00505-1","DOIUrl":"10.1186/s40659-024-00505-1","url":null,"abstract":"<p><p>Obesity, associated with the intake of a high-fat diet (HFD), and anxiety are common among those living in modern urban societies. Recent studies suggest a role of microbiome-gut-brain axis signaling, including a role for brain serotonergic systems in the relationship between HFD and anxiety. Evidence suggests the gut microbiome and the serotonergic brain system together may play an important role in this response. Here we conducted a nine-week HFD protocol in male rats, followed by an analysis of the gut microbiome diversity and community composition, brainstem serotonergic gene expression (tph2, htr1a, and slc6a4), and anxiety-related defensive behavioral responses. We show that HFD intake decreased alpha diversity and altered the community composition of the gut microbiome in association with obesity, increased brainstem tph2, htr1a and slc6a4 mRNA expression, including in the caudal part of the dorsomedial dorsal raphe nucleus (cDRD), a subregion previously associated with stress- and anxiety-related behavioral responses, and, finally, increased anxiety-related defensive behavioral responses. The HFD increased the Firmicutes/Bacteroidetes ratio relative to control diet, as well as higher relative abundances of Blautia, and decreases in Prevotella. We found that tph2, htr1a and slc6a4 mRNA expression were increased in subregions of the dorsal raphe nucleus in the HFD, relative to control diet. Specific bacterial taxa were associated with increased serotonergic gene expression in the cDRD. Thus, we propose that HFD-induced obesity is associated with altered microbiome-gut-serotonergic brain axis signaling, leading to increased anxiety-related defensive behavioral responses in rats.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"57 1","pages":"23"},"PeriodicalIF":6.7,"publicationDate":"2024-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11071217/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140856535","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Rapid development and mass production of SARS-CoV-2 neutralizing chicken egg yolk antibodies with protective efficacy in hamsters. 快速开发和大规模生产对仓鼠具有保护效力的 SARS-CoV-2 中和性鸡卵黄抗体。
IF 6.7 2区 生物学 Q1 BIOLOGY Pub Date : 2024-05-06 DOI: 10.1186/s40659-024-00508-y
Binan Zhao, Haoran Peng, Yanjing Zhang, Jie Zhang, Desheng Kong, Sai Cao, Yan Li, Dan Yang, Chuanwen Sun, Xinyi Pu, Ping Zhao, Yan Xu, Kai Zhao, Liangzhi Xie

Despite the record speed of developing vaccines and therapeutics against the SARS-CoV-2 virus, it is not a given that such success can be secured in future pandemics. In addition, COVID-19 vaccination and application of therapeutics remain low in developing countries. Rapid and low cost mass production of antiviral IgY antibodies could be an attractive alternative or complementary option for vaccine and therapeutic development. In this article, we rapidly produced SARS-CoV-2 antigens, immunized hens and purified IgY antibodies in 2 months after the SARS-CoV-2 gene sequence became public. We further demonstrated that the IgY antibodies competitively block RBD binding to ACE2, neutralize authentic SARS-CoV-2 virus and effectively protect hamsters from SARS-CoV-2 challenge by preventing weight loss and lung pathology, representing the first comprehensive study with IgY antibodies. The process of mass production can be easily implemented in most developing countries and hence could become a new vital option in our toolbox for combating viral pandemics. This study could stimulate further studies, optimization and potential applications of IgY antibodies as therapeutics and prophylactics for human and animals.

尽管针对 SARS-CoV-2 病毒的疫苗和疗法的开发速度创下了纪录,但并不意味着未来的大流行病也能取得这样的成功。此外,在发展中国家,COVID-19 疫苗的接种率和疗法的应用率仍然很低。快速、低成本地大规模生产抗病毒 IgY 抗体可能是疫苗和疗法开发的一个有吸引力的替代或补充方案。在这篇文章中,我们在 SARS-CoV-2 基因序列公开后的 2 个月内快速制备了 SARS-CoV-2 抗原、免疫母鸡并纯化了 IgY 抗体。我们进一步证明了 IgY 抗体能竞争性地阻断 RBD 与 ACE2 的结合,中和真实的 SARS-CoV-2 病毒,并能有效地保护仓鼠免受 SARS-CoV-2 的挑战,防止体重减轻和肺部病变,这是首次用 IgY 抗体进行的全面研究。大规模生产的过程在大多数发展中国家都很容易实现,因此可以成为我们抗击病毒大流行工具箱中的一个新的重要选择。这项研究可以促进进一步研究、优化和潜在应用 IgY 抗体作为人类和动物的治疗和预防药物。
{"title":"Rapid development and mass production of SARS-CoV-2 neutralizing chicken egg yolk antibodies with protective efficacy in hamsters.","authors":"Binan Zhao, Haoran Peng, Yanjing Zhang, Jie Zhang, Desheng Kong, Sai Cao, Yan Li, Dan Yang, Chuanwen Sun, Xinyi Pu, Ping Zhao, Yan Xu, Kai Zhao, Liangzhi Xie","doi":"10.1186/s40659-024-00508-y","DOIUrl":"10.1186/s40659-024-00508-y","url":null,"abstract":"<p><p>Despite the record speed of developing vaccines and therapeutics against the SARS-CoV-2 virus, it is not a given that such success can be secured in future pandemics. In addition, COVID-19 vaccination and application of therapeutics remain low in developing countries. Rapid and low cost mass production of antiviral IgY antibodies could be an attractive alternative or complementary option for vaccine and therapeutic development. In this article, we rapidly produced SARS-CoV-2 antigens, immunized hens and purified IgY antibodies in 2 months after the SARS-CoV-2 gene sequence became public. We further demonstrated that the IgY antibodies competitively block RBD binding to ACE2, neutralize authentic SARS-CoV-2 virus and effectively protect hamsters from SARS-CoV-2 challenge by preventing weight loss and lung pathology, representing the first comprehensive study with IgY antibodies. The process of mass production can be easily implemented in most developing countries and hence could become a new vital option in our toolbox for combating viral pandemics. This study could stimulate further studies, optimization and potential applications of IgY antibodies as therapeutics and prophylactics for human and animals.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"57 1","pages":"24"},"PeriodicalIF":6.7,"publicationDate":"2024-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11071260/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140853130","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Establishment of primary prostate epithelial and tumorigenic cell lines using a non-viral immortalization approach 利用非病毒永生化方法建立原代前列腺上皮细胞系和肿瘤细胞系
IF 6.7 2区 生物学 Q1 BIOLOGY Pub Date : 2024-05-04 DOI: 10.1186/s40659-024-00507-z
Simon Lange, Anna Kuntze, Neele Wüstmann, Theresa Reckers, Verena Humberg, Wilhelm G. Dirks, Sebastian Huss, Julia Vieler, Andres Jan Schrader, Martin Bögemann, Katrin Schlack, Christof Bernemann
Research on prostate cancer is mostly performed using cell lines derived from metastatic disease, not reflecting stages of tumor initiation or early progression. Establishment of cancer cell lines derived from the primary tumor site has not been described so far. By definition, cancer cells are able to be cultured indefinitely, whereas normal epithelial cells undergo senescence in vitro. Epithelial cells can be immortalized, accomplished by using viral integration of immortalization factors. Viral approaches, however, might be impaired by regulatory and safety issues as well as random integration into regulatory genetic elements, modifying precise gene expression. We intend to use surgical specimen of prostate cancer patients to (i) prove for establishment of cancer cell lines, and (ii) perform non-viral, Sleeping Beauty (SB) transposase-based immortalization of prostate epithelial cells. Radical prostatectomy samples of prostate cancer patients (n = 4) were dissociated and cultured in vitro. Cells were cultivated either without or after non-viral, Sleeping-Beauty transposase-based stable transfection with immortalization factors SV40LT and hTERT. Established cell lines were analyzed in vitro and in vivo for characteristics of prostate (cancer) cells. Initial cell cultures without genetic manipulation underwent senescence within ≤ 15 passages, demonstrating inability to successfully derive primary prostate cancer cell lines. By using SB transposase-based integration of immortalization factors, we were able to establish primary prostate cell lines. Three out of four cell lines displayed epithelial characteristics, however without expression of prostate (cancer) characteristics, e.g., androgen receptor. In vivo, one cell line exhibited tumorigenic potential, yet characteristics of prostate adenocarcinoma were absent. Whereas no primary prostate cancer cell line could be established, we provide for the first-time immortalization of primary prostate cells using the SB transposase system, thereby preventing regulatory and molecular issues based on viral immortalization approaches. Although, none of the newly derived cell lines demonstrated prostate cancer characteristics, tumor formation was observed in one cell line. Given the non-prostate adenocarcinoma properties of the tumor, cells have presumably undergone oncogenic transformation rather than prostate cancer differentiation. Still, these cell lines might be used as a tool for research on prostate cancer initiation and early cancer progression.
对前列腺癌的研究大多使用来自转移性疾病的细胞系,不能反映肿瘤的起始阶段或早期进展。迄今为止,还没有人描述过从原发肿瘤部位提取的癌细胞系的建立情况。根据定义,癌细胞可以无限期培养,而正常上皮细胞在体外会衰老。上皮细胞可以通过病毒整合永生化因子实现永生化。然而,病毒方法可能会受到监管和安全问题的影响,也可能会随机整合到调控遗传元件中,从而改变精确的基因表达。我们打算利用前列腺癌患者的手术标本:(i) 证明癌症细胞系的建立;(ii) 对前列腺上皮细胞进行基于睡美人(SB)转座酶的非病毒永生化。对前列腺癌患者的前列腺根治术样本(n = 4)进行离体和体外培养。培养细胞时不使用病毒,也不使用基于睡美人转座酶的稳定转染永生化因子 SV40LT 和 hTERT。对建立的细胞系进行了体外和体内分析,以确定前列腺(癌)细胞的特征。未进行基因操作的初始细胞培养物在传代次数不超过 15 次时就会衰老,这表明无法成功培育出原代前列腺癌细胞系。通过使用基于 SB 转座酶的永生化因子整合,我们建立了原代前列腺细胞系。四个细胞系中有三个显示出上皮特征,但没有前列腺(癌症)特征(如雄激素受体)的表达。在体内,一种细胞系具有致瘤潜能,但没有前列腺腺癌的特征。虽然无法建立原代前列腺癌细胞系,但我们首次利用 SB 转座酶系统实现了原代前列腺细胞的永生化,从而避免了基于病毒永生化方法的调控和分子问题。虽然新获得的细胞系都没有表现出前列腺癌的特征,但在一个细胞系中观察到了肿瘤的形成。鉴于肿瘤的非前列腺腺癌特性,细胞可能发生了致癌转化,而非前列腺癌分化。不过,这些细胞系仍可作为研究前列腺癌发生和早期癌症进展的工具。
{"title":"Establishment of primary prostate epithelial and tumorigenic cell lines using a non-viral immortalization approach","authors":"Simon Lange, Anna Kuntze, Neele Wüstmann, Theresa Reckers, Verena Humberg, Wilhelm G. Dirks, Sebastian Huss, Julia Vieler, Andres Jan Schrader, Martin Bögemann, Katrin Schlack, Christof Bernemann","doi":"10.1186/s40659-024-00507-z","DOIUrl":"https://doi.org/10.1186/s40659-024-00507-z","url":null,"abstract":"Research on prostate cancer is mostly performed using cell lines derived from metastatic disease, not reflecting stages of tumor initiation or early progression. Establishment of cancer cell lines derived from the primary tumor site has not been described so far. By definition, cancer cells are able to be cultured indefinitely, whereas normal epithelial cells undergo senescence in vitro. Epithelial cells can be immortalized, accomplished by using viral integration of immortalization factors. Viral approaches, however, might be impaired by regulatory and safety issues as well as random integration into regulatory genetic elements, modifying precise gene expression. We intend to use surgical specimen of prostate cancer patients to (i) prove for establishment of cancer cell lines, and (ii) perform non-viral, Sleeping Beauty (SB) transposase-based immortalization of prostate epithelial cells. Radical prostatectomy samples of prostate cancer patients (n = 4) were dissociated and cultured in vitro. Cells were cultivated either without or after non-viral, Sleeping-Beauty transposase-based stable transfection with immortalization factors SV40LT and hTERT. Established cell lines were analyzed in vitro and in vivo for characteristics of prostate (cancer) cells. Initial cell cultures without genetic manipulation underwent senescence within ≤ 15 passages, demonstrating inability to successfully derive primary prostate cancer cell lines. By using SB transposase-based integration of immortalization factors, we were able to establish primary prostate cell lines. Three out of four cell lines displayed epithelial characteristics, however without expression of prostate (cancer) characteristics, e.g., androgen receptor. In vivo, one cell line exhibited tumorigenic potential, yet characteristics of prostate adenocarcinoma were absent. Whereas no primary prostate cancer cell line could be established, we provide for the first-time immortalization of primary prostate cells using the SB transposase system, thereby preventing regulatory and molecular issues based on viral immortalization approaches. Although, none of the newly derived cell lines demonstrated prostate cancer characteristics, tumor formation was observed in one cell line. Given the non-prostate adenocarcinoma properties of the tumor, cells have presumably undergone oncogenic transformation rather than prostate cancer differentiation. Still, these cell lines might be used as a tool for research on prostate cancer initiation and early cancer progression.","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"119 1","pages":""},"PeriodicalIF":6.7,"publicationDate":"2024-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140841311","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
General regulatory factors exert differential effects on nucleosome sliding activity of the ISW1a complex 一般调控因子对 ISW1a 复合物的核小体滑动活性产生不同影响
IF 6.7 2区 生物学 Q1 BIOLOGY Pub Date : 2024-05-04 DOI: 10.1186/s40659-024-00500-6
Andrea Oyarzún-Cisterna, Cristián Gidi, Fernanda Raiqueo, Roberto Amigo, Camila Rivas, Marcela Torrejón, José L. Gutiérrez
Chromatin dynamics is deeply involved in processes that require access to DNA, such as transcriptional regulation. Among the factors involved in chromatin dynamics at gene regulatory regions are general regulatory factors (GRFs). These factors contribute to establishment and maintenance of nucleosome-depleted regions (NDRs). These regions are populated by nucleosomes through histone deposition and nucleosome sliding, the latter catalyzed by a number of ATP-dependent chromatin remodeling complexes, including ISW1a. It has been observed that GRFs can act as barriers against nucleosome sliding towards NDRs. However, the relative ability of the different GRFs to hinder sliding activity is currently unknown. Considering this, we performed a comparative analysis for the main GRFs, with focus in their ability to modulate nucleosome sliding mediated by ISW1a. Among the GRFs tested in nucleosome remodeling assays, Rap1 was the only factor displaying the ability to hinder the activity of ISW1a. This effect requires location of the Rap1 cognate sequence on linker that becomes entry DNA in the nucleosome remodeling process. In addition, Rap1 was able to hinder nucleosome assembly in octamer transfer assays. Concurrently, Rap1 displayed the highest affinity for and longest dwell time from its target sequence, compared to the other GRFs tested. Consistently, through bioinformatics analyses of publicly available genome-wide data, we found that nucleosome occupancy and histone deposition in vivo are inversely correlated with the affinity of Rap1 for its target sequences in the genome. Our findings point to DNA binding affinity, residence time and location at particular translational positions relative to the nucleosome core as the key features of GRFs underlying their roles played in nucleosome sliding and assembly.
染色质动态与转录调控等需要访问 DNA 的过程密切相关。参与基因调控区染色质动态的因子包括一般调控因子(GRFs)。这些因子有助于建立和维持核糖体缺失区(NDR)。这些区域通过组蛋白沉积和核小体滑动由核小体填充,后者由包括 ISW1a 在内的多种 ATP 依赖性染色质重塑复合物催化。据观察,GRFs 可作为阻止核小体滑向 NDRs 的屏障。然而,目前还不清楚不同 GRFs 阻碍滑动活动的相对能力。有鉴于此,我们对主要的 GRFs 进行了比较分析,重点分析它们调节 ISW1a 介导的核小体滑动的能力。在核小体重塑实验中测试的 GRFs 中,Rap1 是唯一能够阻碍 ISW1a 活性的因子。这种作用要求 Rap1 的同源序列位于连接体上,而连接体在核小体重塑过程中成为入口 DNA。此外,在八聚体转移试验中,Rap1 还能阻碍核小体的组装。同时,与测试的其他 GRF 相比,Rap1 对其目标序列的亲和力最高,停留时间最长。同样,通过对公开的全基因组数据进行生物信息学分析,我们发现体内核小体占据率和组蛋白沉积与 Rap1 对基因组中靶序列的亲和力成反比。我们的研究结果表明,DNA结合亲和力、停留时间和相对于核小体核心的特定翻译位置是GRFs的关键特征,它们在核小体滑动和组装中发挥着重要作用。
{"title":"General regulatory factors exert differential effects on nucleosome sliding activity of the ISW1a complex","authors":"Andrea Oyarzún-Cisterna, Cristián Gidi, Fernanda Raiqueo, Roberto Amigo, Camila Rivas, Marcela Torrejón, José L. Gutiérrez","doi":"10.1186/s40659-024-00500-6","DOIUrl":"https://doi.org/10.1186/s40659-024-00500-6","url":null,"abstract":"Chromatin dynamics is deeply involved in processes that require access to DNA, such as transcriptional regulation. Among the factors involved in chromatin dynamics at gene regulatory regions are general regulatory factors (GRFs). These factors contribute to establishment and maintenance of nucleosome-depleted regions (NDRs). These regions are populated by nucleosomes through histone deposition and nucleosome sliding, the latter catalyzed by a number of ATP-dependent chromatin remodeling complexes, including ISW1a. It has been observed that GRFs can act as barriers against nucleosome sliding towards NDRs. However, the relative ability of the different GRFs to hinder sliding activity is currently unknown. Considering this, we performed a comparative analysis for the main GRFs, with focus in their ability to modulate nucleosome sliding mediated by ISW1a. Among the GRFs tested in nucleosome remodeling assays, Rap1 was the only factor displaying the ability to hinder the activity of ISW1a. This effect requires location of the Rap1 cognate sequence on linker that becomes entry DNA in the nucleosome remodeling process. In addition, Rap1 was able to hinder nucleosome assembly in octamer transfer assays. Concurrently, Rap1 displayed the highest affinity for and longest dwell time from its target sequence, compared to the other GRFs tested. Consistently, through bioinformatics analyses of publicly available genome-wide data, we found that nucleosome occupancy and histone deposition in vivo are inversely correlated with the affinity of Rap1 for its target sequences in the genome. Our findings point to DNA binding affinity, residence time and location at particular translational positions relative to the nucleosome core as the key features of GRFs underlying their roles played in nucleosome sliding and assembly.","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"41 1","pages":""},"PeriodicalIF":6.7,"publicationDate":"2024-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140841093","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
The effect of diabetes mellitus on differentiation of mesenchymal stem cells into insulin-producing cells 糖尿病对间充质干细胞分化为胰岛素分泌细胞的影响
IF 6.7 2区 生物学 Q1 BIOLOGY Pub Date : 2024-05-02 DOI: 10.1186/s40659-024-00502-4
Omar I. Badr, Mohamed M. Kamal, Shohda A. El-Maraghy, Heba R. Ghaiad
Diabetes mellitus (DM) is a global epidemic with increasing incidences. DM is a metabolic disease associated with chronic hyperglycemia. Aside from conventional treatments, there is no clinically approved cure for DM up till now. Differentiating mesenchymal stem cells (MSCs) into insulin-producing cells (IPCs) is a promising approach for curing DM. Our study was conducted to investigate the effect of DM on MSCs differentiation into IPCs in vivo and in vitro. We isolated adipose-derived mesenchymal stem cells (Ad-MSCs) from the epididymal fat of normal and STZ-induced diabetic Sprague–Dawley male rats. Afterwards, the in vitro differentiation of normal-Ad-MSCs (N-Ad-MSCs) and diabetic-Ad-MSCs (DM-Ad-MSCs) into IPCs was compared morphologically then through determining the gene expression of β-cell markers including neurogenin-3 (Ngn-3), homeobox protein (Nkx6.1), musculoaponeurotic fibrosarcoma oncogene homolog A (MafA), and insulin-1 (Ins-1) and eventually, through performing glucose-stimulated insulin secretion test (GSIS). Finally, the therapeutic potential of N-Ad-MSCs and DM-Ad-MSCs transplantation was compared in vivo in STZ-induced diabetic animals. Our results showed no significant difference in the characteristics of N-Ad-MSCs and DM-Ad-MSCs. However, we demonstrated a significant difference in their abilities to differentiate into IPCs in vitro morphologically in addition to β-cell markers expression, and functional assessment via GSIS test. Furthermore, the abilities of both Ad-MSCs to control hyperglycemia in diabetic rats in vivo was assessed through measuring fasting blood glucose (FBGs), body weight (BW), histopathological examination of both pancreas and liver and immunoexpression of insulin in pancreata of study groups. Our findings reveal the effectiveness of N-Ad-MSCs in differentiating into IPCs in vitro and controlling the hyperglycemia of STZ-induced diabetic rats in vivo compared to DM-Ad-MSCs.
糖尿病(DM)是一种全球性流行病,发病率不断上升。糖尿病是一种与慢性高血糖有关的代谢性疾病。除了传统治疗方法外,迄今为止还没有临床认可的治疗糖尿病的方法。将间充质干细胞(MSCs)分化为胰岛素分泌细胞(IPCs)是治疗DM的一种很有前景的方法。我们的研究旨在探讨DM对间充质干细胞在体内和体外分化为IPCs的影响。我们从正常大鼠和 STZ 诱导的糖尿病 Sprague-Dawley 雄性大鼠的附睾脂肪中分离出脂肪间充质干细胞(Ad-MSCs)。随后,比较了正常-Ad-间充质干细胞(N-Ad-MSCs)和糖尿病-Ad-间充质干细胞(DM-Ad-MSCs)在体外分化成 IPCs 的形态学结果,并通过测定神经原蛋白-3(Ngn-3)、同工酶蛋白(Nkx6.1)、肌肉神经纤维肉瘤癌基因同源物 A(MafA)和胰岛素-1(Ins-1)等β细胞标志物的基因表达,最后通过葡萄糖刺激胰岛素分泌试验(GSIS)进行检测。最后,在 STZ 诱导的糖尿病动物体内比较了 N-Ad-MSCs 和 DM-Ad-MSCs 移植的治疗潜力。结果显示,N-Ad-间充质干细胞和DM-Ad-间充质干细胞的特性没有明显差异。然而,我们发现它们在体外分化成 IPC 的能力除了在形态学上有显著差异外,在 β 细胞标志物表达和通过 GSIS 测试进行功能评估方面也有显著差异。此外,我们还通过测量研究组大鼠的空腹血糖(FBGs)、体重(BW)、胰腺和肝脏的组织病理学检查以及胰岛素在胰腺中的免疫表达,评估了两种 Ad-MSCs 在体内控制糖尿病大鼠高血糖的能力。我们的研究结果表明,与 DM-Ad-MSCs 相比,N-Ad-MSCs 能有效地在体外分化成 IPC,并控制 STZ 诱导的糖尿病大鼠体内的高血糖。
{"title":"The effect of diabetes mellitus on differentiation of mesenchymal stem cells into insulin-producing cells","authors":"Omar I. Badr, Mohamed M. Kamal, Shohda A. El-Maraghy, Heba R. Ghaiad","doi":"10.1186/s40659-024-00502-4","DOIUrl":"https://doi.org/10.1186/s40659-024-00502-4","url":null,"abstract":"Diabetes mellitus (DM) is a global epidemic with increasing incidences. DM is a metabolic disease associated with chronic hyperglycemia. Aside from conventional treatments, there is no clinically approved cure for DM up till now. Differentiating mesenchymal stem cells (MSCs) into insulin-producing cells (IPCs) is a promising approach for curing DM. Our study was conducted to investigate the effect of DM on MSCs differentiation into IPCs in vivo and in vitro. We isolated adipose-derived mesenchymal stem cells (Ad-MSCs) from the epididymal fat of normal and STZ-induced diabetic Sprague–Dawley male rats. Afterwards, the in vitro differentiation of normal-Ad-MSCs (N-Ad-MSCs) and diabetic-Ad-MSCs (DM-Ad-MSCs) into IPCs was compared morphologically then through determining the gene expression of β-cell markers including neurogenin-3 (Ngn-3), homeobox protein (Nkx6.1), musculoaponeurotic fibrosarcoma oncogene homolog A (MafA), and insulin-1 (Ins-1) and eventually, through performing glucose-stimulated insulin secretion test (GSIS). Finally, the therapeutic potential of N-Ad-MSCs and DM-Ad-MSCs transplantation was compared in vivo in STZ-induced diabetic animals. Our results showed no significant difference in the characteristics of N-Ad-MSCs and DM-Ad-MSCs. However, we demonstrated a significant difference in their abilities to differentiate into IPCs in vitro morphologically in addition to β-cell markers expression, and functional assessment via GSIS test. Furthermore, the abilities of both Ad-MSCs to control hyperglycemia in diabetic rats in vivo was assessed through measuring fasting blood glucose (FBGs), body weight (BW), histopathological examination of both pancreas and liver and immunoexpression of insulin in pancreata of study groups. Our findings reveal the effectiveness of N-Ad-MSCs in differentiating into IPCs in vitro and controlling the hyperglycemia of STZ-induced diabetic rats in vivo compared to DM-Ad-MSCs. ","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"1 1","pages":""},"PeriodicalIF":6.7,"publicationDate":"2024-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140841327","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Control of astrocytic Ca2+ signaling by nitric oxide-dependent S-nitrosylation of Ca2+ homeostasis modulator 1 channels 一氧化氮依赖的钙离子平衡调节器 1 通道 S-亚硝基化对星形胶质细胞钙离子信号的控制
IF 6.7 2区 生物学 Q1 BIOLOGY Pub Date : 2024-04-30 DOI: 10.1186/s40659-024-00503-3
Mariela Puebla, Manuel F. Muñoz, Mauricio A. Lillo, Jorge E. Contreras, Xavier F. Figueroa
Astrocytes Ca2+ signaling play a central role in the modulation of neuronal function. Activation of metabotropic glutamate receptors (mGluR) by glutamate released during an increase in synaptic activity triggers coordinated Ca2+ signals in astrocytes. Importantly, astrocytes express the Ca2+-dependent nitric oxide (NO)-synthetizing enzymes eNOS and nNOS, which might contribute to the Ca2+ signals by triggering Ca2+ influx or ATP release through the activation of connexin 43 (Cx43) hemichannels, pannexin-1 (Panx-1) channels or Ca2+ homeostasis modulator 1 (CALHM1) channels. Hence, we aim to evaluate the participation of NO in the astrocytic Ca2+ signaling initiated by stimulation of mGluR in primary cultures of astrocytes from rat brain cortex. Astrocytes were stimulated with glutamate or t-ACPD and NO-dependent changes in [Ca2+]i and ATP release were evaluated. In addition, the activity of Cx43 hemichannels, Panx-1 channels and CALHM1 channels was also analyzed. The expression of Cx43, Panx-1 and CALHM1 in astrocytes was confirmed by immunofluorescence analysis and both glutamate and t-ACPD induced NO-mediated activation of CALHM1 channels via direct S-nitrosylation, which was further confirmed by assessing CALHM1-mediated current using the two-electrode voltage clamp technique in Xenopus oocytes. Pharmacological blockade or siRNA-mediated inhibition of CALHM1 expression revealed that the opening of these channels provides a pathway for ATP release and the subsequent purinergic receptor-dependent activation of Cx43 hemichannels and Panx-1 channels, which further contributes to the astrocytic Ca2+ signaling. Our findings demonstrate that activation of CALHM1 channels through NO-mediated S-nitrosylation in astrocytes in vitro is critical for the generation of glutamate-initiated astrocytic Ca2+ signaling.
星形胶质细胞的 Ca2+ 信号在调节神经元功能方面发挥着核心作用。突触活动增加时释放的谷氨酸会激活代谢型谷氨酸受体(mGluR),从而触发星形胶质细胞中协调的 Ca2+ 信号。重要的是,星形胶质细胞表达依赖 Ca2+ 的一氧化氮(NO)合成酶 eNOS 和 nNOS,它们可能通过激活连接素 43(Cx43)半桥通道、pannexin-1(Panx-1)通道或 Ca2+ 平衡调节器 1(CALHM1)通道触发 Ca2+ 流入或 ATP 释放,从而促进 Ca2+ 信号。因此,我们旨在评估 NO 在大鼠大脑皮层星形胶质细胞原代培养物中参与由 mGluR 刺激引发的星形胶质细胞 Ca2+ 信号转导的情况。用谷氨酸或 t-ACPD 刺激星形胶质细胞,评估[Ca2+]i 和 ATP 释放的 NO 依赖性变化。此外,还分析了 Cx43 半通道、Panx-1 通道和 CALHM1 通道的活性。免疫荧光分析证实了 Cx43、Panx-1 和 CALHM1 在星形胶质细胞中的表达,谷氨酸和 t-ACPD 通过直接 S-亚硝基化诱导了 NO 介导的 CALHM1 通道活化。药理阻断或 siRNA 介导的 CALHM1 表达抑制显示,这些通道的开放为 ATP 释放以及随后嘌呤能受体依赖性激活 Cx43 半ich通道和 Panx-1 通道提供了途径,从而进一步促进了星形胶质细胞的 Ca2+ 信号转导。我们的研究结果表明,在体外星形胶质细胞中通过 NO 介导的 S-亚硝基化激活 CALHM1 通道对于谷氨酸引发的星形胶质细胞 Ca2+ 信号的产生至关重要。
{"title":"Control of astrocytic Ca2+ signaling by nitric oxide-dependent S-nitrosylation of Ca2+ homeostasis modulator 1 channels","authors":"Mariela Puebla, Manuel F. Muñoz, Mauricio A. Lillo, Jorge E. Contreras, Xavier F. Figueroa","doi":"10.1186/s40659-024-00503-3","DOIUrl":"https://doi.org/10.1186/s40659-024-00503-3","url":null,"abstract":"Astrocytes Ca2+ signaling play a central role in the modulation of neuronal function. Activation of metabotropic glutamate receptors (mGluR) by glutamate released during an increase in synaptic activity triggers coordinated Ca2+ signals in astrocytes. Importantly, astrocytes express the Ca2+-dependent nitric oxide (NO)-synthetizing enzymes eNOS and nNOS, which might contribute to the Ca2+ signals by triggering Ca2+ influx or ATP release through the activation of connexin 43 (Cx43) hemichannels, pannexin-1 (Panx-1) channels or Ca2+ homeostasis modulator 1 (CALHM1) channels. Hence, we aim to evaluate the participation of NO in the astrocytic Ca2+ signaling initiated by stimulation of mGluR in primary cultures of astrocytes from rat brain cortex. Astrocytes were stimulated with glutamate or t-ACPD and NO-dependent changes in [Ca2+]i and ATP release were evaluated. In addition, the activity of Cx43 hemichannels, Panx-1 channels and CALHM1 channels was also analyzed. The expression of Cx43, Panx-1 and CALHM1 in astrocytes was confirmed by immunofluorescence analysis and both glutamate and t-ACPD induced NO-mediated activation of CALHM1 channels via direct S-nitrosylation, which was further confirmed by assessing CALHM1-mediated current using the two-electrode voltage clamp technique in Xenopus oocytes. Pharmacological blockade or siRNA-mediated inhibition of CALHM1 expression revealed that the opening of these channels provides a pathway for ATP release and the subsequent purinergic receptor-dependent activation of Cx43 hemichannels and Panx-1 channels, which further contributes to the astrocytic Ca2+ signaling. Our findings demonstrate that activation of CALHM1 channels through NO-mediated S-nitrosylation in astrocytes in vitro is critical for the generation of glutamate-initiated astrocytic Ca2+ signaling.","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"45 1","pages":""},"PeriodicalIF":6.7,"publicationDate":"2024-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140841308","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Identification and expression analysis of two steamer-like retrotransposons in the Chilean blue mussel (Mytilus chilensis) 智利蓝贻贝(Mytilus chilensis)中两个类似蒸汽机的转座子的鉴定和表达分析
IF 6.7 2区 生物学 Q1 BIOLOGY Pub Date : 2024-04-26 DOI: 10.1186/s40659-024-00498-x
Gloria Arriagada, Johan Quezada, Nicolas Merino-Veliz, Fernando Avilés, Diana Tapia-Cammas, Jorge Gomez, Daniela Curotto, Juan A. Valdes, Pablo A. Oyarzún, Cristian Gallardo-Escárate, Michael J. Metzger, Marco Alvarez
Disseminated neoplasia (DN) is a proliferative cell disorder of the circulatory system of bivalve mollusks. The disease is transmitted between individuals and can also be induced by external chemical agents such as bromodeoxyuridine. In Mya arenaria, we have cloned and characterized an LTR-retrotransposon named Steamer. Steamer mRNA levels and gene copy number correlates with DN and can be used as a marker of the disease. So far, the only mollusk where a retrotransposon expression relates to DN is Mya arenaria. On the other hand, it has been reported that the Chilean blue mussel Mytilus chilensis can also suffers DN. Our aim was to identify retrotransposons in Mytilus chilensis and to study their expression levels in the context of disseminated neoplasia. Here we show that 7.1% of individuals collected in August 2018, from two farming areas, presents morphological characteristics described in DN. Using Steamer sequence to interrogate the transcriptome of M. chilensis we found two putative retrotransposons, named Steamer-like elements (MchSLEs). MchSLEs are present in the genome of M. chilensis and MchSLE1 is indeed an LTR-retrotransposon. Neither expression, nor copy number of the reported MchSLEs correlate with DN status but both are expressed at different levels among individual animals. We also report that in cultured M. chilensis haemocytes MchSLEs1 expression can be induced by bromodeoxyuridine. We conclude that SLEs present in Mytilus chilensis are differentially expressed among individuals and do not correlate with disseminated neoplasia. Treatment of haemocytes with a stressor like bromodeoxyuridine induces expression of MchSLE1 suggesting that in Mytilus chilensis environmental stressors can induce activation of LTR-retrotransposon.
播散性肿瘤病(DN)是双壳类软体动物循环系统的一种细胞增生性疾病。这种疾病可在个体间传播,也可由溴脱氧尿苷等外部化学物质诱发。我们克隆并鉴定了一种名为 Steamer 的 LTR 反转座子。Steamer mRNA水平和基因拷贝数与DN相关,可用作该疾病的标记。迄今为止,逆转录转座子的表达与 DN 有关的软体动物只有 Mya arenaria。另一方面,有报道称智利蓝贻贝(Mytilus chilensis)也会感染 DN。我们的目的是鉴定智利贻贝中的逆转录转座子,并研究它们在播散性肿瘤中的表达水平。在此,我们表明,2018 年 8 月从两个养殖区采集的个体中有 7.1%呈现出 DN 所描述的形态特征。利用Steamer序列询问M. chilensis的转录组,我们发现了两个推定的逆转录转座子,命名为Steamer样元件(MchSLEs)。MchSLEs 存在于 M. chilensis 的基因组中,而且 MchSLE1 确实是一个 LTR 逆转录转座子。所报道的 MchSLEs 的表达和拷贝数均与 DN 状态无关,但两者在不同动物个体中的表达水平不同。我们还报告说,在培养的 M. chilensis 血细胞中,溴脱氧尿苷可诱导 MchSLEs1 的表达。我们的结论是,存在于智利贻贝中的 SLEs 在不同个体中的表达量不同,而且与播散性肿瘤无关。用溴脱氧尿苷等应激源处理血细胞可诱导 MchSLE1 的表达,这表明在智利贻贝中,环境应激源可诱导 LTR 反转座子的活化。
{"title":"Identification and expression analysis of two steamer-like retrotransposons in the Chilean blue mussel (Mytilus chilensis)","authors":"Gloria Arriagada, Johan Quezada, Nicolas Merino-Veliz, Fernando Avilés, Diana Tapia-Cammas, Jorge Gomez, Daniela Curotto, Juan A. Valdes, Pablo A. Oyarzún, Cristian Gallardo-Escárate, Michael J. Metzger, Marco Alvarez","doi":"10.1186/s40659-024-00498-x","DOIUrl":"https://doi.org/10.1186/s40659-024-00498-x","url":null,"abstract":"Disseminated neoplasia (DN) is a proliferative cell disorder of the circulatory system of bivalve mollusks. The disease is transmitted between individuals and can also be induced by external chemical agents such as bromodeoxyuridine. In Mya arenaria, we have cloned and characterized an LTR-retrotransposon named Steamer. Steamer mRNA levels and gene copy number correlates with DN and can be used as a marker of the disease. So far, the only mollusk where a retrotransposon expression relates to DN is Mya arenaria. On the other hand, it has been reported that the Chilean blue mussel Mytilus chilensis can also suffers DN. Our aim was to identify retrotransposons in Mytilus chilensis and to study their expression levels in the context of disseminated neoplasia. Here we show that 7.1% of individuals collected in August 2018, from two farming areas, presents morphological characteristics described in DN. Using Steamer sequence to interrogate the transcriptome of M. chilensis we found two putative retrotransposons, named Steamer-like elements (MchSLEs). MchSLEs are present in the genome of M. chilensis and MchSLE1 is indeed an LTR-retrotransposon. Neither expression, nor copy number of the reported MchSLEs correlate with DN status but both are expressed at different levels among individual animals. We also report that in cultured M. chilensis haemocytes MchSLEs1 expression can be induced by bromodeoxyuridine. We conclude that SLEs present in Mytilus chilensis are differentially expressed among individuals and do not correlate with disseminated neoplasia. Treatment of haemocytes with a stressor like bromodeoxyuridine induces expression of MchSLE1 suggesting that in Mytilus chilensis environmental stressors can induce activation of LTR-retrotransposon.","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"39 1","pages":""},"PeriodicalIF":6.7,"publicationDate":"2024-04-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140801070","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Noncoding RNAs in skeletal development and disorders 骨骼发育和骨骼疾病中的非编码 RNA
IF 6.7 2区 生物学 Q1 BIOLOGY Pub Date : 2024-04-22 DOI: 10.1186/s40659-024-00497-y
Qing Yao, Tailin He, Jian-You Liao, Rongdong Liao, Xiaohao Wu, Lijun Lin, Guozhi Xiao
Protein-encoding genes only constitute less than 2% of total human genomic sequences, and 98% of genetic information was previously referred to as “junk DNA”. Meanwhile, non-coding RNAs (ncRNAs) consist of approximately 60% of the transcriptional output of human cells. Thousands of ncRNAs have been identified in recent decades, and their essential roles in the regulation of gene expression in diverse cellular pathways associated with fundamental cell processes, including proliferation, differentiation, apoptosis, and metabolism, have been extensively investigated. Furthermore, the gene regulation networks they form modulate gene expression in normal development and under pathological conditions. In this review, we integrate current information about the classification, biogenesis, and function of ncRNAs and how these ncRNAs support skeletal development through their regulation of critical genes and signaling pathways in vivo. We also summarize the updated knowledge of ncRNAs involved in common skeletal diseases and disorders, including but not limited to osteoporosis, osteoarthritis, rheumatoid arthritis, scoliosis, and intervertebral disc degeneration, by highlighting their roles established from in vivo, in vitro, and ex vivo studies.
编码蛋白质的基因只占人类基因组总序列的不到 2%,98% 的遗传信息以前被称为 "垃圾 DNA"。与此同时,非编码 RNA(ncRNA)约占人类细胞转录输出的 60%。近几十年来,已经发现了数千种 ncRNA,它们在与增殖、分化、凋亡和新陈代谢等基本细胞过程相关的各种细胞通路中调控基因表达的重要作用已得到广泛研究。此外,它们形成的基因调控网络在正常发育和病理条件下调节基因表达。在这篇综述中,我们整合了当前关于 ncRNA 的分类、生物发生和功能的信息,以及这些 ncRNA 如何通过调控体内关键基因和信号通路来支持骨骼发育。我们还总结了与常见骨骼疾病和失调(包括但不限于骨质疏松症、骨关节炎、类风湿性关节炎、脊柱侧弯和椎间盘变性)有关的 ncRNA 的最新知识,重点介绍了这些 ncRNA 在体内、体外和体外研究中发挥的作用。
{"title":"Noncoding RNAs in skeletal development and disorders","authors":"Qing Yao, Tailin He, Jian-You Liao, Rongdong Liao, Xiaohao Wu, Lijun Lin, Guozhi Xiao","doi":"10.1186/s40659-024-00497-y","DOIUrl":"https://doi.org/10.1186/s40659-024-00497-y","url":null,"abstract":"Protein-encoding genes only constitute less than 2% of total human genomic sequences, and 98% of genetic information was previously referred to as “junk DNA”. Meanwhile, non-coding RNAs (ncRNAs) consist of approximately 60% of the transcriptional output of human cells. Thousands of ncRNAs have been identified in recent decades, and their essential roles in the regulation of gene expression in diverse cellular pathways associated with fundamental cell processes, including proliferation, differentiation, apoptosis, and metabolism, have been extensively investigated. Furthermore, the gene regulation networks they form modulate gene expression in normal development and under pathological conditions. In this review, we integrate current information about the classification, biogenesis, and function of ncRNAs and how these ncRNAs support skeletal development through their regulation of critical genes and signaling pathways in vivo. We also summarize the updated knowledge of ncRNAs involved in common skeletal diseases and disorders, including but not limited to osteoporosis, osteoarthritis, rheumatoid arthritis, scoliosis, and intervertebral disc degeneration, by highlighting their roles established from in vivo, in vitro, and ex vivo studies.","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"135 1","pages":""},"PeriodicalIF":6.7,"publicationDate":"2024-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140634361","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
期刊
Biological Research
全部 Acc. Chem. Res. ACS Applied Bio Materials ACS Appl. Electron. Mater. ACS Appl. Energy Mater. ACS Appl. Mater. Interfaces ACS Appl. Nano Mater. ACS Appl. Polym. Mater. ACS BIOMATER-SCI ENG ACS Catal. ACS Cent. Sci. ACS Chem. Biol. ACS Chemical Health & Safety ACS Chem. Neurosci. ACS Comb. Sci. ACS Earth Space Chem. ACS Energy Lett. ACS Infect. Dis. ACS Macro Lett. ACS Mater. Lett. ACS Med. Chem. Lett. ACS Nano ACS Omega ACS Photonics ACS Sens. ACS Sustainable Chem. Eng. ACS Synth. Biol. Anal. Chem. BIOCHEMISTRY-US Bioconjugate Chem. BIOMACROMOLECULES Chem. Res. Toxicol. Chem. Rev. Chem. Mater. CRYST GROWTH DES ENERG FUEL Environ. Sci. Technol. Environ. Sci. Technol. Lett. Eur. J. Inorg. Chem. IND ENG CHEM RES Inorg. Chem. J. Agric. Food. Chem. J. Chem. Eng. Data J. Chem. Educ. J. Chem. Inf. Model. J. Chem. Theory Comput. J. Med. Chem. J. Nat. Prod. J PROTEOME RES J. Am. Chem. Soc. LANGMUIR MACROMOLECULES Mol. Pharmaceutics Nano Lett. Org. Lett. ORG PROCESS RES DEV ORGANOMETALLICS J. Org. Chem. J. Phys. Chem. J. Phys. Chem. A J. Phys. Chem. B J. Phys. Chem. C J. Phys. Chem. Lett. Analyst Anal. Methods Biomater. Sci. Catal. Sci. Technol. Chem. Commun. Chem. Soc. Rev. CHEM EDUC RES PRACT CRYSTENGCOMM Dalton Trans. Energy Environ. Sci. ENVIRON SCI-NANO ENVIRON SCI-PROC IMP ENVIRON SCI-WAT RES Faraday Discuss. Food Funct. Green Chem. Inorg. Chem. Front. Integr. Biol. J. Anal. At. Spectrom. J. Mater. Chem. A J. Mater. Chem. B J. Mater. Chem. C Lab Chip Mater. Chem. Front. Mater. Horiz. MEDCHEMCOMM Metallomics Mol. Biosyst. Mol. Syst. Des. Eng. Nanoscale Nanoscale Horiz. Nat. Prod. Rep. New J. Chem. Org. Biomol. Chem. Org. Chem. Front. PHOTOCH PHOTOBIO SCI PCCP Polym. Chem.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1