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PTN from Leydig cells activates SDC2 and modulates human spermatogonial stem cell proliferation and survival via GFRA1 来自犁地细胞的 PTN 通过 GFRA1 激活 SDC2 并调节人类精原干细胞的增殖和存活
IF 6.7 2区 生物学 Q1 BIOLOGY Pub Date : 2024-09-16 DOI: 10.1186/s40659-024-00546-6
Xueheng Zhao, Lvjun Liu, Zenghui Huang, Fang Zhu, Huan Zhang, Dai Zhou
Spermatogonial stem cells (SSCs) are essential for the maintenance and initiation of male spermatogenesis. Despite the advances in understanding SSC biology in mouse models, the mechanisms underlying human SSC development remain elusive. Here, we analyzed the signaling pathways involved in SSC regulation by testicular somatic cells using single-cell sequencing data (GEO datasets: GSE149512 and GSE112013) and identified that Leydig cells communicate with SSCs through pleiotrophin (PTN) and its receptor syndecan-2 (SDC2). Immunofluorescence, STRING prediction, and protein immunoprecipitation assays confirmed the interaction between PTN and SDC2 in spermatogonia, but their co-localization was observed only in approximately 50% of the cells. The knockdown of SDC2 in human SSC lines impaired cell proliferation, DNA synthesis, and the expression of PLZF, a key marker for SSC self-renewal. Transcriptome analysis revealed that SDC2 knockdown downregulated the expression of GFRA1, a crucial factor for SSC proliferation and self-renewal, and inhibited the HIF-1 signaling pathway. Exogenous PTN rescued the proliferation and GFRA1 expression in SDC2 knockdown SSC lines. In addition, we found downregulation of PTN and SDC2 as well as altered localization in non-obstructive azoospermia (NOA) patients, suggesting that downregulation of PTN and SDC2 may be associated with impaired spermatogenesis. Our results uncover a novel mechanism of human SSC regulation by the testicular microenvironment and suggest a potential therapeutic target for male infertility.
精原干细胞(SSC)对男性精子发生的维持和启动至关重要。尽管在小鼠模型中对SSC生物学的理解取得了进展,但人类SSC发育的内在机制仍然难以捉摸。在这里,我们利用单细胞测序数据(GEO 数据集:GSE149512 和 GSE112013)分析了睾丸体细胞调控 SSC 所涉及的信号通路,发现 Leydig 细胞通过多养蛋白(PTN)及其受体辛迪加-2(SDC2)与 SSC 进行交流。免疫荧光、STRING 预测和蛋白免疫沉淀测定证实了 PTN 和 SDC2 在精原细胞中的相互作用,但它们的共定位只在大约 50% 的细胞中观察到。在人类精原细胞系中敲除 SDC2 会影响细胞增殖、DNA 合成和 PLZF(精原细胞自我更新的关键标志物)的表达。转录组分析显示,SDC2基因敲除会下调GFRA1的表达(GFRA1是SSC增殖和自我更新的关键因子),并抑制HIF-1信号通路。外源性 PTN 挽救了 SDC2 敲除 SSC 株系的增殖和 GFRA1 表达。此外,我们还发现在非梗阻性无精子症(NOA)患者中,PTN和SDC2的下调以及定位发生了改变,这表明PTN和SDC2的下调可能与精子发生障碍有关。我们的研究结果揭示了人类SSC受睾丸微环境调控的新机制,并提出了治疗男性不育症的潜在靶点。
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引用次数: 0
The effect of CGRP and SP and the cell signaling dialogue between sensory neurons and endothelial cells CGRP 和 SP 的作用以及感觉神经元与内皮细胞之间的细胞信号对话
IF 6.7 2区 生物学 Q1 BIOLOGY Pub Date : 2024-09-11 DOI: 10.1186/s40659-024-00538-6
Alice Leroux, Micaela Roque, Elina Casas, Jacques Leng, Christelle Guibert, Beatrice L’Azou, Hugo Oliveira, Joëlle Amédée, Bruno Paiva dos Santos
Increasing evidences demonstrate the role of sensory innervation in bone metabolism, remodeling and repair, however neurovascular coupling in bone is rarely studied. Using microfluidic devices as an indirect co-culture model to mimic in vitro the physiological scenario of innervation, our group demonstrated that sensory neurons (SNs) were able to regulate the extracellular matrix remodeling by endothelial cells (ECs), in particular through sensory neuropeptides, i.e. calcitonin gene-related peptide (CGRP) and substance P (SP). Nonetheless, still little is known about the cell signaling pathways and mechanism of action in neurovascular coupling. Here, in order to characterize the communication between SNs and ECs at molecular level, we evaluated the effect of SNs and the neuropeptides CGRP and SP on ECs. We focused on different pathways known to play a role on endothelial functions: calcium signaling, p38 and Erk1/2; the control of signal propagation through Cx43; and endothelial functions through the production of nitric oxide (NO). The effect of SNs was evaluated on ECs Ca2+ influx, the expression of Cx43, endothelial nitric oxide synthase (eNOS) and nitric oxide (NO) production, p38, ERK1/2 as well as their phosphorylated forms. In addition, the role of CGRP and SP were either analyzed using respective antagonists in the co-culture model, or by adding directly on the ECs monocultures. We show that capsaicin-stimulated SNs induce increased Ca2+ influx in ECs. SNs stimulate the increase of NO production in ECs, probably involving a decrease in the inhibitory eNOS T495 phosphorylation site. The neuropeptide CGRP, produced by SNs, seems to be one of the mediators of this effect in ECs since NO production is decreased in the presence of CGRP antagonist in the co-culture of ECs and SNs, and increased when ECs are stimulated with synthetic CGRP. Taken together, our results suggest that SNs play an important role in the control of the endothelial cell functions through CGRP production and NO signaling pathway.
越来越多的证据表明感觉神经支配在骨代谢、重塑和修复中的作用,但对骨中神经血管耦合的研究却很少。我们的研究小组利用微流体设备作为间接共培养模型,在体外模拟神经支配的生理情景,结果表明感觉神经元(SNs)能够调节内皮细胞(ECs)的细胞外基质重塑,特别是通过感觉神经肽,即降钙素基因相关肽(CGRP)和P物质(SP)。然而,人们对神经血管耦合中的细胞信号传导途径和作用机制仍然知之甚少。在此,为了在分子水平上描述神经元和血管内皮细胞之间的交流,我们评估了神经元和神经肽 CGRP 和 SP 对血管内皮细胞的影响。我们重点研究了已知对内皮功能起作用的不同途径:钙信号、p38 和 Erk1/2;通过 Cx43 控制信号传播;以及通过产生一氧化氮(NO)促进内皮功能。研究评估了 SNs 对 ECs Ca2+ 流入、Cx43 表达、内皮一氧化氮合酶(eNOS)和一氧化氮(NO)产生、p38、ERK1/2 及其磷酸化形式的影响。此外,还在共培养模型中使用各自的拮抗剂或直接添加到单培养的 ECs 上分析了 CGRP 和 SP 的作用。我们的研究表明,辣椒素刺激的SN诱导心血管内 Ca2+ 流入增加。SNs刺激心血管内NO生成的增加,可能涉及抑制性eNOS T495磷酸化位点的减少。SNs产生的神经肽CGRP似乎是心肌产生这种效应的介质之一,因为在心肌和SNs共培养过程中,如果有CGRP拮抗剂存在,NO的产生就会减少;而当用合成CGRP刺激心肌时,NO的产生就会增加。综上所述,我们的研究结果表明,SNs 在通过 CGRP 生成和 NO 信号通路控制内皮细胞功能方面发挥着重要作用。
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引用次数: 0
Mouse testicular macrophages can independently produce testosterone and are regulated by Cebpb 小鼠睾丸巨噬细胞可独立产生睾酮并受 Cebpb 调节
IF 6.7 2区 生物学 Q1 BIOLOGY Pub Date : 2024-09-09 DOI: 10.1186/s40659-024-00544-8
Nengliang Duan, Yuanshuai Ran, Huapei Wang, Ya Luo, Zhixiang Gao, Xingyu Lu, Fengmei Cui, Qiu Chen, Boxin Xue, Xiaolong Liu
Testicular macrophages (TM) have long been recognized for their role in immune response within the testicular environment. However, their involvement in steroid hormone synthesis, particularly testosterone, has not been fully elucidated. This study aims to explore the capability of TM to synthesize and secrete testosterone de novo and to investigate the regulatory mechanisms involved. Transcriptomic analysis revealed significant expression of Cyp11a1, Cyp17a1, Hsd3b1, and Hsd17b3 in TM, which are key enzymes in the testosterone synthesis pathway. qPCR analysis and immunofluorescence validation confirmed the autonomous capability of TM to synthesize testosterone. Ablation of TM in mice resulted in decreased physiological testosterone levels, underscoring the significance of TM in maintaining testicular testosterone levels. Additionally, the study also demonstrated that Cebpb regulates the expression of these crucial genes, thereby modulating testosterone synthesis. This research establishes that TM possess the autonomous capacity to synthesize and secrete testosterone, contributing significantly to testicular testosterone levels. The transcription factor Cebpb plays a crucial role in this process by regulating the expression of key genes involved in testosterone synthesis.
睾丸巨噬细胞(TM)在睾丸环境中的免疫反应作用早已得到公认。然而,它们参与类固醇激素(尤其是睾酮)合成的情况尚未完全阐明。本研究旨在探索睾丸激素从头合成和分泌睾酮的能力,并研究其中的调控机制。转录组分析表明,Cyp11a1、Cyp17a1、Hsd3b1和Hsd17b3在TM中大量表达,它们是睾酮合成途径中的关键酶。在小鼠体内消融 TM 会导致生理睾酮水平下降,这凸显了 TM 在维持睾丸睾酮水平方面的重要作用。此外,该研究还证明,Cebpb 可调节这些关键基因的表达,从而调节睾酮的合成。这项研究证实,睾丸组织具有合成和分泌睾酮的自主能力,对睾丸睾酮水平的提高有重要作用。转录因子 Cebpb 通过调节参与睾酮合成的关键基因的表达,在这一过程中发挥了至关重要的作用。
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引用次数: 0
Fruit sugar hub: gene regulatory network associated with soluble solids content (SSC) in Prunus persica. 果糖中心:与柿子可溶性固形物含量(SSC)相关的基因调控网络。
IF 4.3 2区 生物学 Q1 BIOLOGY Pub Date : 2024-09-06 DOI: 10.1186/s40659-024-00539-5
Gerardo Núñez-Lillo, Victoria Lillo-Carmona, Alonso G Pérez-Donoso, Romina Pedreschi, Reinaldo Campos-Vargas, Claudio Meneses

Chilean peach growers have achieved worldwide recognition for their high-quality fruit products. Among the main factors influencing peach fruit quality, sweetness is pivotal for maintaining the market's competitiveness. Numerous studies have been conducted in different peach-segregating populations to unravel SSC regulation. However, different cultivars may also have distinct genetic conformation, and other factors, such as environmental conditions, can significantly impact SSC. Using a transcriptomic approach with a gene co-expression network analysis, we aimed to identify the regulatory mechanism that controls the sugar accumulation process in an 'O × N' peach population. This population was previously studied through genomic analysis, associating LG5 with the genetic control of the SSC trait. The results obtained in this study allowed us to identify 91 differentially expressed genes located on chromosome 5 of the peach genome as putative new regulators of sugar accumulation in peach, together with a regulatory network that involves genes directly associated with sugar transport (PpSWEET15), cellulose biosynthesis (PpCSLG2), flavonoid biosynthesis (PpPAL1), pectin modifications (PpPG, PpPL and PpPMEi), expansins (PpEXPA1 and PpEXPA8) and several transcription factors (PpC3H67, PpHB7, PpRVE1 and PpCBF4) involved with the SSC phenotype. These results contribute to a better understanding of the genetic control of the SSC trait for future breeding programs in peaches.

智利的桃子种植者以其高品质的水果产品获得了全世界的认可。在影响桃果质量的主要因素中,甜度是保持市场竞争力的关键。为了揭示 SSC 的调节机制,对不同的桃隔离群体进行了大量研究。然而,不同的栽培品种也可能具有不同的遗传构象,而且环境条件等其他因素也会对 SSC 产生重大影响。我们利用转录组方法和基因共表达网络分析,旨在确定控制'O × N'桃群体糖分积累过程的调控机制。以前曾通过基因组分析对该群体进行过研究,发现 LG5 与 SSC 性状的遗传控制有关。这项研究的结果使我们确定了位于桃基因组第 5 号染色体上的 91 个差异表达基因,以及一个涉及与糖转运直接相关的基因(PpSWEET15)的调控网络,这些基因可能是桃糖积累的新调控因子、纤维素生物合成(PpCSLG2)、类黄酮生物合成(PpPAL1)、果胶修饰(PpPG、PpPL 和 PpPMEi)、扩张素(PpEXPA1 和 PpEXPA8)以及与 SSC 表型有关的几个转录因子(PpC3H67、PpHB7、PpRVE1 和 PpCBF4)。这些结果有助于更好地了解 SSC 性状的遗传控制,为今后的桃育种计划提供参考。
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引用次数: 0
Antimicrobial activity of compounds identified by artificial intelligence discovery engine targeting enzymes involved in Neisseria gonorrhoeae peptidoglycan metabolism. 人工智能发现引擎针对参与淋病奈瑟菌肽聚糖代谢的酶鉴定出的化合物的抗菌活性。
IF 4.3 2区 生物学 Q1 BIOLOGY Pub Date : 2024-09-05 DOI: 10.1186/s40659-024-00543-9
Ravi Kant, Hannah Tilford, Camila S Freitas, Dayana A Santos Ferreira, James Ng, Gwennan Rucinski, Joshua Watkins, Ryan Pemberton, Tigran M Abramyan, Stephanie C Contreras, Alejandra Vera, Myron Christodoulides

Background: Neisseria gonorrhoeae (Ng) causes the sexually transmitted disease gonorrhoea. There are no vaccines and infections are treated principally with antibiotics. However, gonococci rapidly develop resistance to every antibiotic class used and there is a need for developing new antimicrobial treatments. In this study we focused on two gonococcal enzymes as potential antimicrobial targets, namely the serine protease L,D-carboxypeptidase LdcA (NgO1274/NEIS1546) and the lytic transglycosylase LtgD (NgO0626/NEIS1212). To identify compounds that could interact with these enzymes as potential antimicrobials, we used the AtomNet virtual high-throughput screening technology. We then did a computational modelling study to examine the interactions of the most bioactive compounds with their target enzymes. The identified compounds were tested against gonococci to determine minimum inhibitory and bactericidal concentrations (MIC/MBC), specificity, and compound toxicity in vitro.

Results: AtomNet identified 74 compounds that could potentially interact with Ng-LdcA and 84 compounds that could potentially interact with Ng-LtgD. Through MIC and MBC assays, we selected the three best performing compounds for both enzymes. Compound 16 was the most active against Ng-LdcA, with a MIC50 value < 1.56 µM and MBC50/90 values between 0.195 and 0.39 µM. In general, the Ng-LdcA compounds showed higher activity than the compounds directed against Ng-LtgD, of which compound 45 had MIC50 values of 1.56-3.125 µM and MBC50/90 values between 3.125 and 6.25 µM. The compounds were specific for gonococci and did not kill other bacteria. They were also non-toxic for human conjunctival epithelial cells as judged by a resazurin assay. To support our biological data, in-depth computational modelling study detailed the interactions of the compounds with their target enzymes. Protein models were generated in silico and validated, the active binding sites and amino acids involved elucidated, and the interactions of the compounds interacting with the enzymes visualised through molecular docking and Molecular Dynamics Simulations for 50 ns and Molecular Mechanics Poisson-Boltzmann Surface Area (MM-PBSA).

Conclusions: We have identified bioactive compounds that appear to target the N. gonorrhoeae LdcA and LtgD enzymes. By using a reductionist approach involving biological and computational data, we propose that compound Ng-LdcA-16 and Ng-LtgD-45 are promising anti-gonococcal compounds for further development.

背景:淋病奈瑟菌(Ng)会导致性传播疾病淋病。目前还没有疫苗,主要使用抗生素治疗感染。然而,淋球菌会迅速对所有抗生素产生抗药性,因此需要开发新的抗菌治疗方法。在这项研究中,我们将淋球菌的两种酶作为潜在的抗菌目标,即丝氨酸蛋白酶 L,D-羧肽酶 LdcA(NgO1274/NEIS1546)和裂解转糖基酶 LtgD(NgO0626/NEIS1212)。为了找出能与这些酶相互作用的化合物作为潜在的抗菌剂,我们使用了 AtomNet 虚拟高通量筛选技术。然后,我们进行了计算建模研究,以检验最具生物活性的化合物与其目标酶之间的相互作用。我们对鉴定出的化合物进行了淋球菌测试,以确定最小抑菌和杀菌浓度(MIC/MBC)、特异性以及化合物的体外毒性:结果:AtomNet发现了74种可能与Ng-LdcA相互作用的化合物和84种可能与Ng-LtgD相互作用的化合物。通过 MIC 和 MBC 检测,我们选出了对这两种酶活性最好的三种化合物。化合物 16 对 Ng-LdcA 的活性最强,其 MIC50 值为结论:我们发现了似乎能靶向淋球菌 LdcA 和 LtgD 酶的生物活性化合物。通过使用涉及生物和计算数据的还原法,我们认为化合物 Ng-LdcA-16 和 Ng-LtgD-45 是有希望进一步开发的抗淋球菌化合物。
{"title":"Antimicrobial activity of compounds identified by artificial intelligence discovery engine targeting enzymes involved in Neisseria gonorrhoeae peptidoglycan metabolism.","authors":"Ravi Kant, Hannah Tilford, Camila S Freitas, Dayana A Santos Ferreira, James Ng, Gwennan Rucinski, Joshua Watkins, Ryan Pemberton, Tigran M Abramyan, Stephanie C Contreras, Alejandra Vera, Myron Christodoulides","doi":"10.1186/s40659-024-00543-9","DOIUrl":"10.1186/s40659-024-00543-9","url":null,"abstract":"<p><strong>Background: </strong>Neisseria gonorrhoeae (Ng) causes the sexually transmitted disease gonorrhoea. There are no vaccines and infections are treated principally with antibiotics. However, gonococci rapidly develop resistance to every antibiotic class used and there is a need for developing new antimicrobial treatments. In this study we focused on two gonococcal enzymes as potential antimicrobial targets, namely the serine protease L,D-carboxypeptidase LdcA (NgO1274/NEIS1546) and the lytic transglycosylase LtgD (NgO0626/NEIS1212). To identify compounds that could interact with these enzymes as potential antimicrobials, we used the AtomNet virtual high-throughput screening technology. We then did a computational modelling study to examine the interactions of the most bioactive compounds with their target enzymes. The identified compounds were tested against gonococci to determine minimum inhibitory and bactericidal concentrations (MIC/MBC), specificity, and compound toxicity in vitro.</p><p><strong>Results: </strong>AtomNet identified 74 compounds that could potentially interact with Ng-LdcA and 84 compounds that could potentially interact with Ng-LtgD. Through MIC and MBC assays, we selected the three best performing compounds for both enzymes. Compound 16 was the most active against Ng-LdcA, with a MIC50 value < 1.56 µM and MBC50/90 values between 0.195 and 0.39 µM. In general, the Ng-LdcA compounds showed higher activity than the compounds directed against Ng-LtgD, of which compound 45 had MIC50 values of 1.56-3.125 µM and MBC50/90 values between 3.125 and 6.25 µM. The compounds were specific for gonococci and did not kill other bacteria. They were also non-toxic for human conjunctival epithelial cells as judged by a resazurin assay. To support our biological data, in-depth computational modelling study detailed the interactions of the compounds with their target enzymes. Protein models were generated in silico and validated, the active binding sites and amino acids involved elucidated, and the interactions of the compounds interacting with the enzymes visualised through molecular docking and Molecular Dynamics Simulations for 50 ns and Molecular Mechanics Poisson-Boltzmann Surface Area (MM-PBSA).</p><p><strong>Conclusions: </strong>We have identified bioactive compounds that appear to target the N. gonorrhoeae LdcA and LtgD enzymes. By using a reductionist approach involving biological and computational data, we propose that compound Ng-LdcA-16 and Ng-LtgD-45 are promising anti-gonococcal compounds for further development.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"57 1","pages":"62"},"PeriodicalIF":4.3,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11375863/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142139279","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
Hormonal influence: unraveling the impact of sex hormones on vascular smooth muscle cells. 荷尔蒙的影响:揭示性荷尔蒙对血管平滑肌细胞的影响。
IF 4.3 2区 生物学 Q1 BIOLOGY Pub Date : 2024-09-04 DOI: 10.1186/s40659-024-00542-w
Keran Jia, Xin Luo, Jingyan Yi, Chunxiang Zhang

Sex hormones play a pivotal role as endocrine hormones that exert profound effects on the biological characteristics and vascular function of vascular smooth muscle cells (VSMCs). By modulating intracellular signaling pathways, activating nuclear receptors, and regulating gene expression, sex hormones intricately influence the morphology, function, and physiological state of VSMCs, thereby impacting the biological properties of vascular contraction, relaxation, and growth. Increasing evidence suggests that abnormal phenotypic changes in VSMCs contribute to the initiation of vascular diseases, including atherosclerosis. Therefore, understanding the factors governing phenotypic alterations in VSMCs and elucidating the underlying mechanisms can provide crucial insights for refining interventions targeted at vascular diseases. Additionally, the varying levels of different types of sex hormones in the human body, influenced by sex and age, may also affect the phenotypic conversion of VSMCs. This review aims to explore the influence of sex hormones on the phenotypic switching of VSMCs and the development of associated vascular diseases in the human body.

性激素作为一种内分泌激素,对血管平滑肌细胞(VSMC)的生物特性和血管功能有着举足轻重的影响。通过调节细胞内信号通路、激活核受体和调控基因表达,性激素错综复杂地影响着血管平滑肌细胞的形态、功能和生理状态,从而影响血管收缩、舒张和生长的生物学特性。越来越多的证据表明,血管内皮细胞表型的异常变化是包括动脉粥样硬化在内的血管疾病的诱因。因此,了解支配血管内皮细胞表型改变的因素并阐明其潜在机制,可为完善针对血管疾病的干预措施提供重要见解。此外,人体内不同类型的性激素水平受性别和年龄的影响,也可能影响 VSMC 的表型转换。本综述旨在探讨性激素对人体 VSMC 表型转换及相关血管疾病发展的影响。
{"title":"Hormonal influence: unraveling the impact of sex hormones on vascular smooth muscle cells.","authors":"Keran Jia, Xin Luo, Jingyan Yi, Chunxiang Zhang","doi":"10.1186/s40659-024-00542-w","DOIUrl":"10.1186/s40659-024-00542-w","url":null,"abstract":"<p><p>Sex hormones play a pivotal role as endocrine hormones that exert profound effects on the biological characteristics and vascular function of vascular smooth muscle cells (VSMCs). By modulating intracellular signaling pathways, activating nuclear receptors, and regulating gene expression, sex hormones intricately influence the morphology, function, and physiological state of VSMCs, thereby impacting the biological properties of vascular contraction, relaxation, and growth. Increasing evidence suggests that abnormal phenotypic changes in VSMCs contribute to the initiation of vascular diseases, including atherosclerosis. Therefore, understanding the factors governing phenotypic alterations in VSMCs and elucidating the underlying mechanisms can provide crucial insights for refining interventions targeted at vascular diseases. Additionally, the varying levels of different types of sex hormones in the human body, influenced by sex and age, may also affect the phenotypic conversion of VSMCs. This review aims to explore the influence of sex hormones on the phenotypic switching of VSMCs and the development of associated vascular diseases in the human body.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"57 1","pages":"61"},"PeriodicalIF":4.3,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11373308/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142124795","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
Unraveling the impact of hyperleptinemia on female reproduction: insights from transgenic pig model. 揭示高瘦血症对雌性生殖的影响:转基因猪模型的启示。
IF 4.3 2区 生物学 Q1 BIOLOGY Pub Date : 2024-09-04 DOI: 10.1186/s40659-024-00545-7
Muhammad Ameen Jamal, Yixiao Cheng, Deling Jiao, Wen Cheng, Di Zou, Xia Wang, Taiyun Wei, Jianxiong Guo, Kaixiang Xu, Heng Zhao, Shaoxia Pu, Chang Yang, Yubo Qing, Baoyu Jia, Honghui Li, Rusong Zhao, Hong-Ye Zhao, Hong-Jiang Wei

Background: Infertility is a growing global health concern affecting millions of couples worldwide. Among several factors, an extreme body weight adversely affects reproductive functions. Leptin is a well-known adipokine that serves as an endocrine signal between adiposity and fertility. However, the exact mechanisms underlying the effects of high leptin level on female reproduction remain unclear.

Methods: Transgenic pigs overexpressing leptin (♀) were produced by backcrossing and screened for leptin overexpression. The growth curve, fat deposition, reproductive performance, apoptosis, serum hormones and cholesterol production, RNA sequencing, and single-nucleus RNA sequencing (snRNA-seq) of the leptin-overexpressing pigs and wild-type group were evaluated.

Results: Transgenic pigs overexpressing leptin (♀) were obtained, which exhibited significantly reduced body weight, body size, and back fat thickness. These pigs manifested a late onset of puberty (330 ± 54.3 vs. 155 ± 14.7 days), irregular estrous behavior characterized by increased inter-estrous interval (29.2 ± 0 vs. 21.3 ± 0.7 days), and more number of matings until pregnancy (at least 3 times). This reproductive impairment in leptin pigs was related to hormonal imbalances characterized by increased levels of FSH, LH, prolactin, E2, P4, and TSH, altered steroidogenesis such as increased levels of serum cholesterol esters along with steroidogenic markers (StAR, CYP19A), and ovarian dysfunctions manifested by neutrophilic infiltration and low expression of caspase-3 positive cells in the ovaries. Moreover, bulk RNA sequencing of the ovaries also revealed neutrophilic infiltration followed by upregulation of inflammation-related genes. Furthermore, snRNA-seq reflected that leptin overexpression triggered immune response, suppressed follicle development and luteinization, resulting in metabolic dysfunction and hormone imbalance in the ovary.

Conclusions: Low body weight in leptin overexpressing pigs adversely affects the reproductive performance, causing delayed puberty, irregular estrous cycles, and reduced breeding efficiency. This is linked to metabolic imbalances, an increased immune response, and altered ovarian functions. This study provides a theoretical basis for the complex mechanisms underlying leptin, and infertility by employing leptin-overexpressing female pigs.

背景:不孕症是一个日益严重的全球性健康问题,影响着全球数百万对夫妇。在多种因素中,体重过重会对生殖功能产生不利影响。瘦素是一种众所周知的脂肪因子,是脂肪与生育之间的内分泌信号。然而,高瘦素水平影响女性生殖的确切机制仍不清楚:方法:通过回交产生过表达瘦素(♀)的转基因猪,并进行瘦素过表达筛选。结果:过表达瘦素的转基因猪和野生型猪的生长曲线、脂肪沉积、繁殖性能、细胞凋亡、血清激素和胆固醇的产生、RNA测序和单核RNA测序(snRNA-seq)都得到了评估:结果:获得了过表达瘦素(♀)的转基因猪,这些猪的体重、体型和背部脂肪厚度显著降低。这些猪的青春期起始较晚(330 ± 54.3 天 vs. 155 ± 14.7 天),发情行为不规律,发情间隔时间延长(29.2 ± 0 天 vs. 21.3 ± 0.7 天),怀孕前的配种次数增加(至少 3 次)。瘦素猪的繁殖障碍与激素失衡有关,表现为 FSH、LH、催乳素、E2、P4 和 TSH 水平升高;类固醇生成发生改变,如血清胆固醇酯和类固醇生成标记物(StAR、CYP19A)水平升高;卵巢功能失调,表现为中性粒细胞浸润和卵巢中 Caspase-3 阳性细胞的低表达。此外,卵巢的大量 RNA 测序也显示了中性粒细胞浸润,随后炎症相关基因上调。此外,snRNA-seq还反映出瘦素过表达会引发免疫反应,抑制卵泡发育和黄体化,导致卵巢代谢功能障碍和激素失衡:结论:过表达瘦素的猪体重过轻会对繁殖性能产生不利影响,导致青春期延迟、发情周期不规则和繁殖效率降低。这与代谢失衡、免疫反应增强和卵巢功能改变有关。本研究通过使用瘦素过表达的母猪,为瘦素与不孕症的复杂机制提供了理论依据。
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引用次数: 0
Deciphering genetic and nongenetic factors underlying tumour dormancy: insights from multiomics analysis of two syngeneic MRD models of melanoma and leukemia. 解密肿瘤休眠的遗传和非遗传因素:对黑色素瘤和白血病两种共生 MRD 模型进行多组学分析的启示。
IF 4.3 2区 生物学 Q1 BIOLOGY Pub Date : 2024-09-03 DOI: 10.1186/s40659-024-00540-y
Marie-Océane Laguillaumie, Sofia Titah, Aurélie Guillemette, Bernadette Neve, Frederic Leprêtre, Pascaline Ségard, Faruk Azam Shaik, Dominique Collard, Jean-Claude Gerbedoen, Léa Fléchon, Lama Hasan Bou Issa, Audrey Vincent, Martin Figeac, Shéhérazade Sebda, Céline Villenet, Jérôme Kluza, William Laine, Isabelle Fournier, Jean-Pascal Gimeno, Maxence Wisztorski, Salomon Manier, Mehmet Cagatay Tarhan, Bruno Quesnel, Thierry Idziorek, Yasmine Touil

Background: Tumour dormancy, a resistance mechanism employed by cancer cells, is a significant challenge in cancer treatment, contributing to minimal residual disease (MRD) and potential relapse. Despite its clinical importance, the mechanisms underlying tumour dormancy and MRD remain unclear. In this study, we employed two syngeneic murine models of myeloid leukemia and melanoma to investigate the genetic, epigenetic, transcriptomic and protein signatures associated with tumour dormancy. We used a multiomics approach to elucidate the molecular mechanisms driving MRD and identify potential therapeutic targets.

Results: We conducted an in-depth omics analysis encompassing whole-exome sequencing (WES), copy number variation (CNV) analysis, chromatin immunoprecipitation followed by sequencing (ChIP-seq), transcriptome and proteome investigations. WES analysis revealed a modest overlap of gene mutations between melanoma and leukemia dormancy models, with a significant number of mutated genes found exclusively in dormant cells. These exclusive genetic signatures suggest selective pressure during MRD, potentially conferring resistance to the microenvironment or therapies. CNV, histone marks and transcriptomic gene expression signatures combined with Gene Ontology (GO) enrichment analysis highlighted the potential functional roles of the mutated genes, providing insights into the pathways associated with MRD. In addition, we compared "murine MRD genes" profiles to the corresponding human disease through public datasets and highlighted common features according to disease progression. Proteomic analysis combined with multi-omics genetic investigations, revealed a dysregulated proteins signature in dormant cells with minimal genetic mechanism involvement. Pathway enrichment analysis revealed the metabolic, differentiation and cytoskeletal remodeling processes involved in MRD. Finally, we identified 11 common proteins differentially expressed in dormant cells from both pathologies.

Conclusions: Our study underscores the complexity of tumour dormancy, implicating both genetic and nongenetic factors. By comparing genomic, transcriptomic, proteomic, and epigenomic datasets, our study provides a comprehensive understanding of the molecular landscape of minimal residual disease. These results provide a robust foundation for forthcoming investigations and offer potential avenues for the advancement of targeted MRD therapies in leukemia and melanoma patients, emphasizing the importance of considering both genetic and nongenetic factors in treatment strategies.

背景:肿瘤休眠是癌细胞采用的一种抵抗机制,是癌症治疗中的一个重大挑战,会导致最小残留病(MRD)和潜在复发。尽管肿瘤休眠具有重要的临床意义,但肿瘤休眠和 MRD 的机制仍不清楚。在这项研究中,我们采用了骨髓性白血病和黑色素瘤的两种合成小鼠模型来研究与肿瘤休眠相关的遗传学、表观遗传学、转录组学和蛋白质特征。我们采用多组学方法阐明了驱动MRD的分子机制,并确定了潜在的治疗靶点:我们进行了深入的全组学分析,包括全外显子组测序(WES)、拷贝数变异(CNV)分析、染色质免疫沉淀测序(ChIP-seq)、转录组和蛋白质组研究。WES分析显示,黑色素瘤和白血病休眠模型的基因突变略有重叠,大量突变基因只存在于休眠细胞中。这些独有的基因特征表明,在MRD期间存在选择性压力,有可能使细胞对微环境或疗法产生抗药性。CNV、组蛋白标记和转录组基因表达特征与基因本体(GO)富集分析相结合,突显了突变基因的潜在功能作用,为了解与MRD相关的通路提供了线索。此外,我们还通过公共数据集将 "小鼠 MRD 基因 "特征与相应的人类疾病进行了比较,并根据疾病的进展突出了共同特征。蛋白质组分析与多组学遗传学研究相结合,揭示了休眠细胞中蛋白质失调的特征,其中遗传机制的参与度极低。通路富集分析揭示了 MRD 所涉及的代谢、分化和细胞骨架重塑过程。最后,我们确定了两种病理休眠细胞中差异表达的11种常见蛋白质:结论:我们的研究强调了肿瘤休眠的复杂性,涉及遗传和非遗传因素。通过比较基因组、转录组、蛋白质组和表观基因组数据集,我们的研究提供了对极小残留病分子图谱的全面了解。这些结果为今后的研究奠定了坚实的基础,并为推进白血病和黑色素瘤患者的 MRD 靶向治疗提供了潜在的途径,强调了在治疗策略中考虑遗传和非遗传因素的重要性。
{"title":"Deciphering genetic and nongenetic factors underlying tumour dormancy: insights from multiomics analysis of two syngeneic MRD models of melanoma and leukemia.","authors":"Marie-Océane Laguillaumie, Sofia Titah, Aurélie Guillemette, Bernadette Neve, Frederic Leprêtre, Pascaline Ségard, Faruk Azam Shaik, Dominique Collard, Jean-Claude Gerbedoen, Léa Fléchon, Lama Hasan Bou Issa, Audrey Vincent, Martin Figeac, Shéhérazade Sebda, Céline Villenet, Jérôme Kluza, William Laine, Isabelle Fournier, Jean-Pascal Gimeno, Maxence Wisztorski, Salomon Manier, Mehmet Cagatay Tarhan, Bruno Quesnel, Thierry Idziorek, Yasmine Touil","doi":"10.1186/s40659-024-00540-y","DOIUrl":"10.1186/s40659-024-00540-y","url":null,"abstract":"<p><strong>Background: </strong>Tumour dormancy, a resistance mechanism employed by cancer cells, is a significant challenge in cancer treatment, contributing to minimal residual disease (MRD) and potential relapse. Despite its clinical importance, the mechanisms underlying tumour dormancy and MRD remain unclear. In this study, we employed two syngeneic murine models of myeloid leukemia and melanoma to investigate the genetic, epigenetic, transcriptomic and protein signatures associated with tumour dormancy. We used a multiomics approach to elucidate the molecular mechanisms driving MRD and identify potential therapeutic targets.</p><p><strong>Results: </strong>We conducted an in-depth omics analysis encompassing whole-exome sequencing (WES), copy number variation (CNV) analysis, chromatin immunoprecipitation followed by sequencing (ChIP-seq), transcriptome and proteome investigations. WES analysis revealed a modest overlap of gene mutations between melanoma and leukemia dormancy models, with a significant number of mutated genes found exclusively in dormant cells. These exclusive genetic signatures suggest selective pressure during MRD, potentially conferring resistance to the microenvironment or therapies. CNV, histone marks and transcriptomic gene expression signatures combined with Gene Ontology (GO) enrichment analysis highlighted the potential functional roles of the mutated genes, providing insights into the pathways associated with MRD. In addition, we compared \"murine MRD genes\" profiles to the corresponding human disease through public datasets and highlighted common features according to disease progression. Proteomic analysis combined with multi-omics genetic investigations, revealed a dysregulated proteins signature in dormant cells with minimal genetic mechanism involvement. Pathway enrichment analysis revealed the metabolic, differentiation and cytoskeletal remodeling processes involved in MRD. Finally, we identified 11 common proteins differentially expressed in dormant cells from both pathologies.</p><p><strong>Conclusions: </strong>Our study underscores the complexity of tumour dormancy, implicating both genetic and nongenetic factors. By comparing genomic, transcriptomic, proteomic, and epigenomic datasets, our study provides a comprehensive understanding of the molecular landscape of minimal residual disease. These results provide a robust foundation for forthcoming investigations and offer potential avenues for the advancement of targeted MRD therapies in leukemia and melanoma patients, emphasizing the importance of considering both genetic and nongenetic factors in treatment strategies.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"57 1","pages":"59"},"PeriodicalIF":4.3,"publicationDate":"2024-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11370043/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142118990","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
Genetic deletion of ITIH5 leads to increased development of adipose tissue in mice. 基因缺失 ITIH5 会导致小鼠脂肪组织发育加快。
IF 4.3 2区 生物学 Q1 BIOLOGY Pub Date : 2024-08-29 DOI: 10.1186/s40659-024-00530-0
Thomas M Sessler, Justus P Beier, Sophia Villwock, Danny Jonigk, Edgar Dahl, Tim Ruhl

Background: Adipocytokines play a pivotal role in maintaining adipose tissue homeostasis by regulating cellular metabolism, proliferation, differentiation, and secretory activity. These soluble factors are relevant components for healthy adipose tissue, while their deficiency is closely associated with the development of obesity and related metabolic diseases, e.g., chronic inflammation. In human adipose tissue, inter-α-trypsin inhibitor heavy chain 5 (ITIH5) is expressed in proportion to the development of adipose tissue, i.e., the individual's BMI. Thus, ITIH5 has been proposed to be an inert marker of human obesity. However, when applied to adipose stem cells in vitro, recombinant (r)ITIH5 protein inhibited proliferation and adipogenesis, suggesting that ITIH5 negatively affects the development of fat mass. We now tested the role of ITIH5 in vivo and compared ITIH5+/+ wildtype with ITIH5-/- knockout mice.

Results: Genetic deletion of ITIH5 significantly increased adipose tissue mass relative to animal bodyweight (p < 0.05). Next, we characterized adipose stem cells (ASCs) from both genotypes in vitro. ITIH5-/- cells exhibited increased proliferation and adipogenic differentiation (p < 0.001), which could explain the increase in adipose tissue in vivo. Furthermore, ASCs from ITIH5-/- animals were more responsive to stimulation with inflammatory mediators, i.e., these cells released greater amounts of IL-6 and MCP-1 (p < 0.001). Importantly, the application of the rITIH5 protein reversed the observed knockout effects in ASCs.

Conclusions: Our data suggest that ITIH5 potently regulates adipose tissue development and homeostasis by modulating ASC biology in mice. In addition, the effect of the rITIH5 protein underscores its potential as a therapeutic agent to correct the adipose tissue dysregulation often associated with obesity and metabolic disorders.

背景:脂肪细胞因子通过调节细胞的新陈代谢、增殖、分化和分泌活性,在维持脂肪组织平衡方面发挥着关键作用。这些可溶性因子是健康脂肪组织的相关成分,而它们的缺乏则与肥胖和相关代谢疾病(如慢性炎症)的发生密切相关。在人体脂肪组织中,α-胰蛋白酶间抑制物重链 5(ITIH5)的表达与脂肪组织的发育(即个人的体重指数)成正比。因此,ITIH5 被认为是人类肥胖的惰性标志物。然而,当应用于体外脂肪干细胞时,重组(r)ITIH5 蛋白会抑制增殖和脂肪生成,这表明 ITIH5 对脂肪量的发展有负面影响。我们现在测试了 ITIH5 在体内的作用,并比较了 ITIH5+/+ 野生型与 ITIH5-/- 基因敲除小鼠:结果:相对于动物体重,遗传性缺失 ITIH5 会显著增加脂肪组织质量(p -/-细胞表现出增殖和成脂分化增加(p -/-动物对炎症介质刺激的反应更强,即这些细胞释放出更多的 IL-6 和 MCP-1(p 结论):我们的数据表明,ITIH5 可通过调节小鼠 ASC 的生物学特性来有效调节脂肪组织的发育和稳态。此外,rITIH5 蛋白的作用还强调了其作为一种治疗剂的潜力,可纠正通常与肥胖和代谢紊乱相关的脂肪组织失调。
{"title":"Genetic deletion of ITIH5 leads to increased development of adipose tissue in mice.","authors":"Thomas M Sessler, Justus P Beier, Sophia Villwock, Danny Jonigk, Edgar Dahl, Tim Ruhl","doi":"10.1186/s40659-024-00530-0","DOIUrl":"10.1186/s40659-024-00530-0","url":null,"abstract":"<p><strong>Background: </strong>Adipocytokines play a pivotal role in maintaining adipose tissue homeostasis by regulating cellular metabolism, proliferation, differentiation, and secretory activity. These soluble factors are relevant components for healthy adipose tissue, while their deficiency is closely associated with the development of obesity and related metabolic diseases, e.g., chronic inflammation. In human adipose tissue, inter-α-trypsin inhibitor heavy chain 5 (ITIH5) is expressed in proportion to the development of adipose tissue, i.e., the individual's BMI. Thus, ITIH5 has been proposed to be an inert marker of human obesity. However, when applied to adipose stem cells in vitro, recombinant (r)ITIH5 protein inhibited proliferation and adipogenesis, suggesting that ITIH5 negatively affects the development of fat mass. We now tested the role of ITIH5 in vivo and compared ITIH5<sup>+/+</sup> wildtype with ITIH5<sup>-/-</sup> knockout mice.</p><p><strong>Results: </strong>Genetic deletion of ITIH5 significantly increased adipose tissue mass relative to animal bodyweight (p < 0.05). Next, we characterized adipose stem cells (ASCs) from both genotypes in vitro. ITIH5<sup>-/-</sup> cells exhibited increased proliferation and adipogenic differentiation (p < 0.001), which could explain the increase in adipose tissue in vivo. Furthermore, ASCs from ITIH5<sup>-/-</sup> animals were more responsive to stimulation with inflammatory mediators, i.e., these cells released greater amounts of IL-6 and MCP-1 (p < 0.001). Importantly, the application of the rITIH5 protein reversed the observed knockout effects in ASCs.</p><p><strong>Conclusions: </strong>Our data suggest that ITIH5 potently regulates adipose tissue development and homeostasis by modulating ASC biology in mice. In addition, the effect of the rITIH5 protein underscores its potential as a therapeutic agent to correct the adipose tissue dysregulation often associated with obesity and metabolic disorders.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"57 1","pages":"58"},"PeriodicalIF":4.3,"publicationDate":"2024-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11360682/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142092180","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
Inhibition of forward and reverse transport of Ca2+ via Na+/Ca2+ exchangers (NCX) prevents sperm capacitation. 通过 Na+/Ca2+ 交换器(NCX)抑制 Ca2+ 的正向和反向运输可阻止精子获能。
IF 4.3 2区 生物学 Q1 BIOLOGY Pub Date : 2024-08-23 DOI: 10.1186/s40659-024-00535-9
Marc Yeste, Adeel Ahmad, Estel Viñolas, Sandra Recuero, Sergi Bonet, Elisabeth Pinart

Background: While calcium is known to play a crucial role in mammalian sperm physiology, how it flows in and out of the male gamete is not completely understood. Herein, we investigated the involvement of Na+/Ca2+ exchangers (NCX) in mammalian sperm capacitation. Using the pig as an animal model, we first confirmed the presence of NCX1 and NCX2 isoforms in the sperm midpiece. Next, we partially or totally blocked Ca2+ outflux (forward transport) via NCX1/NCX2 with different concentrations of SEA0400 (2-[4-[(2,5-difluorophenyl)methoxy]phenoxy]-5-ethoxyaniline; 0, 0.5, 5 and 50 µM) and Ca2+ influx (reverse transport) with SN6 (ethyl 2-[[4-[(4-nitrophenyl)methoxy]phenyl]methyl]-1,3-thiazolidine-4-carboxylate; 0, 0.3, 3 or 30 µM). Sperm were incubated under capacitating conditions for 180 min; after 120 min, progesterone was added to induce the acrosome reaction. At 0, 60, 120, 130, and 180 min, sperm motility, membrane lipid disorder, acrosome integrity, mitochondrial membrane potential (MMP), tyrosine phosphorylation of sperm proteins, and intracellular levels of Ca2+, reactive oxygen species (ROS) and superoxides were evaluated.

Results: Partial and complete blockage of Ca2+ outflux and influx via NCX induced a significant reduction of sperm motility after progesterone addition. Early alterations on sperm kinematics were also observed, the effects being more obvious in totally blocked than in partially blocked samples. Decreased sperm motility and kinematics were related to both defective tyrosine phosphorylation and mitochondrial activity, the latter being associated to diminished MMP and ROS levels. As NCX blockage did not affect the lipid disorder of plasma membrane, the impaired acrosome integrity could result from reduced tyrosine phosphorylation.

Conclusions: Inhibition of outflux and influx of Ca2+ triggered similar effects, thus indicating that both forward and reverse Ca2+ transport through NCX exchangers are essential for sperm capacitation.

背景:众所周知,钙在哺乳动物精子的生理过程中起着至关重要的作用,但钙如何进出雄性配子却并不完全清楚。在此,我们研究了 Na+/Ca2+ 交换器(NCX)在哺乳动物精子获能过程中的参与。以猪为动物模型,我们首先证实了精子中段存在 NCX1 和 NCX2 同工酶。接着,我们用不同浓度的 SEA0400(2-[4-[(2,5-二氟苯基)甲氧基]苯氧基]-5-乙氧基苯胺;0、0.5、5 和 50 µM)部分或完全阻断了通过 NCX1/NCX2 的 Ca2+ 外流(前向运输)。SN6(2-[[4-[(4-硝基苯基)甲氧基]苯基]甲基]-1,3-噻唑烷-4-甲酸乙酯;0、0.3、3 或 30 µM)。精子在获能条件下孵育 180 分钟;120 分钟后,加入黄体酮以诱导顶体反应。在 0、60、120、130 和 180 分钟内,对精子的运动能力、膜脂紊乱、顶体完整性、线粒体膜电位(MMP)、精子蛋白质的酪氨酸磷酸化以及细胞内 Ca2+、活性氧(ROS)和超氧化物的水平进行了评估:结果:部分和完全阻断 Ca2+ 通过 NCX 的外流和内流可导致精子活力在黄体酮添加后显著下降。还观察到精子运动学的早期改变,完全阻断比部分阻断样本的影响更明显。精子运动能力和运动学特征的降低与酪氨酸磷酸化和线粒体活性的缺陷有关,后者与 MMP 和 ROS 水平的降低有关。由于NCX阻断并不影响质膜脂质紊乱,顶体完整性受损可能是酪氨酸磷酸化减少所致:结论:抑制Ca2+的外流和流入会引发相似的效应,因此表明通过NCX交换器进行的Ca2+正向和反向运输对精子获能至关重要。
{"title":"Inhibition of forward and reverse transport of Ca<sup>2+</sup> via Na<sup>+</sup>/Ca<sup>2+</sup> exchangers (NCX) prevents sperm capacitation.","authors":"Marc Yeste, Adeel Ahmad, Estel Viñolas, Sandra Recuero, Sergi Bonet, Elisabeth Pinart","doi":"10.1186/s40659-024-00535-9","DOIUrl":"10.1186/s40659-024-00535-9","url":null,"abstract":"<p><strong>Background: </strong>While calcium is known to play a crucial role in mammalian sperm physiology, how it flows in and out of the male gamete is not completely understood. Herein, we investigated the involvement of Na<sup>+</sup>/Ca<sup>2+</sup> exchangers (NCX) in mammalian sperm capacitation. Using the pig as an animal model, we first confirmed the presence of NCX1 and NCX2 isoforms in the sperm midpiece. Next, we partially or totally blocked Ca<sup>2+</sup> outflux (forward transport) via NCX1/NCX2 with different concentrations of SEA0400 (2-[4-[(2,5-difluorophenyl)methoxy]phenoxy]-5-ethoxyaniline; 0, 0.5, 5 and 50 µM) and Ca<sup>2+</sup> influx (reverse transport) with SN6 (ethyl 2-[[4-[(4-nitrophenyl)methoxy]phenyl]methyl]-1,3-thiazolidine-4-carboxylate; 0, 0.3, 3 or 30 µM). Sperm were incubated under capacitating conditions for 180 min; after 120 min, progesterone was added to induce the acrosome reaction. At 0, 60, 120, 130, and 180 min, sperm motility, membrane lipid disorder, acrosome integrity, mitochondrial membrane potential (MMP), tyrosine phosphorylation of sperm proteins, and intracellular levels of Ca<sup>2+</sup>, reactive oxygen species (ROS) and superoxides were evaluated.</p><p><strong>Results: </strong>Partial and complete blockage of Ca<sup>2+</sup> outflux and influx via NCX induced a significant reduction of sperm motility after progesterone addition. Early alterations on sperm kinematics were also observed, the effects being more obvious in totally blocked than in partially blocked samples. Decreased sperm motility and kinematics were related to both defective tyrosine phosphorylation and mitochondrial activity, the latter being associated to diminished MMP and ROS levels. As NCX blockage did not affect the lipid disorder of plasma membrane, the impaired acrosome integrity could result from reduced tyrosine phosphorylation.</p><p><strong>Conclusions: </strong>Inhibition of outflux and influx of Ca<sup>2+</sup> triggered similar effects, thus indicating that both forward and reverse Ca<sup>2+</sup> transport through NCX exchangers are essential for sperm capacitation.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"57 1","pages":"57"},"PeriodicalIF":4.3,"publicationDate":"2024-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11342557/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142035208","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
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