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Beyond Lux: methods for species and photoreceptor-specific quantification of ambient light for mammals. 超越 Lux:针对哺乳动物物种和感光器的环境光量化方法。
IF 4.4 1区 生物学
BMC Biology Pub Date : 2024-11-14 DOI: 10.1186/s12915-024-02038-1
Richard J McDowell, Altug Didikoglu, Tom Woelders, Mazie J Gatt, Finn Moffatt, Saba Notash, Roelof A Hut, Timothy M Brown, Robert J Lucas
{"title":"Beyond Lux: methods for species and photoreceptor-specific quantification of ambient light for mammals.","authors":"Richard J McDowell, Altug Didikoglu, Tom Woelders, Mazie J Gatt, Finn Moffatt, Saba Notash, Roelof A Hut, Timothy M Brown, Robert J Lucas","doi":"10.1186/s12915-024-02038-1","DOIUrl":"10.1186/s12915-024-02038-1","url":null,"abstract":"<p><strong>Background: </strong>Light is a key environmental regulator of physiology and behaviour. Mistimed or insufficient light disrupts circadian rhythms and is associated with impaired health and well-being across mammals. Appropriate lighting is therefore crucial for indoor housed mammals. Light is commonly measured in lux. However, this employs a spectral weighting function for human luminance and is not suitable for 'non-visual' effects of light or use across species. In humans, a photoreceptor-specific (α-opic) metrology system has been proposed as a more appropriate way of measuring light.</p><p><strong>Results: </strong>Here we establish technology to allow this α-opic measurement approach to be readily extended across mammalian species, accounting for differences in photoreceptor types, photopigment spectral sensitivities, and eye anatomy. We develop a high-throughput method to derive spectral sensitivities for recombinantly expressed mammalian opsins and use it to establish the spectral sensitivity of melanopsin from 13 non-human mammals. We further address the need for simple measurement strategies for species-specific α-opic measures by developing an accessible online toolbox for calculating these units and validating an open hardware multichannel light sensor for 'point and click' measurement. We finally demonstrate that species-specific α-opic measurements are superior to photopic lux as predictors of physiological responses to light in mice and allow ecologically relevant comparisons of photosensitivity between species.</p><p><strong>Conclusions: </strong>Our study presents methods for measuring light in species-specific α-opic units that are superior to the existing unit of photopic lux and holds the promise of improvements to the health and welfare of animals, scientific research reproducibility, agricultural productivity, and energy usage.</p>","PeriodicalId":9339,"journal":{"name":"BMC Biology","volume":"22 1","pages":"257"},"PeriodicalIF":4.4,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11562817/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142615249","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
T4Seeker: a hybrid model for type IV secretion effectors identification. T4Seeker:用于识别 IV 型分泌效应物的混合模型。
IF 4.4 1区 生物学
BMC Biology Pub Date : 2024-11-14 DOI: 10.1186/s12915-024-02064-z
Jing Li, Shida He, Jian Zhang, Feng Zhang, Quan Zou, Fengming Ni
{"title":"T4Seeker: a hybrid model for type IV secretion effectors identification.","authors":"Jing Li, Shida He, Jian Zhang, Feng Zhang, Quan Zou, Fengming Ni","doi":"10.1186/s12915-024-02064-z","DOIUrl":"10.1186/s12915-024-02064-z","url":null,"abstract":"<p><strong>Background: </strong>The type IV secretion system is widely present in various bacteria, such as Salmonella, Escherichia coli, and Helicobacter pylori. These bacteria use the type IV secretion system to secrete type IV secretion effectors, infect host cells, and disrupt or modulate the communication pathways. In this study, type III and type VI secretion effectors were used as negative samples to train a robust model.</p><p><strong>Results: </strong>The area under the curve of T4Seeker on the validation and independent test sets were 0.947 and 0.970, respectively, demonstrating the strong predictive capacity and robustness of T4Seeker. After comparing with the classic and state-of-the-art T4SE identification models, we found that T4Seeker, which is based on traditional features and large language model features, had a higher predictive ability.</p><p><strong>Conclusion: </strong>The T4Seeker proposed in this study demonstrates superior performance in the field of T4SEs prediction. By integrating features at multiple levels, it achieves higher predictive accuracy and strong generalization capability, providing an effective tool for future T4SE research.</p>","PeriodicalId":9339,"journal":{"name":"BMC Biology","volume":"22 1","pages":"259"},"PeriodicalIF":4.4,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11566746/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142615255","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
CRBPSA: CircRNA-RBP interaction sites identification using sequence structural attention model. CRBPSA:利用序列结构关注模型识别 CircRNA-RBP 相互作用位点。
IF 4.4 1区 生物学
BMC Biology Pub Date : 2024-11-14 DOI: 10.1186/s12915-024-02055-0
Chao Cao, Chunyu Wang, Qi Dai, Quan Zou, Tao Wang
{"title":"CRBPSA: CircRNA-RBP interaction sites identification using sequence structural attention model.","authors":"Chao Cao, Chunyu Wang, Qi Dai, Quan Zou, Tao Wang","doi":"10.1186/s12915-024-02055-0","DOIUrl":"10.1186/s12915-024-02055-0","url":null,"abstract":"<p><strong>Background: </strong>Due to the ability of circRNA to bind with corresponding RBPs and play a critical role in gene regulation and disease prevention, numerous identification algorithms have been developed. Nevertheless, most of the current mainstream methods primarily capture one-dimensional sequence features through various descriptors, while neglecting the effective extraction of secondary structure features. Moreover, as the number of introduced descriptors increases, the issues of sparsity and ineffective representation also rise, causing a significant burden on computational models and leaving room for improvement in predictive performance.</p><p><strong>Results: </strong>Based on this, we focused on capturing the features of secondary structure in sequences and developed a new architecture called CRBPSA, which is based on a sequence-structure attention mechanism. Firstly, a base-pairing matrix is generated by calculating the matching probability between each base, with a Gaussian function introduced as a weight to construct the secondary structure. Then, a Structure_Transformer is employed to extract base-pairing information and spatial positional dependencies, enabling the identification of binding sites through deeper feature extraction. Experimental results using the same set of hyperparameters on 37 circRNA datasets, totaling 671,952 samples, show that the CRBPSA algorithm achieves an average AUC of 99.93%, surpassing all existing prediction methods.</p><p><strong>Conclusions: </strong>CRBPSA is a lightweight and efficient prediction tool for circRNA-RBP, which can capture structural features of sequences with minimal computational resources and accurately predict protein-binding sites. This tool facilitates a deeper understanding of the biological processes and mechanisms underlying circRNA and protein interactions.</p>","PeriodicalId":9339,"journal":{"name":"BMC Biology","volume":"22 1","pages":"260"},"PeriodicalIF":4.4,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11566611/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142615252","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
Uncovering bacterial-mammalian cell interactions via single-cell tracking. 通过单细胞追踪揭示细菌与哺乳动物细胞之间的相互作用
IF 4.4 1区 生物学
BMC Biology Pub Date : 2024-11-11 DOI: 10.1186/s12915-024-02056-z
Narendra K Dewangan, Sayed Golam Mohiuddin, Shayne Sensenbach, Prashant Karki, Mehmet A Orman
{"title":"Uncovering bacterial-mammalian cell interactions via single-cell tracking.","authors":"Narendra K Dewangan, Sayed Golam Mohiuddin, Shayne Sensenbach, Prashant Karki, Mehmet A Orman","doi":"10.1186/s12915-024-02056-z","DOIUrl":"10.1186/s12915-024-02056-z","url":null,"abstract":"<p><strong>Background: </strong>The interactions between bacterial pathogens and host cells are characterized by a multitude of complexities, leading to a wide range of heterogeneous outcomes. Despite extensive research, we still have a limited understanding of how bacterial motility in complex environments impacts their ability to tolerate antibiotics and adhere to mammalian cell surfaces. The challenge lies in unraveling the complexity of these interactions and developing quantitative microscopy approaches to predict the behavior of bacterial populations.</p><p><strong>Results: </strong>To address this challenge, we directed our efforts towards Pseudomonas aeruginosa, a pathogenic bacterium known for producing thick films in the lungs of cystic fibrosis patients, and Escherichia coli, used as a proof of concept to develop and demonstrate our single-cell tracking approaches. Our results revealed that P. aeruginosa exhibits diverse and complex interactions on mammalian cell surfaces, such as adhesion, rotational motion, and swimming, unlike the less interactive behavior of Escherichia coli. Our analysis indicated that P. aeruginosa demonstrated lower mean-squared displacement (MSD) values and greater adherence to mammalian cells compared to E. coli, which showed higher MSD slopes and less frequent adherence. Genetic mutations in membrane proteins of P. aeruginosa resulted in altered displacement patterns and reduced adhesion, with the ΔfliD mutant displaying a more Gaussian displacement distribution and significantly less adherence to mammalian cells. Adhesion and tolerance mechanisms are diverse and complex, potentially involving distinct pathways; however, our findings highlight the therapeutic potential of targeting the fliD gene (encoding a critical flagellum protein), as its deletion not only reduced adherence but also antibiotic tolerance.</p><p><strong>Conclusions: </strong>Overall, our findings underscore the importance of single cell tracking in accurately assessing bacterial behavior over short time periods and highlight its significant potential in guiding effective intervention strategies.</p>","PeriodicalId":9339,"journal":{"name":"BMC Biology","volume":"22 1","pages":"256"},"PeriodicalIF":4.4,"publicationDate":"2024-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11552363/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142615259","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
Widespread 3'UTR capped RNAs derive from G-rich regions in proximity to AGO2 binding sites. 广泛存在的 3'UTR 带帽 RNA 来自 AGO2 结合位点附近的富含 G 的区域。
IF 4.4 1区 生物学
BMC Biology Pub Date : 2024-11-07 DOI: 10.1186/s12915-024-02032-7
Nejc Haberman, Holly Digby, Rupert Faraway, Rebecca Cheung, Anob M Chakrabarti, Andrew M Jobbins, Callum Parr, Kayoko Yasuzawa, Takeya Kasukawa, Chi Wai Yip, Masaki Kato, Hazuki Takahashi, Piero Carninci, Santiago Vernia, Jernej Ule, Christopher R Sibley, Aida Martinez-Sanchez, Boris Lenhard
{"title":"Widespread 3'UTR capped RNAs derive from G-rich regions in proximity to AGO2 binding sites.","authors":"Nejc Haberman, Holly Digby, Rupert Faraway, Rebecca Cheung, Anob M Chakrabarti, Andrew M Jobbins, Callum Parr, Kayoko Yasuzawa, Takeya Kasukawa, Chi Wai Yip, Masaki Kato, Hazuki Takahashi, Piero Carninci, Santiago Vernia, Jernej Ule, Christopher R Sibley, Aida Martinez-Sanchez, Boris Lenhard","doi":"10.1186/s12915-024-02032-7","DOIUrl":"10.1186/s12915-024-02032-7","url":null,"abstract":"<p><p>The 3' untranslated region (3'UTR) plays a crucial role in determining mRNA stability, localisation, translation and degradation. Cap analysis of gene expression (CAGE), a method for the detection of capped 5' ends of mRNAs, additionally reveals a large number of apparently 5' capped RNAs derived from locations within the body of the transcript, including 3'UTRs. Here, we provide direct evidence that these 3'UTR-derived RNAs are indeed capped and widespread in mammalian cells. By using a combination of AGO2 enhanced individual nucleotide resolution UV crosslinking and immunoprecipitation (eiCLIP) and CAGE following siRNA treatment, we find that these 3'UTR-derived RNAs likely originate from AGO2-binding sites, and most often occur at locations with G-rich motifs bound by the RNA-binding protein UPF1. High-resolution imaging and long-read sequencing analysis validate several 3'UTR-derived RNAs, showcase their variable abundance and show that they may not co-localise with the parental mRNAs. Taken together, we provide new insights into the origin and prevalence of 3'UTR-derived RNAs, show the utility of CAGE-seq for their genome-wide detection and provide a rich dataset for exploring new biology of a poorly understood new class of RNAs.</p>","PeriodicalId":9339,"journal":{"name":"BMC Biology","volume":"22 1","pages":"254"},"PeriodicalIF":4.4,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11546257/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142602319","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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-resolution chromosome-level genome of Scylla paramamosain provides molecular insights into adaptive evolution in crabs. 高分辨率的副螯蟹(Scylla paramamosain)染色体级基因组为了解螃蟹的适应性进化提供了分子信息。
IF 4.4 1区 生物学
BMC Biology Pub Date : 2024-11-07 DOI: 10.1186/s12915-024-02054-1
Yin Zhang, Ye Yuan, Mengqian Zhang, Xiaoyan Yu, Bixun Qiu, Fangchun Wu, Douglas R Tocher, Jiajia Zhang, Shaopan Ye, Wenxiao Cui, Jonathan Y S Leung, Mhd Ikhwanuddin, Waqas Waqas, Tariq Dildar, Hongyu Ma
{"title":"High-resolution chromosome-level genome of Scylla paramamosain provides molecular insights into adaptive evolution in crabs.","authors":"Yin Zhang, Ye Yuan, Mengqian Zhang, Xiaoyan Yu, Bixun Qiu, Fangchun Wu, Douglas R Tocher, Jiajia Zhang, Shaopan Ye, Wenxiao Cui, Jonathan Y S Leung, Mhd Ikhwanuddin, Waqas Waqas, Tariq Dildar, Hongyu Ma","doi":"10.1186/s12915-024-02054-1","DOIUrl":"10.1186/s12915-024-02054-1","url":null,"abstract":"<p><strong>Background: </strong>Evolutionary adaptation drives organismal adjustments to environmental pressures, exemplified in the diverse morphological and ecological adaptations seen in Decapoda crustaceans, particularly brachyuran crabs. Crabs thrive in diverse ecosystems, from coral reefs to hydrothermal vents and terrestrial habitats. Despite their ecological importance, the genetic mechanisms underpinning their developmental processes, reproductive strategies, and nutrient acquisition remain poorly understood.</p><p><strong>Results: </strong>Here, we report a comprehensive genomic analysis of the green mud crab Scylla paramamosain using ultralong sequencing technologies, achieving a high-quality chromosome-level assembly. The refined 1.21 Gb genome, with an impressive contig N50 of 11.45 Mb, offers a valuable genomic resource. The genome exhibits 33,662 protein-coding genes, enriched in various pathways related to development and environmental adaptation. Gene family analysis shows expansion in development-related pathways and contraction in metabolic pathways, indicating niche adaptations. Notably, investigation into Hox gene regulation sheds light on their role in pleopod development, with the Abd-A gene identified as a linchpin. Post-transcriptional regulation involving novel-miR1317 negatively regulates Abd-A levels. Furthermore, the potential role of fru gene in ovarian development and the identification of novel-miR35 as a regulator of Spfru2 add complexity to gene regulatory networks. Comparative functional analysis across Decapoda species reveals neo-functionalization of the elovl6 gene in the synthesis of long-chain polyunsaturated fatty acids (LC-PUFA), suggesting its importance in environmental adaptation.</p><p><strong>Conclusions: </strong>Our findings shed light on various aspects of crab biology, including genome sequencing, assembly, and annotation, as well as gene family expansion, contraction, and regulatory mechanisms governing crucial developmental processes such as metamorphosis, reproductive strategies, and fatty acid metabolism.</p>","PeriodicalId":9339,"journal":{"name":"BMC Biology","volume":"22 1","pages":"255"},"PeriodicalIF":4.4,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11545969/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142602312","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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 function of single-target regulator NorR involved in swarming motility and biofilm formation revealed in Vibrio alginolyticus. 揭示了藻溶弧菌中参与蜂拥运动和生物膜形成的单目标调节器 NorR 的新功能。
IF 4.4 1区 生物学
BMC Biology Pub Date : 2024-11-06 DOI: 10.1186/s12915-024-02057-y
Tongxian Chen, Xiaoling Zhou, Ruonan Feng, Shuhao Shi, Xiyu Chen, Bingqi Wei, Zhong Hu, Tao Peng
{"title":"Novel function of single-target regulator NorR involved in swarming motility and biofilm formation revealed in Vibrio alginolyticus.","authors":"Tongxian Chen, Xiaoling Zhou, Ruonan Feng, Shuhao Shi, Xiyu Chen, Bingqi Wei, Zhong Hu, Tao Peng","doi":"10.1186/s12915-024-02057-y","DOIUrl":"10.1186/s12915-024-02057-y","url":null,"abstract":"<p><p>NorR, as a single-target regulator, has been demonstrated to be involved in NO detoxification in bacteria under anaerobic conditions. Here, the norR gene was identified and deleted in the genome of Vibrio alginolyticus. The results showed that deletion of norR in Vibrio alginolyticus led to lower swarming motility and more biofilm formation on aerobic condition. Moreover, we proved that NorR from E. coli had a similar function in controlling motility. NorR overexpression led to increased resistance to oxidative stress and tetracycline. We also observed a reduced ability of the NorR-overexpressing strain to adapt to iron limitation condition. Transcriptome analysis showed that the genes responsible for bacterial motility and biofilm formation were affected by NorR. The expressions of several sigma factors (RpoS, RpoN, and RpoH) and response regulators (LuxR and MarR) were also controlled by NorR. Furthermore, Chip-qPCR showed that there is a direct binding between NorR and the promoter of rpoS. Based on these results, NorR appears to be a central regulator involved in biofilm formation and swarming motility in Vibrio alginolyticus.</p>","PeriodicalId":9339,"journal":{"name":"BMC Biology","volume":"22 1","pages":"253"},"PeriodicalIF":4.4,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11542441/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142589391","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
Hibernation reduces GABA signaling in the brainstem to enhance motor activity of breathing at cool temperatures. 冬眠会减少脑干中的 GABA 信号,从而在低温条件下增强呼吸运动。
IF 4.4 1区 生物学
BMC Biology Pub Date : 2024-11-04 DOI: 10.1186/s12915-024-02050-5
Sandy E Saunders, Joseph M Santin
{"title":"Hibernation reduces GABA signaling in the brainstem to enhance motor activity of breathing at cool temperatures.","authors":"Sandy E Saunders, Joseph M Santin","doi":"10.1186/s12915-024-02050-5","DOIUrl":"10.1186/s12915-024-02050-5","url":null,"abstract":"<p><strong>Background: </strong>Neural circuits produce reliable activity patterns despite disturbances in the environment. For this to occur, neurons elicit synaptic plasticity during perturbations. However, recent work suggests that plasticity not only regulates circuit activity during disturbances, but these modifications may also linger to stabilize circuits during future perturbations. The implementation of such a regulation scheme for real-life environmental challenges of animals remains unclear. Amphibians provide insight into this problem in a rather extreme way, as circuits that generate breathing are inactive for several months during underwater hibernation and use compensatory plasticity to promote ventilation upon emergence.</p><p><strong>Results: </strong>Using ex vivo brainstem preparations and electrophysiology, we find that hibernation in American bullfrogs reduces GABA<sub>A</sub> receptor (GABA<sub>A</sub>R) inhibition in respiratory rhythm generating circuits and motor neurons, consistent with a compensatory response to chronic inactivity. Although GABA<sub>A</sub>Rs are normally critical for breathing, baseline network output at warm temperatures was not affected. However, when assessed across a range of temperatures, hibernators with reduced GABA<sub>A</sub>R signaling had greater activity at cooler temperatures, enhancing respiratory motor output under conditions that otherwise strongly depress breathing.</p><p><strong>Conclusions: </strong>Hibernation reduces GABA<sub>A</sub>R signaling to promote robust respiratory output only at cooler temperatures. Although frogs do not ventilate lungs during underwater hibernation, we suggest this would be beneficial for stabilizing breathing when the animal passes through a large temperature range during emergence in the spring. More broadly, these results demonstrate that compensatory synaptic plasticity can increase the operating range of circuits in harsh environments, thereby promoting adaptive behavior in conditions that suppress activity.</p>","PeriodicalId":9339,"journal":{"name":"BMC Biology","volume":"22 1","pages":"251"},"PeriodicalIF":4.4,"publicationDate":"2024-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11533357/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142574836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
A powerful and versatile new fixation protocol for immunostaining and in situ hybridization that preserves delicate tissues. 用于免疫染色和原位杂交的功能强大、用途广泛的新固定方案,可保护脆弱的组织。
IF 4.4 1区 生物学
BMC Biology Pub Date : 2024-11-04 DOI: 10.1186/s12915-024-02052-3
Carlos Guerrero-Hernández, Viraj Doddihal, Frederick G Mann, Alejandro Sánchez Alvarado
{"title":"A powerful and versatile new fixation protocol for immunostaining and in situ hybridization that preserves delicate tissues.","authors":"Carlos Guerrero-Hernández, Viraj Doddihal, Frederick G Mann, Alejandro Sánchez Alvarado","doi":"10.1186/s12915-024-02052-3","DOIUrl":"10.1186/s12915-024-02052-3","url":null,"abstract":"<p><strong>Background: </strong>Understanding how genes function to heal wounds and restore lost tissue is essential for studying regeneration. Whole-mount in situ hybridization (WISH) is a powerful and widely used technique to visualize the expression patterns of genes in different biological systems. Yet, existing methods to permeabilize samples for WISH can damage or destroy fragile regenerating tissues, thereby preventing such experiments.</p><p><strong>Results: </strong>Here, we describe a new protocol for in situ hybridization (ISH) and immunostaining in the highly regenerative planarian Schmidtea mediterranea. This new Nitric Acid/Formic Acid (NAFA) protocol is compatible with both the assays and prevents degradation of the epidermis and regeneration blastema. The NAFA protocol achieves this without the use of proteinase K digestion which likely leads to better preservation of antigen epitopes. We show that the NAFA protocol successfully permits development of chromogenic and fluorescent signals in situ, while preserving the anatomy of the animal. Furthermore, the immunostaining of different proteins was compatible with the NAFA protocol following fluorescent in situ hybridization. Additionally, the tissue fixation protocol was easily adapted for regenerating killifish tail fin, which yielded better ISH signal with minimal background.</p><p><strong>Conclusions: </strong>Thus, the NAFA protocol robustly preserves the delicate wounded tissues while also facilitating probe and antibody penetration into internal tissues. Furthermore, the fixation protocol is compatible for WISH on regenerating teleost fins suggesting that it will be a valuable technique for studying the processes of wounding response and regeneration in multiple species.</p>","PeriodicalId":9339,"journal":{"name":"BMC Biology","volume":"22 1","pages":"252"},"PeriodicalIF":4.4,"publicationDate":"2024-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11533299/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142574917","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
Evolutionary divergent clusters of transcribed extinct truncated retroposons drive low mRNA expression and developmental regulation in the protozoan Leishmania. 原生动物利什曼原虫中转录已灭绝的截短反转录子的进化分化群驱动低 mRNA 表达和发育调控。
IF 4.4 1区 生物学
BMC Biology Pub Date : 2024-10-29 DOI: 10.1186/s12915-024-02051-4
Gabriel Reis Ferreira, Jean-Guillaume Emond-Rheault, Lysangela Alves, Philippe Leprohon, Martin A Smith, Barbara Papadopoulou
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