Matrix Biology Plus最新文献

{"title":"Laminin-derived peptide, IKVAV, modulates macrophage phenotype through integrin mediation","authors":"Aakanksha Jha ,&nbsp;Erika Moore","doi":"10.1016/j.mbplus.2024.100143","DOIUrl":"10.1016/j.mbplus.2024.100143","url":null,"abstract":"<div><p>Macrophages are highly plastic immune cells known to exist on a spectrum of phenotypes including pro-inflammatory (M1) or pro-healing (M2). Macrophages interact with extracellular matrix (ECM) ligands, such as fragments of collagen and laminin. Interaction of macrophages with ECM ligands is mediated through integrin receptors. However, the role of ECM ligands in directing macrophage function through integrins is not yet fully understood. Particularly, α2β1 has been implicated in modulating macrophage function, but complexity in mechanisms employed for integrin-ligation especially with laminin-derived peptides makes it challenging to understand macrophage-ECM interactions. We hypothesize that targeting α2β1 through laminin-derived peptide, IKVAV, will modulate macrophage phenotype. In this work we: i) investigated macrophage response to IKVAV in 2D and in a 3D platform, and ii) identified α2β1′s role as it pertains to macrophage modulation via IKVAV. Soluble IKVAV treatment significantly reduced M1 markers and increased M2 markers via immunocytochemistry and gene expression. While the 3D ECM-mimicking PEG-IKVAV hydrogels did not have significant effects in modulating macrophage phenotype, we found that macrophage modulation via IKVAV is dependent on the concentration of peptide used and duration of exposure. To investigate integrin-ligand interactions for macrophages, α2β1 signaling was modulated by antagonists and agonists. We observed that blocking α2β1 reduces M1 activation. To understand integrin-ligand interactions and leveraging the therapeutic ability of macrophages in designing immunomodulatory solutions, it is critical to elucidate IKVAV’s role in mediating macrophage phenotype.</p></div>","PeriodicalId":52317,"journal":{"name":"Matrix Biology Plus","volume":"22 ","pages":"Article 100143"},"PeriodicalIF":0.0,"publicationDate":"2024-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590028524000036/pdfft?md5=9fc044be5e1e4dc4a1f095bb96b9b049&pid=1-s2.0-S2590028524000036-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139882274","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A preliminary study into the emergence of tendon microstructure during postnatal development","authors":"Helena Raymond-Hayling ,&nbsp;Yinhui Lu ,&nbsp;Tom Shearer ,&nbsp;Karl Kadler","doi":"10.1016/j.mbplus.2024.100142","DOIUrl":"10.1016/j.mbplus.2024.100142","url":null,"abstract":"<div><p>Tendons maintain mechanical function throughout postnatal development whilst undergoing significant microstructural changes. We present a study of postnatal tendon growth and characterise the major changes in collagen fibril architecture in mouse tail tendon from birth to eight weeks by analysing the geometries of cross-sectional transmission electron microscopy images. This study finds that a bimodal distribution of fibril diameters emerges from a unimodal distribution of narrow fibrils as early as the eighth day postnatal, and three distinct fibril populations are visible at around 14 days. The tendons in this study do not show evidence of precise hexagonal packing, even at birth, and the spaces between the fibrils remain constant throughout development. The fibril number in the tissue stabilises around day 28, and the fibril area fraction stabilises around day 26. This study gives coarse-grained insight into the transition periods in early tendon development.</p></div>","PeriodicalId":52317,"journal":{"name":"Matrix Biology Plus","volume":"21 ","pages":"Article 100142"},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590028524000024/pdfft?md5=45dd56b1c29032ce3e1911900a9f3479&pid=1-s2.0-S2590028524000024-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139637217","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Proteomic analysis of the extracellular matrix of human atherosclerotic plaques shows marked changes between plaque types","authors":"Lasse G. Lorentzen ,&nbsp;Karin Yeung ,&nbsp;Nikolaj Eldrup ,&nbsp;Jonas P. Eiberg ,&nbsp;Henrik H. Sillesen ,&nbsp;Michael J. Davies","doi":"10.1016/j.mbplus.2024.100141","DOIUrl":"10.1016/j.mbplus.2024.100141","url":null,"abstract":"<div><p>Cardiovascular disease is the leading cause of death, with atherosclerosis the major underlying cause. While often asymptomatic for decades, atherosclerotic plaque destabilization and rupture can arise suddenly and cause acute arterial occlusion or peripheral embolization resulting in myocardial infarction, stroke and lower limb ischaemia. As extracellular matrix (ECM) remodelling is associated with plaque instability, we hypothesized that the ECM composition would differ between plaques. We analyzed atherosclerotic plaques obtained from 21 patients who underwent carotid surgery following recent symptomatic carotid artery stenosis. Plaques were solubilized using a new efficient, single-step approach. Solubilized proteins were digested to peptides, and analyzed by liquid chromatography-mass spectrometry using data-independent acquisition. Identification and quantification of 4498 plaque proteins was achieved, including 354 ECM proteins, with unprecedented coverage and high reproducibility. Multidimensional scaling analysis and hierarchical clustering indicate two distinct clusters, which correlate with macroscopic plaque morphology (soft/unstable versus hard/stable), ultrasound classification (echolucent versus echogenic) and the presence of hemorrhage/ulceration. We identified 714 proteins with differential abundances between these groups. Soft/unstable plaques were enriched in proteins involved in inflammation, ECM remodelling, and protein degradation (e.g. matrix metalloproteinases, cathepsins). In contrast, hard/stable plaques contained higher levels of ECM structural proteins (e.g. collagens, versican, nidogens, biglycan, lumican, proteoglycan 4, mineralization proteins). These data indicate that a single-step proteomics method can provide unique mechanistic insights into ECM remodelling and inflammatory mechanisms within plaques that correlate with clinical parameters, and help rationalize plaque destabilization. These data also provide an approach towards identifying biomarkers for individualized risk profiling of atherosclerosis.</p></div>","PeriodicalId":52317,"journal":{"name":"Matrix Biology Plus","volume":"21 ","pages":"Article 100141"},"PeriodicalIF":0.0,"publicationDate":"2024-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590028524000012/pdfft?md5=0cf5c67aa0b3306c6014e70a8011d47f&pid=1-s2.0-S2590028524000012-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139456080","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of an in vitro engineered ligament model","authors":"Alec M. Avey ,&nbsp;Omar Valdez ,&nbsp;Keith Baar","doi":"10.1016/j.mbplus.2023.100140","DOIUrl":"https://doi.org/10.1016/j.mbplus.2023.100140","url":null,"abstract":"<div><p><em>In vivo</em> tendon and ligament research can be limited by the difficultly of obtaining tissue samples that can be biochemically analyzed. In this study, we characterize the most widely used <em>in vitro</em> engineered ligament model. Despite previous works suggesting multiple passages change gene expression in 2D primary tenocytes, we found no relationship between passage number and expression of classical tendon fibroblast markers across different biological donors. When engineered into 3D ligaments, there was an increase in maximal tensile load between 7 and 14 days in culture, that corresponded with an increase in collagen content. By contrast, percent collagen increased logarithmically from Day 7 to Day 14, and this was similar to the increase in the modulus of the tissue. Importantly, there was no relationship between passage number and mechanical function or collagen content in the two independent donors tested. These results suggest that the model develops quickly and is reliable across differing passage numbers. This provides the field with the ability to 1) consistently determine functional changes of interventions out to passage number 10; and 2) to time interventions to the appropriate developmental stage: developing/regenerating (Day 7) or mature (Day 14) tissue.</p></div>","PeriodicalId":52317,"journal":{"name":"Matrix Biology Plus","volume":"21 ","pages":"Article 100140"},"PeriodicalIF":0.0,"publicationDate":"2023-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590028523000133/pdfft?md5=b6d704769d86e1e0c11806088a61213c&pid=1-s2.0-S2590028523000133-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139100373","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Designing collagens to shed light on the multi-scale structure–function mapping of matrix disorders","authors":"Sonal Gahlawat ,&nbsp;Vikas Nanda ,&nbsp;David I. Shreiber","doi":"10.1016/j.mbplus.2023.100139","DOIUrl":"10.1016/j.mbplus.2023.100139","url":null,"abstract":"<div><p>Collagens are the most abundant structural proteins in the extracellular matrix of animals and play crucial roles in maintaining the structural integrity and mechanical properties of tissues and organs while mediating important biological processes. Fibrillar collagens have a unique triple helix structure with a characteristic repeating sequence of (Gly-X-Y)_n. Variations within the repetitive sequence can cause misfolding of the triple helix, resulting in heritable connective tissue disorders. The most common variations are single-point missense mutations that lead to the substitution of a glycine residue with a bulkier amino acid (Gly → X). In this review, we will first discuss the importance of collagen’s triple helix structure and how single Gly substitutions can impact its folding, structure, secretion, assembly into higher-order structures, and biological functions. We will review the role of “designer collagens,” i.e., synthetic collagen-mimetic peptides and recombinant bacterial collagen as model systems to include Gly → X substitutions observed in collagen disorders and investigate their impact on structure and function utilizing <em>in vitro</em> studies. Lastly, we will explore how computational modeling of collagen peptides, especially molecular and steered molecular dynamics, has been instrumental in probing the effects of Gly substitutions on structure, receptor binding, and mechanical stability across multiple length scales.</p></div>","PeriodicalId":52317,"journal":{"name":"Matrix Biology Plus","volume":"21 ","pages":"Article 100139"},"PeriodicalIF":0.0,"publicationDate":"2023-12-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590028523000121/pdfft?md5=f665a8dedce3173e618017d91ffbebfd&pid=1-s2.0-S2590028523000121-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139017085","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Spatial gene expression in the adult rat patellar tendon","authors":"Danielle Steffen ,&nbsp;Michael Mienaltowski ,&nbsp;Keith Baar","doi":"10.1016/j.mbplus.2023.100138","DOIUrl":"https://doi.org/10.1016/j.mbplus.2023.100138","url":null,"abstract":"<div><p>Tendons are dense connective tissues with relatively few cells which makes studying the molecular profile of the tissue challenging. There is not a consensus on the spatial location of various cell types within a tendon, nor the accompanying transcriptional profile. In the present study, we used two male rat patellar tendon samples for sequencing-based spatial transcriptomics to determine the gene expression profile. We integrated our data with a mouse Achilles single cell dataset to predict the cell type composition of the patellar tendon as a function of location within the tissue. The spatial location of the predicated cell types suggested that there were two populations of tendon fibroblasts, one located in the tendon midsubstance, while the other localized with red blood cells, pericytes, and immune cells to the tendon peripheral connective tissue. Of the highest expressed spatially variable genes, there were multiple genes with known function in tendon: Col1a1, Col1a2, Dcn, Fmod, Sparc, and Comp. Further, a novel spatially regulated gene (AABR07000398.1) with no known function was identified. The spatial gene expression of tendon associated genes (Scx, Thbs4, Tnmd, Can, Bgn, Lum, Adamts2, Lox, Ppib, Col2a1, Col3a1, Col6a2) was also visualized. Both patellar tendon samples had similar expression patterns for all these genes. This dataset provides new spatial insights into gene expression in a healthy tendon.</p></div>","PeriodicalId":52317,"journal":{"name":"Matrix Biology Plus","volume":"19 ","pages":"Article 100138"},"PeriodicalIF":0.0,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S259002852300011X/pdfft?md5=92f789b40475e66f6e1aa25a6e729868&pid=1-s2.0-S259002852300011X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138475136","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Omentum-derived matrix enables the study of metastatic ovarian cancer and stromal cell functions in a physiologically relevant environment","authors":"Lisa J. Neilson ,&nbsp;Douglas Cartwright ,&nbsp;Maija Risteli ,&nbsp;Elina M. Jokinen ,&nbsp;Lynn McGarry ,&nbsp;Toni Sandvik ,&nbsp;Konstantina Nikolatou ,&nbsp;Kelly Hodge ,&nbsp;Samuel Atkinson ,&nbsp;Maria Vias ,&nbsp;Emily J. Kay ,&nbsp;James D. Brenton ,&nbsp;Leo M. Carlin ,&nbsp;David M. Bryant ,&nbsp;Tuula Salo ,&nbsp;Sara Zanivan","doi":"10.1016/j.mbplus.2023.100136","DOIUrl":"https://doi.org/10.1016/j.mbplus.2023.100136","url":null,"abstract":"<div><p>High-grade serous (HGS) ovarian cancer is the most lethal gynaecological disease in the world and metastases is a major cause. The omentum is the preferential metastatic site in HGS ovarian cancer patients and <em>in vitro</em> models that recapitulate the original environment of this organ at cellular and molecular level are being developed to study basic mechanisms that underpin this disease. The tumour extracellular matrix (ECM) plays active roles in HGS ovarian cancer pathology and response to therapy. However, most of the current <em>in vitro</em> models use matrices of animal origin and that do not recapitulate the complexity of the tumour ECM in patients.</p><p>Here, we have developed omentum gel (OmGel), a matrix made from tumour-associated omental tissue of HGS ovarian cancer patients that has unprecedented similarity to the ECM of HGS omental tumours and is simple to prepare. When used in 2D and 3D <em>in vitro</em> assays to assess cancer cell functions relevant to metastatic ovarian cancer, OmGel performs as well as or better than the widely use Matrigel and does not induce additional phenotypic changes to ovarian cancer cells. Surprisingly, OmGel promotes pronounced morphological changes in cancer associated fibroblasts (CAFs). These changes were associated with the upregulation of proteins that define subsets of CAFs in tumour patient samples, highlighting the importance of using clinically and physiologically relevant matrices for <em>in vitro</em> studies. Hence, OmGel provides a step forward to study the biology of HGS omental metastasis. Metastasis in the omentum are also typical of other cancer types, particularly gastric cancer, implying the relevance of OmGel to study the biology of other highly lethal cancers.</p></div>","PeriodicalId":52317,"journal":{"name":"Matrix Biology Plus","volume":"19 ","pages":"Article 100136"},"PeriodicalIF":0.0,"publicationDate":"2023-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590028523000091/pdfft?md5=db15a375c67dbdf83c9f1115a1abae05&pid=1-s2.0-S2590028523000091-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138439427","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Urokinase-type plasminogen activator (uPA) regulates invasion and matrix remodelling in colorectal cancer","authors":"Auxtine Micalet ,&nbsp;Luke J. Tappouni ,&nbsp;Katarzyna Peszko ,&nbsp;Despoina Karagianni ,&nbsp;Ashley Lam ,&nbsp;John R. Counsell ,&nbsp;Sergio A. Quezada ,&nbsp;Emad Moeendarbary ,&nbsp;Umber Cheema","doi":"10.1016/j.mbplus.2023.100137","DOIUrl":"https://doi.org/10.1016/j.mbplus.2023.100137","url":null,"abstract":"<div><h3>Background</h3><p>Cancer cells remodel their local physical environment through processes of matrix reorganisation, deposition, stiffening and degradation. Urokinase-type plasminogen activator (uPA), which is encoded by the <em>PLAU</em> gene, is an extracellular proteolytic enzyme known to be involved in cancer progression and tumour microenvironment (TME) remodelling. Perturbing uPA therefore has a strong potential as a mechano-based cancer therapy. This work is a bioengineering investigation to validate whether 1) uPA is involved in matrix degradation and 2) preventing matrix degradation by targeting uPA can reduce cancer cell invasion and metastasis.</p></div><div><h3>Methods</h3><p>To this aim, we used an engineered 3D <em>in vitro</em> model, termed the tumouroid, that appropriately mimics the tumour’s native biophysical environment (3 kPa). A CRISPR-Cas9 mediated uPA knockout was performed to introduce a loss of function mutation in the gene coding sequence. Subsequently, to validate the translational potential of blocking uPA action, we tested a pharmacological inhibitor, UK-371,801. The changes in matrix stiffness were measured by atomic force microscopy (AFM). Invasion was quantified using images of the tumouroid, obtained after 21 days of culture.</p></div><div><h3>Results</h3><p>We showed that uPA is highly expressed in invasive breast and colorectal cancers, and these invasive cancer cells locally degrade their TME. <em>PLAU</em> (uPA) gene knock-out (KO) completely stopped matrix remodelling and significantly reduced cancer invasion. Many invasive cancer gene markers were also downregulated in the <em>PLAU</em> KO tumouroids. Pharmacological inhibition of uPA showed similarly promising results, where matrix degradation was reduced and so was the cancer invasion.</p></div><div><h3>Conclusion</h3><p>This work supports the role of uPA in matrix degradation. It demonstrates that the invasion of cancer cells was significantly reduced when enzymatic breakdown of the TME matrix was prevented. Collectively, this provides strong evidence of the effectiveness of targeting uPA as a mechano-based cancer therapy.</p></div>","PeriodicalId":52317,"journal":{"name":"Matrix Biology Plus","volume":"19 ","pages":"Article 100137"},"PeriodicalIF":0.0,"publicationDate":"2023-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590028523000108/pdfft?md5=47553b59807ff2b369969b99375d6266&pid=1-s2.0-S2590028523000108-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138412731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Temporal expression and spatial distribution of the proteoglycan versican during cardiac fibrosis development","authors":"Athiramol Sasi ,&nbsp;Andreas Romaine ,&nbsp;Pugazendhi Murugan Erusappan ,&nbsp;Arne Olav Melleby ,&nbsp;Almira Hasic ,&nbsp;Christen Peder Dahl ,&nbsp;Kaspar Broch ,&nbsp;Vibeke Marie Almaas ,&nbsp;Rosa Doñate Puertas ,&nbsp;H. Llewelyn Roderick ,&nbsp;Ida Gjervold Lunde ,&nbsp;Ivar Sjaastad ,&nbsp;Maria Vistnes ,&nbsp;Geir Christensen","doi":"10.1016/j.mbplus.2023.100135","DOIUrl":"10.1016/j.mbplus.2023.100135","url":null,"abstract":"<div><p>Cardiac fibrosis is a central pathological feature in several cardiac diseases, but the underlying molecular players are insufficiently understood. The extracellular matrix proteoglycan versican is elevated in heart failure and suggested to be a target for treatment. However, the temporal expression and spatial distribution of versican and the versican cleavage fragment containing the neoepitope DPEAAE in cardiac fibrosis remains to be elucidated. In this study, we have examined versican during cardiac fibrosis development in a murine pressure overload model and in patients with cardiomyopathies. We found that versican, mainly the V1 isoform, was expressed immediately after induction of pressure overload, preceding collagen accumulation, and versican protein levels extended from the perivascular region into the cardiac interstitium. In addition, we found increased production of versican by collagen expressing fibroblasts, and that it was deposited extensively in the fibrotic extracellular matrix during pressure overload. In cardiac cell cultures, the expression of versican was induced by the pro-fibrotic transforming growth factor beta and mechanical stretch. Furthermore, we observed that the proteolytic cleavage of versican (DPEAAE fragment) increased in the late phase of fibrosis development during pressure overload. In patients with hypertrophic and dilated cardiomyopathies, we found elevated levels of versican and a positive correlation between versican and collagen mRNA in the heart, as well as increased cleavage of full-length protein. Taken together, the temporal expression profile and the spatial distribution of both the full-length versican and the DPEAAE fragment observed in this study indicates a role for versican in development of cardiac fibrosis.</p></div>","PeriodicalId":52317,"journal":{"name":"Matrix Biology Plus","volume":"19 ","pages":"Article 100135"},"PeriodicalIF":0.0,"publicationDate":"2023-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S259002852300008X/pdfft?md5=2f40143c76be0e607a9220491b8dfec2&pid=1-s2.0-S259002852300008X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135566416","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Visualization of basement membranes by a nidogen-based fluorescent reporter in mice","authors":"Sugiko Futaki ,&nbsp;Ayano Horimoto ,&nbsp;Chisei Shimono ,&nbsp;Naoko Norioka ,&nbsp;Yukimasa Taniguchi ,&nbsp;Hitomi Hamaoka ,&nbsp;Mari Kaneko ,&nbsp;Mayo Shigeta ,&nbsp;Takaya Abe ,&nbsp;Kiyotoshi Sekiguchi ,&nbsp;Yoichi Kondo","doi":"10.1016/j.mbplus.2023.100133","DOIUrl":"10.1016/j.mbplus.2023.100133","url":null,"abstract":"<div><p>Basement membranes (BMs) are thin, sheet-like extracellular structures that cover the basal side of epithelial and endothelial tissues and provide structural and functional support to adjacent cell layers. The molecular structure of BMs is a fine meshwork that incorporates specialized extracellular matrix proteins. Recently, live visualization of BMs in invertebrates demonstrated that their structure is flexible and dynamically rearranged during cell differentiation and organogenesis. However, the BM dynamics in mammalian tissues remain to be elucidated. We developed a mammalian BM imaging probe based on nidogen-1, a major BM-specific protein. Recombinant human nidogen-1 fused with an enhanced green fluorescent protein (Nid1-EGFP) retains its ability to bind to other BM proteins, such as laminin, type IV collagen, and perlecan, in a solid-phase binding assay. When added to the culture medium of embryoid bodies derived from mouse ES cells, recombinant Nid1-EGFP accumulated in the BM zone of embryoid bodies, and BMs were visualized <em>in vitro</em>. For <em>in vivo</em> BM imaging, a knock-in reporter mouse line expressing human nidogen-1 fused to the red fluorescent protein mCherry (R26-CAG-Nid1-mCherry) was generated. R26-CAG-Nid1-mCherry showed fluorescently labeled BMs in early embryos and adult tissues, such as the epidermis, intestine, and skeletal muscles, whereas BM fluorescence was unclear in several other tissues, such as the lung and heart. In the retina, Nid1-mCherry fluorescence visualized the BMs of vascular endothelium and pericytes. In the developing retina, Nid1-mCherry fluorescence labeled the BM of the major central vessels; however, the BM fluorescence were hardly observed in the peripheral growing tips of the vascular network, despite the presence of endothelial BM. Time-lapse observation of the retinal vascular BM after photobleaching revealed gradual recovery of Nid1-mCherry fluorescence, suggesting the turnover of BM components in developing retinal blood vessels. To the best of our knowledge, this is the first demonstration of <em>in vivo</em> BM imaging using a genetically engineered mammalian model. Although R26-CAG-Nid1-mCherry has some limitations as an <em>in vivo</em> BM imaging model, it has potential applications in the study of BM dynamics during mammalian embryogenesis, tissue regeneration, and pathogenesis.</p></div>","PeriodicalId":52317,"journal":{"name":"Matrix Biology Plus","volume":"18 ","pages":"Article 100133"},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10149278/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9773433","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}