Distribution Of Extracellular Matrix Molecules Research Articles

Mapping the molecular and structural specialization of the skin basement membrane for inter-tissue interactions

Inter-tissue interaction is fundamental to multicellularity. Although the basement membrane (BM) is located at tissue interfaces, its mode of action in inter-tissue interactions remains poorly understood, mainly because the molecular and structural details of the BM at distinct inter-tissue interfaces remain unclear. By combining quantitative transcriptomics and immunohistochemistry, we systematically identify the cellular origin, molecular identity and tissue distribution of extracellular matrix molecules in mouse hair follicles, and reveal that BM composition and architecture are exquisitely specialized for distinct inter-tissue interactions, including epithelial–fibroblast, epithelial–muscle and epithelial–nerve interactions. The epithelial–fibroblast interface, namely, hair germ–dermal papilla interface, makes asymmetrically organized side-specific heterogeneity in the BM, defined by the newly characterized interface, hook and mesh BMs. One component of these BMs, laminin α5, is required for hair cycle regulation and hair germ–dermal papilla anchoring. Our study highlights the significance of BM heterogeneity in distinct inter-tissue interactions.

Distribution of extracellular matrix molecules in human uterine tubes during the menstrual cycle: a histological and immunohistochemical analysis.

The uterine tube (UT) is an important and complex organ of the women's reproductive system. In general, the anatomy and basic histology of this organ are well-known. However, the composition and function of the extracellular matrix (ECM) of the UT is still poorly understood. The ECM is a complex supramolecular material produced by cells which is commonly restricted to the basement membrane and interstitial spaces. ECM molecules play not only a structural role, they are also important for cell growth, survival and differentiation in all tissues. In this context, the aim of this study was to evaluate the deposition and distribution of type I and III collagens and proteoglycans (decorin, biglycan, fibromodulin and versican) in human UT during the follicular and luteal phases by using histochemical and immunohistochemical techniques. Our results showed a broad synthesis of collagens (I and III) in the stroma of the UT. The analysis by regions showed, in the mucosa, a specific distribution of versican and fibromodulin in the epithelial surface, whereas decorin and fibromodulin were observed in the lamina propria. Versican and decorin were found in the stroma of the muscular layer, whereas all studied proteoglycans were identified in the serosa. Curiously, biglycan was restricted to the wall of the blood vessels of the serosa and muscular layers. Furthermore, there was an immunoreaction for collagens, decorin, versican and fibromodulin in the UT peripheral nerves. The differential distribution of these ECM molecules in the different layers of the UT could be related to specific structural and/or biomechanical functions needed for the oviductal transport, successful fertilization and early embryogenesis. However, further molecular studies under physiological and pathological conditions are still needed to elucidate the specific role of each molecule in the human UT.

Regulated metabolism of Vitamin A by retinol dehydrogenase 10 is critical for embryonic development of the heart

Congenital heart disease is a serious human health issue, affecting as many as 1% of live births. For most cases of cardiac birth defect the cause is unknown, reflecting the limitations of our understanding of which environmental agents, genetic factors, and gene‐environment interactions contribute to cardiac abnormalities. The purpose of the current study is to investigate the mode whereby an embryo converts Vitamin A into the potent morphogen retinoic acid and to understand how disturbances in the production of retinoic acid may contribute to developmental malformations of the heart. Precise regulation of retinoic acid production is critical ‐ either excessive or insufficient levels result in cardiac birth defects ‐ yet very little is known about the mechanism whereby this process is controlled. We have demonstrated that the short chain dehydrogenase/reductase RDH10 is the primary enzyme responsible for the first step of embryonic Vitamin A metabolism. We show that embryos with mutations in Rdh10 have severe cardiac developmental abnormalities, including defects in heart looping, myocardial growth, aorticopulmonary septation and morphogenesis of the atria. Using transgenic reporter mice and RNA in situ hybridization we have determined that the Rdh10−/− cardiac phenotypes are associated with abnormal expression of Hox family A‐P patterning genes and also with defects in cell adhesion and abnormal distribution of extracellular matrix molecules. Finally, utilizing quantitative reverse transcriptase PCR we have demonstrated that Rdh10 gene transcription is a point for feedback control in regulation of retinoic acid production. These data suggest that Rdh10 may be a candidate gene for association with human cardiac abnormalities. Moreover, the data shed light on the molecular mechanism whereby disruptions in Vitamin A metabolism may contribute to congenital heart disease.Grant Funding Source: NIH‐COBRE

Manipulations in hydrogel chemistry control photoencapsulated chondrocyte behavior and their extracellular matrix production.

In engineering a cell-carrier to support cartilage growth, hydrogels provide a unique, largely aqueous environment for 3-dimensional chondrocyte culture that facilitates nutrient transport yet provides an elastic framework dictating tissue shape and supporting external loads. Although the gel environment is often >90% water, we demonstrate that slight variations in hydrogel chemistry control gel degradation, evolving macroscopic properties, and ultimately the secretion and distribution of extracellular matrix molecules. Specifically, biodegradable poly(ethylene glycol)-co-poly(lactic acid) hydrogels were fabricated via photopolymerization. When chondrocytes were photoencapsulated in these gels, changes in the poly(ethylene glycol)-co-poly(lactic acid) repeat unit ratio from 19 to 7 increased total collagen synthesis 2.5-fold after 6 weeks in vitro. Furthermore, the ratio of collagen to glycosaminoglycans varied from glycosaminoglycan-rich, 0.33 +/- 0.13, to collagen-rich, 4.58 +/- 1.21, depending on gel chemistry and in vitro versus in vivo culture environment. By tuning scaffold chemistry, and subsequently, gel structure and degradation behavior, we can better guide tissue evolution and development.

Controlling the spatial distribution of ECM components in degradable PEG hydrogels for tissue engineering cartilage.

In developing a scaffold to support new tissue growth, the degradation rate and mass loss profiles of the scaffold are important design parameters. In this study, hydrogels were prepared by copolymerizing a degradable macromer, poly(lactic acid)-b-poly(ethylene glycol)-b-poly(lactic acid) endcapped with acrylate groups (PEG-LA-DA) with a nondegradable macromer, poly(ethylene glycol) dimethacrylate (PEGDM). The resulting hydrogels exhibited a range of degradation behavior and mass loss profiles. Chondrocytes were photoencapsulated in gels formulated with 50:50, 25:75, and 15:85 (mol % PEGDM: mol % PEG-LA-DA) and cultured for 6 weeks in vitro. The neocartilaginous tissue formed was examined biochemically and histologically. After 6 weeks, the DNA content in gels with 75 and 85% degradable crosslinks was nearly twice that of the DNA content in the 50% gels. The total collagen content was significantly higher in the 85% gel [2.4 +/- 0.8% wet weight (ww)] compared to the 50% gel (0.22 +/- 0.29% ww). In examining the neocartilaginous tissue with immunohistochemistry, type II collagen was localized in the pericellular region in the 50% gel; however, when increased degradation was incorporated into the gel, type II collagen was found throughout the neotissue. In summary, the important role of hydrogel degradation in controlling and influencing the deposition and distribution of extracellular matrix molecules was demonstrated and quantified.

Directional specificity and patterning of sensory axons in trigeminal ganglion–whisker pad cocultures

In the rodent trigeminal pathway, trigeminal axons invade the developing whisker pad from a caudal to rostral direction. We investigated directional specificity of embryonic day (E) 15 rat trigeminal axons within this peripheral target field using explant cocultures. E15 trigeminal axons readily grow into the same age whisker pad explants and form follicle-related patterns along a caudal to rostral direction. They also can grow into this target from its lateral aspects. In contrast, they are unable to invade the whisker pad from the rostral (nasal) pole. We did not find any correlation between the distribution of extracellular matrix molecules and trigeminal axon growth preferences. We also examined age-related changes in trigeminal axon responsiveness to directional cues. E19 trigeminal axons readily grew into E15 whisker pad explants from either the caudal or the rostral pole. These results suggest the presence of growth permissive and repulsive cues that guide sensory axons in the whisker pad. Furthermore, trigeminal axons lose their responsiveness to growth inhibitory cues at later stages of development.

Spatial and temporal changes in chondroitin sulfate distribution in the sclerotome play an essential role in the formation of migration patterns of mouse neural crest cells.

We have examined the roles of pertinent extracellular matrix molecules in the formation of the neural crest cell migration patterns in the sclerotome of the mouse embryo. The present data indicate that permissiveness for migration is inversely correlated with chondroitin sulfate content. Experimental removal of chondroitin sulfate proteoglycans in the embryo causes neural crest cells to migrate even within the posterior half of the somite, which they do not invade ordinarily. Moreover, three different sclerotomal regions defined by the presence or absence of the ventromedial and/or ventrolateral pathways are present along the anteroposterior axis and undergo systematic temporal changes that affect migration patterns. The most anterior portion of the sclerotome is conducive to both ventromedial and ventrolateral migration (Anterior Region). The intermediate portion is conducive to ventromedial migration only (Intermediate Region). No neural crest cells are seen within the posterior portion of the sclerotome (Posterior Region). At this level, they are observed exclusively in the dorsolateral space adjacent to the roof of the neural tube. With advancing embryonic development, the rostrocaudal length of the Anterior Region decreases and is accompanied by a corresponding enlargement of the Intermediate Region. These results suggest that temporal and regional differences in the sclerotome contribute to the neural crest cell migration patterns in the mouse. To refine our understanding of the underlying mechanisms, regional differences and temporal changes in the distribution of extracellular matrix molecules have been examined during migration. In the sclerotome, chondroitin sulfate displays distinct distribution patterns that are closely correlated with the migration patterns of mouse neural crest cells. Furthermore, their migration patterns are altered in embryos treated with the inhibitors of chondroitin sulfate proteoglycan biosynthesis, sodium chlorate, and beta-D-xyloside. In inhibitor-treated embryos, neural crest cell migration occurs even in the posterior portion of the sclerotome. The metameric organization of dorsal root ganglia is disturbed in these embryos. Our observations provide novel evidence for the importance of sclerotomal chondroitin sulfate distribution patterns in mouse crest cell migration patterns. We conclude that systematic spatiotemporal changes in the distribution of chondroitin sulfate proteoglycans are a key requisite for the formation of migration patterns of mouse neural crest cells in the sclerotome.

Light-microscopical investigation of the distribution of extracellular matrix molecules and calcifications in human dental pulps of various ages.

The distribution of extracellular matrix molecules, especially collagen types I, III, V, and VI, in the extracellular matrix of the connective tissue of human dental pulp of various ages was studied by polarization and indirect immunofluorescence microscopy by using a conventional fluorescence microscope and a confocal laser scanning microscope. Polarization and immunofluorescence microscopy of paraffin sections showed thick fibers of collagen type I, which represented the main component of the connective tissue matrix of the dental pulp. By indirect immunofluorescence, thin fibers and small bundles of collagen type III were determined to be one of the main fibrillar elements present in the dental pulp matrix. Collagen type IV was detected by a clear intense staining of the basement membrane of blood vessels at all ages examined. Collagens type V and VI formed a dense meshwork of thin microfibrils throughout the stroma of the connective tissue of the dental pulp. These fibers were localized around blood vessels and appeared to be enriched in the subodontoblastic layer. Investigations by means of confocal laser scanning microscopy revealed fibers of collagen type VI spiralling between fully differentiated odontoblasts toward the predentin layer. With advancing age, the connective tissue matrix appeared to be condensed and aggregates of thick fiber bundles could be observed. Furthermore, the participation of various collagen types in the composition of pulp stones was shown. These calcifications and diffuse calcifications increased in frequency with advancing age in a statistically significant manner.

Immunohistological and ultrastructural study of the developing tendons of the avian foot.

The aim of the present report is to provide a detailed description of the morphogenesis and initial differentiation of the long tendons of the chick foot, the long autopodial tendons (LAT), from day 6 to day 11 of development. The fine structure of the developing LAT was studied by light and transmission electron microscopy. The characterization by immunofluorescent techniques of the extracellular matrix was performed using laser scanning confocal (tenascin, elastin, fibrillin, emilin, collagen type I, II, III, IV and VI) or routine fluorescence (tenascin, 13F4) microscopy. In addition, cell proliferation in pretendinous blastemas was analyzed by the detection of BrdU incorporation by immunofluorescence. The light microscopic analysis permitted the identification of different stages during LAT morphogenesis. The first stage is the formation of a thick ectoderm-mesenchyme interface along the digital rays, followed by the differentiation of the "mesenchyme lamina", an extracellular matrix tendon precursor, and ending with the formation and differentiation of the cellular condensation that forms the tendon blastema around this lamina. The immunofluorescence study revealed the presence and arrangement of the different molecules analyzed. Tenascin and collagen type VI are precocious markers of the developing tendons and remain present during the whole process of tendon formation. Collagen type I becomes mainly restricted to the developing tendons from day 7.5. Collagens type II and IV are never detected in the developing tendons, while a faint labeling for collagen type III is first detected at day 7. The analysis of the distribution of the elastic matrix components in the developing tendons is a major contribution of our study. Elastin was detected in the periphery of the tendons from day 8 and also in fibrils anchoring the tendons to the skeletal elements. At the same stage, emilin strongly stains the core of the tendon rods, while fibrillin is detected a little later. Our study indicates the existence of an ectoderm-mesoderm interaction at the first stage of the tendon formation. In addition, our results show the different spatial and temporal pattern of distribution of extracellular matrix molecules in developing tendons. Of special importance are the findings concerning the tendinous elastic matrix and its possible role in tendon maturation and stabilization.

Distribution of Extracellular Matrix Molecules in Human Hair Follicles

Annals of the New York Academy of SciencesVolume 642, Issue 1 p. 253-262 Distribution of Extracellular Matrix Molecules in Human Hair Follicles A. G. MESSENGER, A. G. MESSENGER Department of Dermatology Royal Hallamshire Hospital Sheffield S10 2JF, EnglandSearch for more papers by this authorKATHERINE ELLIOTT, KATHERINE ELLIOTT Department of Dermatology Royal Hallamshire Hospital Sheffield S10 2JF, EnglandSearch for more papers by this authorGILLIAN E. WESTGATE, GILLIAN E. WESTGATE Unilever Research Colworth House Sharnbrook, Bedford MK44 ILQ, EnglandSearch for more papers by this authorW. T. GIBSON, W. T. GIBSON Unilever Research Colworth House Sharnbrook, Bedford MK44 ILQ, EnglandSearch for more papers by this author A. G. MESSENGER, A. G. MESSENGER Department of Dermatology Royal Hallamshire Hospital Sheffield S10 2JF, EnglandSearch for more papers by this authorKATHERINE ELLIOTT, KATHERINE ELLIOTT Department of Dermatology Royal Hallamshire Hospital Sheffield S10 2JF, EnglandSearch for more papers by this authorGILLIAN E. WESTGATE, GILLIAN E. WESTGATE Unilever Research Colworth House Sharnbrook, Bedford MK44 ILQ, EnglandSearch for more papers by this authorW. T. GIBSON, W. T. GIBSON Unilever Research Colworth House Sharnbrook, Bedford MK44 ILQ, EnglandSearch for more papers by this author First published: December 1991 https://doi.org/10.1111/j.1749-6632.1991.tb24392.xCitations: 37AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume642, Issue1The Molecular and Structural Biology of HairDecember 1991Pages 253-262 RelatedInformation

Immunolocalization of Fibronectin and Laminin in the Amphibian Intestine During Spontaneous and Triiodothyronine-Induced Metamorphosis: (laminin/fibronectin/extraepithelial matrix/intestine/amphibian).

Fibronectin and laminin were detected by indirect immunofluorescence in the intestine of Alytes obstetricans (anuran amphibian) during triiodothyronine (T3)-induced metamorphosis and spontaneous post-embryonic development. Fibronectin was first detected between a small number of connective tissue cells. As T3-treatment and spontaneous development progressed, fibronectin became detectable as a fine network extending throughout the whole thickness of the connective tissue and particularly in the core of the developing epithelial folds. During the first week of T3-treatment and throughout the spontaneous larval period, laminin was present as a linear band within the basement membrane. Between day 6 and 12 of hormonal treatment, an increase in the laminin fluorescent staining was noted. After hormonal treatment for two weeks and at the end of spontaneous metamorphosis, laminin staining was localized within the basement membrane of the folded epithelium and around muscle fibers. These observations indicate that variations in the density and distribution of extracellular matrix molecules are closely related spatiotemporarily to the structural changes occurring in the connective and muscle tissues of the intestine during metamorphosis.

Immunogold localization of extracellular matrix molecules in Bruch's membrane of the rat.

The distribution of extracellular matrix molecules in Bruch's membrane of the rat was studied by a post-embedding immunogold labeling method. These molecules included laminin, types I, III and IV collagen, fibronectin and heparan sulfate proteoglycan. Laminin and type IV collagen were localized in the basal lamina of the pigment epithelium and choriocapillaris; types I and III collagen, to the inner and outer collagenous zones; fibronectin, to the elastic layer; heparan sulfate proteoglycan was found diffusely throughout the entire Bruch's membrane. These results not only agree with previous biochemical and immunofluorescence studies of these molecules in Bruch's membrane of other species but also provide a more precise localization of these molecules in the various layers of Bruch's membrane.

Neuronal pathfinding in developing wings of the moth Manduca sexta

The neural pattern of the moth wing is a simple two-dimensional network nestled between the two epithelial monolayers that form the upper and lower surfaces of the wing. All neural elements within the wing blade are sensory and their axons grow proximally toward the mesothoracic ganglion. The sensory nerves of the wing are intimately associated with the basal lamina of the upper epithelial layer; and the molding of neural pattern is coupled with cues in the basal lamina. The global landscape of the basal lamina can be altered by exchange of epithelial grafts. Axons generally cross control grafts as well as grafts that have been displaced distally. However, axons generally avoid grafts that have been transposed proximally. This asymmetric response of growing axons implies that directional cues in the substratum are also asymmetric along the length of the wing. The asymmetric, graded distribution of extracellular matrix molecules associated with the basal lamina of the wing's upper epithelium could provide the short-range cues that guide sensory axons in a particular direction.