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R R Markwald

Publications and source records attributed to R R Markwald.

At least 55 records · Page 3Linked to original sources
PubMed1992

Multiple glycoproteins localize to a particulate form of extracellular matrix in regions of the embryonic heart where endothelial cells transform into mesenchyme.

Cells derived from an epithelial-mesenchymal transformation within the atrioventricular canal and outflow tract are involved in the partitioning of the early embryonic heart into a four-chambered organ. This transformation process has been shown to proceed from an inductive interaction between the myocardium and competent, target endothelial cells within these regions of the heart. Interestingly, immunohistochemistry revealed the presence of fibronectin-positive particulates within the matrix of mesenchyme-forming regions (Mjaatvedt et al., 1987). This particulate matrix is extractable by EDTA and can elicit the epithelial-mesenchymal transformation in culture (Mjaatvedt and Markwald, 1989). Analysis of EDTA extracts of embryonic heart tissue revealed the presence of fibronectin and about 40 unidentified proteins, 6 of which appeared to be enriched in the biologically active 100,000g pellet fraction (Mjaatvedt and Markwald, 1989). Based on these and other data we have proposed that the particulate matrix is composed of a multicomponent complex of fibronectin and one or more of the low-molecular-weight proteins in this pellet. The purpose of the present study was to begin a biochemical characterization of the nonfibronectin proteins thought to be present in the matrix particulates. Given that many matrix constituents are glycoproteins, lectins were used to initially characterize the particulate constituents. Of the lectins tested, soybean agglutinin (SBA) was found to be specific only for matrix particulates. Histochemical analyses showed that SBA and antibodies against fibronectin colocalized regionally and temporally to the same matrix particulates in embryonic heart tissue.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

An antiserum (ES1) against a particulate form of extracellular matrix blocks the transition of cardiac endothelium into mesenchyme in culture.

The epithelial-mesenchymal transition of cardiac endothelium is a critical developmental event in the formation of valvular and septal anlagen. We have demonstrated previously that this event can be mimicked in culture by treating atrioventricular canal (AV) endothelium with EDTA-soluble proteins extracted from embryonic heart tissue. This activity was fractionated by ultracentrifugation of the EDTA extract, indicating that the critical proteins existed as a multicomponent complex. Based on these results we propose that: (1) the in vitro particulates in EDTA extracts correspond to an observed particulate form of extracellular matrix within the myocardial basement membrane (MBM) of mesenchyme-forming regions and (2) one or more of the proteins in the MBM particulates function to elicit the epithelial-mesenchymal transition. To test these hypotheses we utilized an antiserum, termed ES1, prepared against EDTA-extractable particulates from embryonic chick hearts. Both ES1 and an anti-fibronectin monoclonal antibody (M3H) co-localized in situ to particles within the MBM; however, no ES1 reactivity towards fibronectin could be detected by ELISA or immunoblot analysis. The ES1-positive MBM particulates were removed by extraction with EDTA, but not with PBS, indicating a divalent cation-mediated association of the constituent proteins. ES1 antibodies recognized two major (28 and 46 kDa) and three minor (93, 109, and 180 kDa) proteins on immunoblots of EDTA-extractable proteins. When tested in culture, ES1 antiserum inhibited the formation of mesenchyme from AV endothelium in a dose-dependent manner, while M3H did not. These results are consistent with an active role for one or more of the ES1 antigens in initiating the formation of AV mesenchyme. The localization of ES1 antigens to the extracellular matrix at other dynamic interfaces, e.g., ectoderm/neural tube and limb bud ectoderm/mesoderm, point to a potentially general importance of ES1 antigens in mediating similar developmental interactions.

Animals↗
PubMed1990

Morphogenesis of precursor subpopulations of chicken limb mesenchyme in three dimensional collagen gel culture.

Although homogeneous in appearance, several lines of evidence suggest early (stage 17-19) limb mesenchymal cells are committed to particular cell lineages, e.g., myogenic or chondrogenic. However, subsequent expression of cell or tissue phenotype in the developing limb does not occur in a randomized process but rather in a spatially specific pattern. The potential regulatory mechanisms controlling the "patterned" expression of tissue phenotype in the limb have not been resolved. The purpose of this study was to determine if, prior to the formation of an apical ectodermal ridge, nondissociated limb mesenchyme has inherent morphogenetic potential to form nonrandomized patterns of tissue organization. The hypotheses to be tested were that, if provided a spatially permissive culture environment, 1) mesenchymal cells committed to a particular lineage would segregate into precursor (sub)populations prior to overt expression of phenotype and 2) the ultimate expression of a tissue phenotype may be regulated, in part, by histogenic interactions between the precursor cell groups. For these studies, mesoblasts (intact mesenchyme minus ectoderm) from stage 17-19 hindlimb buds were explanted intact to the surface of a 1-3 mm thick hydrated lattice of repolymerized type I collagen and incubated for 2-11 days. Examination of cultures at variable intervals revealed three distinct temporal sequences (periods) which were arbitrarily termed early morphogenesis (0-3 days), cytodifferentiation (3-5.5 days), and primitive tissue formation (5.5-11 days) based on similarities to in situ limb development. By the end of the first period, the mesenchymal cells had sorted into three distinct precursor populations: 1) an epithelial-like outgrowth of premyogenic and prefibrogenic cells at the surface of the gel lattice (termed the "surface subset") which circumscribed, 2) a centrally positioned prechondrogenic condensate ("central subset"), and overlaid 3) a dispersed, population of free cells that invaded the collagen lattice ("seeded subset"). Subsequent cytodifferentiation led to the appearance of multinucleated myotubes within the surface subset and chondrification of the central subset. Cells of the seeded subset remained dispersed within the collagen lattice. Primitive histogenic events were initiated during the final period of development including 1) at sites where surface cells established boundaries with the central subset, collectives or "bundles" of variable sized myotubes were formed which became partially ensheathed by the attenuated processes of fibroblastlike cells; and 2) a secondary site of chondrogenic activity was initiated within the gel lattice at the boundary between the central and seeded cell populations. Transformation of seeded fibroblasts into chondroblasts accompanied expansion of the secondary chondrogenic element within the gel lattice.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1990

NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase: immunochemical characterization of the lung enzyme from pregnant rabbits.

A polyclonal antibody was produced in guinea pig against the lung NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase (PGDH) purified from pregnant rabbits. Western blot analysis demonstrated that the protein identified by this antibody in the 105,000g supernatant fraction of lung tissue from pregnant rabbits had a molecular mass of 30 kDa and comigrated with the purified PGDH. The specific activity of the lung PGDH in pregnant rabbits (25- to 28-day gestations) was 36.7 nmol NADH formed/min/mg protein compared to 0.3 nmol NADH formed/min/mg protein in nonpregnant rabbits. Although the PGDH activity in the lung cytosol of nonpregnant rabbits was inhibited by the anti-lung PGDH antibody, the 30-kDa protein was not detected by Western blot analysis. An examination of this 30-kDa protein during the gestational period indicated that the protein was present after 10 days and the amount of the protein increased from Day 10 to Day 28. This increase in the immunochemically reactive protein correlated with the marked increase in PGDH specific activity between 10 and 28 days. An immunochemically reactive protein also was observed in the ovary of 25- to 28-day pregnant rabbits and the specific activity of the ovary PGDH was 19.3 nmol NADH formed/min/mg protein. Only trace levels of the PGDH activity were detected in the ovaries of nonpregnant rabbits. A 30-kDa protein was not detected by the anti-rabbit lung PGDH in brain, kidney, bladder, uterus, liver, and heart tissue of pregnant or nonpregnant rabbits. When rabbit or human placental cytosol was examined with the anti-rabbit lung PGDH only faint 30-kDa bands were observed by Western blot analysis. A monoclonal antibody prepared against human placental PGDH did not recognize the 30-kDa band in the pregnant rabbit lung. Localization studies indicated a marked increase in immunochemical staining in pulmonary epithelial cells of pregnant rabbits as compared to nonpregnant rabbits. Lung epithelial cells but not endothelial cells were identified as containing the PGDH.

Alprostadil↗
PubMed1990

Actin and lamin comprised filaments in the nuclei of Chinese hamster ovary cells affected with Clostridium difficile enterotoxin A.

The major change in the ultrastructure of Chinese hamster ovary (CHO) cells treated with Clostridium difficile enterotoxin A was the formation of bundles of filaments in the nucleoplasm. The filaments appeared after 2.5 h and disappeared by the fourth hour of incubation with the toxin. To partially characterize these filaments, laser diffractometry and ultrastructural immunocytochemistry were employed. The bundles consist of coiled filaments of about 11 nm and 16 nm diameter and a main long axial periodicity of about 26 nm. Postembedding immunogold labelling with monoclonal antibodies demonstrated that these intranuclear filaments are comprised mainly of actin, but also contain lamin and vinculin. Between 2.5 and 3 h of incubation with enterotoxin A, the lamins were located close to the nuclear envelope, but they were also consistently found scattered in the cytoplasm and nucleoplasm. Such an unusual distribution of lamins might mediate the adverse effects of enterotoxin A on nuclear organization and function and thus cause the eventual death of CHO cells.

Actin Cytoskeleton↗
PubMed1989

Effect of selected denervations on glycoconjugate composition and tissue morphology during the initiation phase of limb regeneration in adult Ambystoma.

This study was undertaken to assess the effects of various quantities of neural tissue on the temporal relationship of matrix glycoconjugates to the regeneration morphology. 1) Denervation before amputation revealed that a threshold level of nervous tissue was necessary to activate a regeneration response from the tissue, i.e., appearance of regeneration-specific morphologies and glycoconjugates. 2) Denervation after amputation demonstrated that the level of neural tissue necessary to maintain these responses was below the level necessary to activate the regeneration response. If neural tissue was completely removed there was a concomitant loss of regenerate morphologies and glycoconjugates. 3) Bilateral amputation of a neurogenically intact limb and its completely denervated contralateral limb revealed that the regeneration response was a localized phenomenon during the first 30 days after amputation. After 30 days the regeneration response appeared within the previously degenerated denervating limb. The results suggest that the factors controlling the regenerative response in adult Ambystoma are large diffusible substances that can be transported by the circulation and can affect the regenerative response in remote, previously activated, tissues.

Ambystoma↗
PubMed1989

Glycoconjugates in normal wound tissue matrices during the initiation phase of limb regeneration in adult Ambystoma.

The present study identifies, localizes, and reports the relative composition of specific glycosaminoglycans within tissue matrices during the initiation phase of limb regeneration. The regenerate tissues were harvested and assayed morphologically, histochemically, and chemically. We observed 1) a population of cells interspersed among the cells of the dermis, epimysium, perimysium, perichondrium, and periosteum. 2) This population was distinguishable by a unique pattern of glycoconjugate staining, i.e., intracellular and pericellular heparan sulfate and glycoproteins and extracellularly associated hyaluronate and glycoproteins. 3) Cells with these staining characteristics aggregated to a position directly beneath the apical epidermal cap. 4) Extracellular hyaluronate and glycoproteins colocalized with undifferentiated tissues. And 5) extracellular chondroitin sulfate, dermatan sulfate, and keratan sulfate glycosaminoglycans colocalized with differentiated tissues. The correlations of distinct glycoconjugate compositions with specific regeneration morphologies suggest the possibility that these components may be related to the phenotypic expression of tissues during regeneration.

Ambystoma↗
PubMed1989

Induction of an epithelial-mesenchymal transition by an in vivo adheron-like complex.

The embryonic vertebrate heart consists of two epithelia: the myocardium and endothelium, separated by the myocardial basement membrane (MBM). The myocardium has been shown to induce endothelial transformation into prevalvular mesenchyme in a temporally and site restricted manner. Previously, we hypothesized that the myocardial-endothelial interaction is mediated in vivo by aggregates of 30-nm particles in the MBM which can be removed by EDTA extraction. These MBM extracts contain fibronectin and other lower Mr proteins and can initiate an epithelial-mesenchymal transition in the AV (atrioventricular canal) endothelium of embryonic chick heart in collagen gel culture. These and other data suggested that the 30-nm multicomponent particles are similar, structurally and compositionally, to multimolecular complexes, termed adherons, secreted by L6 muscle cells in culture. The purpose of this study was to (1) test whether the removal of the 30-nm particles from MBM extracts of embryonic chick hearts would remove the in vitro biological activity and (2) determine if the fractionated MBM extracts can cause AV endothelial cells to follow the same differentiation pathway observed in vivo by monitoring immunohistochemically the cell surface expression of N-CAM. Results showed that centrifugation of extract at 100,000g for 1 hr produced a supernatant fraction that was unable to initiate mesenchyme formation from AV endothelium. However, the resuspended pellet fraction did initiate differentiation of endothelium into mesenchyme. Conditioned medium from L6 skeletal muscle cultures could not substitute for the EDTA extract of embryonic heart. Endothelial cells undergoing the transition to form mesenchyme, both in vivo and in vitro, showed a concomitant decrease in N-CAM staining. This suggested that the pellet-induced formation of migrating cells in the collagen gels is not the result a novel in vitro phenomenon.

Animals↗
PubMed1988

Specific configurations of fibronectin-containing particles correlate with pathways taken by neural crest cells at two axial levels.

Although neural crest (NC) cells can potentially enter a number of intertissue spaces, they select a particular pathway that varies depending on the axial level. In the cranial region, NC cells enter the dorsal-lateral pathway (i.e., immediately subjacent to the ectoderm) and avoid the ventral pathway (i.e., pathway between the mesoderm and neural tube and within the mesodermal cell population), whereas in the trunk region, the majority of the NC cells enter the ventral pathway (i.e., between the somite and neural tube) and not the dorsal-lateral pathway. Our working hypothesis is that one determining factor in directing NC cell migration is the composition and/or intermolecular associations of the extracellular matrix (ECM) in these pathways. Histochemical staining, immunostaining, and lectin-binding studies on cryofixed and conventionally fixed tissue were conducted to initially characterize the ECM found in potential NC cell pathways prior to and during initial NC cell migration at two different axial levels. We found that, regardless of the axial level, the pathways into which NC cells eventually enter possessed a characteristic ECM arrangement. This arrangement included: 1) the presence of multicomponent, glycoprotein-containing spherical particles (0.1-0.5 micron in diameter); and 2) a low-sulfated ECM content. Although all particles contained fibronectin, only those in specific regions were able to bind to a monoclonal antibody directed to the cell-binding domain of fibronectin, suggesting that the conformation of fibronectin may be important in the expression of any in situ function of the molecule.

Animals↗
PubMed1988

Initial expression of type I procollagen in chick cardiac mesenchyme is dependent upon myocardial stimulation.

Formation of the atrioventricular (AV) mesenchyme is a critical step in early heart development. Endothelial cells are activated and transformed into a mesenchymal population that invades the cell-free myocardial basement membrane. This process can be duplicated in collagen gel culture, where it has been established that myocardium or its secretory products activate the endothelium. The purpose of the present study was to determine when these activated endothelial and/or mesenchymal cells start producing type I collagen in situ. These results were compared to those obtained from a culture model of mesenchyme formation. The production of type I collagen was monitored using a monoclonal antibody (M38) that recognizes the carboxy-terminal propeptide of human type I procollagen. The initial expression of the latter within activated AV endothelial and mesenchymal cells in ovo was 48 hr following activation. Prior to this time, only the myocardium was reactive with M38. AV explants of early hearts on collagen gels revealed staining of activated endothelial and mesenchymal cells with M38 after 48 hr in coculture with myocardial tissue. Explants that were prevented from activating (myocardium removed) never expressed the M38 antigen. Similarly, AV endothelial monolayers grown in the presence of myocardial conditioned medium activated and expressed type I collagen after 48 hr in culture, whereas those grown in standard medium did not. These results establish the initial expression of type I collagen within activated AV endothelium and mesenchyme. In addition, the data suggest that the expression of type I collagen within the AV mesenchyme may be dependent on extrinsic influences that induce the AV endothelium to transform into mesenchyme.

Age Factors↗
PubMed1987

Distribution of basement membrane antigens in cryopreserved early embryonic hearts.

The early embryonic heart is composed of two cylindrical epithelial layers, an inner endothelium and an outer myocardium. The cardiac jelly (CJ), an acellular accumulation of extracellular matrix (ECM), fills the space between the two epithelia. During development of the heart, a portion of the endothelial cells of the atrioventricular (AV) region differentiate into mesenchyme cells in a temporally and spacially specific manner. Although contiguous with those in the AV region, endothelial cells lining the ventricle never form mesenchyme in situ. At present, the mechanisms controlling the biphasic differentiation of the endothelium and the subsequent migration of cardiac mesenchymal cells are poorly understood. Although the CJ lies between two epithelial and is spatially equivalent to a basement membrane (BM), it has not traditionally been considered to be organized into a BM-like structure. The potential significance of this observation to developmental biology lies in the possibility that BM or their individual components (i.e., fibronectin (FN), laminin (LM), type IV collagen, and heparin sulfate proteoglycan (HSPG] may function as the regulatory site of epithelial differentiation and morphogenesis. A cryofixation technique was developed in order to determine the in situ immunohistochemical distribution of the BM components in the CJ. Results indicated that the CJ exists as the fusion between a larger myocardially derived BM having a lamina densa and an extended reticular lamina and an attenuated, endothelial-associated BM composed only of a lamina densa. Except for FN, the individual BM components were not all present during early stages, but instead appeared in a sequential manner, suggesting that all components of an adult-type BM are not required to initiate the assembly of a structural and functional BM during development. In the AV canal and outflow tract (OT), FN appeared as a progressively expanding gradient of material with the greatest density nearer the myocardium.

Animals↗
PubMed1987

Attachment of neural crest cells to endogenous extracellular matrices.

Newly emerging neural crest (NC) cells will enter either the lateral pathway under the surface ectoderm or the vental pathway along the neural tube depending on the axial level (Pratt et al.: Dev. Biol., 44:298-305, 1975; Thiery et al.: Dev. Biol., 93:324-343, 1982; Newgreen et al.: Cell Tissue Res., 221:521-549, 1982; LeDouarin et al.: In: The Role of Extracellular Matrix in Development. Alan R. Liss, Inc., New York, pp. 373-398, 1984; Brauer et al.: Anat. Rec., 211:57-68, 1985). A number of studies have shown a correlation between the type of extracellular matrix (ECM) associated with adjacent tissues (e.g., ectoderm, neural tube, and mesoderm) and the initial pathway taken by NC cells. Our working hypothesis is that the direction of NC cell migration (ventral vs. lateral pathway) depends on the composition of the ECM associated with the surface ectoderm and its ability to support NC cell attachment. In this study, we tested this hypothesis by isolating endogenous ECM associated with the ectoderm of each region and examining the ability of each endogenous ECM to support cranial and trunk NC cell attachment in vitro. Results indicated that both cranial and trunk NC cells preferentially attached to cranial ectodermal ECM as compared to trunk ectodermal ECM. The differences in NC cell attachment were not due to a preferential adsorption of cranial ectodermal ECM onto the ECM-conditioned plastic substrate over trunk ectodermal since approximately equal amounts of ECM bound to the plastic. These results supported the hypothesis and provide evidence that endogenous ectodermal ECM may be one factor potentially responsible for directing the NC cells along a ventral or a lateral pathway.

Adsorption↗
PubMed1987

Myocardial specificity for initiating endothelial-mesenchymal cell transition in embryonic chick heart correlates with a particulate distribution of fibronectin.

The early chick heart tube consists of myocardium and endothelium separated by a myocardially derived basement membrane (MBM). As development proceeds, the endothelium undergoes a transition into mesenchyme in a regionally specific manner; only the atrioventricular (AV) and outflow tract, but not the ventricular endothelium, is transformed into mesenchyme, the progenitor of heart septa and valves. Recent experiments have shown that an EDTA extract of MBM can initiate AV endothelium to form mesenchyme in an in vitro collagen gel culture system. Two-dimensional gel electrophoresis of AV region EDTA extracts showed potentially three isoelectric forms of fibronectin (Fn), while extracts from ventricle contained only two forms. The purpose of the present study was to further investigate the significance of these regional differences by testing of specific myocardial regions (AV vs ventricle) for their ability to induce endothelium to form mesenchyme in vitro, and to immunohistochemically determine if a regionally specific distribution of Fn exists in the MBM that can be correlated with previous electrophoretic data. Embryonic heart regions cultured on three-dimensional collagen gels showed that AV endothelium could only form mesenchyme if cocultured with AV myocardium. Coculture with ventricular myocardial explants did not initiate differentiation of AV endothelium. In contrast, ventricular endothelial cells did not form mesenchyme when cocultured with AV or ventricle myocardium. Immunohistochemical localization of Fn revealed three distinct morphological patterns of distribution in the AV-MBM, i.e., an intense lamina densa staining, diffuse staining in fibrils, and as particles. The Fn localized in particles (0.1 to 0.5 micron in diameter) appeared as a gradient of decreasing concentration extending from the myocardium toward the endothelium. In contrast, no particulate Fn staining was observed in the ventricular region. EDTA extraction selectively depleted the particulate form of Fn. Previous work has shown that this extract, which contains several lower Mr proteins in addition to Fn, is biologically active in initiating mesenchyme formation from AV endothelium in vitro. These results show that a regionally specific interaction of the myocardium with the endothelium is required to initiate the formation of prevalvular mesenchyme. This interaction may be mediated by a multicomponent complex involving Fn and other proteins which appear as a regionally distinct particulate only in areas of endothelial differentiation.

Animals↗
PubMed1987

Extracellular matrix from embryonic myocardium elicits an early morphogenetic event in cardiac endothelial differentiation.

A critical step in early cardiac morphogenesis can be faithfully duplicated in culture using a hydrated collagen substratum, and thereby serves as a useful model system for studying the molecular mechanisms of cell differentiation. Results from previous work suggested that the myocardium in the atrioventricular canal (AV) region of the developing chick heart secretes extracellular proteins into its associated basement membrane, which may function to promote an epithelial-mesenchymal transition of endothelium to form prevalvular fibroblasts (E. L. Krug, R. B. Runyan, and R. R. Markwald, 1985, Dev. Biol. 112, 414-426; C. H. Mjaatvedt, R. C. Lepera, and R. R. Markwald, 1987, Dev. Biol., in press). In the present study we show that an EDTA-soluble extract of embryonic chick hearts can substitute for the presence of myocardium, the presumptive stimulator tissue, in initiating mesenchyme formation from AV endothelium in culture. Ventricular endothelium was unresponsive to this material in keeping with observed in situ behavior. AV endothelial cells did not survive beyond 4-5 days when cultured in the absence of either the EDTA-soluble heart extract, myocardial conditioned medium, or the myocardium itself. Antibody prepared against a particulate fraction of the EDTA-solubilized heart extract immunohistochemically localized this material to the myocardial basement membrane. In addition, conditioned medium from embryonic myocardial cultures effectively induced mesenchyme formation. Neither a variety of growth factors nor a sarcoma basement membrane preparation were effective in promoting mesenchyme formation indicating a selectivity of the responding embryonic AV endothelial cells to myocardial basement membrane. These observations reflect a truly inductive phenomenon as there was an absolute dependence on the presence of the stimulating substance/tissue and retention, in culture, of both the temporal and regional characteristics observed in situ. This is in contrast to the results of others investigating the cytodifferentiation of committed cells whose phenotypic expression can be either accelerated or diminished but not obligatorily regulated by a specific agent, thus making the interpretation of data difficult, if not irrelevant, to the study of differentiation. The results of this study provide direct experimental support for the hypothesis that extracellular matrix can indeed serve as a direct stimulator or "secondary inducer" of cytodifferentiation.

Animals↗
PubMed1986

Conditioning of native substrates by chondroitin sulfate proteoglycans during cardiac mesenchymal cell migration.

It is generally proposed that embryonic mesenchymal cells use sulfated macromolecules during in situ migration. Attempts to resolve the molecular mechanisms for this hypothesis using planar substrates have been met with limited success. In the present study, we provide evidence that the functional significance of certain sulfated macromolecules during mesenchyme migration required the presence of the endogenous migratory template; i.e., native collagen fibrils. Using three-dimensional collagen gel lattices and whole embryo culture procedures to produce metabolically labeled sulfated macromolecules in embryonic chick cardiac tissue, we show that these molecules were primarily proteoglycan (PG) in nature and that their distribution was class specific; i.e., heparan sulfate PG, the minor labeled component (15%), remained pericellular while chondroitin sulfate (CS) PG, the predominately labeled PG (85%), was associated with collagen fibrils as "trails" of 50-60-nm particles when viewed by scanning electron microscopy. Progressive "conditioning" of collagen with CS-PG inhibited the capacity of the template to support subsequent cell migration. Lastly, metabolically labeled, PG-derived CS chains were compared with respect to degree of sulfation in either the C-6 or C-4 position by chromatographic separation of chondroitinase AC digestion products. Results from temporal and regional comparisons of in situ-labeled PGs indicated a positive correlation between the presence of mesenchyme and an enrichment of disaccharide-4S relative to that from regions lacking mesenchyme (i.e., principally myocardial tissue). The suggestion of a mesenchyme-specific CS-PG was substantiated by similarly examining the PGs synthesized solely by cardiac mesenchymal cells migrating within hydrated collagen lattice in culture. These data were incorporated into a model of "substratum conditioning" which provides a molecular mechanism by which secretion of mesenchyme-specific CS-PGs not only provides for directed and sustained cell movement, but ultimately inhibits migration of the cell population as a whole.

Animals↗