Developmental and Cell-Type-Specific Expression of Cell Surface Heparan Sulfate Proteoglycans in the Rat Heart

doi:10.1006/excr.1996.3400

Experimental Cell Research

Volume 230, Issue 1, 10 January 1997, Pages 145-153

https://doi.org/10.1006/excr.1996.3400 Get rights and content

Abstract

The expression of cell surface heparan sulfate proteoglycans in rat heart was investigated by Northern blot analysis with specific cDNA probes. In adult heart syndecan-3 and glypican mRNAs were abundantly expressed. Lower levels of syndecan-2 mRNA and very low levels of syndecan-1 mRNA were also detected. Analysis of RNA isolated from hearts of rats of various ages revealed that syndecan-3 and glypican mRNAs levels increased dramatically at birth, and continued to be expressed at high levels in adult animals. To determine which of these proteoglycans was expressed in cardiomyocytes, primary cultures of cardiomyocytes and nonmyocytes isolated from neonatal rat hearts were analyzed for proteoglycan expression. Glypican mRNA was localized almost exclusively to cardiomyocytes. Syndecan-3 mRNA was not detected in myocytes, but was detected in the nonmyocyte cells. Biochemical characterization of cardiomyocyte glypican revealed that it was a phosphatidylinositol-anchored heparan sulfate proteoglycan. Results of immunofluorescent staining of rat hearts with anti-glypican antibodies were consistent with the Northern blot data, and localized glypican to the lateral regions of myocyte plasma membrane that contact the basement membrane, as well as sites of myocyte adhesion junctions. At the latter site glypican colocalized with vinculin. Visualization of basic fibroblast growth factor binding sites by means of a tissue slice overlay assay also revealed colocalization with glypican. These results demonstrate developmental and cell-type-specific expression of membrane heparan sulfate proteoglycans in the heart. They also show that glypican is a major heparan sulfate proteoglycan expressed on the cardiomyocyte plasma membrane.

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Cited by (27)

Increased syndecan-1 and glypican-3 predict poor perinatal outcome and treatment resistance in intrahepatic cholestasis
2020, Hepatobiliary and Pancreatic Diseases International
Citation Excerpt :
Szabo et al. [36] showed the increased density of syndecan-1 in the placentas of patients with preeclampsia. Glypican-3 is expressed in the cardiomyocytes of neonatal rats [37]. The SBA was accused of accumulating in the fetal cardiomyocytes and fetal lungs, causing IUFD in ICP patients [38,39].
Intrahepatic cholestasis of pregnancy (ICP) increases the risk of adverse pregnancy outcomes. This study aimed to explore the association between serum syndecan-1 and glypican-3 levels and the adverse perinatal outcome as well as the responses to the treatment of ursodeoxycholic acid (UDCA).
This prospective, case control study included 88 pregnant women (44 women with ICP and 44 healthy controls). The primary end points were the perinatal outcome and the response to UDCA therapy. A logistic regression model was used to identify the independent risk factors of adverse pregnancy outcomes and reduced response to UDCA therapy.
Women with ICP had significantly higher serum syndecan-1 (1.27 ± 0.36 ng/mL vs. 0.98 ± 0.50 ng/mL; P = 0.003), glypican-3 (1.78 ± 0.13 ng/mL vs.1.69 ± 0.16 ng/mL; P = 0.004), AST (128.59 ± 1.44 vs. 13.29 ± 1.32 U/L; P < 0.001), and ALT (129.84 ± 1.53 vs. 8.00 ± 3.67 U/L; P < 0.001) levels compared with the controls. The increased levels of syndecan-1 (OR = 4.715, 95% CI: 1.554–14.310; P = 0.006), glypican-3 (OR = 8.465, 95% CI: 3.372–21.248; P = 0.007), ALT (OR = 1.382, 95% CI: 1.131–1.690; P = 0.002), and postprandial bile acid (PBA) (OR = 3.392, 95% CI: 1.003–12.869; P = 0.026) were correlated to ICP. The adverse neonatal outcome was related to increased glypican-3 (OR = 4.275, 95% CI: 2.726–5.635; P = 0.039), and PBA (OR = 3.026, 95% CI: 1.069–13.569; P = 0.037). Increases of syndecan-1 (OR = 7.464, 95% CI: 2.130–26.153, P = 0.017) and glypican-3 (OR = 6.194, 95% CI: 2.951–13.002; P = 0.025) were the risk factors of decreased response to UDCA treatment.
Syndecan-1 and glypican-3 might be powerful determinants in predicting adverse perinatal outcome in patients with ICP, and they can be used to predict the response to the UDCA treatment.
Syndecan-1 Mediates Sorting of Soluble Lipoprotein Lipase with Sphingomyelin-Rich Membrane in the Golgi Apparatus
2019, Developmental Cell
Citation Excerpt :
In adult tissues, SDC1 is chiefly expressed in epithelial cells, where it localizes to the basolateral domain. Nevertheless, SDC1 expression has been observed at low levels in heart tissue (Asundi et al., 1997) and adipocytes (Zaragosi et al., 2015), where it is tempting to speculate that low SDC1 expression levels could be responsible for limiting the rate of LPL secretion. Indeed, heparin treatment is known to induce a massive exocytosis of LPL from cultured adipocytes (Vannier and Ailhaud, 1989).
In the secretory pathway, budding of vesicular transport carriers from the trans-Golgi network (TGN) must coordinate specification of lipid composition with selection of secreted proteins. We elucidate a mechanism of soluble protein cargo sorting into secretory vesicles with a sphingomyelin-rich membrane; the integral membrane proteoglycan Syndecan-1 (SDC1) acts as a sorting receptor, capturing the soluble enzyme lipoprotein lipase (LPL) during export from the TGN. Sorting of LPL requires bivalent interactions between LPL and SDC1-linked heparan sulfate chains and between LPL and the Golgi membrane. Physical features of the SDC1 transmembrane domain, rather than a specific sequence, confer targeting of SDC1 and bound LPL into the sphingomyelin secretion pathway. This study establishes that physicochemical properties of a protein transmembrane domain that drive lateral heterogeneity of the plasma membrane also operate at the TGN to confer sorting of an integral membrane protein and its ligand within the biosynthetic secretory pathway.
Furosemide modifies heart hypertrophy and glycosaminoglycan myocardium content in a rat model of neurogenic hypertension
2016, European Journal of Pharmacology
Citation Excerpt :
However, the presence of heparan sulphate, heparin and chondroitin sulphates B and C has been documented, but to a lesser extent to that of hyaluronic acid. Aside from the histochemical method used for localization of GAGs populations in the rat myocardium and aorta, several studies confirm that hyaluronic acid is an essential component in the myocardium of newborn and adult rats (Papakonstantinou et al., 1998b) and also all principal types of GAGs are present in the matrix surrounding the cardiomyocytes in the heart of rats (Gonzalez et al., 1992; Asundi et al., 1997;Kumar et al., 2006;Sreeja and Leelamma, 2002). In the present study, evidence was provided that neurogenic hypertension due to bAD, significantly reduced the content of total GAGs in the heart 15 days postoperatively, indicating that GAGs homeostasis in the myocardium is influenced, at least in part, by increased pressure stimuli.
Hypertension is a major risk factor for atherogenesis and heart hypertrophy, both of which are associated with specific morphological and functional changes of the myocardium. Glycosaminoglycans (GAGs) are complex molecules involved both in tissue morphology and function. In the present study, we investigated the effects of neurogenic hypertension and subsequent antihypertensive treatment with furosemide, on heart hypertrophy and the content of GAGs in the myocardium. Neurogenic hypertension was achieved in male Wistar rats by bilateral aortic denervation (bAD). At days 2, 7 and 15 after surgery, animals were sacrificed and the hearts were dissected away, weighted, and homogenized. Total GAGs were assessed by measuring the uronic acid content colorimetrically and individual GAGs were isolated and characterized by enzymatic treatment, with GAG-degrading enzymes, using electrophoresis on polyacrylamide gradient gels and cellulose acetate membranes. In bAD-animals blood pressure, blood pressure lability, heart rate and heart weight were significantly increased 15 days postoperatively. These effects were prevented by treatment with furosemide. Major GAGs identified in the heart were chondroitin sulphates, heparin (H), heparan sulphate (HS) and hyaluronic acid. The content of uronic and the relative content of H and HS in the heart in bAD animals significantly decreased from day 2 to day 15 postoperatively. Furosemide prevented the bAD induced decrease in GAG content. Considering that H and HS are potent inhibitors of cardiomyocyte hypertrophy, our results indicate that heart hypertrophy induced by neurogenic hypertension may be associated with decreases in the relative content of heparin and heparan sulphate in the heart.
The kinetics of FGF-2 binding to heparan sulfate proteoglycans and MAP kinase signaling
2005, Journal of Theoretical Biology
Binding of growth factors to specific cell surface receptors is the first step in initiating cell signaling cascades that ultimately result in diverse activities such as proliferation, differentiation, and apoptosis. Dimerization and phosphorylation of tyrosine kinase transmembrane receptors is the typical paradigm for this activation but, for many growth factors, cell surface interactions are not limited to a single receptor type. In particular, heparin-binding growth factors, such as fibroblast growth factor-2 (FGF-2), bind to heparan sulfate proteoglycans (HSPG) on the cell surface and within the extracellular matrix (ECM), and these molecules have been viewed as accessory co-receptors serving to facilitate tyrosine kinase receptor binding. Recent studies, however, have indicated that HSPG can directly participate in signal transduction in response to FGF-2 binding. Thus, in the present study, we used mathematical modeling to examine whether the kinetics of formation of the various FGF-2 bound complexes on the cell surface correlate with the activation of the downstream mediators of FGF-2 response, Erk1/2. We find that FGF-2 binding to its receptor correlates well with Erk1/2 activation and that HSPG can modulate this response through its ability to stabilize these ligand receptor complexes. Moreover, we also observed that FGF-2 binding to HSPG correlates strongly with Erk1/2 activation under conditions where there is a loss of receptor activity, and we demonstrate that the relative amounts of signaling and non-signaling HSPG on the cell surface, as well as the presence of competing HSPG in the ECM, can impact the signal potential via this pathway. Thus, the selective regulation of specific HSPG might provide a mechanism for fine tuned modulation of heparin-binding growth factor signaling in cells where signal intensity and duration could direct cellular response toward growth, migration or differentiation.
The role of glypicans in mammalian development
2002, Biochimica et Biophysica Acta - General Subjects
Glypicans are a family of heparan sulfate proteoglycans that are bound to the cell surface by a glycosyl-phosphatidylinositol anchor. Six members of this family have been identified in mammals. In general, glypicans are highly expressed during development, and their expression pattern suggests that they are involved in morphogenesis. One member of this family, glypican-3, is mutated in the Simpson–Golabi–Behmel syndrome. This syndrome is characterized by overgrowth and various developmental abnormalities that indicate that glypican-3 inhibits proliferation and cell survival in the embryo. It has consequently been proposed that glypicans can regulate the activity of several growth factors that play a critical role in morphogenesis.
Schwann cells synthesize type v collagen that contains a novel α4 chain: Molecular cloning, biochemical characterization, and high affinity heparin binding of α4(v) collagen
2000, Journal of Biological Chemistry
Citation Excerpt :
Aliquots of column fractions were analyzed on 4–15% SDS-polyacrylamide gradient gels followed by silver staining and immunoblotting with anti-p200 or anti-α1(V) collagen antibodies. Aliquots were also analyzed by immunoblotting with antibodies against the heparan sulfate proteoglycans syndecan-3 (7) and glypican-1 (8). Anti-proteoglycan immunoreactivity was not detected in the purified collagen preparation (data not shown).
Previously, we reported the isolation of a heparan sulfate-binding collagenous protein, p200, that is expressed by Schwann cells in developing peripheral nerves ((1996) J. Biol. Chem. 271, 13844–13853; (1999) J. Neurosci. Res. 56, 284–294). Here, we report the cloning of p200 cDNA from a Schwann cell cDNA library. The deduced amino acid sequence identifies p200 as a novel member of the collagen type V gene family. This polypeptide, which we have named α4 type V (α4(V)) collagen, contains an uninterrupted Gly-X-Xcollagen domain of 1011 amino acids that shows 82% sequence identity to human α3(V) collagen and 71% identity to rat α1(V) collagen. α4(V) is secreted by Schwann cells as a collagen heterotrimer that also contains α1(V) chains. α4(V)-containing collagen molecules synthesized by Schwann cells retain their amino-terminal non-collagenous domains. α4(V) mRNA was detected by reverse transcriptase-linked polymerase chain reaction amplification in neonatal and adult brain and neonatal peripheral nerve. α4(V) mRNA and protein were not detected in most other tissues, including the placenta and heart, which are known to contain α3(V). This pattern of α4(V) expression contrasted with that of α1(V) mRNA and protein, which were ubiquitously expressed. The isolated α4(V) chain demonstrated an unusually high affinity for heparin. The restricted expression and unusual properties of α4(V)-containing collagen type V molecules suggest a unique and important role for these molecules in peripheral nerve development.

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Regular ArticleDevelopmental and Cell-Type-Specific Expression of Cell Surface Heparan Sulfate Proteoglycans in the Rat Heart

Abstract

Regular Article
Developmental and Cell-Type-Specific Expression of Cell Surface Heparan Sulfate Proteoglycans in the Rat Heart