Development of a specific immunological assay for tropoelastin and its application to tissue culture studies

Foster, Jeffrey S.; Knaack, David; Faris, Barbara; Moscaritolo, Rosemarie; Salcedo, Lily L.; Skinner, Martha; Franzblau, Carl

doi:10.1016/0005-2795(76)90096-9

Cited by 19 publications

(5 citation statements)

References 13 publications

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“…Fra-1 and c-Jun rabbit polyclonal antibodies and horseradish peroxidase-conjugated anti-rabbit immunoglobulin G (IgG) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). The polyclonal elastin antibody, ST1 clone, was previously described by Foster et al (1976). PD-98059 was purchased from New England Biolabs (Beverly, MA).…”

Section: Reagentsmentioning

confidence: 99%

Basic fibroblast growth factor decreases elastin gene transcription in aortic smooth muscle cells

Carreras

Rich

Panchenko

et al. 2002

J of Cellular Biochemistry

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The extracellular matrix (ECM) protein elastin plays an essential role in the cardiovascular system by imparting elasticity to blood vessel wall. In this study, we examined the effect of basic fibroblast growth factor (bFGF) on the expression of elastin in aortic smooth muscle cells (SMC) to gain insight into events associated with cardiovascular diseases. The results show that bFGF treatment of SMC causes a significant decrease in elastin mRNA and secreted tropoelastin levels. Nuclear run-on analyses demonstrate that the downregulation is due to a decrease in the level of elastin gene transcription. Transient transfections of SMC with wild-type and mutated elastin gene promoter/chloramphenicol acetyl transferase (CAT) constructs show that a previously identified activator protein-1-cAMP response element (AP1/CRE) (-564 to -558-bp) within the elastin promoter mediates the bFGF-dependent downregulation of elastin gene transcription in SMC. Addition of bFGF to SMC activates the extracellular signal-regulated kinases 1/2 (ERK1/2) resulting in their translocation into the nucleus and subsequent induction of Fra-1. The addition of PD-98059, an inhibitor of ERK1/2 kinase, abrogates the bFGF-dependent decrease of elastin mRNA in SMC. The described inhibitory effect of bFGF on elastin gene expression in SMC may significantly contribute to the inefficient repair of elastin in early stages of vascular wall injury.

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Section: Reagentsmentioning

confidence: 99%

Basic fibroblast growth factor decreases elastin gene transcription in aortic smooth muscle cells

Carreras

Rich

Panchenko

et al. 2002

J of Cellular Biochemistry

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“…Immunohistochemical analyses was conducted according to Gruskin-Lerner and Trinkaus-Randall (1991). The chick tropoelastin polyclonal antibody, ST1, which is specific for newly synthesized elastin and does not react with insoluble elastin (Foster et al, 1975(Foster et al, , 1976Pollock et al, 1990), was diluted 1:20 (1% bovine serum albumin [BSA] in phosphate-buffered saline [PBS]) and its location detected with affinity purified goat antirabbit IgG antibody conjugated to fluorescein isothiocyanate (FITC; Boehringer-Mannheim, Indianapolis, IN) at a dilution of 1:100 (1% BSA in PBS). The monoclonal antibody directed against SMC ␣-actin (Boehringer Mannheim) was diluted to 1:100 in 3% BSA/PBS.…”

Section: Immunohistochemistry and In Situ Hybridizationmentioning

confidence: 99%

Elastogenesis in the developing chick lung is transcriptionally regulated

et al. 1998

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The overall goals of this study were to establish the level at which elastin gene expression is regulated during chick lung embryogenesis and to identify the temporal and spatial relationships among elastogenesis, smooth muscle cell differentiation, and cell proliferation. A comparison of lung elastin mRNA and transcriptional levels during embryogenesis shows that elastin expression is developmentally regulated at the transcriptional level. The increase in elastogenic activity occurs during the late stages of lung embryogenesis and coincides with terminal maturation of the tertiary bronchi. In situ hybridization analysis demonstrates that the increase in elastin mRNA expression is confined to the tertiary bronchial respiratory subunits, connective tissue septa, and supporting vasculature of the lung parenchyma. Immunohistochemical localization of smooth muscle cell α‐actin and tropoelastin suggests that α‐actin–immunoreactive cells of the lung parenchyma are a major contributor to the increase in elastin expression during embryogenesis. This observation is also reflected by Northern blot analysis, which demonstrates a temporal coincidence in the increase of both α‐actin and elastin mRNA levels. Histone mRNA expression, which was used as an index of cellular proliferation, reveals a level and spatial pattern inversely related to that of the elastin transcript. Tissue transfections of chick lungs isolated from 18‐day embryos with various elastin gene deletion/reporter constructs illustrate that the elastin promoter is not promiscuous within a tissue environment and that sequences spanning the −500 to +2 region are capable of directing promoter activity spatially comparable to the endogenous elastin gene. Dev. Dyn. 1998;213:170–181. © 1998 Wiley‐Liss, Inc.

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“…Immunological Techniques. Antibody to purified tropoelastin was raised in rabbits by a slight modification of a procedure we have previously published (Foster et al, 1976b). The only difference in the procedure was that the rabbits were injected in hind toepads.…”

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Translation of chick aortic elastin messenger ribonucleic acid. Comparison to elastin synthesis in chick aorta organ culture

Foster¹,

Rich²,

Fletcher³

et al. 1980

Biochemistry

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Studies were undertaken to define the molecular size of the elastin primary gene product. Translation of chick aortic messenger ribonucleic acid (mRNA) in an mRNA-dependent reticulocyte lysate resulted in the synthesis of two major proteins of 70 000 and 73 000 molecular weights. Both proteins were shown to be soluble forms of elastin by isotope incorporation, immunoprecipitation, collagenase and cyanogen bromide sensitivity, and two-dimensional gel electrophoresis. The 70 000-dalton protein behaves similarly to authentic tropoeleastin in sodium dodecyl sulfate gel electrophoresis. There was no evidence for a high molecular weight form of soluble elastin, although procollagen chains were indirectly identified among the aortic mRNA-directed translation products. The same molecular size proteins were also seen in organ cultures of chick embryonic aortas labeled with [3H]valine. However, the 73 000-dalton protein was not extractable in a neutral salt buffer but was found only if the aortas were extracted with urea in the presence of reducing and alkylating reagents. The results from these studies suggest that elastin is first synthesized as two distinct polypeptide chains which differ slightly in size and overall charge. The possibility that these two proteins may associate posttranslationally to form a dimer prior to secretion is postulated to explain the existence of a putative proelastin molecule seen in other systems.

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Development of a specific immunological assay for tropoelastin and its application to tissue culture studies

Cited by 19 publications

References 13 publications

Basic fibroblast growth factor decreases elastin gene transcription in aortic smooth muscle cells

Basic fibroblast growth factor decreases elastin gene transcription in aortic smooth muscle cells

Elastogenesis in the developing chick lung is transcriptionally regulated

Translation of chick aortic elastin messenger ribonucleic acid. Comparison to elastin synthesis in chick aorta organ culture

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