In order to study miR-29 function in collagen synthesis, we inserted the 3′-UTR sequences downstream of a luciferase reporter (). During progression of liver fibrosis, collagen IV is most prominently upregulated among ECM components . Therefore, in addition to collagen-1, the collagen-4 mRNA could be an important target of miR-29 in HSC after HGF stimulation. Indeed, insertion of the 3′-UTR of col4A1 and col4A5 downstream of the luciferase reporter gene lead to a reduction in luciferase expression after treatment of HSC with ago-miR-29a, mimicking miR-29a ().

Ranking list of putative collagen targets from miR-29*.

Enzymatic degradation of collagen-guided tissue regeneration membranes by periodontal bacteria.

The beginning of collagen synthesis begins with turning on ..

The invention is applicable in cosmetics, preferably in an anti-wrinkle treatment, and in pharmacy in the treatment of diseases associated with a deficiency of the dermo-epidermal junction resulting from an insufficient amount of collagen IV.

Skin and Collagen IV - IV For Life

Collagen, Type IV should be particularly useful in the development of HTS assays for screening Gelatinase A (MMP-2) and Gelatinase A inhibitors, as well as for other gelatinases and collagenases that specifically degrade (Type IV) Collagen.

Lack of association between morphological phenotype and collagen biosynthesis.

In vitro synthesis of type IV procollagen - ResearchGate

In the present study, we investigate the role of the members of the miR-29 family in HGF mediated repression of collagen synthesis. We demonstrate that miR-29 is not only involved in collagen type I but also in type IV synthesis of myofibroblastic HSC. The importance of miR-29 in hepatic collagen homeostasis is underlined by our in vivo data that shows the lack of miR-29 in severe experimental fibrosis after bile duct obstruction. This loss of miR-29 is suggested to be due to the response of HSC to exposure to profibrogenic mediators as shown by our in vitro findings on TGF-β stimulated HSC. Whereas TGF-β stimulation leads to decreased miR-29 levels, but to pronounced upregulation of collagen synthesis, HGF stimulation leads to elevated miR-29 expression, but to repression of collagen synthesis. Thus, our data provide detailed evidence for the antifibrotic action of miR-29 in response to HGF signalling that is counteracted by the profibrotic growth factor TGF-β.

Increased Collagen IV Excretion in Diabetes | Diabetes Care

The synthesis of extracellular matrix proteins is modulated by microRNA-29 (miR-29) in extrahepatic tissue , , , . Recent reports suggest that miR-29 is also involved in the synthesis of collagen type I in liver fibrosis , . The miR-29 family consists of miR-29a, miR-29b (b1, b2), and miR-29c, which differ in only two or three nucleotides, respectively. The genes for miR-29a and miR-29b1 are both located on chromosome 7, whereas the genes for miR-29c and miR-29b2 are located on chromosome 1. Each gene pair is transcribed in tandem resulting in a common pri-miRNA from which the mature miR-29 members are released after further processing , .

Putative binding sites of the members of the miR-29 family to the 3′-UTR of different collagens.

Collagen, type IV, alpha 3 - Wikipedia

The 3′-UTR of collagen 4A1 (col4A1) and collagen 4A5 (col4A5) transcripts bearing putative miRNA-binding sites for miR-29 were amplified from genomic rat DNA using the primer shown in the . Furthermore, the putative binding sites of collagen 1A1 (col1A1), collagen 1A2 (col1A2), collagen 4A1(col4A1), and collagen 4A5 (col4A5) as well as the corresponding mutants containing two point mutations were cloned by insertion of the dimerized oligonucleotides shown in the . Amplicons and oligonucleotide dimers were cloned downstream of the renilla luciferase reporter of the psiCHECKTM-2 vector (Ambion, Austin, USA) ().

Collagen Synthesis and Disorders Flashcards | Quizlet

Total protein concentrations were determined using the BCA protein assay (Thermo Scientific, Bonn, Germany). Equal amounts of protein (10 µg) were resolved on 4–12% SDS-polyacrylamid gels (Biorad, München, Germany), electrotransferred to Hybond ECL nitrocellulose membranes (Amersham Biosciences, München, Germany) and immunostained with the anti-collagen I (11000), anti-collagen IV (15000) antibodies, both from Abcam (Cambridge, UK), or with anti-Met antibodies (1500) from Santa Cruz (Heidelberg, Germany), or anti-actin antibodies (15000) from Sigma-Aldrich (Hamburg, Germany). Signals were visualized and quantified using enhanced chemiluminescence (ECL) substrate and a FLUORCHEM®FC2 Alpha Ease scanner (Biozym, Hess. Oldendorf, Germany).

Collagen IV is essential for basement ..

However, in four other cell lines, even though total synthesis was increased, the type of proteins secreted by these cells was not altered.(ABSTRACT TRUNCATED AT 250 WORDS)