A 25.7 × 10(3) M(r) hydra metalloproteinase (HMP1), a member of the astacin family, localizes to the extracellular matrix of Hydra vulgaris in a head-specific manner and has a developmental function

Development ◽  
1995 ◽  
Vol 121 (6) ◽  
pp. 1591-1602 ◽  
Author(s):  
L. Yan ◽  
G.H. Pollock ◽  
H. Nagase ◽  
M.P. Sarras
Keyword(s):  
Extracellular Matrix ◽  
Gelatin Zymography ◽  
Longitudinal Axis ◽  
The Body ◽  
Cdna Clones ◽  
Head Region ◽  
Differential Distribution ◽  
Functional Studies ◽  
Type Iv ◽  
Sds Page

Hydra extracellular matrix (ECM) is composed of a number of components seen in vertebrate ECM such as laminin, type IV collagen, fibronectin, and heparan sulfate proteoglycan. A number of functional studies have shown that hydra ECM plays an important role in pattern formation and morphogenesis of this simple metazoan. The present study was designed to identify matrix degrading proteinases in hydra and determine their potential function in hydra morphogenesis. Using SDS-PAGE gelatin-zymography, five gelatinolytic bands were identified with relative molecular masses of 67 × 10(3), 51–58 × 10(3) (a triplet) and 25–29 × 10(3), respectively. Inhibition studies indicated that all of these gelatinases were metalloproteinases. Gelatin-zymography indicated that there was a differential distribution of these gelatinases along the longitudinal axis of hydra, with the 67 × 10(3) M(r) gelatinase being concentrated in the body column, while the 51–58 × 10(3) M(r) gelatinase triplet and the 25–29 × 10(3) M(r) gelatinase concentrated in the head region. Purification procedures were successfully developed for the 25–29 × 10(3) M(r) metalloproteinase which has been termed hydra metalloproteinase 1 (HMP1) and which appeared as a single band with a SDS-PAGE mobility of 25.7 × 10(3) M(r). The N-terminal sequence of purified HMP1 indicated that it has structural homology with metalloproteinases that belong to the astacin family. Subsequent cloning and sequencing of cDNA clones confirmed the identification of HMP1 as an astacin-like metalloproteinase. Immunocytochemical studies with antibodies generated against the purified enzyme and to a synthetic peptide indicated that HMP1 was localized to the ECM of tentacles. Functional studies were performed in which purified HMP1, anti-HMP1 IgG, or suspected substrates of HMP1 (e.g. growth factors such as TGF-beta 1) were introduced into the interepithelial compartment of hydra using a ‘DMSO loading’ procedure. These studies indicated that HMP1 has a functional role during a number of developmental processes such as head regeneration and cell differentiation/transdifferentiation of tentacle battery cells.

A novel hydra matrix metalloproteinase (HMMP) functions in extracellular matrix degradation, morphogenesis and the maintenance of differentiated cells in the foot process

Development ◽  
2000 ◽  
Vol 127 (4) ◽  
pp. 907-920 ◽  
Author(s):  
A.A. Leontovich ◽  
J. Zhang ◽  
K. Shimokawa ◽  
H. Nagase ◽  
M.P. Sarras
Keyword(s):  
Extracellular Matrix ◽  
Matrix Metalloproteinase ◽  
Longitudinal Axis ◽  
The Body ◽  
Mmp Inhibitors ◽  
Functional Studies ◽  
Cell Transdifferentiation ◽  
Basal Disk ◽  
Foot Process ◽  
Hemopexin Domain

As a member of Cnidaria, the body wall of hydra is structurally reduced to an epithelial bilayer with an intervening extracellular matrix (ECM). Biochemical and cloning studies have shown that the molecular composition of hydra ECM is similar to that seen in vertebrates and functional studies have demonstrated that cell-ECM interactions are important to developmental processes in hydra. Because vertebrate matrix metalloproteinases (MMPs) have been shown to have an important role in cell-ECM interactions, the current study was designed to determine whether hydra has homologues of these proteinases and, if so, what function these enzymes have in morphogenesis and cell differentiation in this simple metazoan. Utilizing a PCR approach, a single hydra matrix metalloproteinase, named HMMP was identified and cloned. The structure of HMMP was similar to that of vertebrate MMPs with an overall identity of about 35%. Detailed structural analysis indicated some unique features in (1) the cysteine-switch region of the prodomain, (2) the hinge region preceding the hemopexin domain, and (3) the hemopexin domain. Using a bacterial system, HMMP protein was expressed and folded to obtain an active enzyme. Substrate analysis studies indicated that recombinant HMMP could digest a number of hydra ECM components such as hydra laminin. Using a fluorogenic MMP substrate assay, it was determined that HMMP was inhibited by peptidyl hydroxamate MMP inhibitors, GM6001 and matlistatin, and by human recombinant TIMP-1. Whole-mount in situ studies indicated that HMMP mRNA was expressed in the endoderm along the entire longitudinal axis of hydra, but at relatively high levels at regions where cell-transdifferentiation occurred (apical and basal poles). Functional studies using GM6001 and TIMP-1 indicated that these MMP inhibitors could reversibly block foot regeneration. Blockage of foot regeneration was also observed using antisense thio-oligo nucleotides to HMMP introduced into the endoderm of the basal pole using a localized electroporation technique. Studies with adult intact hydra found that GM6001 could also cause the reversible de-differentiation or inhibition of transdifferentiation of basal disk cells of the foot process. Basal disk cells are adjacent to those endoderm cells of the foot process that express high levels of HMMP mRNA. In summary, these studies indicate that hydra has at least one MMP that is functionally tied to morphogenesis and cell transdifferentiation in this simple metazoan.

The Role of Extracellular Matrix in the Morphogenesis and Differentiation of Salivary Glands

1990 ◽  
Vol 4 (1) ◽  
pp. 27-33 ◽  
Author(s):  
L.S. Cutler
Keyword(s):  
Extracellular Matrix ◽  
Salivary Gland ◽  
Type Iv Collagen ◽  
The Body ◽  
Secretory Cells ◽  
Type Iv ◽  
Preliminary Information ◽  
Extracellular Matrix Molecules ◽  
Gland Morphogenesis

The processes of morphogenesis and cytodifferentiation are partially linked, independently regulated processes. The full expression of both processes is modulated or controlled, at least in part, by components of the extracellular matrix. This paper reviews the body of work that demonstrates a role for epithelial-mesenchymal interactions and various extracellular matrix molecules in the induction, control, and maintenance of salivary gland morphogenesis and cytodifferentiation. In addition, new, preliminary information which further elucidates the role of laminin and type IV collagen in the processes of morphogenesis and cytodifferentiation is presented. With regard to the role of extracellular matrix molecules in the regulation of salivary gland morphogenesis and cytodifferentiation, it appears that types I, III, and IV collagen, laminin, and chondroitin sulfate proteoglycan play roles in the control of glandular morphogenesis. With the exception of type IV collagen, these molecules do not appear to be involved in the regulation of cytodifferentiation of salivary gland secretory cells. On the other hand, of the extracellular matrix molecules tested so far, only type IV collagen appears to play a role in the regulation of salivary gland secretory cell differentiation.

Abstract 361: Epistasis Analysis Identifies Extracellular Matrix Genes Interacting With the COL4A1/COL4A2 Coronary Artery Disease Locus

2014 ◽  
Vol 34 (suppl_1) ◽  
Author(s):  
Adam Turner ◽  
Majid Nikpay ◽  
Paulina Lau ◽  
Sebastien Soubeyrand ◽  
Ruth McPherson
Keyword(s):  
Coronary Artery Disease ◽  
Extracellular Matrix ◽  
Coronary Artery ◽  
Association Studies ◽  
Cleveland Clinic ◽  
Genome Wide Association Studies ◽  
Functional Studies ◽  
Epistasis Analysis ◽  
Type Iv ◽  
Artery Disease

The COL4A1/COL4A2 locus on chromosome 13q34 was significantly associated with coronary artery disease (CAD) in the CARDIoGRAM (Nature Genetics, 2011) and the CARDIoGRAMPlusC4D (Nature Genetics, 2013) meta analyses of several large genome-wide association studies (GWAS). Determination of causative SNPs stemming from GWAS data requires a number of bioinformatic and lab based approaches. Functional analysis of GWAS loci can be aided by epistasis analysis, which interrogates synergistic associations with a given trait between pairs of SNPs, either at the same locus or at different loci. We investigated 4 CAD GWAS cohorts in this study (Ottawa Heart Genomics Study A, Ottawa Heart Genomics Study B, Cleveland Clinic Gene Bank, Duke CATHGEN Study). For epistasis analysis we tested whether CAD-associated SNPs at the COL4A1/COL4A2 locus displayed interaction with other CAD-associated SNPs across the genome and used 2 lists of SNPs. The first list of SNPs were ones at the COL4A1/COL4A2 locus significant for CAD association and the second list of SNPs were ones significantly associated with CAD from across the genome. Using PLINK we then generated a list of SNP pairs showing interaction (Pinteraction<0.05) for CAD association within 1 or more cohorts. Epistasis analysis generated over a dozen loci that demonstrated interaction with COL4A1/COL4A2 SNPs for CAD association, many of which have putative functional roles with respect to CAD. Genes integral to pathways involving COL4A1/COL4A2 were of particular interest in understanding the role of type IV collagen in CAD pathogenesis. We identified several extracellular matrix genes showing interaction with COL4A1/COL4A2 including fibronectin (FN1), COL18A1 as well as transmembrane proteins including ITGA2, encoding integrin α2 and SERPINF1, a member of the serpin peptidase family that strongly inhibits angiogenesis. Other interesting loci include NOTCH4, genes involved in angiogenesis, and mediators of TGFβ signalling. Functional studies are currently being conducted to decipher the roles of COL4A1/COL4A2 and these interacting extracellular matrix genes in CAD.

Functional analysis of chondroitin 4 sulfate constituting osseointegration

Impact ◽  
2019 ◽  
Vol 2019 (8) ◽  
pp. 18-20
Author(s):  
Shuhei Tsuchiya
Keyword(s):  
Functional Analysis ◽  
Extracellular Matrix ◽  
Dental Implants ◽  
Maxillofacial Surgery ◽  
The Body ◽  
Oral And Maxillofacial Surgery ◽  
Direct Connection ◽  
Tooth Replacement ◽  
Living Bone ◽  
Natural Tooth

Osseointegration can be defined as a direct connection, both structural and functional, between living bone and the surface of an artificial implant. Indeed, the word comes from the Greek term for 'bone' and 'to make whole'. In dentistry, once dental implants are placed, the body will react with osseointegration, enabling the implants to become a permanent part of the jaw. There are many benefits to this type of implant, compared with traditional tooth replacement options, not least that dental implants mimic the strength and functionality of a natural tooth. Dr Shuhei Tsuchiya is a researcher based in the Division of Oral and Maxillofacial Surgery at Nagoya University, Japan, who is interested in a range of areas, including regenerative medicine and the extracellular matrix. One of his key preoccupations, though, is shedding light on osseointegration. He and his team are working to unravel the mysteries of the mechanism.

scholarly journals Quantitative Evaluation of Osteocyte Morphology and Bone Anisotropic Extracellular Matrix in Rat Femur

2021 ◽  
Author(s):  
Takuya Ishimoto ◽  
Keita Kawahara ◽  
Aira Matsugaki ◽  
Hiroshi Kamioka ◽  
Takayoshi Nakano
Keyword(s):  
Extracellular Matrix ◽  
Longitudinal Axis ◽  
Rat Femur ◽  
Male Rat ◽  
Mechanical Stimuli ◽  
Axis Orientation ◽  
Principal Stress Direction ◽  
Mechanical Integrity ◽  
Osteocyte Lacunae ◽  
The Relationship

AbstractOsteocytes are believed to play a crucial role in mechanosensation and mechanotransduction which are important for maintenance of mechanical integrity of bone. Recent investigations have revealed that the preferential orientation of bone extracellular matrix (ECM) mainly composed of collagen fibers and apatite crystallites is one of the important determinants of bone mechanical integrity. However, the relationship between osteocytes and ECM orientation remains unclear. In this study, the association between ECM orientation and anisotropy in the osteocyte lacuno-canalicular system, which is thought to be optimized along with the mechanical stimuli, was investigated using male rat femur. The degree of ECM orientation along the femur longitudinal axis was significantly and positively correlated with the anisotropic features of the osteocyte lacunae and canaliculi. At the femur middiaphysis, there are the osteocytes with lacunae that highly aligned along the bone long axis (principal stress direction) and canaliculi that preferentially extended perpendicular to the bone long axis, and the highest degree of apatite c-axis orientation along the bone long axis was shown. Based on these data, we propose a model in which osteocytes can change their lacuno-canalicular architecture depending on the mechanical environment so that they can become more susceptible to mechanical stimuli via fluid flow in the canalicular channel.

scholarly journals Optical Microscopy and the Extracellular Matrix Structure: A Review

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1760
Author(s):  
Joshua J. A. Poole ◽  
Leila B. Mostaço-Guidolin
Keyword(s):  
Extracellular Matrix ◽  
Confocal Microscopy ◽  
Laser Scanning ◽  
Biological Tissues ◽  
The Body ◽  
Stimulated Emission ◽  
Substantial Part ◽  
Fluorescence Lifetime Imaging ◽  
Structured Illumination ◽  
Structured Illumination Microscopy

Biological tissues are not uniquely composed of cells. A substantial part of their volume is extracellular space, which is primarily filled by an intricate network of macromolecules constituting the extracellular matrix (ECM). The ECM serves as the scaffolding for tissues and organs throughout the body, playing an essential role in their structural and functional integrity. Understanding the intimate interaction between the cells and their structural microenvironment is central to our understanding of the factors driving the formation of normal versus remodelled tissue, including the processes involved in chronic fibrotic diseases. The visualization of the ECM is a key factor to track such changes successfully. This review is focused on presenting several optical imaging microscopy modalities used to characterize different ECM components. In this review, we describe and provide examples of applications of a vast gamut of microscopy techniques, such as widefield fluorescence, total internal reflection fluorescence, laser scanning confocal microscopy, multipoint/slit confocal microscopy, two-photon excited fluorescence (TPEF), second and third harmonic generation (SHG, THG), coherent anti-Stokes Raman scattering (CARS), fluorescence lifetime imaging microscopy (FLIM), structured illumination microscopy (SIM), stimulated emission depletion microscopy (STED), ground-state depletion microscopy (GSD), and photoactivated localization microscopy (PALM/fPALM), as well as their main advantages, limitations.

Differential distribution of type IV collagen chains in the developing rat testis and ovary

1998 ◽  
Vol 63 (3) ◽  
pp. 125-130 ◽  
Author(s):  
Kim Fröjdman ◽  
Lauri J. Pelliniemi ◽  
Ismo Virtanen
Keyword(s):  
Type Iv Collagen ◽  
Differential Distribution ◽  
Rat Testis ◽  
Type Iv ◽  
Developing Rat

scholarly journals Insights into the Genome Sequence ofChromobacterium amazonenseIsolated from a Tropical Freshwater Lake

2018 ◽  
Vol 2018 ◽  
pp. 1-10
Author(s):  
Alexandre Bueno Santos ◽  
Patrícia Silva Costa ◽  
Anderson Oliveira do Carmo ◽  
Gabriel da Rocha Fernandes ◽  
Larissa Lopes Silva Scholte ◽  
...  
Keyword(s):  
De Novo ◽  
Genomic Diversity ◽  
Protein Coding ◽  
Biotechnological Potential ◽  
Draft Assembly ◽  
Functional Studies ◽  
Alpha Hemolysin ◽  
Type Iv ◽  
Nudix Hydrolases ◽  
Colicin V

Members of the genusChromobacteriumhave been isolated from geographically diverse ecosystems and exhibit considerable metabolic flexibility, as well as biotechnological and pathogenic properties in some species. This study reports the draft assembly and detailed sequence analysis ofChromobacterium amazonensestrain 56AF. The de novo-assembled genome is 4,556,707 bp in size and contains 4294 protein-coding and 95 RNA genes, including 88 tRNA, six rRNA, and one tmRNA operon. A repertoire of genes implicated in virulence, for example, hemolysin, hemolytic enterotoxins, colicin V, lytic proteins, and Nudix hydrolases, is present. The genome also contains a collection of genes of biotechnological interest, including esterases, lipase, auxins, chitinases, phytoene synthase and phytoene desaturase, polyhydroxyalkanoates, violacein, plastocyanin/azurin, and detoxifying compounds. Importantly, unlike otherChromobacteriumspecies, the 56AF genome contains genes for pore-forming toxin alpha-hemolysin, a type IV secretion system, among others. The analysis of theC. amazonensestrain 56AF genome reveals the versatility, adaptability, and biotechnological potential of this bacterium. This study provides molecular information that may pave the way for further comparative genomics and functional studies involvingChromobacterium-related isolates and improves our understanding of the global genomic diversity ofChromobacteriumspecies.

Metalloproteases and egg-hatching in Pediculus humanus, the body (clothes) louse of humans (Phthiraptera: Insecta)

Parasitology ◽  
2007 ◽  
Vol 135 (1) ◽  
pp. 125-130 ◽  
Author(s):  
V. M. BOWLES ◽  
A. R. YOUNG ◽  
S. C. BARKER
Keyword(s):  
Protease Activity ◽  
The Body ◽  
Egg Hatching ◽  
Egg Hatch ◽  
Pediculus Humanus ◽  
Sds Page ◽  
Metalloprotease Inhibitors ◽  
Egg Shell ◽  
Ph Range

SUMMARYTo investigate the biochemical components of egg-hatch in the body louse, Pediculus humanus, egg-shell-washings (ESW) were collected during the first 2 h post-hatching and analysed by gelatin SDS-PAGE. These ESW contained proteases with molecular mass in the range of 25–100 kDa; the most abundant proteases were ~25 kDa. The 3 main regions of protease activity in the one-dimensional gelatin SDS-PAGE gels resolved to at least 23 distinct regions of protease activity when analysed by two-dimensional gelatin SDS-PAGE, with iso-electric points spread over the entire 3 to 10 pH range. Mechanistic characterization indicated that the ESW contained proteases of the metallo-class, inhibited by both 1,10-phenanthroline and EDTA. Several protease inhibitors were tested for their ability to inhibit louse egg-hatch in vitro. The metalloprotease inhibitor 1,10-phenanthroline and the aminopeptidase inhibitor bestatin significantly inhibited (P<0·05) louse egg-hatch (100% and 58%, respectively). The presence of metalloproteases at the time of egg-hatch and the inhibition of egg-hatch in P. humanus by metalloprotease inhibitors suggests a crucial role for these proteases in the hatching of this medically important parasite.

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