Extracellular Matrix 2

From CellBiology

Extracellular Matrix 2

Basal Lamina

Introduction

This second lecture on ECM will cover the glycoprotein components of ECM and specialized epithelial ECM. Finally I will discuss some key experiments exploring the role and function of the ECM of epitheilia (basement membrane) and connective tissues. Basement membranes are mainly composed of laminins, collagen IVs and proteoglycans.

With the epithelial ECM the term "basement membrane" is used with light microscopy and "basal lamina" is used with electron microscopy.


Lecture Slides: 2017 Lecture PDF

Note

Take a look at the 2015 student group projects designed around ECM - 2015 Extracellular Matrix: Group 1 - Small Leucine-Rich Proteoglycans | Group 2 - Integrins | Group 3 - Elastic Fibres | Group 4 - Fibronectin | Group 5 - Laminin | Group 6 - Collagen | Group 7 - Basement Membrane

The projects Small Leucine-Rich Proteoglycans, Fibronectin, Laminin and Basement Membrane relate specifically to this lecture content.

Lab 7 Individual Assessment  
The following peer assessment exercise should be completed before next lab (Lab 8 - 2 May) as your individual assessment for this week (lab missed due to public holiday).

Your answer should be pasted in 2 places

  1. onto each project discussion page (Note you should add anonymously to the discussion page)
  2. your own individual student page for my assessment.

Each individual will provide a brief assessment of the other groups projects. This should take the form of a brief critical (balanced) assessment identifying both the positive (good) and negative (bad) aspects of the project page as it currently exists online.

You may if you choose, use the final project assessment criteria as a guide. Though you are also welcome to use your own criteria.

Group Assessment Criteria

  1. The key points relating to the topic that your group allocated are clearly described.
  2. The choice of content, headings and sub-headings, diagrams, tables, graphs show a good understanding of the topic area.
  3. Content is correctly cited and referenced.
  4. The wiki has an element of teaching at a peer level using the student's own innovative diagrams, tables or figures and/or using interesting examples or explanations.
  5. Evidence of significant research relating to basic and applied sciences that goes beyond the formal teaching activities.
  6. Relates the topic and content of the Wiki entry to learning aims of cell biology.
  7. Clearly reflects on editing/feedback from group peers and articulates how the Wiki could be improved (or not) based on peer comments/feedback. Demonstrates an ability to review own work when criticised in an open edited wiki format. Reflects on what was learned from the process of editing a peer's wiki.
  8. Evaluates own performance and that of group peers to give a rounded summary of this wiki process in terms of group effort and achievement.
  9. The content of the wiki should demonstrate to the reader that your group has researched adequately on this topic and covered the key areas necessary to inform your peers in their learning.
  10. Develops and edits the wiki entries in accordance with the above guidelines.
Recent Basement Membrane Reviews  
Basement membranes.

Curr Biol. 2017 Mar 20;27(6):R207-R211. doi: 10.1016/j.cub.2017.02.006.

Jayadev R1, Sherwood DR2.

Abstract

Basement membranes (BMs) are thin, dense sheets of specialized, self-assembled extracellular matrix that surround most animal tissues (Figure 1, top). The emergence of BMs coincided with the origin of multicellularity in animals, suggesting that they were essential for the formation of tissues. Their sheet-like structure derives from two independent polymeric networks - one of laminin and one of type IV collagen (Figure 1, bottom). These independent collagen and laminin networks are thought to be linked by several additional extracellular matrix proteins, including nidogen and perlecan (Figure 1, bottom). BMs are usually associated with cells and are anchored to cell surfaces through interactions with adhesion receptors and sulfated glycolipids (Figure 1, bottom). Various combinations of other proteins, glycoproteins, and proteoglycans - including fibulin, hemicentin, SPARC, agrin, and type XVIII collagen - are present in BMs, creating biochemically and biophysically distinct structures that serve a wide variety of functions. BMs have traditionally been viewed as static protein assemblies that provide structural support to tissues. However, recent studies have begun to uncover dynamic, active roles for BMs in many developmental processes. Here, we discuss established and emerging roles of BMs in development, tissue construction, and tissue homeostasis. We also explore how cells traverse BM barriers, the roles of BMs in human diseases, and future directions for the field. Copyright © 2017 Elsevier Ltd. All rights reserved.

PMID 28324731

Basement membrane ultrastructure and component localization data from uterine tissues during early mouse pregnancy.

Data Brief. 2016 Nov 5;9:931-939. eCollection 2016.

Jones-Paris CR1, Paria S2, Berg T2, Saus J3, Bhave G4, Paria BC5, Hudson BG6.

Abstract

Basement membranes (BMs) are specialized extracellular scaffolds that provide architecture and modulate cell behaviors in tissues, such as fat, muscle, endothelium, endometrium, and decidua. Properties of BMs are maintained in homeostasis for most adult tissues. However, BM ultrastructure, composition, and localization are rapidly altered in select uterine tissues that are reprogrammed during pregnancy to enable early maternal-embryo interactions. Here, our data exhibit both static and dynamic BMs that were tracked in mouse uterine tissues during pre-, peri-, and postimplantation periods of pregnancy. The data exhibit spatial-temporal patterns of BM property regulation that coincide with the progression of adapted physiology. Further interpretation and discussion of these data in this article are described in the associated research article titled, "Embryo implantation triggers dynamic spatiotemporal expression of the basement membrane toolkit during uterine reprogramming" (C.R. Jones-Paris, S. Paria, T. Berg, J. Saus, G. Bhave, B.C. Paria, B.G. Hudson, 2016) [1].

PMID 27896299

The bi-functional organization of human basement membranes.

PLoS One. 2013 Jul 3;8(7):e67660. doi: 10.1371/journal.pone.0067660. Print 2013.

Halfter W1, Monnier C, Müller D, Oertle P, Uechi G, Balasubramani M, Safi F, Lim R, Loparic M, Henrich PB.

Abstract

The current basement membrane (BM) model proposes a single-layered extracellular matrix (ECM) sheet that is predominantly composed of laminins, collagen IVs and proteoglycans. The present data show that BM proteins and their domains are asymmetrically organized providing human BMs with side-specific properties: A) isolated human BMs roll up in a side-specific pattern, with the epithelial side facing outward and the stromal side inward. The rolling is independent of the curvature of the tissue from which the BMs were isolated. B) The epithelial side of BMs is twice as stiff as the stromal side, and C) epithelial cells adhere to the epithelial side of BMs only. Side-selective cell adhesion was also confirmed for BMs from mice and from chick embryos. We propose that the bi-functional organization of BMs is an inherent property of BMs and helps build the basic tissue architecture of metazoans with alternating epithelial and connective tissue layers.

PMID 23844050

Archive

MH - note that content listed below will not match exactly current lecture structure but has been selected as having similar content

Objectives

  • Understanding of the localisation and origin of extracellular matrix
  • Understanding of the 3 major components
    • fibers, proteoglycans (matrix), adhesive glycoproteins
  • Broad understanding of structure and function
    • fibronectin
    • laminin
    • basement membrane
  • Broad understanding of some key extracellular matrix experiments


Glycosaminoglycans (Gags)

  • five types historically called "mucopolysaccharides"
  • mostf GAGs linked to core proteins, forming proteoglycans
  • Hyaluronan (or hyaluronic acid) main glycosaminoglycan in connective tissue
  • high molecular weight (~ MW 1,000,000 )
  • length of about 2.5 µm hyaluronan
  • "backbone" for the assembly of other glycosaminoglycans
    • Hyaluronan is also a major component of the synovial fluid, which fills joint cavities, and the vitreous body of the eye.
  • Other 4 major glycosaminoglycans
    • chondroitin sulphate, dermatan sulphate, keratan sulphate and heparan sulphate (UK sulphate, US sulfate)
    • attach through core and link proteins to hyaluronic acid backbone

Proteoglycans

MBoC - Proteoglycans in the extracellular matrix of rat cartilage MBoC - Examples of a small (decorin) and a large (aggrecan) proteoglycan found in the extracellular matrix

  • consist of protein (~5%) and polysaccharide chain (~95%)
  • form a gel to embed the fibril network
  • Golgi apparatus - GAG disaccharides are added to protein cores to form proteoglycans
  • 10% by weight but fill most of space
  • unbranched polysaccharide chains
  • disaccharide subunits
  • amino sugar

Proteoglycan Function

  • trap water
  • resistant to compression
  • return to original shape
  • occupy space
  • link to collagen fibers
  • form network
    • in bone combined with calcium hydroxyapatite, calcium carbonate

Development

  • produce a “cell-free” space
  • for cell proliferation and migration into

Adult

  • in areas of compression
  • tissues, joints

hyaluronan-binding proteins

Hyaluronan Synthesis

  • differs from other GAG synthesis
    • synthesized at plasma membranes
    • nascent chains directly extruded into ECM

Cell adhesion

embryonic migration

Proteoglycan - Disease

Mucopolysaccharidosis type I (MPS I) - Hurler disease

  • deficiency of alpha-L iduronidase => lysosomal storage disease, associated with an altered elastic matrix
  • excess heparan sulphate and dermatan sulphate

Cancer development

  • altered types and kinds of proteoglycans formed by cells
  • normal cells -> malignant

Arthritis

  • Cartilage breakdown (cartilage erosion)
  • chondrocytes elicit a catabolic response which exceeds anabolism of new matrix molecules
  • Degrade proteoglycan (aggrecan)
  • Also a mouse model generates antibodies to proteogycan

Fibronectin

Fibronectin binding domains

Fibronectin Structure

  • dimer connected at C-terminus
    • Mr 550 kDa
    • nearly identical subunits composed of types I (F1), II (F2), and III (F3) fibronectin modules
  • S-S linkages
  • rigid and flexible domains
  • fibronectin fibrils have elastic properties and can stretch fibrils up to four-fold their relaxed length.
  • fibrillogenesis - transformation from the compact (soluble) form to the extended fibrillar (insoluble) form of fibronectin, requires application of mechanical forces generated by cells.


Binding Domains

fibronectin and syndecan binding model
  • cell binding segment RGDS
    • arg-gly-asp-ser
  • binds integrin receptor in membrane
    • then mechanically couples to the actin cytoskeleton
  • domains bind
    • heparin sulphate
    • collagen
    • hyaluronic acid
    • Gangliosides
    • fibronectin

Figure 19-54. Coalignment of extracellular fibronectin fibrils and intracellular actin filament bundles

fibronectin stretch

Fibronectin Function

Fibronectin (fl)
  • soluble protein in blood plasma (200–250 kDa monomer)
    • blood clotting process, link to fibrin
  • insoluble protein in extracellular matrix (ECM)
    • ECM fibronectin differs from plasma fibronectin by the presence of additional polypeptide segments and in altering morphology of transformed cells and hemagglutination.

Cell Adhesion

Migration Pathways

  • blocking fibronectin with antibody
  • prevents neural crest migration
  • extension of axons and dendrites
  • branching

Fetal fibronectin (fFN)

  • produced by fetal cells and found acting as an adhesive at the interface of the chorion and the decidua ( fetal membrane and uterine lining).
    • has been used as a diagnostic for preterm birth.

Laminin

Laminin Structure

  • cross-shaped glycoprotein
  • 3 polypeptides a, b1, b2
  • carbohydrate (13% by weight)
  • Mr 900K
  • separate binding domains
    • collagen IV
    • heparin
    • heparin sulphate
    • cell binding
    • cell specific binding - liver, nerve
    • cell surface receptor

Figure 19-57. The structure of laminin

ECM laminin model.jpg

Laminin molecular structure

ECM laminin domains.jpeg

Laminin domains

Laminin Function

  • cell adhesion
  • migration pathways
  • stimulates growth of axons
  • development and regeneration
  • differentiation
  • basal laminae
  • most abundant linking glycoprotein

Integrin- Structure Integrin Function cell membrane receptor for ECM linkers binds RGDS motif 2 subunits alpha (α) and beta (β) transmembrane linked to cell cytoskeleton actin microfilaments via talin and vinculin focal contacts For Review see Integrin signaling revisited. Schwartz MA.Trends Cell Biol 2001 Dec;11(12):466-70

Adhesive Signalling

Integrin and Laminin - Several integrin heterodimers act as laminin receptors on a variety of cell types alpha 1 beta 1 alpha 2 beta 1 alpha 3 beta 1 alpha 6 beta 1 alpha 7 beta 1 alpha 6 beta 4 Microsc Res Tech 2000 Nov 1;51(3):280-301

Integrin and Laminin

  • Roles of laminin-binding integrins in adhesion-mediated events in vertebrates
  • embryonic development, cell migration and tumor cell invasiveness, cell proliferation, differentiation and basement membrane assembly
  • essential role for receptors in maintaining cell polarity and tissue architecture

Basement Membrane

basement membrane

The epithelial ECM the term "basement membrane" is used with light microscopic observation and "basal lamina" is used with electron microscopy.

The basement membrane is composed of two sublayers.

basal lamina

  • (about 40–120 nm thick) consists of fine protein filaments embedded in an amorphous matrix.
  • Membrane proteins of the epithelial cells are anchored in the basal lamina, which is also produced by the epithelial cells.
  • major component of the basal lamina are two glycoproteins - laminin and (usually type IV) collagen

Figure 19-58. A model of the molecular structure of a basal lamina

reticular lamina

  • consists of reticular fibres embedded in ground substance.
  • fibres of the reticular lamina connect the basal lamina with the underlying conective tissue.
  • components of the reticular lamina are synthesised by cells of the connective tissue underlying the epithelium.

Basal Lamina Experiment

Figure 19-60. Regeneration experiments demonstrating the special character of the junctional basal lamina at a neuromuscular junction this link currently not functional. temp link

Neuromuscular Junction

  • Basal lamina directs acetylcholinesterase (AChE) accumulation at synaptic sites in regenerating muscle
  • skeletal muscle damaged such that basal lamina sheaths of the muscle fibers spared
  • new myofibers develop within sheaths and neuromuscular junctions form at original synaptic sites
  • regenerated neuromuscular junctions have junctional folds and accumulations of acetylcholine receptors and AChE
Neuromuscular Junction Expt
Sanes1978 basal lamina 01.jpg Sanes1978 basal lamina 02.jpg

Integrin- Function

  • Activate members of Rho-family of small GTPases
  • Conversely, Rho- and Ras-family proteins can influence the ability of integrins to bind their ligands
    • control of cell motility, and therefore of invasive and metastatic behavior
  • Integrin binding ECM has effects on cell survival, particularly for cells of epithelial origin
    • specific integrins have selective effects on efficiency of signal transduction in cell survival pathways

Text modified from: New aspects of integrin signaling in cancer. Semin Cancer Biol 2000 Dec;10(6):407-14

ECM Reorganisation

Reorganisation can occur through proteolytic degradation changes to ECM proteins (collagen, laminin, and fibronectin). Their activity can be regulated locally by inhibitors.

The proteases form 2 main classes:

Matrix Metalloproteases (MMPs)

  • dependent upon bound Ca2+ or Zn2+ for activity.
  • family of enzymes
    • MMP-2 (Gelatinase A, 72 kDa type IV collagenase) is the most widely distributed
  • collagenases can specifically cleave proteins at a small number of sites.
  • inhibited by tissue inhibitors of metalloproteases (TIMPs).
    • MMP-2 appears to be associated with early breast carcinoma and cervical neoplasia

Serine Proteases

  • have a highly reactive serine in their active site.
  • inhibited by serpins.
  • role in metastasis

Links: SOMS Ocular Immunology Group | Expression of MMPs and TIMPs in breast tumours

History

Below are some example historical research finding related to cell junctions from the JCB Archive.

1978 Basal lamina instructs innervation Joshua Sanes and Jack McMahan show that regenerating nerve axons take their cues for new synapse formation from the extracellular matrix (ECM) of muscle cells and not from the muscle cells themselves.


ECM Scaffold - Tissue Engineering

“Decellularized tissues and organs have been successfully used in a variety of tissue engineering/regenerative medicine applications, …Each of these treatments affect the biochemical composition, tissue ultrastructure, and mechanical behavior of the remaining extracellular matrix (ECM) scaffold, which in turn, affect the host response to the material.”

Reference: Decellularization of tissues and organs. Biomaterials. 2006 Jul;27(19):3675-83. Epub 2006 Mar 7.

References

Recent Reviews

Karin Wang, Bo Ri Seo, Claudia Fischbach, Delphine Gourdon Fibronectin Mechanobiology Regulates Tumorigenesis. Cell Mol Bioeng: 2015, 9;1-11 PubMed 26900407

Gernot Walko, Maria J Castañón, Gerhard Wiche Molecular architecture and function of the hemidesmosome. Cell Tissue Res.: 2015, 360(2);363-78 PubMed 25487405

Wilson Savino, Daniella Arêas Mendes-da-Cruz, Daiane Cristina Ferreira Golbert, Ingo Riederer, Vinicius Cotta-de-Almeida Laminin-Mediated Interactions in Thymocyte Migration and Development. Front Immunol: 2015, 6;579 PubMed 26635793


Textbooks

Essential Cell Biology

  • Essential Cell Biology Chapter 19 p594-604

Molecular Biology of the Cell

Alberts, Bruce; Johnson, Alexander; Lewis, Julian; Raff, Martin; Roberts, Keith; Walter, Peter New York and London: Garland Science; c2002 --Mark Hill (talk) 08:58, 1 May 2014 (EST) These library links currently not functional.

Molecular Cell Biology

Lodish, Harvey; Berk, Arnold; Zipursky, S. Lawrence; Matsudaira, Paul; Baltimore, David; Darnell, James E. New York: W. H. Freeman & Co.; c1999

The Cell- A Molecular Approach

Cooper, Geoffrey M. Sunderland (MA): Sinauer Associates, Inc.; c2000

  • The Cell- A Molecular Approach
  • The Cell - A Molecular Approach - III. Cell Structure and Function Chapter 12. The Cell Surface
  • The Extracellular Matrix

Essentials of Glycobiology, 2nd ed.

Varki, A.; Cummings, R.D.; Esko, J.D.; Freeze, H.H.; Stanley, P.; Bertozzi, C.R.; Hart, G.W.; Etzler, M.E., editors Plainview (NY): Cold Spring Harbor Laboratory Press; 2008

Search Online Textbooks

Books

PubMed

  • PubMed is a service of the U.S. National Library of Medicine that includes over 18 million citations from MEDLINE and other life science journals for biomedical articles back to 1948. PubMed includes links to full text articles and other related resources. PubMed
  • PubMed Central (PMC) is a free digital archive of biomedical and life sciences journal literature at the U.S. National Institutes of Health (NIH) in the National Library of Medicine (NLM) allowing all users free access to the material in PubMed Central. PMC
  • Online Mendelian Inheritance in Man (OMIM) is a comprehensive compendium of human genes and genetic phenotypes. The full-text, referenced overviews in OMIM contain information on all known mendelian disorders and over 12,000 genes. OMIM
  • Entrez is the integrated, text-based search and retrieval system used at NCBI for the major databases, including PubMed, Nucleotide and Protein Sequences, Protein Structures, Complete Genomes, Taxonomy, and others Entrez

Search Pubmed

Reviews

  • Hay ED. The extracellular matrix in development and regeneration. An interview with Elizabeth D. Hay. Int J Dev Biol. 2004;48(8-9):687-94. No abstract available. PMID: 15558460
  • Hay ED. Extracellular matrix. J Cell Biol. 1981 Dec;91(3 Pt 2):205s-223s. Review. No abstract available. PMID: 6172429
  • Sasaki T, Fässler R, Hohenester E. Laminin: the crux of basement membrane assembly. J Cell Biol. 2004 Mar 29;164(7):959-63. Epub 2004 Mar 22. Review.

PMID: 15037599 | JCB



Images


2017 Course Content

Moodle

Lectures: Cell Biology Introduction | Cells Eukaryotes and Prokaryotes | Cell Membranes and Compartments | Cell Nucleus | Cell Export - Exocytosis | Cell Import - Endocytosis | Cytoskeleton Introduction | Cytoskeleton - Microfilaments | Cytoskeleton - Microtubules | Cytoskeleton - Intermediate Filaments | Cell Mitochondria | Cell Junctions | Extracellular Matrix 1 | Extracellular Matrix 2 | Cell Cycle | Cell Division | Cell Death 1 | Cell Death 2 | Signal 1 | Signal 2 | Stem Cells 1 | Stem Cells 2 | Development | 2017 Revision

2017 Laboratories: Introduction to Lab | Fixation and Staining |


2017 Projects: Group 1 - Delta | Group 2 - Duct | Group 3 - Beta | Group 4 - Alpha

Dr Mark Hill 2015, UNSW Cell Biology - UNSW CRICOS Provider Code No. 00098G