Loss of the Extracellular Matrix Protein DIG-1 Causes Glial Fragmentation, Dendrite Breakage and Dendrite Extension Defects

The extracellular matrix (ECM) guides and constrains the shape of the nervous system. In C. elegans, DIG-1 is a giant ECM component that is required for fasciculation of sensory dendrites during development and for maintenance of axon positions throughout life. We identified four novel alleles of dig-1 in three independent screens for mutants affecting disparate aspects of neuronal and glial morphogenesis. First, we find that disruption of DIG-1 causes fragmentation of the amphid sheath glial cell in larvae and young adults. Second, it causes severing of the BAG sensory dendrite from its terminus at the nose tip, apparently due to breakage of the dendrite as animals reach adulthood. Third, it causes embryonic defects in dendrite fasciculation in inner labial (IL2) sensory neurons, as previously reported, as well as rare defects in IL2 dendrite extension that are enhanced by loss of the apical ECM component DYF-7, suggesting that apical and basolateral ECM contribute separately to dendrite extension. Our results highlight novel roles for DIG-1 in maintaining the cellular integrity of neurons and glia, possibly by creating a barrier between structures in the nervous system.

Download Full-text

Initial characterization of anosmin-1, a putative extracellular matrix protein synthesized by definite neuronal cell populations in the central nervous system

Journal of Cell Science ◽

10.1242/jcs.109.7.1749 ◽

1996 ◽

Vol 109 (7) ◽

pp. 1749-1757 ◽

Cited By ~ 5

Author(s):

N. Soussi-Yanicostas ◽

J.P. Hardelin ◽

M.M. Arroyo-Jimenez ◽

O. Ardouin ◽

R. Legouis ◽

...

Keyword(s):

Central Nervous System ◽

Extracellular Matrix ◽

Nervous System ◽

Cell Membrane ◽

Heparan Sulfate ◽

Matrix Protein ◽

Neuronal Cell ◽

Extracellular Matrix Protein ◽

Cell Populations

The KAL gene is responsible for the X-chromosome linked form of Kallmann's syndrome in humans. Upon transfection of CHO cells with a human KAL cDNA, the corresponding encoded protein, KALc, was produced. This protein is N-glycosylated, secreted in the cell culture medium, and is localized at the cell surface. Several lines of evidence indicate that heparan-sulfate chains of proteoglycan(s) are involved in the binding of KALc to the cell membrane. Polyclonal and monoclonal antibodies to the purified KALc were generated. They allowed us to detect and characterize the protein encoded by the KAL gene in the chicken central nervous system at late stages of embryonic development. This protein is synthesized by definite neuronal cell populations including Purkinje cells in the cerebellum, mitral cells in the olfactory bulbs and several subpopulations in the optic tectum and the striatum. The protein, with an approximate molecular mass of 100 kDa, was named anosmin-1 in reference to the deficiency of the sense of smell which characterizes the human disease. Anosmin-1 is likely to be an extracellular matrix component. Since heparin treatment of cell membrane fractions from cerebellum and tectum resulted in the release of the protein, we suggest that one or several heparan-sulfate proteoglycans are involved in the binding of anosmin-1 to the membranes in vivo.

Download Full-text

Restrictin: a chick neural extracellular matrix protein involved in cell attachment co-purifies with the cell recognition molecule F11

Development ◽

10.1242/dev.113.1.151 ◽

1991 ◽

Vol 113 (1) ◽

pp. 151-164 ◽

Cited By ~ 2

Author(s):

F.G. Rathjen ◽

J.M. Wolff ◽

R. Chiquet-Ehrismann

Keyword(s):

Extracellular Matrix ◽

Nervous System ◽

Matrix Protein ◽

Cell Attachment ◽

Extracellular Matrix Protein ◽

Relative Molecular Mass ◽

Adhesion Assay ◽

Cell Recognition ◽

Neural Extracellular Matrix ◽

Recognition Molecule

We report here the characterization of restrictin, a novel chick neural extracellular matrix glycoprotein associated with the cell recognition molecule F11. Immunoaffinity chromatography using monoclonal antibody 23–13 directed to restrictin yield a major relative molecular mass band at 170 × 10(3) and minor bands at 160, 180, 250 and 320 × 10(3) which are immunologically related to each other. Neural cells attach on immobilized restrictin in a short-term adhesion assay. This adhesion can be blocked specifically by monoclonal or polyclonal antibodies to restrictin but not by antibodies to F11 or by the peptide GRGDSP. Antibodies to restrictin do not interfere with the fasciculation of retinal axons and the isolated restrictin does not stimulate the outgrowth of axons. In the developing nervous system, restrictin is localized in very restricted regions and is found within areas of F11 expression. The timing and pattern of expression of restrictin and its cell attachment activity suggest that it participates in developmental events of the nervous system.

Download Full-text

Mechanotransduction Coordinates Inter-Tissue Extracellular Matrix Protein Homeostasis Promoting Longevity in C. Elegans

SSRN Electronic Journal ◽

10.2139/ssrn.3881355 ◽

2021 ◽

Author(s):

Alina C. Teuscher ◽

Cyril Statzer ◽

Seraina A. Domenig ◽

Ingmar Schoen ◽

Viola Vogel ◽

...

Keyword(s):

Extracellular Matrix ◽

Matrix Protein ◽

Extracellular Matrix Protein ◽

Protein Homeostasis ◽

C Elegans

Download Full-text

Tenascin (cytotactin): an extracellular matrix protein involved in morphogenesis of the nervous system

Seminars in Neuroscience ◽

10.1016/1044-5765(91)90051-o ◽

1991 ◽

Vol 3 (4) ◽

pp. 341-350 ◽

Cited By ~ 14

Author(s):

Matthias Chiquet ◽

Bernhard Wehrle-Haller ◽

Manuel Koch

Keyword(s):

Extracellular Matrix ◽

Nervous System ◽

Matrix Protein ◽

Extracellular Matrix Protein

Download Full-text

Definition of an extracellular matrix protein in rostral portions of the human central nervous system

Brain Research ◽

10.1016/0006-8993(88)91355-8 ◽

1988 ◽

Vol 438 (1-2) ◽

pp. 315-322 ◽

Cited By ~ 5

Author(s):

Wolfgang J. Rettig ◽

Pilar Garin Chesa ◽

H. Richard Beresford ◽

Myron R. Melamed ◽

Lloyd J. Old

Keyword(s):

Central Nervous System ◽

Extracellular Matrix ◽

Nervous System ◽

Matrix Protein ◽

Extracellular Matrix Protein ◽

Human Central Nervous System ◽

Definition Of

Download Full-text

1280: Increased Susceptibility to Genitovaginal Prolapse Associated with a Polymorphism in the Promoter of the Extracellular Matrix Protein LAMC1

The Journal of Urology ◽

10.1016/s0022-5347(18)31494-0 ◽

2007 ◽

Vol 177 (4S) ◽

pp. 421-422

Author(s):

Ganka Nikolova ◽

Christian O. Twiss ◽

Hane Lee ◽

Nelson Stanley ◽

Janet Sinsheimer ◽

...

Keyword(s):

Extracellular Matrix ◽

Matrix Protein ◽

Extracellular Matrix Protein ◽

Increased Susceptibility

Download Full-text

Homogenous overexpression of the extracellular matrix protein Netrin-1 in a hollow fiber bioreactor

Applied Microbiology and Biotechnology ◽

10.1007/s00253-021-11438-0 ◽

2021 ◽

Author(s):

Aniel Moya-Torres ◽

Monika Gupta ◽

Fabian Heide ◽

Natalie Krahn ◽

Scott Legare ◽

...

Keyword(s):

Extracellular Matrix ◽

Hollow Fiber ◽

Mammalian Cells ◽

Matrix Protein ◽

Continuous Production ◽

Extracellular Matrix Protein ◽

Close Monitoring ◽

High Quality ◽

Biolayer Interferometry ◽

Hollow Fiber Bioreactor

Abstract The production of recombinant proteins for functional and biophysical studies, especially in the field of structural determination, still represents a challenge as high quality and quantities are needed to adequately perform experiments. This is in part solved by optimizing protein constructs and expression conditions to maximize the yields in regular flask expression systems. Still, work flow and effort can be substantial with no guarantee to obtain improvements. This study presents a combination of workflows that can be used to dramatically increase protein production and improve processing results, specifically for the extracellular matrix protein Netrin-1. This proteoglycan is an axon guidance cue which interacts with various receptors to initiate downstream signaling cascades affecting cell differentiation, proliferation, metabolism, and survival. We were able to produce large glycoprotein quantities in mammalian cells, which were engineered for protein overexpression and secretion into the media using the controlled environment provided by a hollow fiber bioreactor. Close monitoring of the internal bioreactor conditions allowed for stable production over an extended period of time. In addition to this, Netrin-1 concentrations were monitored in expression media through biolayer interferometry which allowed us to increase Netrin-1 media concentrations tenfold over our current flask systems while preserving excellent protein quality and in solution behavior. Our particular combination of genetic engineering, cell culture system, protein purification, and biophysical characterization permitted us to establish an efficient and continuous production of high-quality protein suitable for structural biology studies that can be translated to various biological systems. Key points • Hollow fiber bioreactor produces substantial yields of homogenous Netrin-1 • Biolayer interferometry allows target protein quantitation in expression media • High production yields in the bioreactor do not impair Netrin-1 proteoglycan quality Graphical abstract

Download Full-text