Dorsal root ganglia neurite extension is inhibited by mechanical and chondroitin sulfate‐rich interfaces

Yu, Xiaojun; Bellamkonda, Ravi V.

doi:10.1002/jnr.1225

Cited by 87 publications

(83 citation statements)

References 43 publications

(55 reference statements)

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“…While it is well established that neurons can respond to multiple molecular cues during neurite outgrowth and branching [27,28], only a few studies have investigated effects of the physical environment that neurons encounter [20,29]. The results of this study suggest that mechanical parameters have profound effects on neuronal morphology, specifically branching, and may be one of the cues that neurons integrate when making critical pathfinding decisions.…”

Section: Resultsmentioning

confidence: 81%

Neurite branching on deformable substrates

et al. 2002

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The mechanical properties of substrates underlying cells can have profound effects on cell structure and function. To examine the effect of substrate deformability on neuronal cell growth, proteinlaminated polyacrylamide gels were prepared with differing amounts of bisacrylamide to generate substrates of varying deformability with elastic moduli ranging from 500 to 5500 dyne/cm 2 . Mouse spinal cord primary neuronal cells were plated on the gels and allowed to grow and extend neurites for several weeks in culture. While neurons grew well on the gels, glia, which are normally cocultured with the neurons, did not survive on these deformable substrates even though the chemical environment was permissive for their growth. Substrate flexibility also had a significant effect on neurite branching. Neurons grown on softer substrates formed more than three times as many branches as those grown on stiffer gels. These results show that mechanical properties of the substrate specifically direct the formation of neurite branches, which are critical for appropriate synaptic connections during development and regeneration.

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Section: Resultsmentioning

confidence: 81%

Neurite branching on deformable substrates

et al. 2002

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“…Agarose was chosen as the matrix backbone for this system because it lacks cell surface receptor binding domains, its physical properties may be tailored based on concentration, 5,63 and procedures have been established for the controlled coupling of bioactive ligands. 6,17 Collagen has previously been used for successful 3-D cortical neuronal culture, 46,47 integrin receptors that recognize collagen have been demonstrated for a range of neuronal sub-types, 3,4,8,9,41,56 and it is a major component in our positive control matrix, Matrigel.…”

Section: Preparation Of Protein-conjugated Agarose Gelsmentioning

confidence: 99%

“…In agaroseonly gels, the complex modulus increased proportionally with the AG percentage (data not shown), with values similar to those previously reported. 5,63 For a fixed AG percentage (1.5%), the complex modulus was determined based on [Col] i (ranging from 30 to 300 lg/ mL), revealing that complex modulus increased as a function of [Col] i , with significant differences between all groups with the exception of the Col(30 lg/mL)-AG vs. AG, which were statistically equivalent. Thus, over the frequency range evaluated, the order of complex moduli was Col(300 lg/mL)-AG > -Col(150 lg/mL)-AG > Col(30 lg/mL)-AG = AG, thus revealing a dependence between [Col] i and complex modulus within the range of [Col] evaluated.…”

Section: Col-ag Matrix Characterizationmentioning

confidence: 99%

Collagen-Dependent Neurite Outgrowth and Response to Dynamic Deformation in Three-Dimensional Neuronal Cultures

2007

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Abstract-In vitro models of brain injury that use thick 3-D cultures and control extracellular matrix constituents allow evaluation of cell-matrix interactions in a more physiologically relevant configuration than traditional 2-D cultures. We have developed a 3-D cell culture system consisting of primary rat cortical neurons distributed throughout thick (>500 lm) gels consisting of type IV collagen (Col) conjugated to agarose. Neuronal viability and neurite outgrowth were examined for a range of agarose (AG) percentages (1.0-3.0%) and initial collagen concentrations ([Col] i ; 0-600 lg/ mL). In unmodified AG, 1.5% gels supported viable cultures with significant neurite outgrowth, which was not found at lower (£1.0%) concentrations. Varying [Col] 30, 150, and 300 lg/mL, which supported cultures with similar baseline viability and neurite outgrowth. Conjugation of Col to AG also increased the complex modulus of the hydrogel. Following high rate deformation, neuronal viability significantly decreased with increasing [Col] i , implicating cell-matrix adhesions in acute mechanotransduction events associated with traumatic loading. These results suggest interrelated roles for matrix mechanical properties and receptor-mediated cell-matrix interactions in neuronal viability, neurite outgrowth, and transduction of high rate deformation. This model system may be further exploited for the elucidation of mechanotransduction mechanisms and cellular pathology following mechanical insult.

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“…To improve the understanding of the competing signals between growth promoting molecules such as laminin and inhibitory proteoglycans we and other labs have developed in vitro assays to test how the chondroitin sulfate containing proteoglycan signals presented in a mixed CSPG-laminin surface-attached layer are affecting neuronal cell adhesion and growth cone pathfinding [120][121][122][123][124][125]. It is known that neurons possess the ability to integrate multiple signals in their environment [120,[125][126][127][128], and it has been demonstrated that the ratio of substratum bound laminin and CSPGs such as aggrecan can be varied to yield net neurite outgrowth promoting or inhibiting signals [120,[129][130][131].…”

Section: Protein Patterns and Cell Choice Assays In Studying Neuronalmentioning

confidence: 99%

The effects of proteoglycan surface patterning on neuronal pathfinding

Hlady

Hodgkinson

2007

Materialwissenschaft Werkst

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Protein micropatterning techniques are increasingly applied in cell choice assays to investigate fundamental biological phenomena that contribute to the host response to implanted biomaterials, and to explore the effects of protein stability and biological activity on cell behavior for in vitro cell studies. In the area of neuronal regeneration the protein micropatterning and cell choice assays are used to improve our understanding of the mechanisms directing nervous system during development and regenerative failure in the central nervous system (CNS) wound healing environment. In these cell assays, protein micropatterns need to be characterized for protein stability, bioactivity, and spatial distribution and then correlated with observed mammalian cell behavior using appropriate model system for CNS development and repair. This review provides the background on protein micropatterning for cell choice assays and describes some novel patterns that were developed to interrogate neuronal adaptation to inhibitory signals encountered in CNS injuries. Rationale for studying the role of proteoglycans in CNS injuriesNeurons from the central nervous system (CNS) possess a limited capacity to regenerate beyond scar tissue formation barriers in injuries even in the presence of minimal trauma to tissue organization [1,2]. A major culprit in CNS neuronal regenerative failure are the inihibitory proteoglycans (PGs), which are upregulated at the site of CNS injuries. PGs are composed of a core protein with varying numbers of attached glycosaminoglycan (GAG) side chains [3,4]. Proteoglycans are first translated intracellularly into proteins in the endoplasmic reticulum, and then GAG chains are later attached in the Golgi apparatus before secretion into the extracellular space [5][6][7][8]. The study of PGs has particularly been pronounced in the field of neurobiology and development of the nervous system. Numerous studies have concluded that PGs act as barriers that direct the migration of neural crest cells and the outgrowth direction of pathfinding neurons during development [9][10][11][12][13][14][15][16][17]. A number of these studies have additionally shown that cell guidance by PGs is based on an inhibitory signal located within the chondroitin sulfate GAG chains [9,18,19], with the result that the artificial addition of chondroitin sulfate (CS) sugars to the developing nervous system disrupts normal pathfinding [20] as well as does the removal of chondroitin sulfate chains through enzymatic treatment [9,19]. Davies et al. found that chondroitin sulfate proteoglycans (CSPG) are a major constituent of the glial scar and that dorsal root ganglion (DRG) outgrowth termination coincided with an increase in CSPG expression density [21]. Other sources have also shown that CSPGs are upregulated after CNS injuries [22][23][24][25].Interestingly, native adult CNS neurons actually posses the intrinsic ability to regenerate when the wound healing environment is prepared by eliminating inhibitory factors [23,26,27]. In additi...

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Dorsal root ganglia neurite extension is inhibited by mechanical and chondroitin sulfate‐rich interfaces

Cited by 87 publications

References 43 publications

Neurite branching on deformable substrates

Neurite branching on deformable substrates

Collagen-Dependent Neurite Outgrowth and Response to Dynamic Deformation in Three-Dimensional Neuronal Cultures

The effects of proteoglycan surface patterning on neuronal pathfinding

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