A quantitative method for analysis of in vitro neurite outgrowth

Mitchell, Pamela J.; Hanson, Jeff; Quets-Nguyen, A.T.; Bergeron, Maxime; Smith, Rosamund C.

doi:10.1016/j.jneumeth.2007.04.021

Cited by 50 publications

(42 citation statements)

References 32 publications

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“…Neurite outgrowth assays were performed as described previously (8). Briefly, assays were carried out in 24-well plates after treatment with drugs for 48 h. Random fields were photographed with an Olympus IX71 inverted research microscope.…”

Section: Identification Of Neuronsmentioning

confidence: 99%

Nogo-66 receptor activation inhibits neurite outgrowth and increases β-amyloid protein secretion of cortical neurons

et al. 2011

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Abstract.A Nogo-A to Nogo-66 receptor (NgR) pathway is well known to contribute to the inhibition of the neurite regeneration of adult central nervous system neurons after traumatic injuries. Recent evidence suggests that Nogo-A and NgR are involved in the pathology of Alzheimer's disease (AD), as evidenced by the fact that Nogo-A is overexpressed by hippocampal neurons in patients with AD and is associated with β-amyloid protein (Aβ) deposits in senile plaques. In the present experiments, we investigated the potential role of Nogo-A in both neurite outgrowth and Aβ generation in cortical neurons. Our results showed that activation of NgR not only inhibited neurite outgrowth in cortical neurons by activating the rho-associated coiled coil-containing protein kinase (ROCK) and protein kinase C, but also promoted their Aβ secretion, which was at least in part activated by ROCK. These findings suggest that the overexpression of Nogo-A and the activation of NgR inhibit neurite outgrowth and alter neuronal metabolism, resulting in overproduction and/or release of Aβ, which in turn may trigger the onset and development of AD. Inhibition of ROCK can promote neurite outgrowth and reduce Aβ production of cortical neuron, which suggests that ROCK appears to be a good target for AD therapy.

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Section: Identification Of Neuronsmentioning

confidence: 99%

Nogo-66 receptor activation inhibits neurite outgrowth and increases β-amyloid protein secretion of cortical neurons

et al. 2011

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“…To measure the lengths of neurite from neurons, b-tubulin III positive neurite lengths were measured as previously described [48,49]. Briefly, 15 fields in each well were chosen randomly and photographed through the fluorescent microscope.…”

Section: Measurement Of Neurite Lengthsmentioning

confidence: 99%

The Promotive Effects of Thymosin β4 on Neuronal Survival and Neurite Outgrowth by Upregulating L1 Expression

et al. 2008

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Thymosin beta(4) (Tbeta4) is a major actin-sequestering peptide widely distributed in mammalian tissues including the nervous system. The presence of this peptide in the nervous system likely plays a role in synaptogensis, axon growth, cell migration, and plastic changes in dendritic spine. However, the effects of Tbeta4 on the survival of neurons and axonal outgrowth have still not been fully understood. So far it is not clear if the effects of Tbeta4 are associated with L1 functions. In the present study, we hypothesized that Tbeta4-induced up-regulation of L1 synthesis could be involved in the survival and axon outgrowth of cultured spinal cord neurons. To test this hypothesis, primarily cultured neurons were prepared from the mouse spinal cord and treated with various concentrations of Tbeta4 ranging from 0.1 to 10 microg/ml. The analysis of L1 mRNA expression and protein synthesis in neurons was then carried out using RT-PCR and western blot assays, respectively. After the addition of Tbeta4 to cultures, cells were then treated with antibodies against distinct domains of L1-Fc. Subsequently, beta-tubulin III and L1 double-labeled indirect immunofluorescence was carried out. Meanwhile, L1 immunofluorescent reactivity was analyzed and compared in cells treated with Tbeta4. Furthermore, the number of beta-tubulin III-positive cells and neurite lengths were measured. We found that Tbeta4 enhanced L1 expression in a dose-dependent manner, and the highest L1 mRNA and protein synthesis in cells increased by more than 2.1- and 2.3-fold in the presence of Tbeta4 at identical concentrations, respectively. Moreover, it also dose dependently enhanced neurite outgrowth and neuronal survival. Compared to conditions without Tbeta4, the length of neurite and neuronal survival increased markedly in presence of 0.5, 1, and 5 microg/ml Tbeta4, respectively, whereas the effects of Tbeta4 were significantly attenuated or inhibited in the process of L1-Fc antibodies treatment. These above results indicate that the promotive effect of Tbeta4 on the survival and neurite outgrowth of cultured spinal cord neurons might be mediated, at least in part via a stimulation of the production of L1 in the neurons.

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“…A analise quantitativa em ensaio in vitro através da marcação com DAPI também foi empregada por Mitchell et al [37] , os quais observaram o crescimento de neurites em cultura de neurônios granulares do cerebelo e da uma linhagem específica NG108-15.…”

Section: Resultsunclassified

Estudo das células Neuro2A sobre os biomateriais PCL e PLLA

et al. 2014

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Resumo: Os biomateriais poli L-ácido lático (PLLA) e o poli caprolactona (PCL) são os polímeros mais estudadas na área dos materiais bioreabsorvíveis. Dentre as suas principais características que contribuem para a interação celular, temos a especificidade química da superfície, elétrica, hidrofobicidade e topografia. Ainda, observa-se o tempo de degradação, porosidade, biocompatibilidade com o tecido biológico, bem como, a confecção com as mais variadas formas e dimensões. Já a prática da cultura celular, tem como objetivo estudar a adesão, migração, diferenciação e a proliferação celular utilizando-se um determinado material ou substância. Contudo, poucos trabalhos utilizando os biomateriais ora supracitados e a aplicação em células neuro2A foram realizados. Sabe-se que este tipo celular é derivado de células embrionárias da crista neural, as quais originam em neurônios simpáticos e apresentam como característica a imortalidade, portanto, são excelentes modelos em ensaios in vitro. Nesse sentido, o presente estudo avalia a adesão e a proliferação desta linhagem celular sobre os biopolímeros poli caprolactona (PCL) e poli L-ácido lático (PLLA). Palavras-chave: Biomateriais, PLLA, PLC, neuroblastoma, cultura. Study of Neuro2a Cells on the Biomaterials Poly L-lactic Acid and Poly CaprolactoneAbstract: Biomaterials poly L-lactic acid (PLLA) and poly caprolactone (PCL) are the most studied in the area of bioresorbable materials. Among the main features that contribute to cell interaction, we have the specific surface chemistry, electrical, hydrophobicity and topography. Also, is observed the degradation time, porosity, biocompatibility with biological tissue, as well as the preparation of the most varied shapes and sizes. The practice of cell culture, aims to study the adhesion, migration, differentiation and cell proliferation using a given material or substance. However, few studies were performed using these biomaterials and the application of neuro2A cells. It is known that this cell type is derived from the embryonic neural crest cells, which originate in sympathetic neurons and have the characteristic of immortality, therefore, are excellent models in vitro assays. Accordingly, the present study evaluates the adhesion and proliferation of this cell line on the biopolymers poly caprolactone (PCL) and poly L-lactic acid (PLLA). Glória, 187, Centro, Diamantina, MG, Brasil, Keywords: Biomaterials, PLLA, PLC, neuroblastoma, culture. Autor para correspondência: Amauri Pierucci, Departamento de Ciências Básicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri -UFVJM, Rua da IntroduçãoCom o advento da bioengenharia intensificaram os estudos sobre a aplicação e ação dos biomateriais em diferentes tipos celulares. Sabe-se que a adesão, migração, diferenciação e a proliferação celular sobre determinado substrato depende em parte das propriedades intrínsecas de cada tipo celular, bem como, o tipo de material utilizado [1][2][3][4] . Dentre os produtos celulares, pode-se ressaltar a síntese de moléculas de adesã...

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A quantitative method for analysis of in vitro neurite outgrowth

Cited by 50 publications

References 32 publications

Nogo-66 receptor activation inhibits neurite outgrowth and increases β-amyloid protein secretion of cortical neurons

Nogo-66 receptor activation inhibits neurite outgrowth and increases β-amyloid protein secretion of cortical neurons

The Promotive Effects of Thymosin β4 on Neuronal Survival and Neurite Outgrowth by Upregulating L1 Expression

Estudo das células Neuro2A sobre os biomateriais PCL e PLLA

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