Differential neurite outgrowth is required for axon specification by cultured hippocampal neurons

Yamamoto, Hideaki; Demura, Takanori; Morita, Mayu; Banker, Gary; Takenaka, Takashi; Nakamura, Shun

doi:10.1111/jnc.12001

Cited by 58 publications

(64 citation statements)

References 46 publications

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“…A similar series of dynamic changes in the accumulation of KIF5C have been observed during axon specification in cultured retinal ganglion cells (Randlett, Poggi, Zolessi, & Harris, 2011). The stable accumulation of KIF5C motor domain in a single neurite is among the earliest indicators of axon specification, both in cell culture and in vivo (Distel, Hocking, Volkmann, & Köster, 2010; Jacobson et al, 2006; Randlett et al, 2011; Yamamoto et al, 2012). …”

Section: Introductionmentioning

confidence: 99%

Analyzing kinesin motor domain translocation in cultured hippocampal neurons

Yang¹,

Bentley²,

Huang³

et al. 2016

Methods in Cell Biology

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Neuronal microtubules are subject to extensive posttranslational modifications and are bound by MAPs, tip-binding proteins, and other accessory proteins. All of these features, which are difficult to replicate in vitro, are likely to influence the translocation of kinesin motors. Here we describe assays for evaluating the translocation of a population of fluorescently labeled kinesin motor domains, based on their accumulation in regions of the cell enriched in microtubule plus ends. Neurons lend themselves to these experiments because of their microtubule organization. In axons, microtubules are oriented with their plus ends out; dendrites contain a mixed population of microtubules, but those near the tips are also plus end out. The assays involve the expression of constitutively active kinesins that can walk processively, but that lack the autoinhibitory domain in the tail that normally prevents their binding to microtubules until they attach to vesicles. The degree to which such motor domains accumulate at neurite tips serves as a measure of the efficiency of their translocation. Although these assays cannot provide the kind of quantitative kinetic information obtained from in vitro assays, they offer a simple way to examine kinesin translocation in living neurons. They can be used to compare the translocation efficiency of different kinesin motors and to evaluate how mutations or posttranslational modifications within the motor domain influence kinesin translocation. Changes to motor domain accumulation in these assays can also serve as readout for changes in the microtubule cytoskeleton that affect kinesin translocation.

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Section: Introductionmentioning

confidence: 99%

Analyzing kinesin motor domain translocation in cultured hippocampal neurons

Yang¹,

Bentley²,

Huang³

et al. 2016

Methods in Cell Biology

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“…[13,14] Several methods have been developed to construct in vitro neural network models. However, although in 2D culture systems cell-patterning approaches can easily control neuronal morphologies such as aligned nerve fibers, [16][17][18][19] these culture systems cannot recapitulate the topological complexity of in vivo neural networks because of their low cell density and population. [17,18] Recently, 3D cell patterning approaches that utilize components such as microchambers, [20] building blocks, [21] fiber scaffolds, [22][23][24] colloidal support, [19,25] and microfluidic [+] Present address:…”

Section: Rod-shaped Neural Units For Aligned 3d Neural Network Connecmentioning

confidence: 99%

“…However, although in 2D culture systems cell-patterning approaches can easily control neuronal morphologies such as aligned nerve fibers, [16][17][18][19] these culture systems cannot recapitulate the topological complexity of in vivo neural networks because of their low cell density and population. [17,18] Recently, 3D cell patterning approaches that utilize components such as microchambers, [20] building blocks, [21] fiber scaffolds, [22][23][24] colloidal support, [19,25] and microfluidic medium and cut them into 1 or 3 mm sections using a disposable microtome blade ( www.advancedsciencenews.com www.advhealthmat.de process of the microfiber-shaped neural tissues did not affect cell growth at the edges. We placed the mNSC units on a poly(dimethylsiloxane) (PDMS) guide to investigate whether one connecting end can adhere to the connecting end of an adjacent neural unit (Figure 3a).…”

mentioning

confidence: 99%

Rod‐Shaped Neural Units for Aligned 3D Neural Network Connection

Kato-Negishi

Onoe

Ito

et al. 2017

Adv Healthcare Materials

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COMMUNICATION (1 of 7)© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim Rod-Shaped Neural Units for Aligned 3D Neural Network ConnectionMidori Kato-Negishi, Hiroaki Onoe, Akane Ito, and Shoji Takeuchi* DOI: 10.1002/adhm.201700143 guidance [12,26] have also been developed as alternative means of controlling cell patterning with high cell density and/or aligned nerve fibers. However, coculturing different types of neural tissues is still difficult because culture conditions such as medium, growth factors, and extracellular matrix (ECM) differ according to the type of neural tissues. [27][28][29][30][31][32][33] Neurons also have the characteristic of axon overproduction and, consequently, axons extend over a wide area during early development. [34][35][36] Therefore, selective connectivity between specific, spatially separated neural tissues is technically difficult.This paper proposes neural tissue units with aligned nerve fibers, called "rodshaped neural units," that can connect spatially separate neural regions (Figure 1). To fabricate these units, we employ a meterlong microfiber-shaped neural tissue [22,23] as starting material, in which stem-cell-derived or primary neural cells are encapsulated and aligned within an alginate tubular hydrogel. This tissue is cut into millimeter-sized sections and, after further cell culturing, both edges of the sections bulge out and form spherical ends (Figure 1b). The proposed rod-shaped neural units have the following characteristics: (1) nerve fibers are aligned in the longitudinal direction of the rod, (2) each unit has an insulated region covered with a thin alginate hydrogel layer to prevent the encapsulated neural tissues from connecting to surrounding cells within the regions, and (3) the edges of the unit are connectable, with the spherical ends functioning as glue to connect with other neural tissues or units. In this study, we designed and fabricated the rod-shaped neural units and characterized their connectivity in terms of neuronal morphologies and network formation. In addition, we constructed a neural network from different types of neural tissues in different culture conditions, with connections to specific, spatially neural regions. Therefore, our neural units assembly can coculture different types of neural tissues with aligned nerve fibers that recapitulate the topological complexity of in vivo neural networks with high cell density and cell population.Rod-shaped neural units were prepared by cutting microfibershaped neural tissues, as reported previously. [22,23,37,38] We used three types of neural cells to fabricate microfiber-shaped neural tissues: primary mouse neural stem cells (mNSCs) dissected from the midbrain of mice (mNSC units), primary cortical cells (cortical units), and primary hippocampal cells (hippocampal units). To prepare the rod-shaped neural units in an accurate and reproducible manner, we developed a fiber-cutting device that can fix the microfiber-shaped neural tissues in a culture In the brain, 3D neural networks such as the hippoc...

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“…2F). This might reflect some differences between 417 the two cellular models in the neuronal polarization capability 418 (Yamamoto et al, 2012). 419 We sought to better characterize the neuronal subtypes present in 420 our differentiated cell cultures.…”

mentioning

confidence: 99%

Development of a pluripotent stem cell derived neuronal model to identify chemically induced pathway perturbations in relation to neurotoxicity: Effects of CREB pathway inhibition

Pistollato

Louisse

Scelfo

et al. 2014

Toxicology and Applied Pharmacology

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According to the advocated paradigm shift in toxicology, acquisition of knowledge on the mechanisms underlying the toxicity of chemicals, such as perturbations of biological pathways, is of primary interest. Pluripotent stem cells (PSCs), such as human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), offer a unique opportunity to derive physiologically relevant human cell types to measure molecular and cellular effects of such pathway modulations. Here we compared the neuronal differentiation propensity of hESCs and hiPSCs with the aim to develop novel hiPSC-based tools for measuring pathway perturbation in relation to molecular and cellular effects in vitro. Among other fundamental pathways, also, the cAMP responsive element binding protein (CREB) pathway was activated in our neuronal models and gave us the opportunity to study time-dependent effects elicited by chemical perturbations of the CREB pathway in relation to cellular effects. We show that the inhibition of the CREB pathway, using 2-naphthol-AS-E-phosphate (KG-501), induced an inhibition of neurite outgrowth and synaptogenesis, as well as a decrease of MAP2(+) neuronal cells. These data indicate that a CREB pathway inhibition can be related to molecular and cellular effects that may be relevant for neurotoxicity testing, and, thus, qualify the use of our hiPSC-derived neuronal model for studying chemical-induced neurotoxicity resulting from pathway perturbations.

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Differential neurite outgrowth is required for axon specification by cultured hippocampal neurons

Cited by 58 publications

References 46 publications

Analyzing kinesin motor domain translocation in cultured hippocampal neurons

Analyzing kinesin motor domain translocation in cultured hippocampal neurons

Rod‐Shaped Neural Units for Aligned 3D Neural Network Connection

Development of a pluripotent stem cell derived neuronal model to identify chemically induced pathway perturbations in relation to neurotoxicity: Effects of CREB pathway inhibition

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