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Enteric neuron culture

N Kaji, S Nakayama, K Horiguchi, S Iino, H Ozaki, M Hori
BACKGROUND: Interstitial cells of Cajal (ICC) serve as intestinal pacemakers. Postoperative ileus (POI) is a gastrointestinal motility disorder that occurs following abdominal surgery, which is caused by inflammation-induced dysfunction of smooth muscles and enteric neurons. However, the participation of ICC in POI is not well understood. In this study, we investigated the functional changes of ICC in a mouse model of POI. METHODS: Intestinal manipulation (IM) was performed to induce POI...
March 15, 2018: Neurogastroenterology and Motility: the Official Journal of the European Gastrointestinal Motility Society
Toshio Takahashi, Akira Shiraishi, Jun Murata
Cholinergic signaling, which modulates cell activities via nicotinic and muscarinic acetylcholine receptors (n- and mAChRs) in response to internal or external stimuli, has been demonstrated in mammalian non-neuronal cells that synthesize acetylcholine (ACh). One of the major pathways of excitatory transmission in the enteric nervous system (ENS) is mediated by cholinergic transmission, with the transmitter ACh producing excitatory potentials in postsynaptic effector cells. In addition to ACh-synthesizing and ACh-metabolizing elements in the ENS, the presence of non-neuronal ACh machinery has been reported in epithelial cells of the small and large intestines of rats and humans...
March 5, 2018: International Journal of Molecular Sciences
Chun-Chih Hu, Gong-Her Wu, Tzu-En Hua, Oliver I Wagner, Ta-Jen Yen
We employ the model organism Caenorhabditis elegans to effectively study the toxicology of anatase and rutile phase titanium dioxide (TiO2) nanoparticles (NPs). The experimental results show that the nematode C. elegans can take up fluorescein isothiocyanate (FITC)-labeled TiO2 NPs and that both anatase and rutile TiO2 NPs can be detected in the cytoplasm of cultured primary neurons imaged by transmission electron microscopy (TEM). After TiO2 NPs exposure, these neurons also grow shorter axons, which may be related to the detected impeded worm locomotion behavior...
February 21, 2018: ACS Applied Materials & Interfaces
Sung G Ji, John H Weiss
Excitotoxic Zn2+ and Ca2+ accumulation contributes to neuronal injury after ischemia or prolonged seizures. Synaptically released Zn2+ can enter postsynaptic neurons via routes including voltage sensitive Ca2+ channels (VSCC), and, more rapidly, through Ca2+ permeable AMPA channels. There are also intracellular Zn2+ binding proteins which can either buffer neuronal Zn2+ influx or release bound Zn2+ into the cytosol during pathologic conditions. Studies in culture highlight mitochondria as possible targets of Zn2+; cytosolic Zn2+ can enter mitochondria and induce effects including loss of mitochondrial membrane potential (ΔΨm), mitochondrial swelling, and reactive oxygen species (ROS) generation...
January 17, 2018: Experimental Neurology
Eva Maria Kugler, Klaus Michel, David Kirchenbüchler, Georg Dreissen, Agnes Csiszár, Rudolf Merkel, Michael Schemann, Gemma Mazzuoli-Weber
Within the enteric nervous system, the neurons in charge to control motility of the gastrointestinal tract reside in a particular location nestled between two perpendicular muscle layers which contract and relax. We used primary cultured myenteric neurons of male guinea pigs to study mechanosensitivity of enteric neurons in isolation. Ultrafast Neuroimaging with a voltage sensitive dye technique was used to record neuronal activity in response to shear stress and strain. Strain was induced by locally deforming the elastic cell culture substrate next to a neuron...
January 6, 2018: Neuroscience
Debmalya Bhunia, Prasenjit Mondal, Gaurav Das, Abhijit Saha, Pallabi Sengupta, Jagannath Jana, Saswat Mohapatra, Subhrangsu Chatterjee, Surajit Ghosh
Identification of key amino acids is required for development of efficient cell penetrating peptides (CPPs) and has tremendous implications in medicine. Extensive research work enlightened us about the importance of two amino acids, arginine and tryptophan in the cell penetration. Here, we present a top-down approach to show how spatial po-sitions of two tryptophans regulate the cellular entry and nuclear localization. This enables us to develop short nontoxic tetrapeptides with excellent potential of cell penetration and nuclear localization...
December 28, 2017: Journal of the American Chemical Society
Sumiko Mochida
For reliable transmission at chemical synapses, neurotransmitters must be released dynamically in response to neuronal activity in the form of action potentials. Stable synaptic transmission is dependent on the efficacy of transmitter release and the rate of resupplying synaptic vesicles to their release sites. Accurate regulation is conferred by proteins sensing Ca2+ entering through voltage-gated Ca2+ channels opened by an action potential. Presynaptic Ca2+ concentration changes are dynamic functions in space and time, with wide fluctuations associated with different rates of neuronal activity...
2017: Proceedings of the Japan Academy. Series B, Physical and Biological Sciences
Paola Brun, Hamid I Akbarali
In the enteric nervous system, neurons make contact with smooth muscle cells to regulate gastrointestinal functions. Since neuronal cell alterations and intestinal motor dysfunctions are strictly linked, in vitro models based on the culture of neurons or smooth muscle cells are of great advantage to elucidate the functions of the enteric nervous system under physiological and pathological conditions. In this chapter, we provide the protocol for isolation of neurons and smooth muscle cells from the myenteric plexus of adult mice...
2018: Methods in Molecular Biology
Varuzhan Balasanyan, Kaori Watanabe, William P Dempsey, Tommy L Lewis, Le A Trinh, Don B Arnold
The essential organization of microtubules within neurons has been described; however, less is known about how neuronal actin is arranged and the functional implications of its arrangement. Here, we describe, in live cells, an actin-based structure in the proximal axon that selectively prevents some proteins from entering the axon while allowing the passage of others. Concentrated patches of actin in proximal axons are present shortly after axonal specification in rat and zebrafish neurons imaged live, and they mark positions where anterogradely traveling vesicles carrying dendritic proteins halt and reverse...
December 5, 2017: Cell Reports
Shoichi Nishida, Hisayoshi Yoshizaki, Yoshitomo Yasui, Tsuyoshi Kuwahara, Etsuko Kiyokawa, Miyuki Kohno
The enteric nervous system (ENS) is a network of neurons and glia that are derived from enteric neural crest cells (ENCCs) and essential for regulating peristaltic activity of the colon. ENCCs migrate along the gastrointestinal tract to form the ENS, and disruption of ENCC motility leads to ENS disorders, such as Hirschsprung's disease. Previous ENCC-transplant experiments show that ENCCs can invade into isolated mouse intestines by age E13.5, but not after E15.5. We hypothesized that altered age-specific micro-environments in the intestine are responsible for ENCC invasion/migration...
January 1, 2018: Biochemical and Biophysical Research Communications
Emilie Duchalais, Christophe Guilluy, Steven Nedellec, Melissa Touvron, Anne Bessard, Yann Touchefeu, Céline Bossard, Hélène Boudin, Guy Louarn, Michel Neunlist, Laurianne Van Landeghem
Background & Aims: In several types of cancers, tumor cells invade adjacent tissues by migrating along the resident nerves of the tumor microenvironment. This process, called perineural invasion , typically occurs along extrinsic nerves, with Schwann cells providing physical guidance for the tumor cells. However, in the colorectal cancer microenvironment, the most abundant nervous structures belong to the nonmyelinated intrinsic enteric nervous system (ENS). In this study, we investigated whether colon cancer cells interact with the ENS...
2018: Cellular and Molecular Gastroenterology and Hepatology
Sayantanee Biswas, Katherine Kalil
Proper organization and dynamics of the actin and microtubule (MT) cytoskeleton are essential for growth cone behaviors during axon growth and guidance. The microtubule associated protein tau is known to mediate actin/MT interactions in cell free systems but the role of tau in regulating cytoskeletal dynamics in living neurons is unknown. We used cultures of cortical neurons from postnatal day 0 (P0)-P2 golden Syrian hamsters (Mesocricetus auratus) of either sex to study the role of tau in the organization and dynamics of the axonal growth cone cytoskeleton...
November 22, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
Marco Pirazzini, Domenico Azarnia Tehran, Giulia Zanetti, Ornella Rossetto, Cesare Montecucco
Botulinum (BoNTs) and tetanus (TeNT) neurotoxins are the most toxic substances known and form the growing family of Clostridial neurotoxins (CNT), the etiologic agents of botulism and tetanus. CNT are composed of a metalloprotease light chain (L), linked via a disulfide bond to a heavy chain (H). H mediates the binding to nerve terminals and the membrane translocation of L into the cytosol, where its substrates, the three SNARE proteins, are localized. L translocation is accompanied by unfolding and, once delivered on the cytosolic side of the endosome membrane, it has to be reduced and reacquire the native fold to be active...
October 27, 2017: Toxicon: Official Journal of the International Society on Toxinology
Manuela Lahne, David R Hyde
Sensing and responding to our environment requires functional neurons that act in concert. Neuronal cell loss resulting from degenerative diseases cannot be replaced in humans, causing a functional impairment to integrate and/or respond to sensory cues. In contrast, zebrafish (Danio rerio) possess an endogenous capacity to regenerate lost neurons. Here, we will focus on the processes that lead to neuronal regeneration in the zebrafish retina. Dying retinal neurons release a damage signal, tumor necrosis factor α, which induces the resident radial glia, the Müller glia, to reprogram and re-enter the cell cycle...
August 2017: Neural Regeneration Research
Patrick S Stumpf, Rosanna C G Smith, Michael Lenz, Andreas Schuppert, Franz-Josef Müller, Ann Babtie, Thalia E Chan, Michael P H Stumpf, Colin P Please, Sam D Howison, Fumio Arai, Ben D MacArthur
Pluripotent stem cells can self-renew in culture and differentiate along all somatic lineages in vivo. While much is known about the molecular basis of pluripotency, the mechanisms of differentiation remain unclear. Here, we profile individual mouse embryonic stem cells as they progress along the neuronal lineage. We observe that cells pass from the pluripotent state to the neuronal state via an intermediate epiblast-like state. However, analysis of the rate at which cells enter and exit these observed cell states using a hidden Markov model indicates the presence of a chain of unobserved molecular states that each cell transits through stochastically in sequence...
September 27, 2017: Cell Systems
Xiaomeng Xu, Zhuoyu Wen, Nan Zhao, Xiaohui Xu, Fang Wang, Jie Gao, Yongjun Jiang, Xinfeng Liu
Toll-like receptor 4 (TLR4) is a proinflammatory cascade initiator in poststroke inflammation. In this study, miR-1906, a novel regulator of TLR4, was identified via in silico analysis and microRNA profiling in male adult mice and its expression was then quantitated in the ischemic hemisphere. We found miR-1906 to be significantly brain enriched in the ischemic hemisphere and even more drastically enriched in the peri-infarct regions. Furthermore, in vitro experiments demonstrated that, during oxygen-glucose deprivation, miR-1906 expression was increased in glial cells but decreased in neurons...
October 25, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
F Soufflet, M Biraud, M Rolli-Derkinderen, B Lardeux, C Trang, E Coron, S Bruley des Varannes, A Bourreille, M Neunlist
BACKGROUND: Neuroplastic changes in the enteric nervous system (ENS) observed during IBD might participate in physiopathological processes. Vasoactive intestinal polypeptide has been shown to be involved in intestinal inflammation and barrier functions. We aimed to investigate the modulation of VIP expression in colonic biopsies of IBD patient, the ability of soluble factors from biopsies to reproduce in vitro these modulations and identify soluble factors responsible. METHODS: VIP and cytokines mRNA expressions were assessed in colonic biopsies of healthy subjects (HS) and IBD patients from inflamed (I) and non-inflamed areas (NI)...
August 31, 2017: Neurogastroenterology and Motility: the Official Journal of the European Gastrointestinal Motility Society
Viktoriya Borisevich, Mehmet Hakan Ozdener, Bilal Malik, Barry Rockx
Henipaviruses are emerging zoonotic viruses and causative agents of encephalitis in humans. However, the mechanisms of entry into the central nervous system (CNS) in humans are not known. Here, we evaluated the possible role of olfactory epithelium in virus entry into the CNS. We characterized Hendra virus (HeV) and Nipah virus (NiV) infection of primary human olfactory epithelial cultures. We show that henipaviruses can infect mature olfactory sensory neurons. Henipaviruses replicated efficiently, resulting in cytopathic effect and limited induction of host responses...
May 2017: MSphere
Misawa Niki Ishii, Koji Yamamoto, Masanobu Shoji, Asano Asami, Yuji Kawamata
Accurate risk assessment for drug-induced seizure is expected to be performed before entering clinical studies because of its severity and fatal damage to drug development. Induced pluripotent stem cell (iPSC) technology has allowed the use of human neurons and glial cells in toxicology studies. Recently, several studies showed the advantage of co-culture system of human iPSC (hiPSC)-derived neurons with rodent/human primary astrocytes regarding neuronal functions. However, the application of hiPSC-derived neurons for seizure risk assessment has not yet been fully addressed, and not at all when co-cultured with hiPSC-derived astrocytes...
August 15, 2017: Toxicology
M Zeeshan Qaiser, Diana E M Dolman, David J Begley, N Joan Abbott, Mihaela Cazacu-Davidescu, Delia I Corol, Jonathan P Fry
Little is known about the origin of the neuroactive steroids dehydroepiandrosterone sulphate (DHEAS) and pregnenolone sulphate (PregS) in the brain or of their subsequent metabolism. Using rat brain perfusion in situ, we have found (3) H-PregS to enter more rapidly than (3) H-DHEAS and both to undergo extensive (> 50%) desulphation within 0.5 min of uptake. Enzyme activity for the steroid sulphatase catalysing this deconjugation was enriched in the capillary fraction of the blood-brain barrier and its mRNA expressed in cultures of rat brain endothelial cells and astrocytes...
September 2017: Journal of Neurochemistry
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