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Enteric neuron culture

Lily S Cheng, Hannah K Graham, Wei Hua Pan, Nandor Nagy, Alfonso Carreon-Rodriguez, Allan M Goldstein, Ryo Hotta
BACKGROUND: Enteric neurospheres derived from postnatal intestine represent a promising avenue for cell replacement therapy to treat Hirschsprung disease and other neurointestinal diseases. We describe a simple method to improve the neuronal yield of spontaneously formed gut-derived neurospheres. MATERIALS AND METHODS: Enteric neurospheres were formed from the small and large intestines of mouse and human subjects. Neurosphere size, neural crest cell content, cell migration, neuronal differentiation, and neuronal proliferation in culture were analyzed...
December 2016: Journal of Surgical Research
Stephen R Adams, Mason R Mackey, Ranjan Ramachandra, Sakina F Palida Lemieux, Paul Steinbach, Eric A Bushong, Margaret T Butko, Ben N G Giepmans, Mark H Ellisman, Roger Y Tsien
Electron microscopy (EM) remains the primary method for imaging cellular and tissue ultrastructure, although simultaneous localization of multiple specific molecules continues to be a challenge for EM. We present a method for obtaining multicolor EM views of multiple subcellular components. The method uses sequential, localized deposition of different lanthanides by photosensitizers, small-molecule probes, or peroxidases. Detailed view of biological structures is created by overlaying conventional electron micrographs with pseudocolor lanthanide elemental maps derived from distinctive electron energy-loss spectra of each lanthanide deposit via energy-filtered transmission electron microscopy...
November 17, 2016: Cell Chemical Biology
Abby R Kroken, Faith C Blum, Madison Zuverink, Joseph T Barbieri
Botulinum neurotoxins (BoNTs) are the most toxic proteins for humans, but also common therapies for neurological diseases. BoNTs are di-chain toxins, comprising an N-terminal catalytic domain (LC) disulfide bond linked to a C-terminal heavy chain (HC) which includes a translocation domain (HN) and a receptor-binding domain (HC). Recently, the BoNT/A subtypes A1 and A2 were reported to possess similar potency, but different rates of cellular intoxication and pathology in a mouse model of botulism. The current study measured HCA1 and HCA2 entry into rat primary neurons and cultured Neuro2A cells...
October 17, 2016: Infection and Immunity
W Li, L Huang, J Zeng, W Lin, K Li, J Sun, W Huang, J Chen, G Wang, Q Ke, J Duan, X Lai, R Chen, M Liu, Y Liu, T Wang, X Yang, Y Chen, H Xia, A P Xiang
The enteric nervous system (ENS) is recognized as a second brain because of its complexity and its largely autonomic control of bowel function. Recent progress in studying the interactions between the ENS and the central nervous system (CNS) has implicated alterations of the gut/brain axis as a possible mechanism in the pathophysiology of autism spectrum disorders (ASDs), Parkinson's disease (PD) and other human CNS disorders, whereas the underlying mechanisms are largely unknown because of the lack of good model systems...
October 25, 2016: Molecular Psychiatry
Jairo A Diaz, Mauricio F Murillo, Jhonan A Mendoza, Ana M Barreto, Lina S Poveda, Lina K Sanchez, Laura C Poveda, Katherine T Mora
Emergent biological responses develop via unknown processes dependent on physical collision. In hypoxia, when the tissue architecture collapses but the geometric core is stable, actin cytoskeleton filament components emerge, revealing a hidden internal order that identifies how each molecule is reassembled into the original mold, using one common connection, i.e., a fractal self-similarity that guides the system from the beginning in reverse metamorphosis, with spontaneous self-assembly of past forms that mimics an embryoid phenotype...
2016: American Journal of Stem Cells
Azusa Yoneshige, Man Hagiyama, Takao Inoue, Tomonori Tanaka, Aritoshi Ri, Akihiko Ito
Internal pressure is often involved in neurodegeneration; intraocular and intraventricular pressure elevations over 20-30 cmH2O cause glaucoma and hydrocephalus, respectively. Here, we investigated enteric nerve degeneration in colon segments having tumor-induced stenosis and dilation and examined the mechanism of intraluminal pressure involvement. Histological examination revealed that the enteric ganglion neurons and neurites decreased in density in the dilated colons proportionate to the degree of dilation...
October 8, 2016: Molecular Neurobiology
Marta Domínguez-Prieto, Ana Velasco, Lourdes Vega, Arantxa Tabernero, José M Medina
Amyloid-β (Aβ), Aβ40, Aβ42, and, recently, Aβ25-35 have been directly implicated in the pathogenesis of Alzheimer's disease. We have studied the effects of Aβ on neuronal death, reactive oxygen species (ROS) production, and synaptic assembling in neurons in primary culture. Aβ25-35, Aβ40, and Aβ42 significantly decreased neuronal viability, although Aβ25-35 showed a higher effect. Aβ25-35 showed a more penetrating ability to reach mitochondria while Aβ40 did not enter the neuronal cytosol and Aβ42 was scarcely internalized...
November 1, 2016: Journal of Alzheimer's Disease: JAD
Rodolphe Soret, Nicolas Pilon
Hirschsprung disease (HSCR), also named aganglionic megacolon, is a severe congenital malformation characterized by a lack of enteric nervous system (ENS) in the terminal regions of the bowel (Bergeron et al., 2013). As the ENS notably regulates motility in the whole gastrointestinal track, the segment without neurons remains tonically contracted, resulting in functional intestinal obstruction and accumulation of fecal material (megacolon). HSCR occurs when enteric neural progenitors of vagal neural crest origin fail to fully colonize the developing intestines...
September 5, 2016: Bio-protocol
Anne-Gaëlle Corbillé, Michel Neunlist, Pascal Derkinderen
Since the observation that aggregated α-synuclein, the pathological hallmark of Parkinson's disease (PD), is found in the gut in almost all patients, it has been suggested that the enteric nervous system (ENS) could be a starting point for α-synuclein pathology. α-synuclein has long been thought to occur as a monomer in living cells, but recent studies reported that it instead exists as a tetramer in non-neuronal cells and in neurons. Given the possible key role of the ENS in PD pathophysiology, we undertook the current research to characterize the native state of α-synuclein in rat primary culture of ENS and in adult human healthy ENS...
September 17, 2016: Journal of Neurochemistry
Paulo L C Coelho, Mona N Oliveira, Alessandra B da Silva, Bruno P S Pitanga, Victor D A Silva, Giselle P Faria, Geraldo P Sampaio, Maria de Fatima D Costa, Suzana Braga-de-Souza, Silvia L Costa
This study aimed to investigate the antitumor and immunomodulatory properties of the flavonoid apigenin (5,7,4'-trihydroxyflavone), which was extracted from Croton betulaster Mull, in glioma cell culture using the high-proliferative rat C6 glioma cell line as a model. Apigenin was found to have the ability to reduce the viability and proliferation of C6 cells in a time-dependent and dose-dependent manner, with an IC50 of 22.8 µmol/l, 40 times lower than that of temozolomide (1000 µmol/l), after 72 h of apigenin treatment...
November 2016: Anti-cancer Drugs
Xiaowen Cheng, Antonio Boza-Serrano, Michelle Foldschak Turesson, Tomas Deierborg, Eva Ekblad, Ulrikke Voss
In addition to brain injury stroke patients often suffer gastrointestinal complications. Neuroimmune interactions involving galectin-3, released from microglia in the brain, mediates the post-stroke pro-inflammatory response. We investigated possible consequences of stroke on the enteric nervous system and the involvement of galectin-3. We show that permanent middle cerebral artery occlusion (pMCAO) induces loss of enteric neurons in ileum and colon in galectin-3(+/+), but not in galectin-3(-/-), mice. In vitro we show that serum from galectin-3(+/+), but not from galectin-3(-/-), mice subjected to pMCAO, caused loss of C57BL/6J myenteric neurons, while myenteric neurons derived from TLR4(-/-) mice were unaffected...
2016: Scientific Reports
Anna K Wójtowicz, Konrad A Szychowski, Agnieszka Wnuk, Małgorzata Kajta
Dibutyl phthalate (di-n-butyl phthalate, DBP) is one of the most commonly used phthalate esters. DBP is widely used as a plasticizer in a variety of household industries and consumer products. Because phthalates are not chemically bound to products, they can easily leak out to enter the environment. DBP can pass through the placental and blood-brain barriers due to its chemical structure, but little is known about its mechanism of action in neuronal cells. This study demonstrated the toxic and apoptotic effects of DBP in mouse neocortical neurons in primary cultures...
August 31, 2016: Neurotoxicity Research
Joan F Burgueño, Albert Barba, Elena Eyre, Carolina Romero, Michel Neunlist, Ester Fernández
BACKGROUND: Accumulating evidence suggest that the enteric nervous system (ENS) plays important roles in gastrointestinal inflammatory responses, which could be in part mediated by Toll-like receptor (TLR) activation. The aim of this study was to characterise the expression and functionality of TLR2/4/9 in the ENS. METHODS: TLR2/4/9 expression was assessed in the plexuses of adult rats and embryonic ENS cultures by immunofluorescence and quantitative PCR. Following stimulation with TLR2/4/9 ligands or their combinations, activation of NF-kB, production of TNF-α, IL-6 and MCP-1 and chemoattraction of RAW264...
2016: Journal of Neuroinflammation
Thomas Schikorski
Functional imaging is the measurement of structural changes during an ongoing physiological process over time. In many cases, functional imaging has been implemented by tracking a fluorescent signal in live imaging sessions. Electron microscopy, however, excludes live imaging which has hampered functional imaging approaches on the ultrastructural level. This barrier was broken with the introduction of superfast fixation. Superfast fixation is capable of stopping and fixing membrane traffic at sufficient speed to capture a physiological process at a distinct functional state...
2016: Methods in Molecular Biology
Giuseppe Martano, Luca Murru, Edoardo Moretto, Laura Gerosa, Giulia Garrone, Vittorio Krogh, Maria Passafaro
INTRODUCTION: Neurons have a very high energy requirement, and their metabolism is tightly regulated to ensure delivery of adequate substrate to sustain neuronal activity and neuroplastic changes. The mechanisms underlying the regulation of neuronal metabolism, however, are not completely clear. OBJECTIVE: The objective of this study was to investigate the central carbon metabolism in neurons, in order to identify the regulatory pathways governing neuronal anabolism and catabolism...
2016: Metabolomics: Official Journal of the Metabolomic Society
Catherine Le Berre-Scoul, Julien Chevalier, Elena Oleynikova, François Cossais, Sophie Talon, Michel Neunlist, Hélène Boudin
In the nervous system, the formation of neuronal circuitry results from a complex and coordinated action of intrinsic and extrinsic factors. In the CNS, extrinsic mediators derived from astrocytes have been shown to play a key role in neuronal maturation, including dendritic shaping, axon guidance and synaptogenesis. In the enteric nervous system (ENS), the potential role of enteric glial cells (EGCs) in the maturation of developing enteric neuronal circuit is currently unknown. A major obstacle in addressing this question is the difficulty to obtain a valuable experimental model in which enteric neurons could be isolated and maintained without EGCs...
July 20, 2016: Journal of Physiology
Wei-Kang Pan, Hui Yu, A-Li Wu, Ya Gao, Bai-Jun Zheng, Peng Li, Wei-Li Yang, Qiang Huang, Huai-Jie Wang, Xin Ge
Human enteric neural stem cells (hENSCs) proliferate and differentiate into neurons and glial cells in response to a complex network of neurotrophic factors to form the enteric nervous system. The primary aim of this study was to determine the effect of basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) on in-vitro expansion and differentiation of postnatal hENSCs-containing enteric neurosphere cells. Enteric neurosphere cells were isolated from rectal polyp specimens of 75 children (age, 1-13 years) and conditioned with bFGF, EGF, bFGF+EGF, or plain culture media...
August 3, 2016: Neuroreport
Nina Westphal, Ralf Kleene, David Lutz, Thomas Theis, Melitta Schachner
In the mammalian nervous system, the neural cell adhesion molecule NCAM is the major carrier of the glycan polymer polysialic acid (PSA) which confers important functions to NCAM's protein backbone. PSA attached to NCAM contributes not only to cell migration, neuritogenesis, synaptic plasticity, and behavior, but also to regulation of the circadian rhythm by yet unknown molecular mechanisms. Here, we show that a PSA-carrying transmembrane NCAM fragment enters the nucleus after stimulation of cultured neurons with surrogate NCAM ligands, a phenomenon that depends on the circadian rhythm...
July 2016: Molecular and Cellular Neurosciences
Thomas I-H Park, Vaughan Feisst, Anna E S Brooks, Justin Rustenhoven, Hector J Monzo, Sheryl X Feng, Edward W Mee, Peter S Bergin, Robyn Oldfield, E Scott Graham, Maurice A Curtis, Richard L M Faull, P Rod Dunbar, Mike Dragunow
The human brain is a highly vascular organ in which the blood-brain barrier (BBB) tightly regulates molecules entering the brain. Pericytes are an integral cell type of the BBB, regulating vascular integrity, neuroinflammation, angiogenesis and wound repair. Despite their importance, identifying pericytes amongst other perivascular cell types and deciphering their specific role in the neurovasculature remains a challenge. Using primary adult human brain cultures and fluorescent-activated cell sorting, we identified two CD73(+)CD45(-) mesenchymal populations that showed either high or low CD90 expression...
2016: Scientific Reports
Jiwoo Bae, Nayeon Lee, Wankyu Choi, Suji Lee, Jung Jae Ko, Baek Soo Han, Sang Chul Lee, Noo Li Jeon, Jihwan Song
Microfluidics forms the basis of unique experimental approaches that visualize the development of neural structure using micro-scale devices and aids the guidance of neurite growth in an axonal isolation compartment. We utilized microfluidics technology to monitor the differentiation and migration of neural cells derived from human embryonic stems cells (hESC). We cocultured hESC with PA6 stromal cells and isolated neural rosette-like structures, which subsequently formed neurospheres in a suspension culture...
2016: Methods in Molecular Biology
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