Ca2+ transients in submucous neurons during the colonic migrating motor complex in the isolated murine large intestine

Okamoto, Takanobu; Bayguinov, Peter O.; Broadhead, Matthew J.; Smith, Terence K.

doi:10.1111/j.1365-2982.2012.01934.x

Cited by 17 publications

(30 citation statements)

References 32 publications

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“…Cells can be efficiently stained in a minimally invasive manner by using membrane-permeable fluorescent indicators and optimized staining protocols 21 . This offers the opportunity to monitor a large number of neurons and enteric glia in functionally preserved preparations [14][15][16]22 , as well as in vivo 23 . Whole-mount tissue preparations are bulk loaded with a high-affinity Ca 2+ indicator dye such as Fluo-4 that increases its fluorescence when bound to Ca…”

Section: +mentioning

confidence: 99%

“…Imaging Ca 2+ transients in neurons and glia with fluorescent dyes is an established and widely used technique to study the functional organization and dynamics of the ENS [13][14][15][16][17] . Ca 2+ imaging has been shown to be an important tool in studying intact GI tissue segments to elucidate the spread of excitability through ICC pacemaker networks 18 and gut smooth muscle 19,20 .…”

Section: +mentioning

confidence: 99%

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In Situ Ca2+ Imaging of the Enteric Nervous System

Fried

Gulbransen

2015

JoVE

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Reflex behaviors of the intestine are controlled by the enteric nervous system (ENS). The ENS is an integrative network of neurons and glia in two ganglionated plexuses housed in the gut wall. Enteric neurons and enteric glia are the only cell types within the enteric ganglia. The activity of enteric neurons and glia is responsible for coordinating intestinal functions. This protocol describes methods for observing the activity of neurons and glia within the intact ENS by imaging intracellular calcium (Ca 2+ ) transients with fluorescent indicator dyes. Our technical discussion focuses on methods for Ca 2+ imaging in whole-mount preparations of the myenteric plexus from the rodent bowel.

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Section: +mentioning

confidence: 99%

Section: +mentioning

confidence: 99%

In Situ Ca2+ Imaging of the Enteric Nervous System

Fried

Gulbransen

2015

JoVE

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“…14,22 How the direct release of 5-HT in blood vessels coordinates with the output of vasodilator neurons in the submucosal plexus has not been determined. Blocking 5-HT 3 receptors can lead to ischemic colitis; 4 this could be explained by reduced vasodilator output from submucosal neurons leading to constriction of submucosal arteries.…”

Section: Diverse Projections Of Serotonergic Neurons In the Colonmentioning

confidence: 99%

“…(1) Colonic migrating motor complexes (CMMCs) 8 (2) Tonic inhibition of the muscle 21 (3) Secretomotor activity and blood flow 14,22 CMMCs and high amplitude propagating contractions (HAPCs) are probably a similar integrated motor events ( Fig. 2A and 2B) normally involving both mucosal 5-HT and myenteric 5-HT neurons.…”

Section: Introductionmentioning

confidence: 99%

Colonic Migrating Motor Complexes, High Amplitude Propagating Contractions, Neural Reflexes and the Importance of Neuronal and Mucosal Serotonin

Smith¹,

Park²,

Hennig³

2014

J Neurogastroenterol Motil

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“…5 Also, by using ChAT-GCaMP3 and nNOS-GCaMP3 mice we can begin monitoring Ca 2+ transient activity in broadly defined cholinergic and nitrergic neurons in myenteric and submucosal ganglia, without the necessity of time-consuming post-staining immunohistochemistry, as was previously necessary. 6 In conclusion the new technique should be develop the emerging new calcium imaging technique.…”

Section: Hyun Jin Kimmentioning

confidence: 99%

The New Calcium Imaging Trend (Front Cell Neurosci 2015;9:436)

Kim

2016

J Neurogastroenterol Motil

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SummaryCalcium imaging is a technique for intracellular calcium measurement, and it is important to understand the neural and glial functions. Genetically encoded Ca 2+ indicators have been used extensively in many body systems to detect Ca 2+ transients associated with neuronal activity. The aim of this article is to utilize a number of cell-specific promoters that express the Ca 2+ indicator GCaMP3 in different classes of neurons and glia to determine their effectiveness in measuring activity in enteric neural networks during colonic motor behaviors. 1The authors found that the activity in glial networks appeared to follow neural activity but continued long after neural activity had waned. With these new tools an unprecedented level of detail can be recorded from the enteric nervous system (ENS) with minimal manipulation of tissue. These techniques can be extended in order to better understand the roles of particular enteric neurons and glia during normal and disordered motility. CommentsENS is composed of neurons and glial cells. Enteric glial cells are located in close contact with enteric neurons within the ganglia, along inter-ganglionic connectives of the myenteric and submucosal plexus, and can also be found in the extra-ganglionic layers of the gut wall, 2 glial cells shows dynamic responses to neuronal inputs and have the apparatus to sequester and release the neuro-active factors. Nonetheless, whether these cells indeed regulate synaptic transmission in a physiological context such as gut motility pattern is not well known. 3The live imaging of neuron-glia interaction is very important in understanding these functions. The measuring of intracellular calcium can be done using fluorescent indicators and it has become the most popular technique. It consists of 2 steps, in the first step primary ENS cultures are loaded with the Ca 2+ indicator dye, Fluo4-AM or Fura-2, and the second step is stimulated by 75 mM K + depolarization. 4However, traditional Ca 2+ imaging techniques have limitations.Calcium imaging bleaching changed indicators cannot cross the cell membrane, and the intracellular concentration might decrease during the experiment. This means that long-term recording is difficult, and the recording of drug response is also limitated. The new calcium imaging uses GCaMP3 mice. There are several kind of mice: Wnt1-Cre, Rosa26-Lox-STOP-LoxGCaMP3 mice (GCaMP3 targeted to neural crest derivatives, including enteric neurons and glia via Wnt1-Cre transgenic mice; Jax #009107; heretofore referred to as Wnt1-GCaMP3); GFAPCre, Rosa26-Lox-STOP-Lox-GCaMP3 (GCaMP3 targeted to glia via mice expressing Cre from the human GFAP promoter; Jax #004600; heretofore referred to as GFAP-GCaMP3); ChAT-Cre, Rosa26-Lox-STOP-Lox-GCaMP3 (GCaMP3 targeted to cholinergic neurons via ChAT-Cre BAC transgenic mice; MMRRC #37336; heretofore referred to as ChAT-GCaMP3); and nNOS-

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Ca²⁺ transients in submucous neurons during the colonic migrating motor complex in the isolated murine large intestine

Cited by 17 publications

References 32 publications

<em>In Situ</em> Ca<sup>2+</sup> Imaging of the Enteric Nervous System

<em>In Situ</em> Ca<sup>2+</sup> Imaging of the Enteric Nervous System

Colonic Migrating Motor Complexes, High Amplitude Propagating Contractions, Neural Reflexes and the Importance of Neuronal and Mucosal Serotonin

The New Calcium Imaging Trend (Front Cell Neurosci 2015;9:436)

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