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Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture
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| Zeitschriftentitel: | American Journal of Physiology-Gastrointestinal and Liver Physiology |
|---|---|
| Personen und Körperschaften: | , , , |
| In: | American Journal of Physiology-Gastrointestinal and Liver Physiology, 305, 2013, 2, S. G129-G138 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
American Physiological Society
|
| Schlagwörter: |
| author_facet |
Dong, Hui Jiang, Yanfen Srinivasan, Shanthi Mittal, Ravinder K. Dong, Hui Jiang, Yanfen Srinivasan, Shanthi Mittal, Ravinder K. |
|---|---|
| author |
Dong, Hui Jiang, Yanfen Srinivasan, Shanthi Mittal, Ravinder K. |
| spellingShingle |
Dong, Hui Jiang, Yanfen Srinivasan, Shanthi Mittal, Ravinder K. American Journal of Physiology-Gastrointestinal and Liver Physiology Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture Physiology (medical) Gastroenterology Hepatology Physiology |
| author_sort |
dong, hui |
| spelling |
Dong, Hui Jiang, Yanfen Srinivasan, Shanthi Mittal, Ravinder K. 0193-1857 1522-1547 American Physiological Society Physiology (medical) Gastroenterology Hepatology Physiology http://dx.doi.org/10.1152/ajpgi.00040.2013 <jats:p>The enteric nervous system of the esophagus plays an important role in its sensory and motor functions. Although the esophagus contains enteric neurons, they have never been isolated and characterized in primary culture. We isolated and cultured enteric neurons of the rat esophagus and determined their morphological appearance, chemical coding for neurotransmitters, and functional characteristics. After primary culture for 2 wk, dendrites and axons appeared in the enteric neurons, which usually have one axon and several dendrites. Although the size of neuronal bodies varied from Dogiel type I to type II, their average size was 39 ± 1.8 μm in length and 23 ± 1.4 μm in width. Immmunocytochemical studies revealed that over 95% of these cells were positively stained for two general neuronal markers, PGP 9.5 or Milli-Mark Fluoro. Chemical coding showed that the neurons were positively stained for choline acetyltransferease (53 ± 6%) or nNOS (66 ± 13%). In functional studies, membrane depolarization and stimulation of several G protein-coupled receptors (GPCRs) induced Ca<jats:sup>2+</jats:sup>signaling in the esophageal enteric neurons. The GPCR stimulation was found to induce both intracellular Ca<jats:sup>2+</jats:sup>release and extracellular Ca<jats:sup>2+</jats:sup>entry. The functional expressions of Ca<jats:sup>2+</jats:sup>channels (voltage-gated Ca<jats:sup>2+</jats:sup>channels and store-operated channels) and Ca<jats:sup>2+</jats:sup>pump (sarcoplasmic reticulum Ca<jats:sup>2+</jats:sup>-ATPase) were also demonstrated on these neurons. We have grown, for the first time, esophageal enteric neurons in primary culture, and these contain excitatory and inhibitory neurotransmitters. The functional integrity of GPCRs, Ca<jats:sup>2+</jats:sup>channels, and Ca<jats:sup>2+</jats:sup>pump in these neurons makes them a useful cell model for further studies.</jats:p> Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture American Journal of Physiology-Gastrointestinal and Liver Physiology |
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10.1152/ajpgi.00040.2013 |
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| title |
Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| title_unstemmed |
Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| title_full |
Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| title_fullStr |
Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| title_full_unstemmed |
Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| title_short |
Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| title_sort |
morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| topic |
Physiology (medical) Gastroenterology Hepatology Physiology |
| url |
http://dx.doi.org/10.1152/ajpgi.00040.2013 |
| publishDate |
2013 |
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G129-G138 |
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<jats:p>The enteric nervous system of the esophagus plays an important role in its sensory and motor functions. Although the esophagus contains enteric neurons, they have never been isolated and characterized in primary culture. We isolated and cultured enteric neurons of the rat esophagus and determined their morphological appearance, chemical coding for neurotransmitters, and functional characteristics. After primary culture for 2 wk, dendrites and axons appeared in the enteric neurons, which usually have one axon and several dendrites. Although the size of neuronal bodies varied from Dogiel type I to type II, their average size was 39 ± 1.8 μm in length and 23 ± 1.4 μm in width. Immmunocytochemical studies revealed that over 95% of these cells were positively stained for two general neuronal markers, PGP 9.5 or Milli-Mark Fluoro. Chemical coding showed that the neurons were positively stained for choline acetyltransferease (53 ± 6%) or nNOS (66 ± 13%). In functional studies, membrane depolarization and stimulation of several G protein-coupled receptors (GPCRs) induced Ca<jats:sup>2+</jats:sup>signaling in the esophageal enteric neurons. The GPCR stimulation was found to induce both intracellular Ca<jats:sup>2+</jats:sup>release and extracellular Ca<jats:sup>2+</jats:sup>entry. The functional expressions of Ca<jats:sup>2+</jats:sup>channels (voltage-gated Ca<jats:sup>2+</jats:sup>channels and store-operated channels) and Ca<jats:sup>2+</jats:sup>pump (sarcoplasmic reticulum Ca<jats:sup>2+</jats:sup>-ATPase) were also demonstrated on these neurons. We have grown, for the first time, esophageal enteric neurons in primary culture, and these contain excitatory and inhibitory neurotransmitters. The functional integrity of GPCRs, Ca<jats:sup>2+</jats:sup>channels, and Ca<jats:sup>2+</jats:sup>pump in these neurons makes them a useful cell model for further studies.</jats:p> |
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| author | Dong, Hui, Jiang, Yanfen, Srinivasan, Shanthi, Mittal, Ravinder K. |
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| description | <jats:p>The enteric nervous system of the esophagus plays an important role in its sensory and motor functions. Although the esophagus contains enteric neurons, they have never been isolated and characterized in primary culture. We isolated and cultured enteric neurons of the rat esophagus and determined their morphological appearance, chemical coding for neurotransmitters, and functional characteristics. After primary culture for 2 wk, dendrites and axons appeared in the enteric neurons, which usually have one axon and several dendrites. Although the size of neuronal bodies varied from Dogiel type I to type II, their average size was 39 ± 1.8 μm in length and 23 ± 1.4 μm in width. Immmunocytochemical studies revealed that over 95% of these cells were positively stained for two general neuronal markers, PGP 9.5 or Milli-Mark Fluoro. Chemical coding showed that the neurons were positively stained for choline acetyltransferease (53 ± 6%) or nNOS (66 ± 13%). In functional studies, membrane depolarization and stimulation of several G protein-coupled receptors (GPCRs) induced Ca<jats:sup>2+</jats:sup>signaling in the esophageal enteric neurons. The GPCR stimulation was found to induce both intracellular Ca<jats:sup>2+</jats:sup>release and extracellular Ca<jats:sup>2+</jats:sup>entry. The functional expressions of Ca<jats:sup>2+</jats:sup>channels (voltage-gated Ca<jats:sup>2+</jats:sup>channels and store-operated channels) and Ca<jats:sup>2+</jats:sup>pump (sarcoplasmic reticulum Ca<jats:sup>2+</jats:sup>-ATPase) were also demonstrated on these neurons. We have grown, for the first time, esophageal enteric neurons in primary culture, and these contain excitatory and inhibitory neurotransmitters. The functional integrity of GPCRs, Ca<jats:sup>2+</jats:sup>channels, and Ca<jats:sup>2+</jats:sup>pump in these neurons makes them a useful cell model for further studies.</jats:p> |
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| spelling | Dong, Hui Jiang, Yanfen Srinivasan, Shanthi Mittal, Ravinder K. 0193-1857 1522-1547 American Physiological Society Physiology (medical) Gastroenterology Hepatology Physiology http://dx.doi.org/10.1152/ajpgi.00040.2013 <jats:p>The enteric nervous system of the esophagus plays an important role in its sensory and motor functions. Although the esophagus contains enteric neurons, they have never been isolated and characterized in primary culture. We isolated and cultured enteric neurons of the rat esophagus and determined their morphological appearance, chemical coding for neurotransmitters, and functional characteristics. After primary culture for 2 wk, dendrites and axons appeared in the enteric neurons, which usually have one axon and several dendrites. Although the size of neuronal bodies varied from Dogiel type I to type II, their average size was 39 ± 1.8 μm in length and 23 ± 1.4 μm in width. Immmunocytochemical studies revealed that over 95% of these cells were positively stained for two general neuronal markers, PGP 9.5 or Milli-Mark Fluoro. Chemical coding showed that the neurons were positively stained for choline acetyltransferease (53 ± 6%) or nNOS (66 ± 13%). In functional studies, membrane depolarization and stimulation of several G protein-coupled receptors (GPCRs) induced Ca<jats:sup>2+</jats:sup>signaling in the esophageal enteric neurons. The GPCR stimulation was found to induce both intracellular Ca<jats:sup>2+</jats:sup>release and extracellular Ca<jats:sup>2+</jats:sup>entry. The functional expressions of Ca<jats:sup>2+</jats:sup>channels (voltage-gated Ca<jats:sup>2+</jats:sup>channels and store-operated channels) and Ca<jats:sup>2+</jats:sup>pump (sarcoplasmic reticulum Ca<jats:sup>2+</jats:sup>-ATPase) were also demonstrated on these neurons. We have grown, for the first time, esophageal enteric neurons in primary culture, and these contain excitatory and inhibitory neurotransmitters. The functional integrity of GPCRs, Ca<jats:sup>2+</jats:sup>channels, and Ca<jats:sup>2+</jats:sup>pump in these neurons makes them a useful cell model for further studies.</jats:p> Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture American Journal of Physiology-Gastrointestinal and Liver Physiology |
| spellingShingle | Dong, Hui, Jiang, Yanfen, Srinivasan, Shanthi, Mittal, Ravinder K., American Journal of Physiology-Gastrointestinal and Liver Physiology, Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture, Physiology (medical), Gastroenterology, Hepatology, Physiology |
| title | Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| title_full | Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| title_fullStr | Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| title_full_unstemmed | Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| title_short | Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| title_sort | morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| title_unstemmed | Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture |
| topic | Physiology (medical), Gastroenterology, Hepatology, Physiology |
| url | http://dx.doi.org/10.1152/ajpgi.00040.2013 |