PKC-epsilon translocation in enteric neurons and interstitial cells of Cajal in response to muscarinic stimulation.
Interstitial cells of Cajal in the deep muscular plexus (ICC-DMP) of the small intestine express excitatory neurotransmitter receptors. We tested whether ICC-DMP are functionally innervated by cholinergic neurons in the murine intestine. Muscles were stimulated by intrinsic nerves and ACh and processed for immunohistochemistry to determine these effects on PKC-epsilon activation. Under control conditions, PKC-epsilon-like immunoreactivy (PKC-epsilon-LI) was only observed in myenteric neurons within the tunica muscularis. Electrical field stimulation or ACh caused translocation of neural PKC-epsilon-LI from the cytosol to a peripheral compartment. After stimulation, PKC-epsilon-LI was found in spindle-shaped cells in the DMP. These cells were identified as ICC-DMP by Kit-LI and vimentin-LI. PKC-epsilon-LI in ICC-DMP and translocation of PKC epsilon-LI in neurons were blocked by tetrodotoxin or atropine, suggesting that these responses were due to activation of muscarinic receptors. Western blots also confirmed translocation of PKC-epsilon-LI. In conclusion, PKC-epsilon translocation is linked to muscarinic receptor activation in ICC-DMP and a subpopulation of myenteric neurons. These studies demonstrate that ICC-DMP are functionally innervated by excitatory motoneurons.
Wang
Xuan-Yu
XY
Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, NV 89557, USA.
Ward
Sean M
SM
Gerthoffer
William T
WT
Sanders
Kenton M
KM
D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D001285:Atropine / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects; D004558:Electric Stimulation; D004789:Enzyme Activation / Q000187:drug effects; D007421:Intestine, Small / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D018727:Muscarinic Antagonists / Q000494:pharmacology; D009130:Muscle, Smooth / Q000166:cytology / Q000378:metabolism; D009197:Myenteric Plexus / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D051744:Protein Kinase C-epsilon; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000502:physiology*; D013779:Tetrodotoxin / Q000494:pharmacology; D014746:Vimentin / Q000378:metabolism
D018727:Muscarinic Antagonists; D011976:Receptors, Muscarinic; D014746:Vimentin; D013779:Tetrodotoxin; D001285:Atropine; C497256:Prkce protein, mouse; D019009:Proto-Oncogene Proteins c-kit; D011493:Protein Kinase C; D051744:Protein Kinase C-epsilon; D000109:Acetylcholine