Isolation of acini from nasal glands of the guinea-pig.
A procedure for isolating the acinar cells of the serous gland in the mammalian nasal septum has been developed. This technique is characterized by meticulous and selective isolation with minimal contamination by the surface epithelial cells and employs enzymatic treatment with collagenase. The isolated cells were confirmed to be serous gland acini as shown by negative staining with Alcian blue and a high electron density of the granules. The acini were more than 90% viable as judged by trypan blue exclusion. Ultrastructural integrity of the cells was well maintained following the isolation procedure. Application of acetylcholine to the isolated acini induced an inward current in a whole-cell patch clamp and increased intracellular Ca2+ concentration measured by fura-2. These acetylcholine responses were completely blocked by atropine. These physiological findings directly demonstrated that nasal gland acini possess muscarinic-activated receptors as previously suggested. These isolated cells hold promise for the in vitro study of secretory mechanisms in the mammalian nasal gland.
Sunose
H
H
Department of Otolaryngology, Tohoku University School of Medicine, Sendai, Japan.
Zhang
W
W
Ishigaki
M
M
Katori
Y
Y
Suzuki
M
M
Ikeda
K
K
Takasaka
T
T
Saito
Y
Y
Nishiyama
A
A
D000109:Acetylcholine / Q000037:antagonists & inhibitors / Q000494:pharmacology; D000818:Animals; D001285:Atropine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D003592:Cytophotometry; D016257:Fura-2; D006168:Guinea Pigs; D006651:Histocytochemistry; D066298:In Vitro Techniques; D008564:Membrane Potentials / Q000187:drug effects; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D009297:Nasal Mucosa / Q000033:anatomy & histology* / Q000648:ultrastructure; D009300:Nasal Septum / Q000033:anatomy & histology* / Q000648:ultrastructure; D010275:Parasympathetic Nervous System / Q000502:physiology; D018408:Patch-Clamp Techniques; D015398:Signal Transduction / Q000502:physiology; D013194:Staining and Labeling
D001285:Atropine; D000109:Acetylcholine; D002118:Calcium; D016257:Fura-2