Visualizing form and function in organotypic slices of the adult mouse parotid gland.
An organotypic slice preparation of the adult mouse parotid salivary gland amenable to a variety of optical assessments of fluid and protein secretion dynamics is described. The semi-intact preparation rendered without the use of enzymatic treatment permitted live-cell imaging and multiphoton analysis of cellular and supracellular signals. Toward this end we demonstrated that the parotid slice is a significant addition to the repertoire of tools available to investigators to probe exocrine structure and function since there is currently no cell culture system that fully recapitulates parotid acinar cell biology. Importantly, we show that a subpopulation of the acinar cells of parotid slices can be maintained in short-term culture and retain their morphology and function for up to 2 days. This in vitro model system is a significant step forward compared with enzymatically dispersed acini that rapidly lose their morphological and functional characteristics over several hours, and it was shown to be long enough for the expression and trafficking of exogenous protein following adenoviral infection. This system is compatible with a variety of genetic and physiological approaches used to study secretory function.
Warner
Jennifer D
JD
Department of Neurosciences, Health Science Campus, The University of Toledo College of Medicine, 3035 Arlington Ave., Toledo, OH 43614, USA.
Peters
Christian G
CG
Saunders
Rudel
R
Won
Jong Hak
JH
Betzenhauser
Matthew J
MJ
Gunning
William T
WT
Yule
David I
DI
Giovannucci
David R
DR
D000199:Actins / Q000378:metabolism; D000256:Adenoviridae / Q000235:genetics; D000322:Adrenergic Agonists / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis; D020013:Calcium Signaling / Q000187:drug effects; D016764:Cell Polarity; D049109:Cell Proliferation; D048430:Cell Shape; D002470:Cell Survival; D018679:Cholinergic Agonists / Q000494:pharmacology; D004558:Electric Stimulation; D005089:Exocytosis / Q000187:drug effects; D005822:Genetic Vectors; D008297:Male; D051379:Mice; D018613:Microscopy, Confocal; D036641:Microscopy, Fluorescence, Multiphoton; D010306:Parotid Gland / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D022142:Secretory Vesicles / Q000378:metabolism; D013997:Time Factors; D046509:Tissue Culture Techniques; D014161:Transduction, Genetic
D000199:Actins; D000322:Adrenergic Agonists; D018679:Cholinergic Agonists; D011993:Recombinant Fusion Proteins