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Am J Physiol Gastrointest Liver Physiol 295: G629-G640, 2008. First published July 31, 2008; doi:10.1152/ajpgi.90217.2008
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HORMONES AND SIGNALING

Visualizing form and function in organotypic slices of the adult mouse parotid gland

Jennifer D. Warner,1 Christian G. Peters,1 Rudel Saunders,1 Jong Hak Won,2 Matthew J. Betzenhauser,2 William T. Gunning, III,1 David I. Yule,2 and David R. Giovannucci1

1Department of Neurosciences, University of Toledo College of Medicine, Health Science Campus, Toledo, Ohio; and 2Department of Pharmacology and Physiology, University of Rochester Medical Center, Rochester, New York

Submitted 29 February 2008 ; accepted in final form 24 July 2008

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.

Ca2+ signaling; exocytosis; immunofluorescence; electron microscopy; multiphoton; tissue culture


Address for reprint requests and other correspondence: D. R. Giovannucci, Dept. of Neurosciences, Health Science Campus, The Univ. of Toledo, 3035 Arlington Ave., Toledo, OH 43614 (e-mail: david.giovannucci{at}utoledo.edu)






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