Salivary Glands: A Powerful Experimental System to Study Cell Biology in Live Animals by Intravital Microscopy

Šrámková, Monika; Porat-Shliom, Andrius Masedunkas Natalie; Wigand, Timothy; Amornphimoltham, Panomwat; Weigert, Roberto

doi:10.5772/33524

Cited by 4 publications

(4 citation statements)

References 98 publications

(130 reference statements)

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“…The choice of the SGs as a model organ for IVM is due to the fact that the glands are easily accessible by performing a minor surgery, can be externalized without compromising their physiology, and stabilized to reduce the motion artifacts due to heartbeat and respiration. In addition, SGs can be selectively manipulated genetically by injecting either viral or non-viral based vectors through the salivary duct 8,9 . Finally, SGs are exocrine glands composed of polarized epithelial cells, which form the acini and the ducts, myoepithelial cells, and a complex population of stromal cells.…”

Section: Introductionmentioning

confidence: 99%

Intravital Microscopy for Imaging Subcellular Structures in Live Mice Expressing Fluorescent Proteins

Masedunskas

Porat-Shliom

Tora

et al. 2013

JoVE

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Here we describe a procedure to image subcellular structures in live rodents that is based on the use of confocal intravital microscopy. As a model organ, we use the salivary glands of live mice since they provide several advantages. First, they can be easily exposed to enable access to the optics, and stabilized to facilitate the reduction of the motion artifacts due to heartbeat and respiration. This significantly facilitates imaging and tracking small subcellular structures. Second, most of the cell populations of the salivary glands are accessible from the surface of the organ. This permits the use of confocal microscopy that has a higher spatial resolution than other techniques that have been used for in vivo imaging, such as two-photon microscopy. Finally, salivary glands can be easily manipulated pharmacologically and genetically, thus providing a robust system to investigate biological processes at a molecular level.In this study we focus on a protocol designed to follow the kinetics of the exocytosis of secretory granules in acinar cells and the dynamics of the apical plasma membrane where the secretory granules fuse upon stimulation of the beta-adrenergic receptors. Specifically, we used a transgenic mouse that co-expresses cytosolic GFP and a membrane-targeted peptide fused with the fluorescent protein tandem-Tomato. However, the procedures that we used to stabilize and image the salivary glands can be extended to other mouse models and coupled to other approaches to label in vivo cellular components, enabling the visualization of various subcellular structures, such as endosomes, lysosomes, mitochondria, and the actin cytoskeleton. Video LinkThe video component of this article can be found at

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Section: Introductionmentioning

confidence: 99%

Intravital Microscopy for Imaging Subcellular Structures in Live Mice Expressing Fluorescent Proteins

Masedunskas

Porat-Shliom

Tora

et al. 2013

JoVE

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“…Initially, IVM has been used in neuroscience to image synaptic plasticity in live animals, and later its use has been extended to study cell movement during immuno-response and cell migration during invasion and metastasis [ 3 , 4 , 6 , 8 , 9 , 11 , 12 , 13 , 14 , 15 ]. In the last few years, IVM has been used to image subcellular structures, such as nuclei [ 16 ], mitochondria [ 10 ], endosomes [ 17 , 18 , 19 ], and secretory granules [ 20 , 21 ]. The first time lapse images of endocytosis in vivo were performed in the kidney of live rats and mice where the internalization of fluorescently labeled dextrans and folate have been imaged in proximal tubuli [ 22 , 23 ].…”

Section: Introductionmentioning

confidence: 99%

Intravital Microscopy Reveals Differences in the Kinetics of Endocytic Pathways between Cell Cultures and Live Animals

Masedunskas

Porat-Shliom

Rechache

et al. 2012

Cells

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Intravital microscopy has enabled imaging of the dynamics of subcellular structures in live animals, thus opening the door to investigating membrane trafficking under physiological conditions. Here, we sought to determine whether the architecture and the environment of a fully developed tissue influences the dynamics of endocytic processes. To this aim, we imaged endocytosis in the stromal cells of rat salivary glands both in situ and after they were isolated and cultured on a solid surface. We found that the internalization of transferrin and dextran, two molecules that traffic via distinct mechanisms, is substantially altered in cultured cells, supporting the idea that the three dimensional organization of the tissue and the cues generated by the surrounding environment strongly affect membrane trafficking events.

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“…The proteins that will be secreted are synthesized in the endoplasmic reticulum and transported through the Golgi apparatus to the trans-Golgi network where they are stored in secretory granules (SCGs) after which they are released into the cytoplasm and transported to the periphery of the cell [148]. Electron microscopy studies have shown that the secretory cells of salivary glands are ultrastructurally equipped to produce and store large amounts of secretory proteins in secretory granules (SGs) [149].…”

Section: Microscopy Techniquesmentioning

confidence: 99%

The Use of Electrophoresis for the Study of Saliva Involvement in Ingestive Behavior

Lamy¹,

Morzel²,

Rodrigues³

et al. 2015

Field Effect Electroosmosis - A Novel Phenomenon in Electrokinetics and Its Applications in Capillary Electrophoresis

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Salivary Glands: A Powerful Experimental System to Study Cell Biology in Live Animals by Intravital Microscopy

Cited by 4 publications

References 98 publications

Intravital Microscopy for Imaging Subcellular Structures in Live Mice Expressing Fluorescent Proteins

Intravital Microscopy for Imaging Subcellular Structures in Live Mice Expressing Fluorescent Proteins

Intravital Microscopy Reveals Differences in the Kinetics of Endocytic Pathways between Cell Cultures and Live Animals

The Use of Electrophoresis for the Study of Saliva Involvement in Ingestive Behavior

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