A method for co-localization of tubular lysosomes and microtubules in macrophages: fluorescence microscopy of individual cells.

Robinson, John M.; Chiplonkar, J. M.; Luo, Zhi-Pu

doi:10.1177/44.10.8813075

Cited by 13 publications

(10 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These snake-like tubular lysosomes form a complex 3D network within the acinar cell, inter- twined between the cisternae of the ER, around the nuclei, and in the basolateral region, coming into very close contact with the plasma membrane itself. Similar networks were previously demonstrated in several types of cells, including pancreatic cells, and were shown to contain various lysosomal enzymes, such as adenine dinucleotide phosphatase, trimetaphosphatase, aryl sulfatase B, thiolacetic acid, and several other esterases (Oliver 1983;Beaudoin et al 1984;Oliver and Yuasa 1987;Robinson and Karnovsky 1991;Alvares 1994;Robinson et al 1996). However, two populations of lysosomal structures were ascribed to acinar cells (Oliver et al 1989), one corresponding to the classical acid phosphatase-positive globular structures and a second corresponding to the tubular system and considered as acid phosphatase-negative.…”

Section: Discussionsupporting

confidence: 70%

Demonstration of Acetylcholinesterase Molecular Forms in a Continuous Tubular Lysosomal System of Rat Pancreatic Acinar Cells

Bendayan

Gisiger

2001

J Histochem Cytochem.

View full text Add to dashboard Cite

S U M M A R Y By applying the highly sensitive cytochemical Gautron's technique, we were able to reveal AChE activity in rat pancreatic acinar cells, particularly at the level of a complex membrane-bound network formed by tubules with varicosities located around the nuclei and close to the basolateral membrane. The Golgi apparatus was devoid of cytochemical reaction beside the trans -Golgi network cisternae, which showed a positive reaction. The RER of some acinar cells also presented a signal, demonstrating their capability of synthesizing AChE. Immunogold using a specific anti-AChE antibody yielded similar results. Double-labeling experiments corroborated the presence of enzyme cytochemical and immunocytochemical signals in the same lysosomal tubular network. Biochemical sedimentation assays confirmed the presence of AChE in acinar cells, which exists as two globular molecular forms, G 1 and G 4 . These results were obtained with pancreatic tissue in situ as well as with isolated acinar cells maintained in culture and devoid of neural elements. The existence of a continuous tubular lysosomal network containing AChE is in agreement with previous reports on acinar and other cell types, and supports a more general hypothesis on dynamic continuities among cell structures. Whether AChE is being secreted by the acinar cells or internalized through this endo-lysosomal system was not defined. However, the capability of the acinar cells to synthesize AChE and to channel it through a tubular system is a good indication that the cells can modulate their cholinergic stimulation for optimal secretion of digestive enzymes. (J Histochem Cytochem 49: 29 -39, 2001)

show abstract

Section: Discussionsupporting

confidence: 70%

Demonstration of Acetylcholinesterase Molecular Forms in a Continuous Tubular Lysosomal System of Rat Pancreatic Acinar Cells

Bendayan

Gisiger

2001

J Histochem Cytochem.

View full text Add to dashboard Cite

show abstract

“…Conventional immunofluorescence localization of proteins in fixed cells is known to be a poor reporter of tubular membrane compartments, and the fragmented appearance of some Sifs suggested to us that not all tubules are properly preserved during fixation (15) . If true, this could confound Sif identification, particularly at early time-points when tubules tend to be short.…”

Section: Resultsmentioning

confidence: 99%

Dynamic Behavior of Salmonella‐Induced Membrane Tubules in Epithelial Cells

Drecktrah

Levine-Wilkinson

Dam

et al. 2008

Traffic

125

119

View full text Add to dashboard Cite

Salmonella Typhimurium is a facultative intracellular pathogen that causes acute gastroenteritis in man. Intracellular Salmonella survive and replicate within a modified phagosome known as the Salmonella -containing vacuole (SCV). The onset of intracellular replication is accompanied by the appearance of membrane tubules, called Salmonella -induced filaments (Sifs), extending from the SCV. Sifs are enriched in late endosomal/lysosomal membrane proteins such as lysosome-associated membrane protein 1, but their formation and ability to interact with endosomal compartments are not characterized. In this study, we use live cell imaging techniques to define the dynamics of Sif formation in infected epithelial cells. At early time-points, Sifs are simple tubules extending from the surface of SCVs. These tubules are highly dynamic and exhibit bidirectional, microtubule-dependent movement. At the distal ends of individual Sif tubules, furthest from the SCV, a distinct ‘leader’ domain was often observed. At later times, Sifs develop into highly complex tubular networks that extend throughout the cell and appear less dynamic than nascent Sifs; however, individual tubules continue to display bidirectional dynamics. Sifs can acquire endocytic content by fusion, indicating a sustained interaction with the endocytic pathway. Together, these results show that these Salmonella -induced tubules form a highly dynamic network that involves both microtubule-dependent motility and interactions with endosomal compartments.

show abstract

“…There are often situations in which it is desirable to examine specimens by multiple imaging modalities. Correlative microscopy, in which a single sample can be examined by two or more imaging techniques (e.g., fluorescence and electron microscopy) or when multiple structures or molecules are viewed with the same imaging procedure (e.g., fluorescence microscopy) have been important in many studies related to cell structure and function (e.g., Robinson et al 1996;Verkhovsky et al 1995;Luo and Robinson 1992;Conrad et al 1989). Such an integrated approach to microscopy can provide additional insight into biological questions not gained by a single imaging procedure.…”

mentioning

confidence: 99%

Efficient Immunocytochemical Labeling of Leukocyte Microtubules with FluoroNanogold: An Important Tool for Correlative Microscopy

Robinson

Vandré

1997

J Histochem Cytochem.

View full text Add to dashboard Cite

SUMMARYWe tested the immunoprobe FluoroNanogold (FNG) for its utility as an immunocytochemical labeling reagent. This immunoprobe consists of a 1.4-nm gold particle to which a specific Fab Ј fragment and a fluorochrome are conjugated. We employed the microtubules (MTs) of human phagocytic leukocytes as a model system for testing the usefulness of FNG as a secondary antibody for immunocytochemistry. We show that these fluorescently labeled ultrasmall immunogold particles are very efficient for labeling MTs in these cells. The signal from FNG can be detected directly by fluorescence microscopy or indirectly by other modes of optical microscopy and electron microscopy, after silverenhancement of the gold. The spatial resolution of immunolabeled MTs obtained with FNG and silver enhancement was comparable to that of conventional immunofluorescence detection. Colloidal gold (5-nm and 10-nm in diameter), on the other hand, failed to label MTs in cells prepared in a similar manner. This difference in labeling was due in large part to greater penetration of 1.4-nm gold into aldehyde-fixed cells than either 5-nm or 10-nm gold particles. The fluorescent 1.4-nm immunoprobe was shown to be an important new tool for general use in correlative microscopy. (J Histochem Cytochem 45:631-642, 1997)

show abstract

A method for co-localization of tubular lysosomes and microtubules in macrophages: fluorescence microscopy of individual cells.

Cited by 13 publications

References 12 publications

Demonstration of Acetylcholinesterase Molecular Forms in a Continuous Tubular Lysosomal System of Rat Pancreatic Acinar Cells

Demonstration of Acetylcholinesterase Molecular Forms in a Continuous Tubular Lysosomal System of Rat Pancreatic Acinar Cells

Dynamic Behavior of Salmonella‐Induced Membrane Tubules in Epithelial Cells

Efficient Immunocytochemical Labeling of Leukocyte Microtubules with FluoroNanogold: An Important Tool for Correlative Microscopy

Contact Info

Product

Resources

About