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Rhodamine B, a fluorescent probe for acidic organelles in denervated skeletal muscle

Vult von Steyern, Fredrik LU ; Josefsson, Jan-Owe LU and Tagerud, S (1996) In Journal of Histochemistry and Cytochemistry 44(3). p.267-274
Abstract
We describe a very efficient method for fluorescent labeling of acidic structures in denervated skeletal muscle with rhodamine B. Rhodamine B at 50 ng/ml gave selective and distinct segmental labeling of denervated muscle fibers after 5-min incubation at room temperature. Labeling was also achieved at 4 degrees C. The labeling was disrupted by the ionophores monensin and nigericin, suggesting a labeling confined to acidic structures. Rhodamine B co-localized with the lysosomotropic dye Lyso Tracker Green and a marker for endocytosis (fluorescein isothiocyanate-labeled dextran). Rhodamine B, which is highly lipophilic, showed pH-dependent fluorescence emission in saturated aqueous N-octanol. Tetramethylrhodamine showed similar... (More)
We describe a very efficient method for fluorescent labeling of acidic structures in denervated skeletal muscle with rhodamine B. Rhodamine B at 50 ng/ml gave selective and distinct segmental labeling of denervated muscle fibers after 5-min incubation at room temperature. Labeling was also achieved at 4 degrees C. The labeling was disrupted by the ionophores monensin and nigericin, suggesting a labeling confined to acidic structures. Rhodamine B co-localized with the lysosomotropic dye Lyso Tracker Green and a marker for endocytosis (fluorescein isothiocyanate-labeled dextran). Rhodamine B, which is highly lipophilic, showed pH-dependent fluorescence emission in saturated aqueous N-octanol. Tetramethylrhodamine showed similar characteristics for labeling of denervated muscle fibers and pH-dependent fluorescence in N-octanol. The carboxyl group present in these two compounds appears important, because structurally related compounds that either lack this group or have it esterified failed to label denervated muscle fibers and showed no pH-dependent fluorescence in N-octanol. The results suggest that rhodamine B labels acidic organelles belonging to the endosomal/lyosomal system of denervated skeletal muscle fibers. Nevertheless, it failed to label such organelles in a number of mammalian cell types other than denervated skeletal muscle fibers. (Less)
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publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Histochemistry and Cytochemistry
volume
44
issue
3
pages
267 - 274
publisher
Histochemical Society
external identifiers
  • pmid:8648087
  • scopus:0242509380
ISSN
0022-1554
language
English
LU publication?
yes
id
e677bf4f-4315-4a13-a683-29b9d1b14f59 (old id 1110834)
date added to LUP
2016-04-01 16:35:49
date last changed
2022-01-28 20:45:02
@article{e677bf4f-4315-4a13-a683-29b9d1b14f59,
  abstract     = {{We describe a very efficient method for fluorescent labeling of acidic structures in denervated skeletal muscle with rhodamine B. Rhodamine B at 50 ng/ml gave selective and distinct segmental labeling of denervated muscle fibers after 5-min incubation at room temperature. Labeling was also achieved at 4 degrees C. The labeling was disrupted by the ionophores monensin and nigericin, suggesting a labeling confined to acidic structures. Rhodamine B co-localized with the lysosomotropic dye Lyso Tracker Green and a marker for endocytosis (fluorescein isothiocyanate-labeled dextran). Rhodamine B, which is highly lipophilic, showed pH-dependent fluorescence emission in saturated aqueous N-octanol. Tetramethylrhodamine showed similar characteristics for labeling of denervated muscle fibers and pH-dependent fluorescence in N-octanol. The carboxyl group present in these two compounds appears important, because structurally related compounds that either lack this group or have it esterified failed to label denervated muscle fibers and showed no pH-dependent fluorescence in N-octanol. The results suggest that rhodamine B labels acidic organelles belonging to the endosomal/lyosomal system of denervated skeletal muscle fibers. Nevertheless, it failed to label such organelles in a number of mammalian cell types other than denervated skeletal muscle fibers.}},
  author       = {{Vult von Steyern, Fredrik and Josefsson, Jan-Owe and Tagerud, S}},
  issn         = {{0022-1554}},
  language     = {{eng}},
  number       = {{3}},
  pages        = {{267--274}},
  publisher    = {{Histochemical Society}},
  series       = {{Journal of Histochemistry and Cytochemistry}},
  title        = {{Rhodamine B, a fluorescent probe for acidic organelles in denervated skeletal muscle}},
  volume       = {{44}},
  year         = {{1996}},
}