Trypan Blue as a marker of plasma membrane permeability in alloxan-treated mouse islet cells
(1979) In Journal of Endocrinological Investigation: Official Journal of the Italian Society of Endocrinology 2(2). p.139-145- Abstract
Suspensions of pancreatic islet cells from noninbred ob/ob-mice were incubated with Trypan Blue. Microscope photometry showed that apparently viable cells excluded the dye completely, whereas the nuclei of nonviable cells accumulated Trypan Blue by a saturable process. The nucleus-to-medium dye gradient was more then 30∶1 in media containing 0.1% or less Trypan Blue. The apparent affinity constant for nuclear binding of the dye was 3.1 × 104 l/mol. Albumin partially inhibited the nuclear staining. More than 0.5% Trypan Blue in the medium was toxic per se. In the absence of albumin, 0.5 or 20 mmol/l alloxan, 1 mmol/l N-ethylmaleimide, or 0.1 mmol/l chloromercuribenzene- p-sulphonic acid, but not 20 mmol/l streptozotocin,... (More)
Suspensions of pancreatic islet cells from noninbred ob/ob-mice were incubated with Trypan Blue. Microscope photometry showed that apparently viable cells excluded the dye completely, whereas the nuclei of nonviable cells accumulated Trypan Blue by a saturable process. The nucleus-to-medium dye gradient was more then 30∶1 in media containing 0.1% or less Trypan Blue. The apparent affinity constant for nuclear binding of the dye was 3.1 × 104 l/mol. Albumin partially inhibited the nuclear staining. More than 0.5% Trypan Blue in the medium was toxic per se. In the absence of albumin, 0.5 or 20 mmol/l alloxan, 1 mmol/l N-ethylmaleimide, or 0.1 mmol/l chloromercuribenzene- p-sulphonic acid, but not 20 mmol/l streptozotocin, increased the frequency of islet cells stained with 0.1% Trypan Blue. The absorbance of nuclei was also increased in cells treated with alloxan or N-ethylmaleimide, but not in those treated with chloromercuribenzene- p-sulphonic acid. It is concluded that alloxan rapidly increases the permeability of the plasma membrane in mouse β-cells. This action of alloxan appears to be more acute than any such effect of streptozotocin.
(Less)
- author
- Grankvist, K. LU ; Lernmark LU and Täljedal, I. B.
- publishing date
- 1979-01-01
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Alloxan, chloromercuribenzene- p-sulphonic acid, diabetes, islets of Langerhans, N-ethylmaleimide, pancreatic β-cell, plasma membrane permeability, streptozotocin, Trypan Blue
- in
- Journal of Endocrinological Investigation: Official Journal of the Italian Society of Endocrinology
- volume
- 2
- issue
- 2
- pages
- 7 pages
- publisher
- Springer
- external identifiers
-
- scopus:0018634771
- pmid:226577
- ISSN
- 0391-4097
- DOI
- 10.1007/BF03349305
- language
- English
- LU publication?
- no
- id
- c26791a3-8372-4b49-9f1e-011274554b3d
- date added to LUP
- 2019-12-12 23:26:09
- date last changed
- 2024-04-02 21:49:15
@article{c26791a3-8372-4b49-9f1e-011274554b3d, abstract = {{<p>Suspensions of pancreatic islet cells from noninbred ob/ob-mice were incubated with Trypan Blue. Microscope photometry showed that apparently viable cells excluded the dye completely, whereas the nuclei of nonviable cells accumulated Trypan Blue by a saturable process. The nucleus-to-medium dye gradient was more then 30∶1 in media containing 0.1% or less Trypan Blue. The apparent affinity constant for nuclear binding of the dye was 3.1 × 10<sup>4</sup> l/mol. Albumin partially inhibited the nuclear staining. More than 0.5% Trypan Blue in the medium was toxic per se. In the absence of albumin, 0.5 or 20 mmol/l alloxan, 1 mmol/l N-ethylmaleimide, or 0.1 mmol/l chloromercuribenzene- p-sulphonic acid, but not 20 mmol/l streptozotocin, increased the frequency of islet cells stained with 0.1% Trypan Blue. The absorbance of nuclei was also increased in cells treated with alloxan or N-ethylmaleimide, but not in those treated with chloromercuribenzene- p-sulphonic acid. It is concluded that alloxan rapidly increases the permeability of the plasma membrane in mouse β-cells. This action of alloxan appears to be more acute than any such effect of streptozotocin.</p>}}, author = {{Grankvist, K. and Lernmark and Täljedal, I. B.}}, issn = {{0391-4097}}, keywords = {{Alloxan; chloromercuribenzene- p-sulphonic acid; diabetes; islets of Langerhans; N-ethylmaleimide; pancreatic β-cell; plasma membrane permeability; streptozotocin; Trypan Blue}}, language = {{eng}}, month = {{01}}, number = {{2}}, pages = {{139--145}}, publisher = {{Springer}}, series = {{Journal of Endocrinological Investigation: Official Journal of the Italian Society of Endocrinology}}, title = {{Trypan Blue as a marker of plasma membrane permeability in alloxan-treated mouse islet cells}}, url = {{http://dx.doi.org/10.1007/BF03349305}}, doi = {{10.1007/BF03349305}}, volume = {{2}}, year = {{1979}}, }