Nuclear organization studied with the help of a hypotonic shift: its use permits hydrophilic molecules to enter into living cells
Language English Country Austria Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
10525969
DOI
10.1007/s004120050384
PII: 91080325.412
Knihovny.cz E-resources
- MeSH
- Amanitins metabolism MeSH
- Coloring Agents metabolism MeSH
- Cell Nucleus metabolism MeSH
- Chemical Phenomena MeSH
- Diffusion MeSH
- Drosophila melanogaster metabolism MeSH
- Epithelial Cells drug effects metabolism MeSH
- Fibroblasts drug effects metabolism MeSH
- Chemistry, Physical MeSH
- HeLa Cells drug effects metabolism MeSH
- Hypotonic Solutions pharmacology MeSH
- Microscopy, Immunoelectron MeSH
- Immunoglobulin G metabolism MeSH
- Liver metabolism MeSH
- Rats MeSH
- Cells, Cultured MeSH
- Pharmaceutical Preparations metabolism MeSH
- Kidney MeSH
- Humans MeSH
- Microinjections MeSH
- Molecular Weight MeSH
- Nucleotides metabolism MeSH
- Organ Culture Techniques MeSH
- Cell Membrane Permeability drug effects MeSH
- Dogs MeSH
- Gene Expression Profiling MeSH
- Uridine Triphosphate analogs & derivatives metabolism MeSH
- Cell Survival MeSH
- Xenopus laevis MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Humans MeSH
- Male MeSH
- Dogs MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 5-bromouridine triphosphate MeSH Browser
- Amanitins MeSH
- Coloring Agents MeSH
- Hypotonic Solutions MeSH
- Immunoglobulin G MeSH
- Pharmaceutical Preparations MeSH
- Nucleotides MeSH
- Uridine Triphosphate MeSH
A new procedure for introduction of hydrophilic molecules into living cells based on efficient uptake of these molecules into the cells during hypotonic treatment is presented and its use is demonstrated by a variety of applications. Experiments with cultured vertebrate and Drosophila cells and various animal tissues demonstrated that the increase in cell membrane permeability under hypotonic conditions is a general phenomenon in all animal cells tested. The efficiency of the method depends on the composition and temperature of the hypotonic buffer, the duration of the hypotonic treatment and the molecular weight of the molecules introduced into living cells. The versatility of this approach is demonstrated with various types of molecules such as modified nucleotides, nucleotides with conjugated fluorochrome, peptides, phosphatase substrates and fluorescent dyes. The method opens new possibilities for the direct investigation of a variety of biological problems as documented here with data on the functional organization of the cell nucleus.
References provided by Crossref.org
DNA Replication: From Radioisotopes to Click Chemistry
Dr Jekyll and Mr Hyde: a strange case of 5-ethynyl-2'-deoxyuridine and 5-ethynyl-2'-deoxycytidine
Atomic scissors: a new method of tracking the 5-bromo-2'-deoxyuridine-labeled DNA in situ
In situ reverse transcription: the magic of strength and anonymity
Organization of human replicon: singles or zipping couples?
NORs and their transcription competence during the cell cycle
Prespliceosomal assembly on microinjected precursor mRNA takes place in nuclear speckles
