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Biosensor measurement of purine release from cerebellar cultures and slices
We have previously described an action-potential and Ca(2+)-dependent form of adenosine release in the molecular layer of cerebellar slices. The most likely source of the adenosine is the parallel fibres, the axons of granule cells. Using microelectrode biosensors, we have therefore investigated whe...
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Formato: | Texto |
Lenguaje: | English |
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Springer Netherlands
2010
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2947654/ https://www.ncbi.nlm.nih.gov/pubmed/21103217 http://dx.doi.org/10.1007/s11302-010-9185-8 |
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author | Wall, Mark Eason, Robert Dale, Nicholas |
author_facet | Wall, Mark Eason, Robert Dale, Nicholas |
author_sort | Wall, Mark |
collection | PubMed |
description | We have previously described an action-potential and Ca(2+)-dependent form of adenosine release in the molecular layer of cerebellar slices. The most likely source of the adenosine is the parallel fibres, the axons of granule cells. Using microelectrode biosensors, we have therefore investigated whether cultured granule cells (from postnatal day 7–8 rats) can release adenosine. Although no purine release could be detected in response to focal electrical stimulation, purine (adenosine, inosine or hypoxanthine) release occurred in response to an increase in extracellular K(+) concentration from 3 to 25 mM coupled with addition of 1 mM glutamate. The mechanism of purine release was transport from the cytoplasm via an ENT transporter. This process did not require action-potential firing but was Ca(2+)dependent. The major purine released was not adenosine, but was either inosine or hypoxanthine. In order for inosine/hypoxanthine release to occur, cultures had to contain both granule cells and glial cells; neither cellular component was sufficient alone. Using the same stimulus in cerebellar slices (postnatal day 7–25), it was possible to release purines. The release however was not blocked by ENT blockers and there was a shift in the Ca(2+) dependence during development. This data from cultures and slices further illustrates the complexities of purine release, which is dependent on cellular composition and developmental stage. |
format | Text |
id | pubmed-2947654 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Springer Netherlands |
record_format | MEDLINE/PubMed |
spelling | pubmed-29476542010-11-22 Biosensor measurement of purine release from cerebellar cultures and slices Wall, Mark Eason, Robert Dale, Nicholas Purinergic Signal Original Article We have previously described an action-potential and Ca(2+)-dependent form of adenosine release in the molecular layer of cerebellar slices. The most likely source of the adenosine is the parallel fibres, the axons of granule cells. Using microelectrode biosensors, we have therefore investigated whether cultured granule cells (from postnatal day 7–8 rats) can release adenosine. Although no purine release could be detected in response to focal electrical stimulation, purine (adenosine, inosine or hypoxanthine) release occurred in response to an increase in extracellular K(+) concentration from 3 to 25 mM coupled with addition of 1 mM glutamate. The mechanism of purine release was transport from the cytoplasm via an ENT transporter. This process did not require action-potential firing but was Ca(2+)dependent. The major purine released was not adenosine, but was either inosine or hypoxanthine. In order for inosine/hypoxanthine release to occur, cultures had to contain both granule cells and glial cells; neither cellular component was sufficient alone. Using the same stimulus in cerebellar slices (postnatal day 7–25), it was possible to release purines. The release however was not blocked by ENT blockers and there was a shift in the Ca(2+) dependence during development. This data from cultures and slices further illustrates the complexities of purine release, which is dependent on cellular composition and developmental stage. Springer Netherlands 2010-05-25 2010-09 /pmc/articles/PMC2947654/ /pubmed/21103217 http://dx.doi.org/10.1007/s11302-010-9185-8 Text en © The Author(s) 2010 https://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited. |
spellingShingle | Original Article Wall, Mark Eason, Robert Dale, Nicholas Biosensor measurement of purine release from cerebellar cultures and slices |
title | Biosensor measurement of purine release from cerebellar cultures and slices |
title_full | Biosensor measurement of purine release from cerebellar cultures and slices |
title_fullStr | Biosensor measurement of purine release from cerebellar cultures and slices |
title_full_unstemmed | Biosensor measurement of purine release from cerebellar cultures and slices |
title_short | Biosensor measurement of purine release from cerebellar cultures and slices |
title_sort | biosensor measurement of purine release from cerebellar cultures and slices |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2947654/ https://www.ncbi.nlm.nih.gov/pubmed/21103217 http://dx.doi.org/10.1007/s11302-010-9185-8 |
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