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Searching free zinc at the ultrastructural level in cultured astrocytes

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Searching free zinc at the ultrastructural level in cultured astrocytes

dc.contributor.author	Ballestín Hinojosa, Raúl
dc.contributor.author	Molowny Tudela, Asunción
dc.contributor.author	Marín Muela, María Pilar
dc.contributor.author	Esteban Pretel, Guillermo
dc.contributor.author	Romero, Ana
dc.contributor.author	Renau Piqueras, Jaime
dc.contributor.author	López García, Carlos
dc.contributor.author	Ponsoda i Martí, Xavier
dc.date.accessioned	2022-04-07T11:44:16Z
dc.date.available	2022-04-08T04:45:05Z
dc.date.issued	2009	es_ES
dc.identifier.citation	Póster 2009-09-08 IX European Glial Cell Meeting PARIS	es_ES
dc.identifier.uri	https://hdl.handle.net/10550/82217
dc.description.abstract	Zinc is an element that is necessary for many physiological functions in the body but may play an important role in diseases affecting most systems in the body if its balance is altered by environmental, toxicological or idiosyncrasy of subjects. We have centred our investigation in central nervous system, using cultured astrocytes since they are involved in clearance of zinc exocytated to the extracellular medium during synaptic transmission. In previous works we have used a zinc fluorochrome, i.e., the TSQ (6-Methoxy-(8-p-toluenesulfonamido)quinoline) to in vivo zinc uptake in cultured astrocytes and its accumulation in organelles named zincosomes. However, the precise location of these zinc-enriched structures (zincosomes) at the ultrastructural level is a very hard task. In a previous attempt at the electron microscopy level, only topographical approximation by combining light and electron microscopy allowed us to identify selected zincosomes previously marked with TSQ. Now, our objective is to adapt zinc autometallography (Timm’s method) to TSQ labelled cultured astrocytes. For the electron microscopic detection of zincosomes, the first important step is to achieve a good zinc precipitation during or previous to glutaraldehyde fixation. Surprisingly, neither ditizone nor selenite were successful as zinc precipitating agents; only sodium sulphide gave us good results. We also found that while glutaraldehyde is the best option for animal experimentation, paraformaldehyde prefixation gave us best results. Paraformaldehyde prefixation allowed both ultrastructure preservation as well as zinc-precipitated-detection with Timm autometallography in semithin sections. These semithin sections were included again and zincosomes become clearly visible in ultrathin sections.	es_ES
dc.language.iso	en	es_ES
dc.title	Searching free zinc at the ultrastructural level in cultured astrocytes	es_ES
dc.type	conference output	es_ES
dc.subject.unesco	UNESCO::PSICOLOGÍA	es_ES
dc.accrualmethod	-	es_ES
dc.embargo.terms	0 days	es_ES