Reports describing production of reactive oxygen species in neonatal heart are missing. As lysyl oxidase is potentially important source of H(2)O(2), we studied its role during ontogenic development of rat heart. H(2)O(2) was detected in thin sections of developing rat heart by fluorescence microscopy with the use of fluorescence probe 2'-7'-dichlorofluorescin. The experimental design comprised foetuses 21 days after conception, and then the animals sampled on the 1st, 4th, 7th, 10th, 15th, 30th and 60th day after birth. We also used 7-month-old animals as an example of ageing effects. Since the day 4 on, H(2)O(2) was produced only extracellularly up to the day 15, between days 30 and 60 intracellular production was detected as well, and in 7-month-old animals only extracellular production was observed. The specific inhibitors of lysyl oxidase almost completely quenched the H(2)O(2)-dependent fluorescence. Starting from day 7, blue autofluorescence specific to oxidized proteins developed in the vessel wall. Intracellular blue autofluorescence specific to autoxidation products developed after day 30. Chloroform extraction diminished the intracellular blue fluorescence, leaving the extracellular fluorescence intact. This confirmed the protein nature of the fluorophores. Lysyl oxidase is significant source of H(2)O(2) in the heart vessel wall during development and H(2)O(2) oxidatively modifies elastin producing protein blue autofluorescence.

• MeSH
• chloroform MeSH
• elastin metabolismus   MeSH
• fluoresceiny metabolismus   MeSH
• fluorescence MeSH
• fluorescenční barviva metabolismus   MeSH
• krysa rodu Rattus MeSH
• lysyloxidasa metabolismus   MeSH
• myokard enzymologie   metabolismus   MeSH
• oxaziny metabolismus   MeSH
• peroxid vodíku metabolismus   MeSH
• potkani Wistar MeSH
• srdce růst a vývoj   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 2',7'-dichlorofluorescein MeSH Prohlížeč
• chloroform MeSH
• elastin MeSH
• fluoresceiny MeSH
• fluorescenční barviva MeSH
• lysyloxidasa MeSH
• oxaziny MeSH
• PAcein MeSH Prohlížeč
• peroxid vodíku MeSH

Reactive oxygen species (ROS) play several biological roles. We investigated the applicability of fluorescent probes for their detection (i) in rabbit lens epithelial cells during ageing in culture, and (ii) in thin sections of rat heart. We used dihydroethidium (DHE), dichlorofluorescin (DCFH), and dihydrorhodamine 123 (DHR) together with detection of autofluorescence both in cells and in chloroform extracts. Superoxide production was confirmed by a specific histochemical method using Mn(2+). All methods demonstrated higher production of ROS in older cells. All probes revealed different sites of ROS production in young and old cells and could be used for investigation of ROS generation during cell ageing. In the thin sections of rat heart DCFH was not suitable for intracellular ROS detection. The results indicate that the potential of fluorescent dyes in ROS detection is not usually fully exploited, and that blue autofluorescence is associated with oxidative damage.

• MeSH
• epitelové buňky metabolismus   MeSH
• ethidium analogy a deriváty   MeSH
• fluoresceiny MeSH
• králíci MeSH
• krysa rodu Rattus MeSH
• metody MeSH
• molekulární sondy - techniky * MeSH
• myokard metabolismus   MeSH
• oční čočka cytologie   MeSH
• reaktivní formy kyslíku analýza   MeSH
• rhodaminy MeSH
• volné radikály analýza   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 2',7'-dichlorofluorescein MeSH Prohlížeč
• dihydroethidium MeSH Prohlížeč
• dihydrorhodamine 123 MeSH Prohlížeč
• ethidium MeSH
• fluoresceiny MeSH
• reaktivní formy kyslíku MeSH
• rhodaminy MeSH
• volné radikály MeSH