Difference between revisions of "MiPNet20.12 IOC103 Manaus"
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[[File:Tambaqui 4447.JPG|left|300px]] Tambaqui, ''Colossoma macropomum''. | [[File:Tambaqui 4447.JPG|left|300px]] Tambaqui, ''Colossoma macropomum''. | ||
<gallery> | <gallery> | ||
File:IOC103 0289.JPG | File:IOC103 0289.JPG|The O2k is set up in the seminar room for the training and pilot project. | ||
File:IOC103 0411.JPG | File:IOC103 0411.JPG|Preparation of chemicals. | ||
File:IOC103 0350.JPG | File:IOC103 0350.JPG|Oxygen calibration of the O2k with full attention. | ||
File:IOC103 0425.JPG | File:IOC103 0425.JPG|Ramon goes fishing. | ||
File:IOC103 0432.JPG | File:IOC103 0432.JPG|A small specimen of Tambaqui is selected. | ||
File:IOC103-Tambaqui-heart 4449.JPG | File:IOC103-Tambaqui-heart 4449.JPG|The fish is killed and opened. | ||
File:IOC103-Tambaqui-heart 4451.JPG | File:IOC103-Tambaqui-heart 4451.JPG|The heart is immediately immersed in ice cold BIOPS. | ||
File:IOC103-Tambaqui-heart 4457.JPG | File:IOC103-Tambaqui-heart 4457.JPG|A 20 mg portion of the heart is prepared. | ||
File:IOC103 0467.JPG | File:IOC103 0467.JPG|Mechanical separation of cardiac fibres. | ||
File:IOC103 0495.JPG | File:IOC103 0495.JPG|A balance between carefull separation and speed is important, before determination of the wet weight (c. 20 mg). | ||
File:IOC103 0507.JPG | File:IOC103 0507.JPG|The tissue is ready for homogenization with the Shredder. | ||
File:IOC103 0289.JPG | File:IOC103 0289.JPG|The homogenate (5 ml) is distributed at alternating 500 µl steps into the two O2k-Chambers. | ||
</gallery> | </gallery> | ||
Revision as of 20:40, 13 July 2015
MiPNet20.12 IOC103 Manaus
Manaus BR, 2015 Jul 06-07. To the heart of the Amazon. O2k-Workshop on HRR and O2k-Fluorometry, IOC103. |
OROBOROS (2015-07-06) Mitochondr Physiol Network
Abstract:
2015 Jul 06-07. To the heart of the Amazon: comparative cardiac mitochondrial physiology of Amazon fish. O2k-Workshop on HRR and start-up project, IOC103.
• O2k-Network Lab: BR Manaus Val AL, AT Innsbruck OROBOROS
Labels:
HRR: Oxygraph-2k
ORO, IOC, MiPNet, 2015, Next
To the heart of the Amazon
Mitochondrial function is compared in mitochondrial preparations of fish heart. According to the suggestion by Dr. Adalberto Luis Val, one of each of the major groups should be selected: Potamotrygonidae, Osteoglossidae, Characiformes, Siluriformes, Perciformes (Cichlidae). The results obtained with a basic SUIT protocol will be compared with mitochondrial respiratory control patterns in other species, including human and mouse heart mitochondria. This study explores evolutionary variations of mitochondrial respiratory control patterns in different species, to define mitochondrial fitness in terms of adaptations to different environments and life styles. The results may provide a key towards delineating mitochondrial respiratory control patterns in health in different species and tissues, as a basis to classify mitochondrial risk factors as causes and consequences of disease.
Venue and organization
Venue
- Instituto Nacional de Pesquisas de Amazonia (INPA)
- Laboratório de Ecofisiologia e Evolução Molecular (LEEM)
- Ministerio da Ciencia e Tecnologia
- Av. Andre Araujo, 2938
- 69067-375 Manaus, AM, Brazil
Local organizers and contact
O2k-Workshop and pilot project
Day 1: Assembly of the O2k
From left to right: Erich Gnaiger, Reginaldo da Silva Oliveira, Derek Felipe Campos, Wadir Heinrichs Cardas, Ramon B Baptista.
Day 2: SUIT protocol for comparative mitochondrial physiology - a pilot experiment with 20 mg wet weight of cardiac tissue from Tambaqui (Colossoma macropomum). Tissue homogenate was prepared with the PBI Shredder.
Days 3-4: Project: Mitochondrial respiratory control pattern in Amazon fish heart.
High reproducibility of mitochondrial respiratory control in cardiac tissue from Tambaqui.
Tambaqui
Tambaqui, Colossoma macropomum.
Arapaima or Pirarucu
The Arapaima or Pirarucu is native to the Amazon, and is among the largest freshwater fish in the world (>2.5 m). As an adaptation to the seasonal hypoxic conditions, Arapaima has developed a swim bladder with lung-like function associated with air breathing.
Lecturers and tutors
- Gnaiger Erich, CEO, OROBOROS INSTRUMENTS
- Carolina Gnaiger, Assistant, Fotography, OROBOROS INSTRUMENTS
Participants
- Vera M Fonseca de Almeida e Val - [email protected]
- Adalberto L Val - [email protected]
- Aleicia Holland - [email protected]
- Ramon Barros Baptista - [email protected]
- Derek Selipe Compos - [email protected]
- Waldir Heinrichs Cacdas - [email protected]
- Reginaldo da Silva Oliveira - [email protected]
- Marcio Soares Ferrreira - [email protected]
- Andrea Gnaiger - [email protected]
- Carolina Gnaiger - [email protected]
- Erich Gnaiger - [email protected]
SUIT protocol for the cardiac mitochondrial project
Heart homogante from various fish species will be analyzed with a basic SUIT protocol. The experimental programme is planned from July 8 to 11. 800 px 800 px
Trout heart homogenate (15 °C, 4 mg Ww/ml; 2011-10-05 A-02). Very low LEAK and ROX. No ADP limitation at D1 (1 mM) with glutamate&malate (GM), but glutamate&malate&pyruvate (GMP) was not saturated at 2 mM ADP (D2), as shown by the increase of flux when doubling the ADP concentration (4 mM; D4). No oxygen limitation. Low P/E ratio, indicating limitation of OXPHOS by the phosphorylation system. The optimum uncoupler (U; FCCP) concentration is higher than in liver homogenate (2.5 versus 1.0 µM), but 3 µM exerts already a pronounced time-dependent inhibitory effect (not due to limitation by oxygen, as shown by the lower flux after reoxygenation).
Mouse heart homogenate (37 °C) is shown for comparison.
References
- On respiratory control of human heart mitochondria
- Lemieux H, Semsroth S, Antretter H, Höfer D, Gnaiger E (2011) Mitochondrial respiratory control and early defects of oxidative phosphorylation in the failing human heart. Int J Biochem Cell Biol 43:1729–38. »Bioblast link«
Recommended IOC reading
Please click » here«
» MitoPedia: Respiratory states
O2k-Workshops are listed as MitoGlobal Events.
Some impressions
INPA Science forest
Perspectives
Should we get a boat (Vendo - middle) for a MitoLab on the Amazon?