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Difference between revisions of "MiPNet20.12 IOC103 Manaus"

From Bioblast
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[[File:Tambaqui 4447.JPG|left|300px]] Tambaqui, ''Colossoma macropomum''.
[[File:Tambaqui 4447.JPG|left|300px]] Tambaqui, ''Colossoma macropomum''.
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<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.
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Revision as of 20:40, 13 July 2015

                



MiPNet20.12 IOC103 Manaus

Publications in the MiPMap
O2k-Network

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:

Arapaima, Pirarucu, or Paiche

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 

Amazon, Manaus

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

BR Manaus Val AL
Entrance to LEEM

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



BR Manaus Val AL

Local organizers and contact






O2k-Workshop and pilot project

IOC103 0331.JPG

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 4447.JPG

Tambaqui, Colossoma macropomum.


Arapaima or Pirarucu

Arapaima 4515.JPG

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.


IOC103-intro 4444.JPG

Lecturers and tutors


Participants

IOC103 4543.JPG
IOC103 4558.JPG


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«
O2k-Workshops

Recommended IOC reading

Please click » here«

» MitoPedia: Respiratory statesOXPHOS ROUTINE ETS LEAK ROX

MitoGlobal

O2k-Workshops are listed as MitoGlobal Events.

Some impressions

INPA Science forest

Manati project Piranias in a traditional hut Turtles in the forest

Perspectives

Amazon-boat 4384.JPG Amazon-boat 4434.JPG Amazon-boat 4406.JPG Should we get a boat (Vendo - middle) for a MitoLab on the Amazon?