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Beneficial Effects of Nitrite
http://www.vmi.pitt.edu/Gladwin.html
Nitrite bioactivation by Globin X in zebrafish blood and effects on - - PowerPoint PPT Presentation
Nitrite bioactivation by Globin X in zebrafish blood and effects on - - PowerPoint PPT Presentation
Nitrite bioactivation by Globin X in zebrafish blood and effects on heart regeneration Paola Corti, PhD University of Pittsburgh Gladwin lab Beneficial Effects of Nitrite http://www.vmi.pitt.edu/Gladwin.html Globins are Nitrite Reductases At
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SLIDE 3
Globins are Nitrite Reductases At Low Oxygen
Deoxy-Heme+ NO2
- + H+
Met-Heme + NO + OH- Hemoglobin Myoglobin Blood Heart
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Globins evolution
Hemoglobin Myoglobin Cytoglobin Neuroglobin Globin X Erythrocytes Muscles Heart Fibroblasts Neurons Neurons Endocrine ??? Brain ?
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GbX is found in zebrafish blood
23 kDa
TRANSCRIPT PROTEIN 2 dpf 2 dpf blood
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GbX is a fast nitrite reductase
Protein Nitrite reduction rate (M-1s-1) Zf Ngb 0.68 ± 0.04 Zf Cgb1 28.3 ± 3.1 Zf Cgb2 0.94 ± 0.18 Zf GbX 26.7 ± 2.0 hNgb 0.52 ± 0.19 hCgb 1.14 ± 0.07
(Corti et al, submitted to Nitric Oxide)
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GbX in RBCs produces NO from nitrite
Nitric Oxide Analyzer (NOA)
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GbX in RBCs and nitrite inhibit platelets activation
(Corti et al., FRBM 2014)
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GbX and nitrite inhibit platelet activation
NO signaling Nitrite reduction NO production platelets activation GbX is 6-coordinate
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What is the Effect of Hypoxic Nitrite during Heart Regeneration?
O2 O2
(Chablais et al.,Dev Biol 2011)
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Effect of Nitrite at 5dpC on Injured Area
injured area (%)
Control NO2 Hypoxia Hyp+NO2 10 20 30 40
**** **** ***
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5 dpC
PCNA+/Mef2+ cardiomyocytes (%)
controls NO2 hypoxia hyp+NO2 10 20 30
* * *
n=7-10
control Hyp+NO2
Effect of Nitrite at 5dpC on Cell Proliferation
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Nitrite has anticoagulant effects
P<0.01 * *
Hypoxia
Tg(flk:Gfp;Gata1:DsRed), Dapi
Hyp+NO2
Fluorescence intensity Gata1:DsRed
CTCF/area
h y p
- x
i a h y p + N O 2 10 20 30 40
**
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Future Plans
- Investigate the role of GbX in the blood coagulation process during heart
regeneration
- Evaluate the possible involvement of other globins in
mediating the effect of nitrite. Cytoglobin 1 is found in the fish epicardium: important during regeneration?
- Generate Talen and CRISPr knockout
* * * *
uncut uncut
F0
- Initiate studies of cryoinjury in mouse. Effect of Nitrite
- n mouse cryoinjury
cygb1
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Acknowledgements
Mark Gladwin Michael Tsang Azzurra Missinato Daniel Kim-Shapiro Nadeem Wajia Mr Wholey Jesus Tejero Jianmin Xue Courtney Watkins Charly Lai Bin Sun
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Titolo slide
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GbX is a six-coordinate globin
(Jesus Tejero)
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GbX in RBCs produces NO from nitrite
Nitric Oxide Analyzer (NOA)
SLIDE 19
GbX and nitrite inhibit platelets activation
(Corti et al., 2014)
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