Abstract:

At the University of Miami, RSMAS sub-adult (<100 day old) mahi-mahi were exposed to 24h of crude oil or 24h of pristine conditions. Crude oil mixture used in lab was a 10% OFS (oil from source) HEWAF and saltwater mix. After 24h, mahi-mahi were immediately sacrificed and mitochondrial respiration rate and ADP affinity were measured in fresh tissue using an OROBROS HR respirometer. Frozen tissue samples were taken to measure at a later date enzymatic activity in the complexes associated with the electron transport system. This project was conducted from Nov 2015-July 2016.

Suggested Citation:

Amanda Reynolds, Rachael Heuer, Derek Nelson, Ronald Hoenig, Jon Stieglitz, Martin Grosell, Daniel Benetti, Gina Galli, Dane Crossley. 2017. Changes in cardiac mitochondrial respiration rate after 24h of crude oil exposure in sub-adult mahi-mahi. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/N7SX6BMB

Publications:

Kirby, A. R., Cox, G. K., Nelson, D., Heuer, R. M., Stieglitz, J. D., Benetti, D. D., … Crossley, D. A. (2019). Acute crude oil exposure alters mitochondrial function and ADP affinity in cardiac muscle fibers of young adult Mahi-mahi (Coryphaena hippurus). Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 218, 88–95. doi:10.1016/j.cbpc.2019.01.004

Purpose:

To measure change in mitochondrial respiration.

Data Parameters and Units:

Mitochondrial Respiration Rate (umol O2/mg dry tissue/sec); oil loading rate (1g/L); controls (0% oil solution); treatments (10% oil solutions); holding and experimental tank volumes (3000L); Date (DD-MMM-YY); Time (HH:MM); Water/dilutant volume (L); Mass of oil added (mg); Calculated nominal concentration (mg/L); tank temperatures (26 deg C); tank photo period (14h light:10h dark); tank pH (7.4); tank dissolved oxygen (flow rate 1l/min, mg/L); salinity (35 ppt); total ammonia (0 um); mahi body mass (1kg); mahi ventricle mass (1g); Normalized to Oligomycin (OD/min); Protein Concentration (mg/ml or mmol/min); Normalized to protein (nmol/min/mg or mg/ml); multiple by dilution (mmol/min/mg); absolute value (mmol/min/mg); absolute value (umol/min/mg); 1 to 7 dilution (OD/min); Average absorbance (OD/min or mOD/min); normalized to background (OD/min or mOD/min); extinction coefficient (mmol/min or OD/min); average; standard error; standard deviation; enzyme activity (nmol/min/mg); concentration (mg/ml); Max V (OD/min); Oligomycin (OD/min); Oligomycin 2 (0.5 ug/ml); malonate (10 mM); Rotenone subtraction (OD/min); rotenone (0.5um); rotenone after (OD/min); Oxaloacetate (0.5mM); absorbance (OD/min or mOD/min); Before Mean V (OD/min); Mean V (OD/min); Before R-squared (OD/min); Before y-intercept (OD/min); After Mean V (OD/min); After y-intercept (OD/min); After R-squared (OD/min); Leak substrates (umol O2/mg dry tissue/sec); Complex 1 ADP (umol O2/mg dry tissue/sec); Cyt C (umol 02/mg dry tissue/sec); complex 1-2 succ (umol O2/mg dry tissue/sec); Maximal ETS FCCP (umol O2/mg dry tissue/sec); Complex 2 Rot (umol O2/mg dry tissue/sec); Rox anti A (umol O2/mg dry tissue/sec); COX TMPD/anti a(umol O2/mg dry tissue/sec) reduced cytochrome c check was done with the spectrophotometer to ensure that the cyt c was at the appropriate absorbency; CS activity protein homogenate was the absorbency of the protein samples used to measure citrate synthase activity in the tissue samples

Methods:

Fresh and Frozen tissue

Instruments:

OROBOROS Oxygraph 2k HR respirometer and H1 hybrid spectrophotometer

Error Analysis:

ANOVA and student's t-test

Individual Files: