Journal of Bioenergetics and Biomembranes

Supplementary Material

Dietary fat, fatty acid saturationand mitochondrial bioenergetics

Liping Yu1, Brian D. Fink2, Judith A. Herlein2, Christine L. Oltman3, Kathryn G. Lamping4, and William I. Sivitz2

1NMR Core Facilityand Department of Biochemistry, 2Department of Internal Medicine/Endocrinology,3Department of Internal Medicine/Cardiology, 4Department of Pharmacology, University of Iowa and the Iowa City Veterans Affairs Medical Center, Iowa City, IA, 52242

Corresponding author:

Dr. William Sivitz

Department of Internal Medicine, Division of Endocrinology and Metabolism

The University of Iowa Hospitals and Clinics, 422GH

200 Hawkins Drive

Iowa City, IA. 52242.

Phone: (319) 353-7812; Fax: (319) 353-7850

E-mail:

Supplemental material Index:

Table 1. Dietary regimens

Table 2. Dietary fatty acid content

Table 3. Mitochondrial lipid fatty acid composition

Figure 1. 2-deoxyglucose energy clamp graphic representation

Supplementary Table 1. Composition of dietary regimens for the normal fat (control diet),

high fat lard (HF), or high fat with 50% replacement of lard with menhaden oil (MO) diets.

Composition / Control Dieta / HF Dietb / MO Dietc
Protein (% total kcal) / 34 / 20 / 20
Carbohydrate (% total kcal) / 53 / 20 / 20
Fat (% total kcal) / 13 / 60 / 60
Total (%) / 100 / 100 / 100
kcal/gm / 3.0 / 5.2 / 5.2
Fat distribution
Soybean oil (% total kcal) / 6.5d / 5.5 / 5.5
Lard (% total kcal) / 6.5 / 54.5 / 27.2
Menhaden oil (% total kcal) / 0 / 0 / 27.2
Total fat (% total kcal) / 13 / 60 / 60

aTeklad, Diet 7001, Harlan Labs, Madison, WI (standard rodent chow)

bResearch Diets, D12492, New Brunswick, NJ

cResearch Diets, D10122003

dIncludes corn oil and wheat oil mixed within.

Supplementary Table 2. Fatty acid content of the dietary regimens determined by gas chromatography. Numbers refer to % of total fat calories. MUFA, mono-unsaturated fatty acid; PUFA, poly-unsaturated fatty acid; EPA, eicosapentaenoic acid; DHA, docosahexaenoic acid.

Diet / Saturated Fatty Acid / MUFA / PUFA / Others
16:0 / 18:0 / 18:1 / 18:2 / 20:5 / 22:6
Palmitic Acid / Steric
Acid / Oleic
Acid / n-6
Linoleic
Acid / n-3
EPA / n-3
DHA
Control diet / 24.1 / 8.3 / 28.8 / 30.3 / 0 / 0 / 8.5
HF diet / 19.8 / 10.0 / 35.1 / 28.8 / 0 / 0 / 6.3
MO diet / 17.6 / 7.4 / 20.6 / 15.4 / 9.8 / 6.7 / 22.5

Supplementary Table 3. Fatty acid composition of liver (top panel) and heart (lower panel) mitochondrial lipids isolated from mice fed normal fat (control), high fat lard (HF), or 50% high fat lard + 50% Menhaden Oil (MO) diet. Unsaturation indices were compared by one-way ANOVA with Bonferron’s post-test. n= 4 per group, * p < 0.001 compared to control and HF.† p < 0.05 compared to HF. rel% = relative percent

Control dietHF dietMO diet

FA / rel % / SE / FA / rel % / SE / FA / rel % / SE
16:0 / 19.13 / 0.38 / 16:0 / 17.44 / 1.11 / 16:0 / 16.13 / 1.07
18:0 / 12.44 / 0.20 / 18:0 / 18.90 / 0.66 / 18:0 / 18.02 / 1.11
14:1 / 0.04 / 0.01 / 14:1 / 0.02 / 0.01 / 14:1 / 0.06 / 0.01
16:1 / 2.56 / 0.12 / 16:1 / 0.84 / 0.12 / 16:1 / 0.90 / 0.18
18:1 / 14.44 / 0.41 / 18:1 / 10.56 / 0.59 / 18:1 / 8.31 / 0.48
18:2 / 17.41 / 0.67 / 18:2 / 18.00 / 0.61 / 18:2 / 17.35 / 0.72
18:3 / 0.27 / 0.02 / 18:3 / 0.14 / 0.05 / 18:3 / 0.51 / 0.04
20:1 / 0.40 / 0.03 / 20:1 / 0.19 / 0.06 / 20:1 / 0.16 / 0.05
20:2 / 0.72 / 0.37 / 20:2 / 0.49 / 0.06 / 20:2 / 0.44 / 0.05
20:3 / 2.74 / 0.29 / 20:3 / 2.32 / 0.13 / 20:3 / 1.28 / 0.25
20:4 / 19.51 / 0.66 / 20:4 / 21.52 / 0.83 / 20:4 / 10.31 / 0.38
22:1 / 0.15 / 0.05 / 22:1 / 0.05 / 0.02 / 22:1 / 0.22 / 0.07
20:5 / 0.37 / 0.03 / 20:5 / 0.14 / 0.05 / 20:5 / 8.36 / 0.33
22:4 / 0.25 / 0.15 / 22:4 / 0.14 / 0.05 / 22:4 / 0.00 / 0.00
24:1 / 1.04 / 0.25 / 24:1 / 0.68 / 0.24 / 24:1 / 0.29 / 0.13
22:5 / 0.35 / 0.01 / 22:5 / 0.18 / 0.06 / 22:5 / 0.77 / 0.06
22:6 / 6.66 / 0.25 / 22:6 / 8.67 / 0.85 / 22:6 / 16.00 / 0.87
Unsaturation index: / Unsaturation index: / Unsaturation index:
1.89 ± 0.021 / 1.99 ± 0.044 / 2.36 ± 0.027 *

______

Control dietHF dietMO diet

FA / rel % / SE / FA / rel % / SE / FA / rel % / SE
16:0 / 9.18 / 2.31 / 16:0 / 14.13 / 1.13 / 16:0 / 15.32 / 0.37
16:1 / 0.26 / 0.11 / 16:1 / 0.47 / 0.05 / 16:1 / 0.81 / 0.06
18:0 / 16.68 / 0.46 / 18:0 / 18.31 / 0.39 / 18:0 / 17.60 / 0.41
18:1 / 7.62 / 0.53 / 18:1 / 8.63 / 0.24 / 18:1 / 6.33 / 0.48
18:2 / 13.41 / 1.42 / 18:2 / 12.85 / 0.68 / 18:2 / 5.24 / 0.28
18:3 / 0.49 / 0.04 / 18:3 / 0.30 / 0.03 / 18:3 / 0.24 / 0.01
20:2 / 0.42 / 0.03 / 20:2 / 1.04 / 0.08 / 20:2 / 1.11 / 0.13
20:3 / 1.95 / 0.10 / 20:3 / 1.52 / 0.12 / 20:3 / 0.94 / 0.08
20:4 / 9.48 / 0.43 / 20:4 / 9.60 / 0.48 / 20:4 / 3.42 / 0.29
20:5 / 0.01 / 0.01 / 20:5 / 0.02 / 0.01 / 20:5 / 0.52 / 0.06
22:4 / 8.52 / 1.59 / 22:4 / 3.93 / 0.71 / 22:4 / 3.44 / 0.55
24:1 / 2.62 / 0.56 / 24:1 / 2.29 / 0.09 / 24:1 / 2.01 / 0.07
22:6 / 24.49 / 2.42 / 22:6 / 21.54 / 0.79 / 22:6 / 38.74 / 1.40
Unsaturation index: / Unsaturation index: / Unsaturation index:
2.64 ± 0.185 / 2.28 ± 0.095 / 2.89 ± 0.051 †

Supplementary Figure 1. The 2-deoxyglucose (2DOG) energy clamp. Saturating amounts of 2DOG and hexokinase recycle ATP back to ADP by rapidly and irreversibly converting 2DOG into 2-deoxyglucose phosphate (2DOGP). The resulting ADP availability is clamped at levels determined by the amount of ADP added. ETS = electron transport system, IMM = inner mitochondrial membrane.

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