Effect of High Fat Diet-induced Paternal Obesity on Physical Activity, Insulin

Sensitivity and Body Composition of Offspring

Alexis Zontini, Shauni Bobbs, Yuriy Slyvka, Yizhu Zhang, Josh Ozbolt, John Adame, Leslie Consitt, Felicia V. Nowak
Biomedical Sciences, Ohio University, Heritage College of Osteopathic Medicine, Athens, OH 45701
Results
Results and Conclusions
Figure 1. Group 2 Females have higher insulin sensitivity at all
three time points compared to group 1 females. Group 2 males
show higher insulin sensitivity at 6 weeks and 12 months than
Group 1 males. Insulin sensitivity is lower in males. For all three
time points in both groups, insulin sensitivity is higher in females
than in males.

Figure 2. Group 1 females ran more than group 1 males at 6
weeks and 6 months. Group 2 females, at 6 months and 12
months ran more than Group 2 males. Animals in Group 2 ran
more than those in Group 1 at 6 weeks (females), 6 months
(males and females), and 12 months (males).

Figure 3. Group 2 males weighed more than Group 1 males at
all time points. Body weight was higher in male than females in
Group 2 at all time points, and Group 1 at 6 months.

Figure 4. Males in Group 2 have a higher percent fat than males
in Group 1 at 6 months. The same trend is seen at 12 months but
it is not significant. Females have higher percent body fat than
males, in both groups, at 6wk, 12 months.

Figure 3.
Figure 1.
Figure 4.
Figure 2.
Supported by HCOM Research and Scholarly Affairs Committee and Ohio University
Research Council awards to FVN.
Introduction
The global prevalence of childhood obesity is on the rise. Recent estimates are that
>155 million children and adolescents, and ~22 million infants and toddlers are
over weight. Excess weight gain can result in a wide range of disorders during
childhood and adolescence, including emotional and behavioral problems, and can
predispose to the development of type 2 diabetes, hypertension, liver, renal and
cardiovascular diseases, and many cancers including breast and colon. Parental
obesity is a predictor of childhood excess weight. Having a father, mother or
sibling with a BMI of 40 increases the risk of adult obesity 5 fold compared to
individuals who have only normal weight first degree relatives. Genetic and
epigenetic modes of inheritance have been implicated in these associations.
Obesity in both genetic parents more than doubles the risk of obesity in offspring
and environmental and behavioral factors can further increase the risk.

DNA sequence is predicted to account for only 6-11% of the genetic variation in
BMI. Thus it is highly likely that epigenetic mechanisms will prove to play a
substantial role in inherited risk for obesity. Epigenetic modifications in gene
expression occur without changes in DNA sequence. Phenotypic expression of the
modified genes can vary dependent upon the gender of the individual and, unlike
traits that result from DNA-sequence based genetic inheritance, phenotype can also
differ depending on parental gender. Several genes known to be involved in
energy storage and expenditure are epigenetically modified. These modifications
may be transmitted and predispose to obesity in offspring. The maternal
contribution to obesity and abnormal metabolic profile in offspring has been an
active area of investigation. Mechanisms demonstrated to date include direct
transmission of hormonal and chemical signals (metabolic imprinting) to offspring
during gestation and lactation, abnormalities in metabolic profile, appetite and
glucose regulation, and epigenetic factors. In our model, we are looking at the
effects of paternal diet on the metabolic profile of offspring.
Experimental Design and Methods
Animal Care and Use. Starting at 4 weeks of age, C57BL/6N (Harlan
Laboratories, Inc., Indianapolis, IN) male and female F
0
(first generation)
mice were assigned to either low fat diet (LFD) (D12450B, 10 kcal% fat)
or HFD, (D12451, 45 kcal% fat) (Research Diets, New Brunswick, NJ) for
12 weeks. Eight mating pairs were then assigned for each group: LFD/
LFD (Group 1) and HFD/ LFD (Group 2). Only HFD-fed animals that
fit the criteria for obesity (body weight 10-25% greater than age- and sex-
matched animals on LFD) were mated. Pups from litter sizes between 5 and
7 were included to control for prenatal and postnatal nutrition.
Insulin Sensitivity Testing (IST) (Figure 1). At 5-6 weeks, 5-6 months, and
11-12 months of age, 3 offspring of each sex from each mating pair group
were subjected to IST using 1 mU insulin/g body weight.
Voluntary Physical Activity (Figure 2). At 6 weeks, 5-6 months, and 11-12
months of age, mice, 4 males and 4 females were placed in individual
cages where voluntary activity was assessed using an activity wheel with
usage recording capability.
Body Weight and Composition (Figures 3, 4). Weight and percent body fat
determined by NMR (Bruker Minispec) were assessed at four time points,
20 days, 5-6 weeks, 5-6 months, and 11-12 months.
Statistics: Group comparisons were done using unpaired t-tests (p < 0.05).
Abstract
Epidemiological studies clearly show that parental diet and increased body mass
index (BMI) correlate with abnormal metabolic profile and increased risk for
obesity and insulin resistance in offspring. Genetic, gene regulatory (epigenetic),
behavioral and environmental factors mediate this relationship. Factors inherited
from both parents contribute to body mass and risk for metabolic disease, including
diabetes and cardiovascular. Maternal and paternal effects are independent and
additive. Most animal model studies have focused on the maternal, with a smaller
number just recently beginning to address the paternal contribution. To our
knowledge there are no published reports to investigate combined maternal/paternal
effects. To investigate that environmentally-induced heritable genetic and
epigenetic differences are transmitted from parent to offspring, we phenotypically
characterized a mouse model of trans-generational parental transmission of
metabolic differences due to paternal consumption of a high fat diet (HFD). Males
were fed a HFD for 12 weeks to induce obesity and mated with females fed a LFD.
Male and female pups were studied at postnatal day 20 , 5-6 weeks, 5-6 months and
11-12 months of age. We analyzed body weight, length, composition, and insulin
sensitivity. Exercise desire was measured by a voluntary running test. All measures
from offspring of HFD/LFD parents were compared with those from offspring of
LFD/LFD parents. This work has potential to disclose paternal diet-induced
epigenetic modifications which alter gene expression in offspring to predispose to
obesity and metabolic disease. These findings can be used to develop new
preventive and therapeutic strategies that improve human health and well-being.
Figure 4. Effects of paternal HFD on % body fat in offspring.
*significant difference between Group 1 and group 2. **
significant difference between female and male.
Figure 3. Effects of paternal HFD on body weight in
offspring. *significant difference between Groups1 and 2.
**significant difference between female and male.

Figure 1. Effects of paternal HFD on IST in offspring.
*significant difference between Group 1 and Group 2,
**significant difference between female and male. Area
under curve (AUC) represents the glucose level for the
duration of the test.

Figure 2. Effects of paternal HFD on running wheel activity in
offspring. *significant difference between Group 1 and Group
2, **significant difference between female and male. Rev =
revolutions.

Summary
The results confirm that our model of paternal diet induced obesity results in metabolic and
behavioral differential effects in offspring. There are some gender differences in these effects
as well, heightening the possibility that they are due, at least in part, to inherited epigenetic
modifications. Future studies will identify differences in transcription rates for key molecules
in the pathways that program the observed phenotypes, and investigate the responsible
mechanisms.