experimental evaluation of early effects of carotenoids on metabolic
programming, including epigenetics, and their long-term consequences, including the susceptibility to obesity later in life [13–15].
Moreover, the use of rodent models is convenient because
rodents are extensively used to study the mechanisms by which
obesity develops and induces physiological disorders [16–19],
whereas only few studies of this type have been conducted in gerbils
[20] or in ferrets [21]. Currently, numerous genetic and dietinduced experimental rodent models of obesity are described in
the literature. Monogenic animal models of obesity can be useful
because their obesity and adiposity are often severe, resulting in a
clear distinct phenotype [22]. Nevertheless, environmental or
behavioral factors such as excess caloric food intake and physical
inactivity are most likely the main contributors to the obesity
pandemic, rather than genetic changes [4]. Thus, models with
diet-induced obesity have been preferably used in place of monogenic models to make the experimental model as close as possible to
the human situation and the purpose of the study [23].
2 Materials
2.1 Animals
4–6 weeks old C57BL/6 J male mice (e.g., from Charles River
Laboratories, Barcelona, Spain).
2.2 Diets
and Treatments
1. Defined low-fat low-sucrose (LFD) diet with 10% energy as fat
and 7% energy as sucrose (e.g., D12450J from Research Diets
Inc.).
2. Defined high-fat medium-sucrose (HFD) diet with 45% energy
as fat and 17% energy as sucrose (e.g., D12451 from Research
Diets Inc.).
3. Olive oil (vehicle).
2.3 Glucose
Tolerance Test
and Surrogate Insulin
Resistance Indexes
1. Glucose.
2. Saline (NaCl 0.9%).
3. Sodium heparinized capillary tubes to collect blood used to
obtain plasma.
2.4 Blood
and Tissue/Organ
Collection
1. Isoflurane as anesthesic.
2. 23 G needle attached to a 1 mL self-prepared heparinized
syringes (10–15 IU sodium heparin/syringe mL), for cardiac
puncture.
3. Dissection and surgical tools including forceps, spatulas, scissors, and needles.
4. 1.5 mL plastic storage tubs.
406
Joan Ribot et al.
programming, including epigenetics, and their long-term consequences, including the susceptibility to obesity later in life [13–15].
Moreover, the use of rodent models is convenient because
rodents are extensively used to study the mechanisms by which
obesity develops and induces physiological disorders [16–19],
whereas only few studies of this type have been conducted in gerbils
[20] or in ferrets [21]. Currently, numerous genetic and dietinduced experimental rodent models of obesity are described in
the literature. Monogenic animal models of obesity can be useful
because their obesity and adiposity are often severe, resulting in a
clear distinct phenotype [22]. Nevertheless, environmental or
behavioral factors such as excess caloric food intake and physical
inactivity are most likely the main contributors to the obesity
pandemic, rather than genetic changes [4]. Thus, models with
diet-induced obesity have been preferably used in place of monogenic models to make the experimental model as close as possible to
the human situation and the purpose of the study [23].
2 Materials
2.1 Animals
4–6 weeks old C57BL/6 J male mice (e.g., from Charles River
Laboratories, Barcelona, Spain).
2.2 Diets
and Treatments
1. Defined low-fat low-sucrose (LFD) diet with 10% energy as fat
and 7% energy as sucrose (e.g., D12450J from Research Diets
Inc.).
2. Defined high-fat medium-sucrose (HFD) diet with 45% energy
as fat and 17% energy as sucrose (e.g., D12451 from Research
Diets Inc.).
3. Olive oil (vehicle).
2.3 Glucose
Tolerance Test
and Surrogate Insulin
Resistance Indexes
1. Glucose.
2. Saline (NaCl 0.9%).
3. Sodium heparinized capillary tubes to collect blood used to
obtain plasma.
2.4 Blood
and Tissue/Organ
Collection
1. Isoflurane as anesthesic.
2. 23 G needle attached to a 1 mL self-prepared heparinized
syringes (10–15 IU sodium heparin/syringe mL), for cardiac
puncture.
3. Dissection and surgical tools including forceps, spatulas, scissors, and needles.
4. 1.5 mL plastic storage tubs.
406
Joan Ribot et al.