Reed1 project protocol

Survey of adipose depots, body size, and heart and kidney weights in males of 28 inbred strains of mice   (2009)

Reed DR, Bachmanov AA, Tordoff MG




Reed1_Protocol

Project protocol — Contents
Workflow and sampling
Equipment
Reagents, supplies, and solutions
Procedures for adipose depot dissection
Data
References



Workflow and sampling

Workflow

Steps
Procedure performed
Age (wks)
Equipment and reagents
Data collected
1
Male mice aged and acclimated before survey
29-67
-
-
2
Mice weighed and anesthetized
29-67
balance scale, needles and syringes, anesthesia
body weight, body length
3
Mice euthanized
29-67
dissecting kit
-
4
Both left and right retroperitoneal fat pads dissected
29-67
dissecting kit, balance scale
perirenal fat pads weights
5
Both left and right testicular fat pads dissected
29-67
dissecting kit, balance scale
gonadal fat pad weights
6
Both left and right kidneys dissected
29-67
dissecting kit, balance scale
kidney weights
7
Heart dissected
29-67
dissecting kit, balance scale
heart weight

Equipment

• small animal dissecting kit
• analytical balance scale
• ruler or caliper
• dissecting board
• dissecting scope

Reagents, supplies, solutions  

• paper towels
• disinfectant solution
• rodent anesthesia: 70 mg/kg ketamine +10 mg/kg xylazine (5 mL/kg)
• needles and syringes
• 95 % ethanol

Procedures for adipose depot dissection

a. Male mice between the ages of 29-67 wks are initially measured for body weight (to the nearest 0.1 g) then anesthetized, before measuring body length (from the base of the lower incisors to the base of the anus, distance to the nearest 1 mm).
b. While still under deep anesthesia, the mice are euthanized by exanguination.
c. Following complete exanguination, the dissection of both the left and the right retroperitoneal fat pads, adipose tissue from the perirenal capsules as well as adipose tissues attached to the dorsal body wall near the kidneys are conducted, with the removal of adrenal glands embedded in the adipose kidney capsules.
d. No attempt is made to isolate and discard the renal artery and the ureter from the retroperitoneal fat pads.
e. Gonadal fat pads in male mice are dissected according to their defined proximity to the epididymis and vesicular gland (See lower panel in the figure below).
f. Gonadal fat pads in female mice are dissected accordingly with their defined location and direct proximity to the ovaries and uterus (See lower panel in the figure below).

Investigator's note: "Although the epididymal (male) and parametrial (female) fat depots are categorized with a single label (gonadal), these depots are different in structure and possibly differ in function."

g. Other adipose tissues associated with the omental membrane, ileum, jejunum, or duodenum (mesenteric fat pads) are not removed or measured.
h. Following the dissections of the four total fat depots, they are weighed individually to the nearest 0.01 g.

Data collected by investigator

Fat pads:gonadal fat pads weight and perirenal fat pads weight

Whole body metrics: body length and body weight

Organ weight: heart weight and kidneys weight

Definitions and calculations

Normalized weights = (raw weight ÷ body weight) × 100

BMI = body mass index [kg/m2] = (body weight in g x (kg/1000g)) ÷ ((body length in cm x (m/100cm)) x (body length in cm x (m/100cm))



References

    McDaniel AH, Li X, Tordoff MG, Bachmanov AA, Reed DR. A locus on mouse Chromosome 9 (Adip5) affects the relative weight of the gonadal but not retroperitoneal adipose depot. Mamm Genome. 2006 Nov;17(11):1078-92. Epub 2006 Nov 10. PubMed 17103052   FullText