Phenotype Information for B6-ob (000632)

Type 2 diabetes phenotypes in male and female C57BL/6J leptin mutant mice (B6-ob; B6.Cg-Lep^ob/J) include:

Severe obesity (Figure 1 and Table 2)
Transient hyperglycemia: average blood glucose peaks at 8 weeks and returns to normal levels by 14 weeks (Figure 2 and Table 1) due to pancreatic islet cell hypertrophy in the C57BL/6 background.
Elevated serum total cholesterol, HDL cholesterol, triglycerides and glucose at 8 weeks of age; elevated serum total cholesterol, HDL cholesterol and glucose (females) at 16 weeks of age (Table 3)
Glucose intolerance at 8 & 16 weeks (Figure 3)

Homozygous ob/ob mice are fed LabDiet 5K20 (10% fat w/w) and heterozygous ob/+ mice are fed LabDiet 5K52 formulation (6% fat w/w). Complete results, including spleen flow cytometry data, are available from the Mouse Phenome Database.

Figure 1 Body weight growth curve. Weekly weights of male and female mice between the ages of 4 and 16 weeks. Values represent the mean and one standard deviation of at least 40 mice per age.

Figure 2 Non-fasted glucose. Blood glucose levels of male and female mice between the ages of 6 and 16 weeks. Submandibular blood glucose measurements were obtained using a OneTouch Ultra 2 or UltraMini hand-held glucometer that was validated with a control glucose solution on each day of use. Values represent the mean and one standard deviation of at least 40 non-fasted mice.

Figure 3 Glucose tolerance tests. Blood glucose levels during glucose tolerance tests (GTT) at 8 and 16 weeks of age. Following a 16 hour fast, an initial submandibular blood glucose reading was taken and mice were administered glucose by IP injection at 2 g/kg body weight. Blood glucose was measured in 30 minute intervals. Any result exceeding the maximum reading of the meter was recorded as 600 mg/dL. Each data point represents the mean and one standard deviation of 10-20 mice.

Table 1 Distribution of homozygous (ob/ob) blood glucose values. The blood glucose values shown in Figure 2 were grouped into ranges to show the reversion to non-diabetic values as ob/ob mice age. Rows indicate the percentage of mice in the indicated ranges, with the exception of the final row that states the number of mice analyzed.

Glucose Range (mg/dL)	4 Weeks		6 Weeks		8 Weeks		10 Weeks		12 Weeks		14 Weeks		16 Weeks
	Males	Females	Males	Females	Males	Females	Males	Females	Males	Females	Males	Females	Males	Females
< 250	58%	62%	56%	58%	33%	33%	77%	88%	83%	85%	94%	98%	98%	98%
250 - 349	29%	22%	36%	25%	38%	33%	17%	9%	13%	12%	3%	0%	0%	0%
350 - 450	11%	13%	9%	15%	23%	25%	4%	2%	3%	2%	0%	0%	0%	2%
> 450	2%	4%	0%	1%	6%	9%	1%	2%	1%	2%	3%	2%	2%	0%
# Mice	91	79	90	79	69	67	69	65	69	65	68	65	48	46

Table 2 Body Composition. Mice were weighed and then analyzed using a Lunar PIXImus DEXA scanner. Calculations of body composition exclude the head. Values represent mean and one standard deviation of 10 non-fasted mice per diet and age. Results were analyzed separately by age and sex using two-way ANOVA with Sidak’s multiple comparisons test to identify values that differed significantly between genotypes, using GraphPad Prism version 6.07 for Windows (GraphPad Software).

Age & Sex	Genotype	Body Weight (g)	Bone Mineral Density (g/cm²)	Bone Mineral Content (g)	Bone Area (cm²)	Lean Tissue (g)	Fat Tissue (g)	Percent Fat Tissue
8 Week Males	ob/ob	43.2 ± 1.6^a	0.046 ± 0.001	0.45 ± 0.03	9.7 ± 0.6	17.7 ± 0.9^b	24.4 ± 2.3^a	57.9 ± 3.2^a
8 Week Males	ob/+	27.0 ± 1.2^a	0.050 ± 0.002	0.41 ± 0.03	8.4 ± 0.7	20.4 ± 0.8^b	5.9 ± 0.8^a	22.4 ± 2.3^a
8 Week Females	ob/ob	42.3 ± 2.1^a	0.043 ± 0.001	0.37 ± 0.03	8.6 ± 0.7	16.3 ± 1.1^b	24.8 ± 2.6^a	60.1 ± 3.3^a
8 Week Females	ob/+	21.1 ± 1.1^a	0.047 ± 0.001	0.35 ± 0.02	7.3 ± 0.4	15.2 ± 0.9	5.1 ± 0.9^a	24.8 ± 3.5^a
16 Week Males	ob/ob	57.4 ± 2.2^a	0.053 ± 0.003	0.58 ± 0.07	10.8 ± 1.1	21.5 ± 1.5	34.1 ± 2.8^a	61.3 ± 2.9^a
16 Week Males	ob/+	31.8 ± 1.8^a	0.056 ± 0.003	0.51 ± 0.04	9.0 ± 0.6	23.6 ± 1.5	7.4 ± 1.5^a	23.8 ± 4.3^a
16 Week Females	ob/ob	60.7 ± 3.8^a	0.048 ± 0.002	0.45 ± 0.06	9.5 ± 1.3	20.3 ± 1.1^b	38.0 ± 2.8^a	65.1 ± 2.2^a
16 Week Females	ob/+	26.2 ± 2.2^a	0.055 ± 0.002	0.47 ± 0.02	8.5 ± 0.3	18.0 ± 1.1	7.1 ± 1.7^a	28.0 ± 3.8^a
^a P ≤ 0.0001 ^b P ≤ 0.001

Table 3 Clinical chemistry. All values were measured from serum collected from submandibular blood except HbA1c, which was measured from submandibular whole blood. Values represent mean and one standard deviation of 10-20 non-fasted mice per age, sex, and genotype. Results were obtained using a Beckman Coulter AU680 chemistry analyzer. Results were analyzed separately by age and sex using two-way ANOVA with Sidak’s multiple comparisons test to identify values that differed significantly by genotype, using GraphPad Prism version 6.07 for Windows (GraphPad Software).

Age & Sex	Genotype	Total Cholesterol (mg/dL)	HDL Cholesterol (mg/dL)	LDL Cholesterol (mg/dL)	Triglycerides (mg/dL)	Free Fatty Acids (mEq/L)	Glucose (mg/DL)	HbA1c (NGSP %)
8 Week Males	ob/ob	238 ± 22^a	152 ± 13^a	8.0 ± 1.3	218 ± 92^a	1.30 ± 0.29	362 ± 72^a	7.9 ± 1.6
8 Week Males	ob/+	131 ± 20^a	95 ± 12^a	4.4 ± 1.8	153 ± 44^a	1.45 ± 0.35	199 ± 29^a	5.6 ± 0.3
8 Week Females	ob/ob	267 ± 58^a	160 ± 25^a	10.1 ± 2.7	222 ± 47^a	1.62 ± 0.32	261 ± 47^a	6.7 ± 0.8
8 Week Females	ob/+	103 ± 14^a	74 ± 12^a	6.1 ± 1.1	151 ± 27^a	1.61 ± 0.45	168 ± 18^a	5.2 ± 0.3
16 Week Males	ob/ob	274 ± 34^a	170 ± 14^a	16.3 ± 4.4	131 ± 27	0.88 ± 0.10	212 ± 46	8.1 ± 0.7
16 Week Males	ob/+	127 ± 14^a	96 ± 9^a	2.5 ± 0.5	154 ± 30	1.02 ± 0.29	191 ± 24	6.0 ± 0.4
16 Week Females	ob/ob	198 ± 59^a	136 ± 34^a	7.2 ± 4.2	114 ± 42	0.83 ± 0.24	196 ± 51^b	6.4 ± 0.5
16 Week Females	ob/+	95 ± 10^a	72 ± 7^a	4.8 ± 0.6	136 ± 29	1.23 ± 0.34	166 ± 18^b	5.4 ± 0.4
^a P ≤ 0.0001 ^b P ≤ 0.01