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Maternal influence of prolyl endopeptidase on fat mass of adult progeny.

Warden CH, Fisler JS, Espinal G, Graham J, Havel PJ, Perroud B - Int J Obes (Lond) (2009)

Bottom Line: There were no genotype effects on BW or AI in males or females on either diet.Experiment 2-In contrast, reciprocal crosses of heterozygous males or females with WT B6 revealed highly significant parent of origin effects on all traits except body length.Heterozygosity for PREP GT results in highly significant maternal effects, whereas homozygosity for the PREP(gt/gt) mutation has a much more limited direct effect.

View Article: PubMed Central - PubMed

Affiliation: Rowe Program in Genetics, University of California, Davis, CA 95616, USA. chwarden@ucdavis.edu

ABSTRACT

Background: Maternal genotype has lifetime effects on progeny, but few specific genes, and no proteases, are known to underlie maternal effects. Prolyl endopeptidase (PREP) is a serine protease with putative substrates that regulate appetite or milk production.

Objective: To test effects of PREP on obesity phenotypes in mice.

Design: Mice with a gene trap (GT) of PREP (PREP(gt/gt)) on the C57BL/6J (B6) background were generated. Minimal PREP protein was detected by western blot. In Experiment 1, direct effects of PREP were measured in littermate mice derived from intercrosses of heterozygotes (PREP(WT/gt)). In Experiment 2, maternal effects of PREP were measured in reciprocal crosses of heterozygous (PREP(WT/gt)) and wild-type (WT) (PREP(WT/WT)) males and females. DIETS: Mice were fed either low-fat (LF, Experiments 1 and 2) or high-fat (HF, Experiment 1) defined diets.

Measurements: Adiposity index (AI) was calculated from body weight (BW) and weights of four fat depots measured in 120-day-old mice. Fasting plasma glucose, insulin and leptin were measured. In vivo plasma alpha-MSH levels were measured by targeted quantitative peptidomics.

Results: Experiment 1-In intercross mice, there were significant diet effects, but few genotype effects. There were no genotype effects on BW or AI in males or females on either diet. Experiment 2-In contrast, reciprocal crosses of heterozygous males or females with WT B6 revealed highly significant parent of origin effects on all traits except body length. Progeny (WT and heterozygous genotypes and both sexes) born to female PREP(WT/gt) heterozygotes had fat pads that weighed as much as -twofold more at 120 days old than progeny born to male heterozygotes.

Conclusion: Heterozygosity for PREP GT results in highly significant maternal effects, whereas homozygosity for the PREP(gt/gt) mutation has a much more limited direct effect.

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Intercross mice (Experiment 1). Body weights and adiposity index (AI) of female and male PREP wild-type (PREPWT/WT, WT), heterozygote (PREPWT/gt, H), and gene-trap (PREPgt/gt, GT) mice fed low-fat AIN-76A (LF) or high-fat (HF) diets and killed at 120 days of age. AI was calculated as (total adipose depot weight ÷ live body weight × 100). Data are presented as means ± SE. P values were calculated by ANOVA: Bonferroni correction for multiple tests sets significant P value at ≤ 0.001. Post hoc comparisons were by Tukey's HSD: means not sharing a letter are significant at P = 0.05. Genders were analyzed separately. 2-Way ANOVA. Females: body weight effect of diet, p<0.0001, effect of genotype, NS; AI effect of diet p<0.0001, effect of genotype, NS. Males: body weight effect of diet, p<0.0001, effect of genotype, p=0.0003; AI effect of diet, p<0.0001, effect of genotype, NS.
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Figure 2: Intercross mice (Experiment 1). Body weights and adiposity index (AI) of female and male PREP wild-type (PREPWT/WT, WT), heterozygote (PREPWT/gt, H), and gene-trap (PREPgt/gt, GT) mice fed low-fat AIN-76A (LF) or high-fat (HF) diets and killed at 120 days of age. AI was calculated as (total adipose depot weight ÷ live body weight × 100). Data are presented as means ± SE. P values were calculated by ANOVA: Bonferroni correction for multiple tests sets significant P value at ≤ 0.001. Post hoc comparisons were by Tukey's HSD: means not sharing a letter are significant at P = 0.05. Genders were analyzed separately. 2-Way ANOVA. Females: body weight effect of diet, p<0.0001, effect of genotype, NS; AI effect of diet p<0.0001, effect of genotype, NS. Males: body weight effect of diet, p<0.0001, effect of genotype, p=0.0003; AI effect of diet, p<0.0001, effect of genotype, NS.

Mentions: Intercrosses of heterozygous (PREPgt/gt x PREPWT/WT) parents were used to determine direct genotype effects. An intercross colony was maintained by breeding of heterozygous parents. Obesity phenotypes of PREPWT/WT, PREPWT/gt and PREPgt/gt littermates maintained on either LF or HF diet were compared. Few genotype effects of PREP gene-trap were observed in F2 animals. Figure 2 presents body weight (BW) and adiposity index (AI) of female and male littermates of different genotypes and diets from the intercross. Genders were analyzed separately. Both genders show a strong diet effect (p<0.0001) for both BW and AI (Figure 2). In female mice, genotype effects for both BW and AI were not statistically significant, although when analyzed individually, BW was lower in PREPgt/gt than in PREPWT/WT mice, whereas adiposity index was lower in PREPWT/gt than in PREPWT/WT mice. There was a significant genotype effect on body weight for male mice (p=0.0003) with PREPWT/gt mice on the HF diet being heavier than PREPgt/gt. However, there were no genotype effects on BW or AI in males fed the LF diet (Figure 2). We also observed recessive genotype effects on kidney weight in females fed the LF diet (p=0.002) and BMI in males fed the HF diet (p<0.0001), since PREPWT/WT and PREPWT/gt are statistically indistinguishable whereas PREPgt/gt are significantly smaller (data not shown).


Maternal influence of prolyl endopeptidase on fat mass of adult progeny.

Warden CH, Fisler JS, Espinal G, Graham J, Havel PJ, Perroud B - Int J Obes (Lond) (2009)

Intercross mice (Experiment 1). Body weights and adiposity index (AI) of female and male PREP wild-type (PREPWT/WT, WT), heterozygote (PREPWT/gt, H), and gene-trap (PREPgt/gt, GT) mice fed low-fat AIN-76A (LF) or high-fat (HF) diets and killed at 120 days of age. AI was calculated as (total adipose depot weight ÷ live body weight × 100). Data are presented as means ± SE. P values were calculated by ANOVA: Bonferroni correction for multiple tests sets significant P value at ≤ 0.001. Post hoc comparisons were by Tukey's HSD: means not sharing a letter are significant at P = 0.05. Genders were analyzed separately. 2-Way ANOVA. Females: body weight effect of diet, p<0.0001, effect of genotype, NS; AI effect of diet p<0.0001, effect of genotype, NS. Males: body weight effect of diet, p<0.0001, effect of genotype, p=0.0003; AI effect of diet, p<0.0001, effect of genotype, NS.
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Figure 2: Intercross mice (Experiment 1). Body weights and adiposity index (AI) of female and male PREP wild-type (PREPWT/WT, WT), heterozygote (PREPWT/gt, H), and gene-trap (PREPgt/gt, GT) mice fed low-fat AIN-76A (LF) or high-fat (HF) diets and killed at 120 days of age. AI was calculated as (total adipose depot weight ÷ live body weight × 100). Data are presented as means ± SE. P values were calculated by ANOVA: Bonferroni correction for multiple tests sets significant P value at ≤ 0.001. Post hoc comparisons were by Tukey's HSD: means not sharing a letter are significant at P = 0.05. Genders were analyzed separately. 2-Way ANOVA. Females: body weight effect of diet, p<0.0001, effect of genotype, NS; AI effect of diet p<0.0001, effect of genotype, NS. Males: body weight effect of diet, p<0.0001, effect of genotype, p=0.0003; AI effect of diet, p<0.0001, effect of genotype, NS.
Mentions: Intercrosses of heterozygous (PREPgt/gt x PREPWT/WT) parents were used to determine direct genotype effects. An intercross colony was maintained by breeding of heterozygous parents. Obesity phenotypes of PREPWT/WT, PREPWT/gt and PREPgt/gt littermates maintained on either LF or HF diet were compared. Few genotype effects of PREP gene-trap were observed in F2 animals. Figure 2 presents body weight (BW) and adiposity index (AI) of female and male littermates of different genotypes and diets from the intercross. Genders were analyzed separately. Both genders show a strong diet effect (p<0.0001) for both BW and AI (Figure 2). In female mice, genotype effects for both BW and AI were not statistically significant, although when analyzed individually, BW was lower in PREPgt/gt than in PREPWT/WT mice, whereas adiposity index was lower in PREPWT/gt than in PREPWT/WT mice. There was a significant genotype effect on body weight for male mice (p=0.0003) with PREPWT/gt mice on the HF diet being heavier than PREPgt/gt. However, there were no genotype effects on BW or AI in males fed the LF diet (Figure 2). We also observed recessive genotype effects on kidney weight in females fed the LF diet (p=0.002) and BMI in males fed the HF diet (p<0.0001), since PREPWT/WT and PREPWT/gt are statistically indistinguishable whereas PREPgt/gt are significantly smaller (data not shown).

Bottom Line: There were no genotype effects on BW or AI in males or females on either diet.Experiment 2-In contrast, reciprocal crosses of heterozygous males or females with WT B6 revealed highly significant parent of origin effects on all traits except body length.Heterozygosity for PREP GT results in highly significant maternal effects, whereas homozygosity for the PREP(gt/gt) mutation has a much more limited direct effect.

View Article: PubMed Central - PubMed

Affiliation: Rowe Program in Genetics, University of California, Davis, CA 95616, USA. chwarden@ucdavis.edu

ABSTRACT

Background: Maternal genotype has lifetime effects on progeny, but few specific genes, and no proteases, are known to underlie maternal effects. Prolyl endopeptidase (PREP) is a serine protease with putative substrates that regulate appetite or milk production.

Objective: To test effects of PREP on obesity phenotypes in mice.

Design: Mice with a gene trap (GT) of PREP (PREP(gt/gt)) on the C57BL/6J (B6) background were generated. Minimal PREP protein was detected by western blot. In Experiment 1, direct effects of PREP were measured in littermate mice derived from intercrosses of heterozygotes (PREP(WT/gt)). In Experiment 2, maternal effects of PREP were measured in reciprocal crosses of heterozygous (PREP(WT/gt)) and wild-type (WT) (PREP(WT/WT)) males and females. DIETS: Mice were fed either low-fat (LF, Experiments 1 and 2) or high-fat (HF, Experiment 1) defined diets.

Measurements: Adiposity index (AI) was calculated from body weight (BW) and weights of four fat depots measured in 120-day-old mice. Fasting plasma glucose, insulin and leptin were measured. In vivo plasma alpha-MSH levels were measured by targeted quantitative peptidomics.

Results: Experiment 1-In intercross mice, there were significant diet effects, but few genotype effects. There were no genotype effects on BW or AI in males or females on either diet. Experiment 2-In contrast, reciprocal crosses of heterozygous males or females with WT B6 revealed highly significant parent of origin effects on all traits except body length. Progeny (WT and heterozygous genotypes and both sexes) born to female PREP(WT/gt) heterozygotes had fat pads that weighed as much as -twofold more at 120 days old than progeny born to male heterozygotes.

Conclusion: Heterozygosity for PREP GT results in highly significant maternal effects, whereas homozygosity for the PREP(gt/gt) mutation has a much more limited direct effect.

Show MeSH
Related in: MedlinePlus