Figure 1.
Summary of transcription and methylation at the Gnas cluster.
Transcribed exons are shown as filled boxes and nontranscribed exons as empty boxes. Nespas and Exon 1A give rise to noncoding transcripts. The arrows indicate the direction of transcription and a thin arrow indicates low expression. The methylation status of the Exon 1A differentially methylated region (DMR) and the Nespas DMR (the imprinting control region, ICR, for the cluster) is shown as plus signs for methylated and minus signs for unmethylated.
Figure 2.
Transcript expression in PatDp(dist2).
Summary of transcription and methylation at the Gnas cluster in (A) PatDp(dist2)2∶0, (B) PatDp(dist2)2∶1, (C) PatDp(dist2)2∶2 (D) PatDp(dist2)1∶1, (E) PatDp(dist2)1∶2. Transcribed exons are shown as filled boxes and nontranscribed exons or exons giving rise to very markedly reduced levels of transcript as empty boxes. Nesp exon 1 is noncoding and Nespas and Exon 1A give rise to noncoding transcripts. The arrows indicate the direction of transcription and a thin arrow indicates low expression. The methylation status of the Exon 1A DMR and the Nespas DMR is shown as plus signs for methylated and minus signs for unmethylated.
Table 1.
Crosses to generate PatDp(dist2).
Figure 3.
Gnas expression in newborn brown fat measured by northern blotting.
(A) Bar chart showing Gnas levels in PatDp(2∶2) normalised to β-actin mRNA and shown relative to the level in +/ΔEx1A. Mean ± sem calculated for 5 PatDp(dist2)2∶2 and 8+/ΔEx1A. (B) Northern blot of Gnas and β-actin loading control in +/ΔEx1A and PatDp(2∶2) using 5 µg total RNA from newborn brown fat.
Table 2.
Incidence and Survival of PatDp(dist2).
Table 3.
Neonatal Phenotype of PatDp(dist2).
Figure 4.
Metabolic analyses in early postnatal stages.
(A) blood glucose levels. (n = 9–10) (B) plasma insulin levels.(n = 10–14) (C) plasma glucagon levels.(n = 9–13) (D) plasma noradrenaline. (n = 10–38). Error bars show standard errors of the means; ***P<0.001, **P<0.01, *P<0.05.
Figure 5.
(A) Growth curve of +/ΔEx1A and wild-type littermates from 1 day to 12 weeks. The weights of wild-type littermates have been normalised to 1 at each timepoint and the weights of +/ΔEx1A mice have been taken as a percentage of wild-type weights. (n = 5–26). (B) Growth curve of PatDp(dist2)2∶2 and +/ΔEx1A. The weights of +/ΔEx1A littermates have been normalised to 1 at each timepoint and the weights of PatDp(dist2)2∶2 mice have been taken as a percentage of +/Ex1A weights (n = 9–12). Error bars show standard errors of the means.
Figure 6.
Analysis of metabolic rate in 21 week old male +/ΔEx1A and their wild-type littermates, and PatDp(dist2)2∶2 and their +/ΔEx1A littermates. (A) and (B) Rates of oxygen consumption and carbon dioxide output respectively (n = 4–12). Error bars show standard errors of the means; **P<0.01, *P<0.05.