GHR/HindIII Locus Polymorphisms in Intron-2 GHR Gene of Papua Local Chicken

This study aimed to detect single nucleotide polymorphisms (SNPs) in intron-2 on growth hormone receptor (GHR) gene in Papua local chickens using the PCR-RFLP method to study its relationship with growth characteristics. Data on the bodyweight of 49 chickens aged 1, 2, 3, and 4 months (22 males, 27 females) and DNA samples were used for this study. The DNA fragment of size 718 bp in intron-2 of the GHR gene from the study chicken was successfully amplified using a pair of specific primers. The PCR-RFLP/HindIII analysis results found this locus's two genotypes (HindIII++ and HindIII--). HindIII+ and HindIII- alleles were 0.02 and 0.98, respectively.

Mammalian Co-methylation Network Analysis of Species Characteristics

The comparative cross-species analysis is a powerful tool to resolve the mysteries of evolution and phenotypic disparities among animals. This is the first network analysis of 10,000 DNA methylome data from 176 mammalian species to identify co-methylation modules that relate to individual (age, sex, tissue type) and species characteristics (e.g. phylogenetic order, maximum lifespan, adult weight). The unexpected correlation between DNA methylation and species were sufficiently strong to allow the construction of phyloepigenetic trees that parallel the phylogenetic tree. Weighted correlation network analysis identified 55 distinct co-methylation modules, i.e. sets of highly correlated CpGs. 31 of these modules are readily interpretable in terms of their relationship to age, maximum lifespan, tissue type etc. An age-related module was perturbed by gold standard anti-aging interventions in mice such as caloric restriction or growth hormone receptor knock outs. Our module-based analysis greatly enhances our biological understanding of age-related changes in DNA methylation across many species.

High-throughput mediation analysis of human proteome and metabolome identifies mediators of post-bariatric surgical diabetes control

To improve the power of mediation in high-throughput studies, here we introduce High-throughput mediation analysis (Hitman), which accounts for direction of mediation and applies empirical Bayesian linear modeling. We apply Hitman in a retrospective, exploratory analysis of the SLIMM-T2D clinical trial in which participants with type 2 diabetes were randomized to Roux-en-Y gastric bypass (RYGB) or nonsurgical diabetes/weight management, and fasting plasma proteome and metabolome were assayed up to 3 years. RYGB caused greater improvement in HbA1c, which was mediated by growth hormone receptor (GHR). GHR’s mediation is more significant than clinical mediators, including BMI. GHR decreases at 3 months postoperatively alongside increased insulin-like growth factor binding proteins IGFBP1/BP2; plasma GH increased at 1 year. Experimental validation indicates (1) hepatic GHR expression decreases in post-bariatric rats; (2) GHR knockdown in primary hepatocytes decreases gluconeogenic gene expression and glucose production. Thus, RYGB may induce resistance to diabetogenic effects of GH signaling.Trial Registration: Clinicaltrials.gov NCT01073020.

Growth hormone signaling shapes the impact of environmental temperature on transcriptomic profile of different adipose tissue depots in male mice

Growth hormone receptor knockout (GHRKO) mice are smaller, long living and have an increased metabolic rate compared with normal (N) littermates. However, it is known that thermoneutral conditions (30°-32°C) elicit metabolic adaptations in mice, increasing the metabolic rate. Therefore, we hypothesized that environmental temperature would affect the expression profile of different adipose tissue depots in GHRKO mice. For this, N (n=12) and GHRKO (n=11) male mice were maintained at 23°C or 30°C from weaning until 11 months of age. RNA sequencing from adipose tissue depots (epididymal - eWAT, perirenal - pWAT, subcutaneous - sWAT and brown fat - BAT) was performed. Thermoneutrality increased body weight gain in GHRKO but not N mice. Only a few genes were commonly regulated by temperature in N and GHRKO mice. The BAT was the most responsive to changes in temperature in both N and GHRKO mice. BAT Ucp1 and Ucp3 expression were decreased to a similar extent in both N and GHRKO mice under thermoneutrality. In contrast, eWAT was mostly unresponsive to changes in temperature. The response to thermoneutrality in GHRKO mice was most divergent from N mice in sWAT. Relative weight of sWAT was almost four times greater in GHRKO mice. Very few genes were regulated in N mice sWAT when compared to GHRKO mice. This suggest that this WAT depot has a central role in the adaptation of GHRKO mice to changes in temperature.