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.

Detection of GHR|AluI gene polymorphism and its association with body weight of Madura cattle in Indonesian Beef Cattle Research Station

Madura cattle is a one of the native cattle that are genetically tolerant of tropical climates. Madura cattle are one of the beef cattle breeds in Indonesia. Madura cattle can be a superior breed, but improving growth trait in Madura cattle needs to be conducted. The early step that was needed was for detecting the genes which were involved in growth traits. The purpose of this research was for detecting GHR gene polymorphisms and its association with body weight of Madura cattle population in Indonesian Beef Cattle Research Station. This study used 110 DNA samples of Madura cattle that was collected from the experimental barn at Indonesian Beef Cattle Research Station. DNA samples were extracted by DNA extraction kit. Genotype of GHR gene was detected by PCR-RFLP method using AluI restriction enzymes. In this study, association genotype and body weight was analysed by univariate GLM method. The birth weight, weaning weight, yearling weight and 18 months weight of Madura cattle were 16.8±0.3 kg; 82.4±2.3 kg; 124.6±3.7 kg and 166.7±5.0 kg, respectively. The GHR genes of Madura cattle had 3 genotypes AA, AG and GG and genotype frequencies of GHR gene were 0.273 (AA), 0.373 (AG), 0.355 (GG), respectively. Whereas the allele frequencies of A was 0.459 and the allele frequencies of G was 0.541. Based on Chi-square (X2) analysis showed that the population sample was not disequilibrium. The result of association analysis was significant (P <0.05) on weaning weight, yearling weight and 18 month weight. It was concluded that the GHR gene has potential as a genetic marker for growth traits and can be used as MAS in Madura cattle in Indonesian Beef Cattle Research Station.

Determination of the association of <i>GHR</i>/<i>AluI</i> gene polymorphisms with milk yield traits in Holstein and Jersey cattle raised in Turkey

This research was carried out to determine the effect of a specific single nucleotide polymorphism (SNP) region in exon 10 of the growth hormone receptor (GHR) gene on milk production traits in Jersey and Holstein cows raised in Turkey. Milk samples were recorded as a test day milk yield (TDMY) and an adjusted based 305 d milk yield (305-DMY). Also, milk component traits were detected. Based on the scope of this study, a total of 748 dairy cows, including 305 Holsteins raised in the Marmara Region and 163 Holstein and 280 Jersey raised in the Black Sea Region, were genotyped for the GHR gene using the RFLP-PCR technique. Jersey cows carrying the GG genotype (5.24 %) were associated with higher fat content (P<0.05). Jersey cows with GG and AG also had a higher protein content (3.44 % and 3.38 %, respectively) (P<0.05). Similarly, the protein content was the highest in Holstein cows with the GG genotype (3.46 %) (P<0.01), whereas Holstein cows having AA genotypes displayed higher TDMY (24.64 kg/d) (P<0.05) and 305-DMY (8472.4 kg) (P<0.01). The estimated increase in milk protein and fat contents due to the G allele was 0.07 % and 0.22 % in the Jersey breed, respectively. On the other hand, allele A was highly related to an increase in protein yield and 305-DMY of 0.04 and about 675 kg in the Holstein breed, respectively. The GHR gene should be considered as a potential candidate gene in marker-assisted selection programs to improve the performance of milk and related traits in Turkey dairy cattle populations.

Growth Hormone Receptor Gene Disruption in Mature-Adult Mice Improves Glucose Metabolism and Lifespan in Females

Growth hormone (GH) serves an important role in early and adult life. Reduction of GH action has been shown to increase life span in many species of animals. In fact, mice bearing a congenital disruption of GH receptor (GHR) gene (GHRKO) hold the record for the longest-lived laboratory mice. In addition to extended life span, these mice show improved health with lower rates of cancer, increased insulin sensitivity, and resistance to age-associated cognitive decline. Furthermore, humans with decreased GH action due to inactivating mutations in the GHR (Laron Syndrome patients) are resistant to cancer and diabetes. Even though the beneficial effects of congenital Ghr gene disruption are well studied, the consequences of postnatal disruption of GH action were unknown. Previously our laboratory generated a mouse line with disrupted GH action at 1.5 months of age (1.5mGHRKO mice). Results showed that these mice had improved insulin sensitivity and increased maximal lifespan only in females, yet growth retardation was still present. To consider decreased GH action as a possible therapeutic to extend healthy lifespan, it was imperative to elucidate the effects of disrupting Ghr gene at a mature-adult age, well after the developmental and growth period of the mice. To this end, we hypothesized that removal of GH action in adult life would convey some of the same health and life span benefits seen in the GHRKO mice without the reduced body length. To test this hypothesis, we used the cre-lox system to generate mice with a disrupted Ghr gene at a mature-adult age (6 months), referred as 6mGHRKO mice. We then performed a phenotypic and lifespan characterization, and tested for molecular mechanisms known to be associated with extended longevity, namely oxidative stress resistance and mTOR modulation. We found that similar to GHRKO and 1.5mGHRKO mice, disruption of GHR at 6 months of age resulted in mice with increased adipose tissue mass, decreased lean mass, high circulating GH, but decreased insulin growth factor-1 levels compared to control mice. Furthermore, the 6mGHRKO mice displayed significantly improved insulin sensitivity in males, with no changes in glucose tolerance. Also, serum levels of inflammatory markers and liver triglycerides were unchanged in these mice. Experiments to evaluate the status of oxidative damage and mTOR activation in liver, skeletal muscle and subcutaneous adipose tissue of male and female 6mGHRKO mice showed a tissue-specificity and sexual dimorphism in these results. Importantly, male and female 6mGHRKO mice showed no change in body length, but mean, median and maximal lifespan were significantly extended in females. In conclusion, disruption of GH action well past sexual maturation produces beneficial effects on insulin sensitivity and aging in mice. Acknowledgements: This work is supported by the State of Ohio’s Eminent Scholar Program, by NIH grant AG059779 and by the AMVETS.

Expression of IGF-1, IGF-1 Receptor and Growth Hormone Receptor in Hepatic Tissue in Adults Across the Spectrum of Nonalcoholic Fatty Liver Disease (NAFLD)

Background: Obesity is a state of relative growth hormone (GH) and insulin-like growth factor-1 (IGF-1) deficiency, and the GH/IGF-1 axis has been implicated in the pathophysiology of nonalcoholic fatty liver disease (NAFLD) and the progression to steatohepatitis (NASH) in preclinical models and human studies. GH has both lipolytic and anti-inflammatory properties while IGF-1 has been implicated in reducing hepatic fibrosis and promoting hepatic regeneration. The GH/IGF-1 axis may be a therapeutic target in NAFLD/NASH, however, IGF-1, IGF-1 receptor (IGF-1R) and GH receptor (GHR) expression in adult human hepatic tissue has not been studied across the spectrum of disease severity. Methods: We quantified IGF-1, IGF-1R, and GHR gene expression in hepatic tissue from 318 adults with obesity using the Nanostring nCounter assay. Subjects were classified into four categories of disease severity based on histopathology: normal liver histology (NLH) (n=76, 24%), steatosis only (Steatosis) (n=88, 28%), NASH without fibrosis (NASH F0) (n=72, 23%), and NASH with fibrosis (NASH F1-F4) (n=82, 26%). Gene expression analysis is presented as normalized gene counts by group with p-value of the generalized linear model controlled for age, sex and BMI. Results: Mean (±SD) age (whole cohort 44.0±12 years) and BMI (whole cohort 46.8±7.2 kg/m2) did not differ across groups (p=0.2 for both). ALT was higher with increasing disease severity (NLH 30.1±26.7, Steatosis 31.9±15.7, NASH F0 35.7±16.5, NASH F1-4 48.4±34.9, p&lt;0.001). IGF-1 gene expression was lower in all NAFLD/NASH groups compared to the NLH reference group (NLH 485.4±292.7; Steatosis 396.3±238.0, p=0.04; NASH F0 349.8±220.1, p=0.01 and NASH F1-4 341.2±268.6, p=0.03, all p-values vs NLH). There was no difference in IGF-1R or GHR gene expression across disease severity groups (IGF-1R NLH 43.3±10.2, Steatosis 41.4±11.6, NASH F0 38.8±8.8 and NASH F1-4 39.1±8.1, p&gt;0.05 between any disease state; GHR NLH 6382±2366, Steatosis 6544±2699, NASH F0 7220±2542 and NASH F1-4 5997±2352, p&gt;0.05 between any disease state). Conclusion: We demonstrated that IGF-1 gene expression was lower in liver tissue from patients with NAFLD and NASH than healthy controls. This is consistent with our prior finding that histologic NASH and fibrosis are associated with lower serum IGF-1 levels. Moreover, we demonstrated that hepatic IGF-1R and GHR gene expression is not lower in liver tissue from patients with NAFLD and does not decline across disease severity. This reinforces our prior finding that GHR staining intensity and zonality by immunohistochemistry does not change with increasing disease severity in NAFLD/NASH. These data demonstrate that the GH axis is relatively suppressed but that expression of GHR and IGF-1R receptors is stable with worsening disease severity in NAFLD/NASH, suggesting that GH augmentation may be a viable therapeutic target in NAFLD.

Keragaman Gen Growth Hormone Receptor (GHR) Ekson 10 pada itik Sikumbang Janti

Growth Hormone Receptor (GHR) gene play important roles in duck performances due to their crucialfunctions in growth and has been considered as a candidate gene for growth traits. This reasearch wasaimed to indification polymorphism of Growth Hormon Receptor (GHR) gene in Sikumbang Janti duckand this study used 68 blood duck. Polymerase chain reaction (PCR) and sequencing analysis wereconducted to detect SNP existence in exon 10 of GHR gene. The result of this study showed that wasfound two SNP at g.1112 A>G and g.1304 C>T with transision mutation in exon 10 GHR gene. Genotypesfound in the g.1112 A>G were AA and GG with types of Allele A and G. Genotypes found in the g.1304T>C the were TT and CC with types of Allele T and C. Polymorphism of GHR gene were polymorphicand Population of Sikumbang Janti duck in Hardy-Weinberg equilibrium. It could be concluded thatGHR gene has potential to can be marker genetic in Sikumbang Janti ducks.

Association analysis of novel SNPs in exon 10 of the growth hormone receptor gene with growth traits in Indian sheep

The growth hormone receptor (GHR) gene encodes the type I cytokine receptor that helps in joining the growth hormone to this receptor, thus promoting receptor-dimerization, leading to up-regulating growth. The ovine GHR gene located on chromosome 16, which consists of 10 exons and 9 introns, along with untranslated regions on either side, comprise a total size of 178.09 kb. However, earlier reports about polymorphism have mainly dealt with exon 10 which is also a larger fragment of this gene comprising 1102 bp. Hence, this study was carried out to detect polymorphism in exon 10 of the GHR gene and its association with growth traits. Genomic DNA was isolated from blood samples of Madras Red and Mecheri sheep breeds from India. Part of exon 10 (895 bp) of the GHR gene was amplified and sent for sequencing. The sequence analysis revealed transition of nucleotide G>A at loci G177624A and G177878A in both sheep breeds. Populations were screened by Tetra-primer ARMS-PCR. The genotype frequencies of GG, GA and AA were 0.276, 0.519 and 0.205 at 177624 G>A, and 0.307, 0.444 and 0.149 at 177878 G>A in Madras Red sheep; whereas in Mecheri they were 0.476, 0.372 and 0.152 at 177624 G>A, and 0.629, 0.314 and 0.057 at 177878 G>A, respectively. Likewise the estimated allele frequencies of G and A were 0.5355 and 0.4645 at 177624 G>A, and 0.5790 and 0.4210 at 177878 G>A in Madras Red sheep; whereas in Mecheri they were 0.6620 and 0.3380 at 177624 G>A, and 0.7860 and 0.2140 at 177878 G>A, respectively. The effect of sex was significant for birth, six and nine month weight; but non-significant for three and 12 month weight in Mecheri sheep. However, in the Madras Red breed the effect of sex was significant for all body weights except weaning weight. The effect of variations on growth traits, viz., birth weight, weight at weaning, and weight at six, nine and twelve months in both breeds were analysed for their association, and they were found non-significant. Since these SNPs are salient findings of GHR gene polymorphism in Indian sheep breeds, further investigation is required into the significant effects of these novel SNPs, which could be useful for genetic improvement based on marker assisted selection.

The Role of Growth Hormone Receptor Isoforms and Their Effects in Bone Metabolism and Skeletal Fragility

Acromegaly and Growth Hormone Deficiency (GHD) are associated with skeletal fragility and with an increased prevalence of Vertebral Fractures (VFs). In the most recent years, several authors tried to investigate surrogate markers that may predict the risk of bone fragility in these endocrine disorders. The aim of this review is to evaluate the role of GH receptor polymorphisms in skeletal fragility in patients affected by GHD and acromegaly. In fact, until now, two different isoforms of the GH Receptor (GHR) were described, that differ for the presence or the absence of transcription of the exon 3 of the GHR gene. Both the isoforms produce a functioning receptor, but the exon 3-deleted isoforms (d3-GHR) has a higher sensitivity to endogenous and recombinant GH as compared to the full-length isoform (fl-GHR).