Growth Hormone Receptor 1A mRNA Research Articles

Effect of pre- and postpartum supplementation with lipid-encapsulated conjugated linoleic acid on reproductive performance and the growth hormone–insulin-like growth factor-I axis in multiparous high-producing dairy cows

The objective of the study was to evaluate the effect of prepartum and postpartum (PP) supplementation with 2 isomers of conjugated linoleic acid (CLA) on reproductive parameters and some related metabolic factors in dairy cows. High-producing, multiparous Holstein Friesian cows (n = 60) were allotted to 3 treatment groups: the CLA1 group (n = 20) was supplemented with 70 g of lipid-encapsulated CLA providing 7 g each of cis-9,trans-11 and trans-10,cis-12 CLA from d 21 (d 21) before expected calving until d 7 after artificial insemination (AI), that is, until 77 to 91 d PP; the CLA2 group (n = 20) was supplemented with the same amount of CLA beginning at calving until d 7 after AI; and the control group (n = 20) received an isocaloric, isonitrogenous, and isolipidic diet. Blood samples were taken weekly to measure glucose, insulin, insulin-like growth factor-I (IGF-I), and leptin. Liver biopsy was performed in 10 cows per group for growth hormone receptor 1A and IGF-I mRNA analyses. At d 49 to 63 PP, ovulation was synchronized with the Pre-Synch protocol followed by fixed-time AI. Milk progesterone was monitored from calving until d 35 post-AI. Cows returning to estrus following AI were inseminated. Supplementation with CLA before calving improved the recovery of plasma leptin levels in the early PP period (from the day of calving until wk 3 PP; treatment effect). Later PP (wk 5), plasma IGF-I, and leptin remained significantly higher in both CLA1 and CLA2 groups compared with control, although hepatocellular IGF-I mRNA was not different among groups. Plasma IGF-I levels remained higher in both CLA-treated groups on the day of AI. Growth hormone receptor 1A mRNA levels in hepatic tissue decreased in all groups, reaching a nadir in the first week PP. Days to first PP ovulation did not differ between groups; however, both supplemented groups conceived earlier compared with control (d 97 ± 19, d 97 ± 23, and d 113 ± 30 for CLA1, CLA2, and control, respectively). Plasma progesterone concentration was higher in both supplemented groups on d 2 to 5 following the synchronized ovulation than in controls. We concluded that CLA supplementation around calving alters PP metabolic signals as reflected by higher plasma leptin and IGF-I levels. Conjugated linoleic acid stimulated early luteal function and reduced the PP interval to conception.

Variable liver fat concentration as a proxy for body fat mobilization postpartum has minor effects on insulin-induced changes in hepatic gene expression related to energy metabolism in dairy cows

The liver plays a central role in adaptation for energy requirements around calving, and changes in the effects of insulin on hepatic energy metabolism contribute to metabolic adaptation in dairy cows. Hepatic insulin effects may depend on body fat mobilization. The objective of this study was to investigate the effects of insulin on the hepatic gene expression of enzymes involved in energy metabolism and factors related to nutrition partitioning in cows with high and low total liver fat concentration (LFC) after calving. Holstein cows were retrospectively grouped according to their LFC after calving as a proxy for body fat mobilization. Cows were classified as low (LLFC; LFC <24% fat/dry matter; n = 9) and high (HLFC; LFC >24.4% fat/dry matter; n = 10) fat-mobilizing after calving. Euglycemic-hyperinsulinemic clamps [6 mU/(kg × min) of insulin for 6 h] were performed in wk 5 antepartum (ap) and wk 3 postpartum (pp). Before and at the end of the euglycemic-hyperinsulinemic clamps, liver biopsies were taken to measure the mRNA abundance of enzymes involved in carbohydrate and lipid metabolism, expression related to the somatotropic axis, and adrenergic and glucocorticoid receptors. The mRNA abundance of pyruvate carboxylase, cytosolic phosphoenolpyruvate carboxykinase (PEPCK; PCK1), acyl-CoA-dehydrogenase very long chain (ACADVL), and hydroxyl-methyl-glutaryl-CoA-synthase 1 increased, but the mRNA abundance of solute carrier family 2 (SLC2A2 and SLC2A4), growth hormone receptor 1A (GHR1A), insulin-like growth factor 1 (IGF1), sterol regulatory element binding factor 1, adrenoceptor α 1A, and glucocorticoid receptor decreased from ap to pp. Insulin treatment was associated with decreased PCK1, mitochondrial PEPCK, glucose-6-phosphatase, propionyl-CoA-carboxylase α, carnitine-palmitoyl-transferase 1A, ACADVL, and insulin receptor mRNA, but increased IGF1 and SLC2A4 mRNA ap and pp and GHR1A mRNA pp. The mRNA abundance of SLC2A4 was greater, and the mRNA abundance of GHR1A and IGF1 tended to be lower in LLFC than in HLFC. Administration of insulin, albeit at a supraphysiological dose, was associated with inhibition of gene expression related to glucose production and β-oxidation, but we observed variable effects in the degree of insulin depression of individual genes. Insulin status is important for regulation of nutrient partitioning, but different LFC pp had very little influence on changes in hepatic gene expression following administration of insulin.

Effects of Dietary Supplementation with Different Polyunsaturated Fatty Acids on Expression of Genes Related to Somatotropic Axis Function in the Liver, Selected Blood Indicators, Milk Yield and Milk Fatty Acids Profile in Dairy Cows

Abstract The objective of this study was to investigate whether dietary supplementation with different polyunsaturated fatty acids (PUFA s) affects expression of genes related to somatotropic axis and the plasma concentrations of insulin, glucose, non-esterified fatty acids (NEFA), beta hydroxyl butyrate acids (BHBA) and insulin-like growth factor 1 (IGF1) and milk fatty acids profile. Right after calving, Holstein cows (n=45) were randomly assigned to one of three diets supplemented with roasted whole soybean as a source of omega-6 PUFA (omega-6, n=15), linseed as a source of omega-3 PUFA (omega-3, n=15) or palm oil (control, n=15). Each cow was in the study over a period of 70 days. Blood samples were collected every two weeks from day 1 to 70 of lactation and plasma concentrations of insulin, glucose, NEFA, BHBA and IGF1 were determined. Liver samples were taken from a subset of 18 cows (6 per diet) at day 70 postpartum and hepatic mRNA level of total growth hormone-receptor 1A (GHR1A), insulin receptor (INSR), IGF1 and insulinlike growth factor binding protein (IGFBP2) was assessed. Experimental diets did not affect milk yield. Plasma glucose and insulin concentrations were greater for omega-3 treatment compared to omega-6 and control treatments. Cows fed diets enriched in omega-3 exhibited greater INSR and GHR1A mRNA expression, and a tendency for greater IGF1 mRNA expression in the liver compared to omega-6 and control cows. Plasma IGF1 concentration was significantly higher in omega-3 treatment compared with omega-6 and control treatments. Results of this study suggest that feeding omega-3 PUFA s during early postpartum couples with the somatotropic axis, leading to an increase in plasma IGF1 concentration in dairy cows.

Partial Feed Restriction Decreases Growth Hormone Receptor 1A mRNA Expression in Postpartum Dairy Cows

Uncoupling of the growth hormone (GH) axis in early postpartum dairy cows is correlated with a decrease in liver GH receptor (GHR) 1A mRNA and a decrease in liver GH receptor protein. Postpartum recoupling of the GH axis is also correlated with GHR 1A mRNA and GHR protein. We hypothesized that dry matter intake (DMI) partially controls the increase in GHR 1A mRNA postpartum. Prepartum Holstein dairy cows (n = 11) were offered feed ad libitum. After calving, 6 cows were fed 70% of their expected DMI (feed restriction) for 14 d and 5 cows were fed ad libitum (control). Both groups were fed ad libitum after d 14. Liver was biopsied prepartum and on d 1, 7, 14, and 21 postpartum; blood was sampled throughout the experimental period. Rate of increase in postpartum milk production was less for feed-restricted cows. The GHR 1A mRNA decreased from prepartum to d 1 postpartum and subsequently increased. Rate of postpartum increase in GHR 1A mRNA was less in feed-restricted cows. Diminished GHR 1A persisted for at least 7 d after feed-restricted cows returned to ad libitum feeding. Liver insulin-like growth factor-I mRNA concentrations decreased from prepartum to d 1 as well, but were similar for feed restricted and control thereafter. We concluded that DMI partially controls GHR 1A mRNA expression in early postpartum dairy cows and that the decrease in GHR 1A in response to feed restriction persisted for at least 1 wk after ad libitum feeding was restored.

Expression of Growth Hormone Receptor 1A mRNA is Decreased in Dairy Cows but not in Beef Cows at Parturition

The promoter controlling expression of a major bovine growth hormone (GH) receptor (GHR) mRNA variant, GHR 1A, contains a common DNA element for transcription factors hepatocyte nuclear factor 4α (HNF-4α), hepatocyte nuclear factor 4γ (HNF-4γ), and chicken ovalbumin transcription factor II (COUP-TFII). Expression of GHR 1A mRNA is decreased in the liver of dairy cows at parturition. The objective of this study was to determine whether reduced expression of GHR 1A mRNA in dairy cows at parturition is associated with changed expression of HNF-4α, HNF-4γ, or COUP-TFII mRNA. Liver biopsy samples were taken from multiparous Holstein cows 7 to 23 d before parturition, within 24h after parturition (i.e., at parturition), and 8 to 18 d after parturition, and the relative amounts of GHR 1A, insulin-like growth factor-I (IGF-I), HNF-4α, HNF-4γ, and COUP-TFII mRNA in these samples were measured by ribonuclease protection assays. As expected, expression of GHR 1A, total GHR, and IGF-I mRNA was decreased at parturition, compared with that detected prepartum or during the postpartum period. Expression of HNF-4α and COUP-TFII mRNA was unchanged, but that of HNF-4γ mRNA was increased at parturition. The same study was also conducted in multiparous Angus cows 7 to 23 d before parturition, at parturition, and 8 to 18 d after parturition. Neither expression of GHR 1A, total GHR, or IGF-I mRNA, nor expression of HNF-4α, COUP-TFII, or HNF-4γ mRNA was changed in the liver of beef cows at parturition. These results together suggest that, at the molecular level, decreased expression of GHR 1A mRNA in the liver of dairy cows at parturition may involve increased expression of HNF-4γ mRNA and that, at the systemic level, decreased expression of GHR 1A mRNA is not a direct result of the end of pregnancy, parturition, or the initiation of lactation.

Trait‐associated sequence variation in the bovine growth hormone receptor 1A promoter does not affect promoter activity in vitro

Growth hormone (GH) plays a central role in growth and metabolism in cattle by binding to growth hormone receptor (GHR) and stimulating production of insulin-like growth factor 1 (IGF1). Two sequence variations in the promoter transcribing a major GHR mRNA variant, GHR 1A mRNA, have been reported to be associated with quantitative differences in growth rate or blood concentration of IGF1 in cattle. One such variation is in the length of a TG-repeat, being 11 or 16-20; the other variation is in the nucleotide 155 bp upstream from the transcription start site, being G or A. In this study, we determined whether these sequence variations would affect the activity of GHR 1A promoter. We cloned GHR 1A promoters bearing different sequence variations and linked each of them to a reporter gene. Transient transfection analyses revealed that these promoter-reporter constructs did not differ in reporter gene expression. Cotransfection analyses demonstrated that they also did not differ in activation by hepatocyte nuclear factor 4alpha, hepatocyte nuclear factor 4gamma and nuclear receptor subfamily 2 group F member 2, known transcription factors for bovine GHR 1A promoter. These in vitro results, together with a previous observation that neither the nucleotide 155 bp upstream from the transcription start site nor the TG-repeat was part of the GHR 1A promoter region interacting with nuclear proteins from bovine liver, do not support a cause-effect relationship between the reported sequence variations and the associated changes in growth rate or blood IGF1 concentration in cattle.

Plasma Hormones and Expression of Growth Hormone Receptor and Insulin-Like Growth Factor-I mRNA in Hepatic Tissue of Periparturient Dairy Cows

Growth hormone plays a central role in the change in nutrient metabolism that occurs during the initiation of lactation. The actions of growth hormone are mediated by the growth hormone receptor (GHR) whose mRNA is present in three alternatively spliced forms (GHR 1A, 1B, and 1C). Liver-specific GHR 1A mRNA is transiently decreased around parturition, but the exact timing of the decline is not known. Our objective was to generate a daily profile for total GHR (GHRtot; all GHR transcripts), GHR 1A, and IGF-I mRNA expression in liver of periparturient Holstein cows and evaluate these daily mRNA profiles relative to daily profiles for periparturient hormones and metabolites. Liver biopsies and blood samples (n=139) were collected from 65 Holstein cows at the University of Missouri Dairy Farm. At least two cows were sampled on each day from 14 d before to 14 d after parturition. Total cellular RNA was isolated and reverse transcribed to cDNA. Target cDNA were measured by quantitative real-time polymerase chain reaction. Plasma was assayed for progesterone, estradiol, insulin, growth hormone, IGF-I, glucose, and nonesterified fatty acids. The GHR 1A mRNA declined 2 d before parturition, was lowest 3 to 4 d after parturition, and then increased. The IGF-I mRNA declined 1 d after parturition, was lowest 2 to 5 d after parturition and then increased. Total GHR mRNA was not affected by day. The decrease in GHR 1A mRNA was associated with a decrease in progesterone and an increase in estradiol shortly before parturition. A detailed profile of GHR 1A, IGF-I, and GHRtot mRNA expression during the periparturient period was provided. The decreases in GHR 1A and IGF-I during the transition period occurred immediately before (GHR 1A) or shortly after (IGF-I) parturition. Rapid changes in placental and ovarian steroids before parturition were coincident with changes in GHR 1A mRNA.

Effects of BST on Oviductal and Uterine Genes Encoding Components of the IGF System in Lactating Dairy Cows

Lactating Holstein cows, averaging 80 d in milk, were used to examine effects of exogenous bovine somatotropin (bST) on oviductal and uterine genes encoding components of the insulin-like growth factor (IGF) system. About 12h before expected ovulation in an Ovsynch protocol, cows were assigned randomly to receive bST (500mg; n = 11) or serve as untreated controls (n = 10). Cows that ovulated (n = 9 bST, 8 control) were divided within treatment to be sacrificed on d 3 or 7 postovulation. Samples of oviductal and intercaruncular endometrial tissue from oviducts and uterine horns ipsilateral to the corpus luteum (CL) were collected and immediately frozen at−80°C for subsequent mRNA analyses. Northern blots revealed mRNAs for IGF-II, IGF-binding protein-2 (IGFBP-2), and IGFBP-3 in all oviductal and endometrial tissues. Significant amounts of IGF-I and growth hormone receptor-1A (GHR-1A) mRNAs were detected in uteri but not in oviducts. The bST treatment had no effect on amount of IGF-I mRNA transcript in uterine endometrium. The mRNA encoding IGF-II was induced by bST in oviducts collected on both d 3 and 7 but was down-regulated in endometrium on d 7. Transcript of IGFBP-2 mRNA was greater in endometrial than oviductal tissues and did not differ between treatments. Both oviductal and endometrial IGFBP-3 mRNA concentrations increased between d 3 and 7 postovulation, with a tendency for greater endometrial IGFBP-3 mRNA in bST-treated cows on d 7. On d 7, concentrations of endometrial GHR-1A mRNA were 30% lower in bST-treated cows. Results indicate complex and tissue-specific regulation of the uterine IGF system components by exogenous bST. Some of those biological responses to bST may be important in early development of bovine embryos.

Reduced Insulin-Like Growth Factor-I after Acute Feed Restriction in Lactating Dairy Cows is Independent of Changes in Growth Hormone Receptor 1A mRNA

Growth hormone receptor (GHR) and insulin-like growth factor-I (IGF-I) mRNA decrease in the liver of dairy cows at parturition. Epinephrine is released and feed intake is decreased at the same time. The objective of this study was to determine whether feed restriction and (or) administration of epinephrine could recapitulate the changes in the hepatic GHR 1A and IGF-I mRNA that occur at parturition. Eight lactating cows were randomly assigned to one of two rations (ad libitum or restricted). The cows assigned to the restricted ration were fed 75, 60, 60, and 25% of feed consumed by the ad libitum cows on successive days to mimic the changes in feed intake around parturition. Liver samples were collected by needle biopsy before and after feed restriction. Cows received either 0.02 mg/kg of epinephrine or saline approximately 24h before the second liver biopsy so that a 2 × 2 factorial arrangement of treatments was created. Feed restriction increased plasma nonesterified fatty acids and liver triglyceride percentages and decreased plasma IGF-I concentrations. The decrease in plasma IGF-I after feed restriction was associated with a decrease in IGF-I mRNA in feed-restricted cows. The amount of GHR 1A mRNA did not change after feed restriction. Epinephrine treatment did not affect the amount of GHR 1A or IGF-I mRNA. We conclude that the decrease in feed intake at parturition may be partially responsible for the decrease in IGF-I but may not cause the decrease in GHR 1A.

A novel phenotype for Laron dwarfism in miniature Bos indicus cattle suggests that the expression of growth hormone receptor 1A in liver is required for normal growth☆

Mutations within the growth hormone receptor (GHR) gene that lead to an inactivated or truncated GHR protein cause abnormal growth and small adult size in a variety of species (Laron dwarfism). We studied a line of miniature Bos indicus cattle that have phenotypic (small mature size) and endocrine (increased blood growth hormone and decreased blood insulin-like growth factor-I concentrations) similarities to Laron dwarfs. Liver mRNA from miniature and control cattle was used to amplify a cDNA within the coding region of the GHR. The miniature cattle had GHR mRNA size (determined by Northern blot) and cDNA sequence that were similar to control cattle and, therefore, were unlike most Laron dwarf genotypes in which the GHR gene is mutated. Amounts of mRNA from liver as well as muscle (superficial neck and longissimus) were analyzed by ribonuclease protection assay for IGF-I, total GHR, GHR 1A (inducible, liver-specific GHR mRNA), and GHR 1B (constitutive GHR mRNA). Four control and five miniature bulls were tested. As expected, liver IGF-I mRNA was decreased in the miniature cattle (approximately 12% of control; P < 0.01). The amount of the total GHR as well as GHR 1A mRNA were also decreased in liver (17% and 19% of control, respectively; P < 0.01). Other GHR mRNA, including GHR 1B mRNA, were similar for miniature and control cattle. In muscle, there was a tendency (P < 0.10) for decreased IGF-I mRNA and increased GHR mRNA in miniature compared with control cattle. In summary, a novel phenotype for Laron dwarfism in Bos indicus cattle was associated with underexpression of GHR 1A mRNA, but not other GHR mRNA variants in liver. In addition to decreased GHR 1A mRNA, the miniature cattle had decreased liver IGF-I mRNA. Full expression of GHR 1A in liver, therefore, may be required for full liver IGF-I expression and normal growth.

Reduced growth hormone receptor (GHR) messenger ribonucleic acid in liver of periparturient cattle is caused by a specific down-regulation of GHR 1A that is associated with decreased insulin-like growth factor I.

GH receptor (GHR) messenger RNA (mRNA) is transcribed from at least three different promoters within the liver of cattle. The first promoter (P1) is liver specific and alternatively splices exon 1A onto the GHR mRNA (GHR 1A mRNA). The second and third promoters (P2 and P3) have constitutive activity in many tissues and alternatively splice exons 1B and 1C onto the GHR mRNA (GHR 1B and GHR 1C mRNA). The total amount of GHR in the liver partially determines liver insulin-like growth factor I (IGF-I) synthesis in response to GH. Two studies were conducted to characterize the changes in GHR 1A mRNA, alternatively spliced GHR mRNA, and IGF-I mRNA during late pregnancy and early lactation in dairy cattle. Liver RNA was isolated from pregnant Holstein cattle (Bos taurus) on days -14, 0, and 21 relative to parturition (study 1) or days -14, 0, 15, 30, 60, and 90 relative to parturition (study 2). Ribonuclease protection assays were used to quantify total GHR (all GHR variants) as well as liver-specific GHR 1A and alternatively spliced GHR mRNA. Likewise, total IGF-I as well as alternatively spliced IGF-I mRNA (class 1 and class 2 transcripts) were measured. A decrease in total GHR mRNA at parturition (P < 0.01) was associated with a specific decrease in GHR 1A mRNA (P < 0.001). The amount of alternatively spliced GHR mRNA (including GHR 1B and GHR 1C mRNA) did not change at parturition (P > 0.10). Total liver IGF-I mRNA and blood IGF-I concentrations were also decreased at parturition (P < 0.05 and P < 0.01, respectively). However, a decrease in IGF-I mRNA was observed for both class 1 and class 2 IGF-I transcripts (P < 0.01 and P < 0.05, respectively). We conclude that the reduced amount of GHR mRNA during early lactation is caused by a specific down-regulation of GHR 1A mRNA that was associated with decreased liver IGF-I mRNA and decreased blood IGF-I concentrations. These data provide evidence for independent regulation of GHR mRNA by mechanisms that discriminate between GHR P1 (transcribes GHR 1A) and alternative promoters that transcribe constitutive GHR mRNA.

Expression of growth hormone receptor 1A messenger ribonucleic acid in liver of dairy cows during lactation and after administration of recombinant bovine somatotropin.

The mRNA for growth hormone receptor is transcribed from at least three different promoters in cattle. The first promoter (P1) is liver-specific and transcribes growth hormone receptor mRNA containing exon 1A (growth hormone receptor 1A). The second and third promoters (P2 and P3) are active in a variety of tissues and transcribe growth hormone receptor mRNA containing exon 1B and 1C. The objective was to characterize P1 activity by measuring the amount of growth hormone receptor 1A mRNA in liver of dairy cows at different stages of lactation as well as after administration of recombinant bovine somatotropin (rbST). In study 1, liver RNA was isolated from Holstein cows during the dry period (nonlactating, n = 6) and during early (n = 6), mid (n = 6), and late (n = 11) stages of lactation. Six of the late-lactation cows received injections of rbST (25 mg/d) for 7 d prior to collection of liver tissue. In study 2, lactating Holstein cows received either no infusion (control, n = 10) or continuous infusion of rbST (29 mg/d, n = 10) for 63 d. The amount of growth hormone receptor 1A mRNA was decreased in early- and mid-lactation cows compared with late-lactation cows or nonlactating cows (study 1). Administration of rbST increased growth hormone receptor 1A mRNA (studies 1 and 2). The total amount of growth hormone receptor transcribed from alternative promoters (growth hormone receptor P2 and P3) remained unchanged during different stages of lactation or in response to rbST. We conclude that changes in liver growth hormone receptor mRNA in lactating dairy cattle primarily depend on growth hormone receptor P1 activity.