Insulin-like Growth factor-I Gene Research Articles

Nucleotides supplementation (Nucleoforce fish™) in Caspian roach (Rutilus caspicus) diet: Growth performance, skin mucosal immune response, and resistance to salinity stress

In this study, the growth, epidermal mucosal immunity, expression of growth-related genes, cross-protection, and resistance to salinity stress of Caspian roach were scrutinized in response to dietary levels of nucleotides (NT). Accordingly, 1200 fish (0.51 ± 0.01 g) were fed ad libitum with a basal diet (38.88 % crude protein and 10.04 % crude lipid in dry basis) containing incremental levels of NT at 0 (NT-0; control), 0.3 g kg−1 (NT-0.3), 0.6 g kg−1 (NT-0.6), and 1.2 g kg−1 (NT-1.2) for 8 weeks in triplicates. The growth performance was significantly increased in the fish fed with NT-0.6 and NT-1.2 diets compared to the control group (p < 0.05). A significant elevation in the growth hormone and insulin-like growth factor-I gene expression was recorded in NT-added groups at 0.6 and 1.2 g kg−1 compared to the control group (p < 0.05). In contrast to the control group, feeding on NT-0.6 and NT-1.2 diets had a remarkable effect on the skin mucus soluble protein and immunoglobulin levels (p < 0.05). After the feeding trial, we examined how salinity stress (15 g/l salinity) lonely and salinity stress under non-lethal thermal shock (+10 °C) affected heat shock protein (HSP70). Then, the mRNA expression of HSP70 gene from the gill was analyzed at 0, 2, 8, and 24 h post-challenge tests. The HSP70 gene expression level was approximately up-regulated more than 2-fold in NT-6 and NT-1.2 treatments compared to the control group under the salinity stress. Altogether, this research represents that the addition of NT at 0.6 and 1.2 g kg−1 in Caspian roach diet can improve overall performance and resistance to salinity stress.

An investigation of the assumed efficacy of methionine hydroxy analogue compared to DL-methionine by measuring growth performance, carcass traits, and GHR and IGF-I expression in broilers

This experiment was conducted to investigate the assumed efficacy of 65% on the product basis of methionine hydroxy analog (OH-Met, 88% aqueous solution of DL-2-hydroxy-4-(methylthio) butanoic acid) relative to DL-methionine (DL-Met) in broiler chickens. A total of 792-day-old male chickens were randomly allotted to 7 dietary treatments consisting of a methionine deficient-basal diet based on corn-soybean meal and 6 different diets obtained by supplying the basal diet with methionine at 3 levels (25, 100 and 125%; levels of addition relative to the required amount of additional Met to meet SID Met + Cys requirements) from either DL-Met or OH-Met at a ratio of 65:100 (DL-Met to OH-Met on a product basis) in the corresponding treatments. Regardless of the source, methionine supplementation enhanced (p < .05) growth performance, carcase and breast meat yield. Supplemental level and the source of Met had no significant effect on the proximate chemical composition of breast meat (p > .05). Comparison of effects of OH-Met and DL-Met supplemented at 100–65% weight ratio revealed no differences at any Met + Cys level and for performance criterion confirming the applied concept. Expression of growth hormone receptor (GHR) mRNA, which showed a positive correlation with body weight gain, carcase yield and breast percentage, in broiler liver significantly increased with 100% and 125% relative addition of DL-Met, while only 100% of OH-Met addition did (p < .05) when compared to the basal diet. The expression of the insulin-like growth factor-I (IGF-I) gene was not significantly affected by either Met source or supplementation level. In conclusion, our data indicated that DL-Met can be substituted with a 1.54 times higher amount of OH-Met in corn-soybean meal based broiler diets.

Effects of age and seasonal temperatures on cortisol levels and GHR, IGF-I, and IGF-II expressions in rainbow trout (Oncorhynchus mykiss)

The growth hormone (GH)/insulin-like growth factor (IGF) endocrine axis regulates the cellular growth and organ development in related to changing environmental conditions and age. The aim of this study is to determine growth factor genes, which are biological markers of growth in muscle and liver of rainbow trout (Oncorhynchus mykiss) in different seasonal temperatures and age ranges and the reveal of the relationship between serum cortisol (COR) levels from stress parameters. No difference was found in serum COR levels between the groups with respect to temperature and age. Serum GH levels were found to be higher in the summer juvenile trout compared to the winter juvenile and adult trouts. There was no difference in liver growth hormone receptor (GHR) mRNA levels of juvenile and adult trouts in winter while insulin-like growth factor-I (IGF-I) and insulin-like growth factor-II (IGF-II) mRNA levels in liver and muscle tissues were found to be higher in juvenile trout compared to adults. Among trout of different ages, GHR, IGF-I and IGF-II mRNA levels in liver in summer were higher for juvenile trout compared to adults. Muscle IGF-I mRNA levels in summer were higher in adult trout compared to juveniles. IGF-II mRNA levels in liver and muscle tissues of juvenile trout showed an increase in winter compared to in summer. While the GHR and IGF-II mRNA levels in the liver tissue of adult trout were observed to higher in winter compared to summer, IGF-I mRNA levels were found to be higher in summer. GHR, IGF-I and IGF-II mRNA levels in muscle tissue of adult trout were found to be higher in summer than in winter. This study indicated that juvenile and adult rainbow trout (Oncorhynchus mykiss) are adapted to both winter and summer temperature and that GHR, IGF-I and IGF-II genes are highly expressed.

Bone-Derived IGF-I Regulates Radial Bone Growth in Adult Male Mice.

Insulin-like growth factor-I (IGF-I) levels, which are reduced by age, and cortical bone dimensions are major determinants of fracture risk in elderly subjects. Inactivation of liver-derived circulating IGF-I results in reduced periosteal bone expansion in young and older mice. In mice with lifelong depletion of IGF-I in osteoblast lineage cells, the long bones display reduced cortical bone width. However, it has not previously been investigated whether inducible inactivation of IGF-I locally in bone in adult/old mice affects the bone phenotype. Adult tamoxifen-inducible inactivation of IGF-I using a CAGG-CreER mouse model (inducible IGF-IKO mice) substantially reduced IGF-I expression in bone (-55%) but not in liver. Serum IGF-I and body weight were unchanged. We used this inducible mouse model to assess the effect of local IGF-I on the skeleton in adult male mice, avoiding confounding developmental effects. After tamoxifen-induced inactivation of the IGF-I gene at 9 months of age, the skeletal phenotype was determined at 14 months of age. Computed tomography analyses of tibia revealed that the mid-diaphyseal cortical periosteal and endosteal circumferences and calculated bone strength parameters were decreased in inducible IGF-IKO mice compared with controls. Furthermore, 3-point bending showed reduced tibia cortical bone stiffness in inducible IGF-IKO mice. In contrast, the tibia and vertebral trabecular bone volume fraction was unchanged. In conclusion, inactivation of IGF-I in cortical bone with unchanged liver-derived IGF-I in older male mice resulted in reduced radial growth of cortical bone. This suggests that not only circulating IGF-I but also locally derived IGF-I regulates the cortical bone phenotype in older mice.

In Vitro Investigation of Insulin-Like Growth Factor-I and Mechano-Growth Factor on Proliferation of Neural Stem Cells in High Glucose Environment

Objective: High glucose levels cause metabolic and vascular complications by affecting the nervous system with an increased risk of stroke and seizures. There are still no effective treatment modalities for the high morbidity and mortality rates detected in such situations. Neural stem cells (NSCs) provide neurogenesis in the nervous system, but high glucose is detrimental to NSCs. This study investigates the intrinsic and extrinsic effects of the growth factors insulin-like growth factor-I (IGF-I) and mechano-growth factor (MGF) on NSCs when exposed to high glucose levels. Thus, the possibility of new treatment options for diabetes patients is explored. Materials and Methods: Rat NSCs grown in cell culture conditions were exposed to a control glucose concentration of 17.5 mM and high concentrations of 27.75, 41.75, and 83.75 mM for 24 h. The high glucose concentrations were designed to recapitulate the in vivo conditions of diabetes mellitus, diabetic ketoacidosis, and hyperglycemia hyperosmolar status. Then, 0.2 μg/ml IGF-I and MGF growth factors were separately added and their expressions in the NCSs investigated by real-time reverse transcription-polymerase chain reaction. The effects of exogenous IGF-I and MGF administration on NSC proliferation under high glucose conditions were measured by BrdU incorporation assay using flow cytometry analysis. Results: A significant increase was detected in the relative gene expression fold changes of IGF-I and MGF in the NSCs. The MGF relative fold change was greater than the IGF-I for each high glucose condition. NSCs exposed to 27.75 mM glucose revealed a 17-fold and 40-fold increase in the IGF-I and MGF gene expressions, respectively; the 41.75 mM glucose similarly revealed 68-and 161-fold increases and the 83.75 mM glucose 75-and 137-fold increases. Exogenous IGF-I administration increased its expression profile, while the administration of MGF lowered its expression. The NSC was in the growth (G0/G1) phase of the cell cycle during the 24 h culture time. The percentage of proliferated NSC decreased to 89% (17.5 mM), 85% (27.75 mM), 50.30% (41.75 mM), and 28.97% (83.75 mM). Surprisingly, the increase in both IGF-I and MGF saved the NSCs from cell death. Conclusion: Exogenous IGF-I and MGF administrations via high glucose environments increased NSC proliferation at the time of injury and protected the NSCs from cell death. The neuroprotective effect of MGF was greater than that of IGF-I. Thus, due to their neurogenesis potential, exogenous IGF-I and MGF could be applied in the treatment of diabetes patients to relieve neural damage.

The growth performance, antioxidative status and muscle quality of grass carp (Ctenopharyngodon idellus) cultured in the recirculating pond aquaculture system (RPAS)

The recirculating pond aquaculture system (RPAS) consisted of culture ponds, a purification pond, inflow ditch, outflow pipe, ecological ditch, and electrically driven pump. The performance of RPAS on grass carp (Ctenopharyngodon idellus) was explored in this study. In both the RPAS and the traditional pond aquaculture system (TPAS), water quality, fish growth performance, muscle quality, and tissue biochemical properties were assessed. Pond water was exchanged every seven days in RPAS and treated using a purification system that included an ecological ditch and a purification pond with a floating bed and aquatic plant. In RPAS, nitrate‑nitrogen, ammonia‑nitrogen, and nitrite‑nitrogen concentrations were lower than in TPAS. Fish cultured in RPAS exhibited significant higher growth rate and lower feed conversion rate. A significant increase in the expression of growth-related genes of myogenin (MyoG), myofactar-5 (Myf-5), insulin-like growth factor-I gene (igf-1), insulin-like growth factor-II gene (igf-2), and growth hormone receptor gene (ghr) were found in the fish cultured in the RPAS ponds. The content of malondialdehyde in either serum or liver and hepatic protein carbonyls content declined significantly in the RPAS ponds. The activities of T-SOD and GSH in liver and GSH in serum elevated significantly in RPAS fish. Muscular histological characteristics and texture profiles did not alter between two groups. The muscular contents of aromatic amino acids of aspartic acid and glutamic acid were significantly higher in RPAS than those in TPAS. It is inferred that RPAS is beneficial to promote fish growth, decrease oxidative stress, and increase muscle umami flavor.

High association of a novel variant in the adiponectin gene with the litter size in Awassi ewes

Flock productivity is affected by reproductive traits controlled by many factors such as ovulation rate, hormones, and genetics. Many genes are associated with fertility and other features related to it such as melatonin receptor 1A ( MNTR1A ) gene, inhibin βA ( INHBA) gene, insulin-like growth factor-I ( IGF-1 ) gene, and prolactin gene, and many other new candidates, adiponectin gene ( ADIPOQ ) is one of them, which are associated with reproduction and fertility. This study aimed to evaluate the genetic variation of the ADIPOQ gene concerning the litter size of Awassi ewes. Genomic DNA isolated from 114 Awassi ewes produced single lambs and 84 Awassi ewes produced twin lambs. By polymerase chain reaction (PCR), we amplified all three exons (368 bp, 448 bp, and 363 bp) of the ADIPOQ gene (exons 1, 2, and 3). Three genotypes were found in the 368 bp amplicons, the CC, CA, and AA genotypes. Novel mutation c.198473337C > A was discovered in the CA genotype through a sequencing reaction. This single nucleotide polymorphism (SNP) showed a highly significant ( P ≤ 0.01) association with litter size, in that the sheep with this SNP had a larger litter size than those without it. The ewes with the AA genotype had 1.97 lambs more than those with the CC and CA genotypes. Using logistic regression, the c.198473337C > A SNP was confirmed to be responsible for increasing the litter size. In conclusion, the c.198473337C > A SNP appears to have an interesting and positive impact on reproductive traits (especially litter size) in Awassi sheep. In light of these data, the genetic variant of the ADIPOQ gene could be used for developing marker-assisted selection programs to improve litter size in Awassi sheep.

Exploring single nucleotide polymorphisms in GH, IGF-I, MC4R and DGAT1 genes as predictors for growth performance in dromedary camel using multiple linear regression analysis

Understanding the genetic factors influencing the growth traits of animal is very important for an accurate selection and genetic improvement of these quantitative traits. Therefore, the aim of this study was to identify polymorphisms in growth hormone (GH), insulin-like growth factor-I (IGF-I), melanocortin-4 receptor (MC4R) and diacylglycerol O-acyltransferase 1 (DGAT1) genes and their association with body weight and daily gain in dromedary camels. Blood samples were collected from seventy camels using vacutainer tube containing EDTA as an anticoagulant for DNA extraction. PCR-DNA sequencing approach was performed to investigate polymorphisms in fragments for 5’ UTR of GH (286-bp) and part of coding sequence (CDS) of IGF-I (462-bp), MC4R (512-bp), and DGAT1 (390-bp) genes. Results revealed that there were single nucleotide polymorphisms (SNPs) in the investigated genes between the enrolled camels except for IGF-I gene that elicited a monomorphic pattern. The identified SNPs allocated camels into different groups according to which animal harbored the explored SNPs. Using multiple linear regression analysis (MLR), identified SNPs had a significant effect on body weight and weight gain at different age stages. Determination coefficients (R2) value indicated that variability in the phenotypic measurements of growth performance might be correlated with the identified SNPs in growth related genes. The area under the ROC curves was higher in the case of DGAT (0.793-1.0), followed by GH (0.736-0.866) and then MCR (0.410-0.545). DGAT anticipates body weight and weight gain traits better than GH and MCR. As a pilot study, single nucleotide polymorphisms in the investigated genes could be used as proxy predictors and genetic markers that permit a marker assisted selection for growth traits in dromedary camels.

Growth and Hepatic Insulin-Like Growth Factor-I Gene Expression in Dominant and Subordinate Oreochromis Niloticus L Under Limited Period of Physical Interaction

Social stress is known to regulate several aspects of the teleost physiology. This study explored the influence of limited period of physical interaction on growth and hepatic insulin-like growth factor-I (IGF-I) messenger RNA (mRNA) expression of the fish. Twenty all-male Oreochromis niloticus were isolated for 10 days and were used in a social pair study. After the social interaction was settled, dominant and subordinate individuals in a pair were reared separately in one aquarium separated by glass divider. The fish were fed at the same rate daily to remove the possible effect of nutrition. The glass divider was removed 10 min daily for social interaction. Weight was monitored on Days 2, 7 and 14 during the experimental period, then hepatic IGF-I mRNA expression was quantified. During the 14 days social experiment, mean specific growth rate of dominant fish (1.6%·day−1) was significantly higher (P<0.01) than that of the subordinate fish (0.9%·day−1). Dominant fish also had 2.5 fold significantly higher (P<0.05) mean IGF-I mRNA expression than that of subordinate fish. These indicate that even under limited period of physical contact but with period of visual communication, social status regulates growth and hepatic IGF-I gene expression in this species of fish. There was also a significant positive correlation (r = 0.52; P<0.01) between growth and IGF-I mRNA level which supports the previous studies that hepatic IGF-I gene expression has a potential utility as an instantaneous growth rate indicator for O. niloticus.

Potassium diformate in the diet of sterlet sturgeon ( Acipenser ruthenus ): Zootechnical performance, humoral and skin mucosal immune responses, growth‐related gene expression and intestine morphology

This study evaluated the dietary effect of potassium diformate (KDF) on growth performance, body composition, humoral and skin mucosal immune responses, growth-relevant gene expression and intestine histomorphology in sterlet sturgeon. Sterlet fingerlings (N = 270; 6.86 ± 0.14 g) were scattered randomly into 18 tanks (six experimental diets in triplicate groups) and fed with incremental levels of KDF including 0 (control), 3 (KDF3), 6 (KDF6), 9 (KDF9), 12 (KDF12) and 15 (KDF15) g kg−1 for 70 days. The groups fed with KDF6 and KDF9 diets presented significantly higher growth indices and whole-body protein content, whereas the lowest feed conversion ratio was measured in these treatments compared with other groups (p < .05). Total immunoglobulin, lysozyme activity and total protein levels in the serum and skin mucus samples were increased by increasing the levels of KDF, and the highest values were obtained in KDF9. The expression levels of growth hormone, ghrelin, and insulin-like growth factor-I genes were notably upregulated in KDF9 compared with the control, KDF3 and KDF15 groups. The villus length and width of hind-gut were more than doubled in KDF9 treatment compared with the control group (p < .05). These results showed that the application of KDF in sterlet diet had the potential to achieve high overall performance, and also, the optimum inclusion level of KDF ranged from 8.48 to 8.83 g kg−1 based on the second-order polynomial analysis models.

Effect of temperature manipulation during incubation on body weight, plasma parameters, muscle histology, and expression of myogenic genes in breast muscle of embryos and broiler chickens from two commercial strains

ABSTRACT 1. This study evaluated the effect of a higher incubation temperature on body weight, plasma profile, histology and expression of myogenin (MYOG), insulin-like growth factor-I (IGF-I) and vascular endothelial growth factor A (VEGFA) genes in breast muscle of embryos and broilers from two commercial strains. 2. A total of 784 eggs from Ross 308 and Cobb 500 broiler breeder flocks were used. Half of the eggs per strain were incubated at control temperature (37.8°C), whereas the other half were exposed to heat treatment (HT) of 38.8°C between embryonic day (ED) 10 and 14, for 6 h/day. Embryos and chicks were sampled on ED 19 and at hatch. A total of 480, one-day-old chicks per strain and incubation temperature were reared up to 42 d post-hatch. 3. The HT increased hatch weight of Ross chicks and 42-d body weight of broilers from both strains. Lower plasma triacylglycerol levels were measured for HT embryos and broilers on ED 19 and 42 d post-hatch, respectively. HT reduced plasma T3 levels in Ross embryos and broilers for the same periods. Hepatic TBARS concentrations were elevated by HT compared to the control incubation. 4. The HT reduced breast muscle VEGFA gene expression of Cobb embryos on ED 19, whereas expression was stimulated in day-old chicks. At 42 d post-hatch, fibre area was increased by HT regardless of strain. Compared to the control incubation, HT increased the breast yield of Ross broilers and leg yield of Cobb. Ross-HT broilers had a higher pH at 24 h after slaughter and better water holding capacity than Cobb-HT broilers. 5. These results suggested that HT increased body weight, fibre area, IGF-I gene expression and lowered plasma triacylglycerol levels of broiler chickens from both strains at 42 d. However, HT influenced the expression of VEGF-A and MYOG genes and meat quality differently between the broiler strains.

Supplementation of Siberian sturgeon (Acipenser baerii) diet with barberry (Berberis vulgaris) fruit extract: Growth performance, hemato-biochemical parameters, digestive enzyme activity, and growth-related gene expression

In this study, barberry fruit extract (BFE) was used as an herbal feed additive at 0 (control), 150, 300, 600, and 750 mg/kg to evaluate growth performance, physiometabolic responses, and gene expression related to growth and appetite in Siberian sturgeon. The fish (N = 150; 29.7 ± 2.2 g) were fed with the experimental diets for 56 days. Upon the completion of the feeding trial, the fish fed with BFE at 750 mg/kg showed higher growth indices than the control fish and the lowest feed conversion ratio (P < 0.05). The highest levels of red blood cell, hemoglobin, and hematocrit were observed in the group fed with 750 mg/kg BFE (P < 0.05). Different dietary levels of BFE had no significant effect on the levels of alkaline phosphatase and alanine aminotransferase in the serum samples (P > 0.05), while a decreasing trend was seen in aspartate aminotransferase, lactate dehydrogenase, glucose, and triglyceride contents by increasing in the dietary levels of BFE. The activities of lipase, amylase, and protease in the BFE-added groups were higher than the control group (P < 0.05). The highest expression of growth hormone gene was recorded in 600 and 750 mg/kg BFE treatments (P < 0.05). The transcription of insulin-like growth factor-I gene was significantly up-regulated in 750 mg/kg BFE treatment compared to others (P < 0.05). Ghrelin gene expression was significantly elevated in both brain and stomach tissue samples by increasing dietary levels of BFE. Overall, dietary BFE markedly at 750 mg/kg exerts growth-promoting effects in Siberian sturgeon.

Association of growth hormone and insulin-like growth factor I genotype with body weight, dominance of body weight, and mRNA expression in Korat slow-growing chickens.

ObjectiveGrowth hormone (GH) and insulin-like growth factor I (IGF-I) play a critical role in animal growth rates. We aimed to investigate the effect of GH and IGF-I genotypes on body weight (BW), dominance, and gene expression in slow-growing chickens at different ages.MethodsA total of 613 Korat chickens (KRs) were bred and divided into three groups by genotype – A1A1, A1A3, and A3A3 for GH and AA, AC, and CC for IGF-I. Chickens were weighed every two weeks, and liver and breast muscle tissues were collected at 10 weeks of age. Genetic parameters of KRs were estimated using ASReml software. The GH and IGF-I mRNA levels were measured by quantitative polymerase chain reaction. Significant differences between traits were analyzed using the generalized linear model.ResultsA significant effect of GH genotypes on BW was found at most ages, and the A1A1 genotype had the highest value of BW. Compared with the A3A3 genotype, the A1A1 and A1A3 genotypes showed a higher dominance effect at 0 and 2 weeks, and genotype A1A1 had the highest value of dominance at 8 weeks of age. A difference in GH mRNA levels between genotypes was detected in breast muscle at 6 weeks and in the liver tissue at 2 weeks. In the case of IGF-I gene, the AA genotype had the highest BW at the beginning of life. Significant differences in BW dominance were found at 2 weeks. However, IGF-I mRNA levels were not different among genotypes in both breast muscles and liver tissues.ConclusionOur results revealed that GH and IGF-I influence growth, but may not be involved in heterosis. GH can be used as a marker gene in selection programs for growth because the homozygous genotype (A1A1) had the highest BW at all ages. The IGF-I is not a useful marker gene for selection programs.

Selenium nanoparticle and selenomethionine as feed additives: effects on growth performance, hepatic enzymes' activity, mucosal immune parameters, liver histology, and appetite-related gene transcript in goldfish (Carassius auratus).

The aim of this study was to assess the effects of different dietary selenium sources, selenium nanoparticle (nSe), and selenomethionine (SeMet) as feed additives on growth performance, hepatic enzymes' activity, biochemical, mucosal immune parameters, liver histology, and appetite-related gene transcript in goldfish (Carassius auratus). At first, goldfish juveniles (n=480; mean 4.54 g) were fed dietary selenium nanoparticle at 0, 0.3, 0.6, and 0.9 mg nSe/kg diet and SeMet at 0, 0.3, 0.6, and 0.9 mg Se/kg for 9 weeks. Growth performance was evaluated using standard procedures. Blood, skin mucus, and tissue samples (liver and intestine) were collected for biochemical, mucosal immune response, histology, and ghrelin and insulin-like growth factor-I (IGF-I) gene expression. The results showed that fish fed diets fortified with 0.6 mg nSe/kg and 0.6 mg Se/kg had a significant higher weight gain, specific growth rates (SGR), and lower feed conversion ratios (FCR) than fish fed basal diets (p<0.05). Furthermore, dietary nSe and SeMet enhanced blood biochemical profiles especially alkaline phosphatase (ALP) (p < 0.05) and mucosal immunity than the control group in goldfish. Moreover, the liver histological investigation showed that fish fed 0.9 mg of SeMet and nSe kg-1 diets had higher liver lesion scores such as karyolysis, lipidosis, and hyperemia while fish fed 0, 0.3, and 0.6 mg of SeMet and nSe kg-1 diets had small liver changes at 9 weeks. The study further established that inclusion of nSe and SeMet in the diet of goldfish greatly promoted ghrelin and IGF-1genes expressions (p <0.05). Overall, dietary nSe performs better than SeMet and basal diets. The results evoked that nSe and SeMet stimulate the growth, biochemical, and mucosal immunity in goldfish at 0.6 mg/kg.

Identification of the cDNA Encoding the Growth Hormone Receptor (GHR) and the Regulation of GHR and IGF-I Gene Expression by Nutritional Status in Reeves' Turtle (Chinemys reevesii).

Chinemys reevesii (Reeves’ turtle) is a slow-growing reptile that is distributed widely across China. Prior to this study, the cDNA sequence of the growth hormone receptor (GHR) in the Reeve’s turtle, or how periods of starvation might influence the gene expression of GHR and insulin-like growth factor I (IGF-I) in this species, were unknown. Here, we identified the full-length sequence of the cDNA encoding GHR in Reeves’ turtle by using RT-PCR and RACE. The full-length GHR cDNA was identified to be 3936 base-pairs in length, with a 1848 base-pair open reading frame (ORF) that encodes a 615 amino acid protein. Analysis showed that GHR mRNA was detectable in a wide range of tissues; the highest and lowest levels of expression were detected in the liver and the gonad, respectively. IGF-I was also expressed in a range of tissues, but not in the gonad; the highest levels of IGF-I expression were detected in the liver. After 4 weeks of fasting, the expression levels of GHR and IGF-I in the liver had decreased significantly; however, these gradually returned to normal after refeeding. We report the first cloned cDNA sequence for the GHR gene in the Reeve’s turtle. Our findings provide a foundation from which to investigate the specific function of the GHR in Reeve’s turtle, and serve as a reference for studying the effects of different nutrient levels on GHR expression in this species.

Associations between genetic variants of the insulin-like growth factor I (IGF-I) gene and growth traits in European sea bass (Dicentrarchus labrax, L.).

The IGF-I is a significant peptide hormone to cell growth, development, reproduction, and metabolism of teleost fish. This study was conducted to investigate the IGF-I gene polymorphism and to reveal the association between the genotypes of IGF-I gene and growth traits in 80 harvest size European sea bass (Dicentrarchus labrax, L.) reared in İzmir region of Turkey. The 821bp long exon 1 and 5' UTR regions of IGF-I gene were amplified, and PCR products were analyzed via DNA sequencing method. Two novel single nucleotide polymorphisms (SNPs) such as g.46672A > G and g.46749C > T in the 5' UTR region of IGF-I gene in the European sea bass were detected for the first time with this study. Associations between the genotypes of IGF-I g.46749C > T locus and body weight were found statistically significant (p < 0.05). The genotypes of IGF-I g.46749C > T and g.46672A > G loci were found significantly associated with total length (p < 0.05). However, both loci did not show significant associations with body width. The studied gene region sequences were entered to NCBI GenBank database with the accession numbers: MK621680-MK621682. Thus, these polymorphisms have a significant effect on the body weight and total length which could be useful for sea bass selection and breeding for marker-assisted selection (MAS) program.

Supplementation of various zinc sources modify sexual development and testicular IGF family gene expression in pre-pubertal male Japanese quail

Zinc plays an important role in the regulation of insulin-like growth factor-I (IGF-I). IGF system, in turn, has a key role in the development and functions of the reproductive organs. This research was performed to investigate the effects of different sources of zinc on IGF-I gene expression and testicular development in pre-pubertal male Japanese quail. A total of 512 unsexed day-old Japanese quail chicks were randomly divided into 16 groups (4 dietary treatments × 4 replicates) and kept for 35 days. The control group diet was not supplemented with zinc whereas the diets of three groups were supplemented with 25 mg kg−1 zinc oxide (ZnO), zinc oxide nanoparticle (ZnON), and zinc-methionine (Zn-Met). On days 28 and 35, one birds from each subgroup were weighed, bled, and euthanized to evaluate gonado-somatic index (GSI), testicular histology, serum testosterone concentration, cloacal gland index (CGI), and the testicular IGF family gene expression. The results showed that GSI was higher in ZnON (2.307) than control (1.619) on day 35 (P < .05). Germinal epithelium thickness was higher in ZnON (78.88 μm) and Zn-Met (79.73 μm) than control (67.73 μm) on day 35 (P < .05). On day 35, the testosterone concentration was lowest in the control (5.830 ng/ml, P < .05). The CGI of 35-day-old birds was higher in Zn-Met (411.28) than the control (307.59, P < .05). IGF-IR mRNA expression was highest in Zn-Met group on day 28. Therefore, supplementation of diet with Zn-methionine is superior to other sources of zinc for diet supplementation in immature Japanese quail.

Association of polymorphisms in the IGF-I, GHR and STAT5A genes with serum IGF-I concentration and reproductive performance of Holstein dairy cows

This study was conducted to evaluate associations of polymorphisms in the genes for the growth hormone receptor (GHR), insulin-like growth factor I (IGF-I) and signal transducer and activator of transcription 5A (STAT5A) with serum concentrations of IGF-I, reproductive performance and milk production of postpartum Holstein dairy cows. Days from calving to first ovulation (DTO) and calving to conception interval (CCI) were evaluated in 95 Holstein cows. Serum concentrations of IGF-I and β-hydroxybutyrate (BHBA) were quantified in samples collected in sequential blood collections. Genotyping of the IGF-I and STAT5A genes was performed. The IGF-I polymorphism distribution was 35.9% CC, 46.1% CT and 18% TT. The IGF-I concentrations in circulation were greater in cows of the TT compared with both the CT and CC groups (P < 0.05). Genotype had a linear association (P < 0.05) with DTO and CCI, which were less for cows of the TT group. There was no association of STAT5A BstEII on serum IGF-I or reproductive variables (P> 0.05). When combining the GHR AluI T allele, obtained in a previous study, and the IGF-I SnaBI T allele from the current study, for the same cows, there were additive associations of both with serum IGF-I, BHBA, number of services per conception, DTO and CCI (P < 0.05). Thus, the IGF-1 SnaBI TT appears to be associated with fewer DTO and lesser CCI of lactating dairy cows and had an additive association with the GHR AluI T allele on indicators for improvement of fertility.

Characterization of a novel alternatively-spliced 5′ exon in the human insulin-like growth factor I (IGF-I) gene, expressed in liver and some cancers

In mammals, the large IGF-I gene comprises 6 exons, which are subject to alternative splicing. All transcripts contain exons 3 and 4, encoding mature IGF-I, but the other exons are included in various combinations, giving at least 6 possible mature mRNAs. At the 5′ end, exons 1 and 2 are spliced alternatively to exon 3, giving different leader/signal sequences. It is shown in this study that in human an additional exon (designated exon 0) is present, upstream of exon 1. This can be spliced directly to exon 3 or, less frequently, into exon 1. Exon 0 is utilized in liver, in about 24% of IGF-I transcripts, to a minor extent in prostate and endometrium (<1% of transcripts), but not in any of 29 other normal human tissues examined. The exon 0 sequence includes an in-frame ATG/AUG, potentially providing a translation start point giving an IGF-I precursor with a very long signal peptide. However, this ATG is very close to the 5′ end, and may not be included in all transcripts; an in-frame ATG in exon 3 could provide an alternative start point. Utilization of exon 0 was detected in other apes, and to a small extent in Old World monkeys, but not in New World monkeys, prosimians or various non-primate mammals. Exon 0 was not expressed in most human tumours, but was utilized in many prostate tumours, at levels much greater than seen in normal prostate, and in liver tumours, at a lower level than in normal liver.

Effects of In Ovo Methionine-Cysteine Injection on Embryonic Development, Antioxidant Status, IGF-I and TLR4 Gene Expression, and Jejunum Histomorphometry in Newly Hatched Broiler Chicks Exposed to Heat Stress during Incubation.

Simple SummaryIn the current study, we hypothesize that the In ovo concurrent injection of methionine (Met) and cysteine (Cys) may have positive effects on embryonic development, insulin-like growth factor-I (IGF-I) and toll-like receptor-4 (TLR4) gene expression, antioxidant status, biochemical profile, and jejunum histomorphometry of newly hatched broiler chicks exposed to heat stress during the incubation period. As indicated, the In ovo injection of Met and Cys resulted in increased hatch weight, higher levels of total antioxidant capacity, and glutathione activity in the serum and tissues. At the same time, a decrease in the level of heat shock protein-90 and an increase in TLR4 and IGF-I expression in the tissues was observed. In addition, the In ovo injection of Met and Cys improved jejunum histomorphometric parameters of newly hatched broiler chicks. Generally, the concurrent In ovo injection of Met and Cys improved broiler embryonic development, antioxidant status, TLR4, and IGF-I expression and jejunum histomorphometric parameters in newly hatched broiler chicks exposed to heat stress conditions during incubation. Sulfur amino acids are typically the first-limiting amino acids (AA) used in protein metabolism in poultry. Therefore, we hypothesized that their utilization in the pre-hatch period would affect embryonic development, IGF-I and TLR4 gene expression, antioxidant status, serum biochemical profile, and jejunum histomorphometry of newly hatched Ross broiler chicks incubated under heat stress conditions. A total of 150 fertile broiler eggs were subjected to heat stress (39.6 °C for 6 h/d) from d10 until d18 and injected at d 17.5 of incubation with methionine and cysteine (Met-Cys) at a dose of 5.90 mg l-methionine plus 3.40 mg l-cysteine. The effects of Met-Cys administration were examined and compared with the control (Non-injected group) and 0.75% NaCl injected group. The results showed that no significant differences among all groups in serum protein profiles (total protein, albumin, globulin, and albumin/globulin ratio) and creatine kinase were observed. The level of heat shock protein-90 was decreased with Met-Cys In ovo injection. The In ovo injection of Met-Cys also improved the values of total antioxidants capacity and glutathione in examined tissues. At the same time, an increase in fold change mRNA abundance of IGF-I and TLR4 was observed after Met-Cys injection in tested tissues. Finally, an increase of 29% in villus area was found after Met-Cys injection compared to the control group. In conclusion, the In ovo injection of Met-Cys resulted in improved embryonic development, IGF-I and TLR4 gene expression, antioxidant status and jejunum histomorphometry of newly hatched broiler chicks exposed to heat stress during incubation.