Insulin-like Growth Factor-1 mRNA Research Articles

IGF-1 rs6218 polymorphisms modulate the susceptibility to age-related cataract.

Single nucleotide polymorphisms (SNPs), as the most abundant form of DNA variation in the human genome, contribute to age-related cataracts (ARC) development. Apoptosis of lens epithelial cells (LECs) is closely related to ARC formation. Insulin-like growth factor 1 (IGF1) contributes to cell apoptosis regulation. Moreover, IGF1 was indicated to exhibit a close association with cataract formation. Afterward, an investigation was conducted to examine the correlation between polymorphisms in IGF1 and the susceptibility to ARC. The present investigation was a case-control study. Venous blood draws were collected from the participants for DNA genotyping. Lens capsule samples were collected to detect mRNA and apoptosis. TaqMan RT-PCR was used to detect IGF1 polymorphism genotypes and qRT PCR was used to detect IGF1 mRNA levels in LECs. LEC apoptosis was evaluated through flow cytometry. The chi-square test was used to compare differences between ARCs and controls of each SNP. We found that the G allele frequency in the IGF1-rs6218 was higher in the ARCs than in the controls. Furthermore, it was observed that the rs6218 GG genotype exhibited a positive correlation to elevated levels of IGF1 mRNA in LECs. The IGF1 mRNA in the LECs and the apoptosis of LECs in nuclear type of ARCs (ARNC) was higher than the controls. The susceptibility to ARC was related to IGF1-rs6218 polymorphism, and this polymorphism is associated with IGF1 expression at the mRNA level. Moreover, apoptosis in LECs of ARNCs was found to be increased.

Identification of Insulin-like Growth Factor (IGF) Family Genes in the Golden Pompano, Trachinotus ovatus: Molecular Cloning, Characterization and Gene Expression.

Insulin-like growth factors (IGFs) are hormones that primarily stimulate and regulate animal physiological processes. In this study, we cloned and identified the open reading frame (ORF) cDNA sequences of IGF family genes: the insulin-like growth factor 1 (IGF1), insulin-like growth factor 2 (IGF2), and insulin-like growth factor 3 (IGF3). We found that IGF1, IGF2, and IGF3 have a total length of 558, 648, and 585 base pairs (bp), which encoded a predicted protein with 185, 215, and 194 amino acids (aa), respectively. Multiple sequences and phylogenetic tree analysis showed that the mature golden pompano IGFs had been conserved and showed high similarities with other teleosts. The tissue distribution experiment showed that IGF1 and IGF2 mRNA levels were highly expressed in the liver of female and male fish. In contrast, IGF3 was highly expressed in the gonads and livers of male and female fish, suggesting a high influence on fish reproduction. The effect of fasting showed that IGF1 and mRNA expression had no significant difference in the liver but significantly decreased after long-term (7 days) fasting in the muscles and started to recover after refeeding. IGF2 mRNA expression showed no significant difference in the liver but had a significant difference in muscles for short-term (2 days) and long-term fasting, which started to recover after refeeding, suggesting muscles are more susceptible to both short-term and long-term fasting. In vitro incubation of 17β-estradiol (E2) was observed to decrease the IGF1 and IGF3 mRNA expression level in a dose- (0.1, 1, and 10 μM) and time- (3, 6, and 12 h) dependent manner. In addition, E2 had no effect on IGF2 mRNA expression levels in a time- and dose-dependent manner. The effect of 17α-methyltestosterone (MT) in vitro incubation was observed to significantly increase the IGF3 mRNA expression level in a time- and dose-dependent manner. MT had no effect on IGF2 mRNA but was observed to decrease the IGF1 mRNA expression in the liver. Taken together, these data indicate that E2 and MT may either increase or decrease IGF expression in fish; this study provides basic knowledge and understanding of the expression and regulation of IGF family genes in relation to the nutritional status, somatic growth, and reproductive endocrinology of golden pompano for aquaculture development.

Effects of glycyrrhiza polysaccharides on growth performance, meat quality, serum parameters and growth/meat quality-related gene expression in broilers.

With growing restrictions on the use of antibiotics in animal feed, plant extracts are increasingly favored as natural feed additive sources. Glycyrrhiza polysaccharide (GP), known for its multifaceted biological benefits including growth promotion, immune enhancement, and antioxidative properties, has been the focus of recent studies. Yet, the effects and mechanisms of GP on broiler growth and meat quality remain to be fully elucidated. This study aimed to investigate the effects of GP on growth, serum biochemistry, meat quality, and gene expression in broilers. The broilers were divided into five groups, each consisting of five replicates with six birds. These groups were supplemented with 0, 500, 1,000, 1,500, and 2,000 mg/kg of GP in their basal diets, respectively, for a period of 42 days. The results indicated that from day 22 to day 42, and throughout the entire experimental period from day 1 to day 42, the groups receiving 1,000 and 1,500 mg/kg of GP showed a significant reduction in the feed-to-gain ratio (F:G) compared to the control group. On day 42, an increase in serum growth hormone (GH) levels was shown in groups supplemented with 1,000 mg/kg GP or higher, along with a significant linear increase in insulin-like growth factor-1 (IGF-1) concentration. Additionally, significant upregulation of GH and IGF-1 mRNA expression levels was noted in the 1,000 and 1,500 mg/kg GP groups. Furthermore, GP significantly elevated serum concentrations of alkaline phosphatase (AKP) and globulin (GLB) while reducing blood urea nitrogen (BUN) levels. In terms of meat quality, the 1,500 and 2,000 mg/kg GP groups significantly increased fiber density in pectoral muscles and reduced thiobarbituric acid (TBA) content. GP also significantly decreased cooking loss rate in both pectoral and leg muscles and the drip loss rate in leg muscles. It increased levels of linoleic acid and oleic acid, while decreasing concentrations of stearic acid, myristic acid, and docosahexaenoic acid. Finally, the study demonstrated that the 1,500 mg/kg GP group significantly enhanced the expression of myogenin (MyoG) and myogenic differentiation (MyoD) mRNA in leg muscles. Overall, the study determined that the optimal dosage of GP in broiler feed is 1,500 mg/kg.

Minoxidil delivered via a stem cell membrane delivery controlled release system promotes hair growth in C57BL/6J mice.

Objective: Umbilical cord-derived mesenchymal stem cell membrane-loaded minoxidil (MXD) nanoparticles (STCM-MXD-NPs) were prepared to investigate their effects on hair growth in C57BL/6J mice. Methods: STCM-MXD-NPs were obtained by freeze-thawing and differential centrifugation, and their effects on hair growth were evaluated using C57BL/6J mice. The mRNA and protein expression levels of vascular endothelial growth factor (VEGF) and insulin-like growth factor-1 (IGF-1) were detected by real-time polymerase chain reaction and enzyme-linked immunosorbent assays, respectively. Protein expression levels of marker of proliferation Ki-67 (MKI67) and β-catenin (CTNNB) in skin tissue were detected by immunohistochemistry. Results: STCM-MXD-NPs improved MXD solubility. They released the drug slowly, increasing its transdermal properties, accumulation in the skin, and content in the hair bulb tissues with a better efficacy than that of ordinary MXD. Moreover, STCM-MXD-NPs significantly upregulated the mRNA and protein levels of VEGF and IGF-1 and promoted the protein expression of MKI67 and CTNNB in mouse skin tissues, promoting mouse hair growth. Conclusion: Stem cell membrane-loaded MXD nanoparticles with slow-release properties increased MXD accumulation in the skin by improving its transdermal properties, increasing VEGF, IGF-1, MKI67, and CTNNB expression levels and promoting hair growth in C57BL/6J mice.

Dietary Effect of S-Methylmethionine Sulfonium Chloride on Growth, Serum Biochemical Parameters, Body Composition, and Expression of Some Related Genes in Oreochromis niloticus

Abstract The main objective of the current trial was to investigate the impacts of tilapia diets supplemented with S-methylmethionine sulfonium chloride (MMSC) on the growth performance, feed efficiency, body analysis, blood biochemistry and regulation of myostatin (MSTN) and insulin-like growth factor-1 (IGF-1) genes of nile tilapia (Oreochromis niloticus). The experimental fish (n=180) weighing 22.4±0.17 g were allocated randomly into three equal groups in triplicate as follows; the control group received an un-supplemented diet; the other two groups received two different levels of MMSC (0.2 and 0.4 g/kg) for eight weeks. The obtained findings demonstrated that tilapia diets enriched with small dosages of MMSC considerably (P<0.05) enhanced all assessed growth performance and feed efficiency indicators when compared with the control group. When compared to the control group, tilapia meals supplemented with 0.2 g MMSC significantly (P<0.05 or 0.01) raised blood protein profile, particularly total protein and globulin levels. Contrarily, blood creatinine levels were significantly (P<0.01) reduced in the group of tilapia fed diets containing MMSC (0.2 or 0.4 g/kg) compared to the group without receiving any supplementation. Whereas, fish body analysis showed higher significant increases in crude protein and ether extract levels (P<0.01) as well as reduced ash content (P<0.05) in the fish group that received lower MMSC doses in comparison to other treated and control groups. In addition, MMSC dietary supplementation significantly downregulated the expression of MSTN and upregulated IGF-1 mRNA expression compared with the control group. Additionally, both dosages of MMSC supplementation modestly enhanced the intestinal villus histomorphometric score indices with observed tall, thick, and various broad tips in comparison to the control group. In conclusion, it could be recommended that fish diets supplemented with 0.2 g MMSC per kg diet may promote the growth and general health status of Nile Tilapia.

Involvement of α-klotho in growth hormone (GH) signaling.

Growth hormone (GH) exerts multiple effects on different organs directly or via its main mediator, insulin-like growth factor1 (IGF1). In this study, we focused on the novel relationship between GH action and the antiaging hormone α-klotho. Immunofluorescent staining of α-klotho was observed in the renal distal tubules and pituitary glands of somatostatin- and GH-positive cells in wild-type (WT) mice. Treatment of 4-week-old WT mice with GH increased IGF1 mRNA expression in the pituitary gland, liver, heart, kidney, and bone but increased α-klotho mRNA expression only in the pituitary gland, kidney, and bone. Increased α-klotho protein levels were observed in the kidney but not in the pituitary gland. No induction of α-klotho RNA expression by GH was observed in juvenile mice with kidney disease, indicating GH resistance. Furthermore, GH and α-klotho supplementation in HEK293 cells transfected with GHR increased Janus kinase 2 mRNA (a GH downstream signal) expression compared to supplementation with GH alone. In conclusion, we suggest that 1) the kidney is the main source of secreted α-klotho, which is detected in blood by the downstream action of GH, 2) α-klotho induction by GH is resistant in kidney disease, and 3) α-klotho might be an enhanced regulator of GH signaling.

Hypocalcemia in combination with hyperphosphatemia impairs muscle cell differentiation in vitro

PurposeHypoparathyroidism is a rare endocrine disorder characterized by low or absent secretion of parathyroid hormone (PTH), which leads to decreased calcium and increased phosphorus levels in the serum. The diagnosis of hypoparathyroidism is based on the identification of the aforementioned biochemical abnormalities, which may be accompanied by clinical manifestations. Symptoms of hypoparathyroidism, primarily attributed to hypocalcemia, include muscle cramps or spasms, facial, leg, and foot pain, seizures, and tingling in the lips or fingers. The treatment of hypoparathyroidism depends on the severity of symptoms and the underlying pathology. Over the long term, calcium supplements, active vitamin D analogs, and thiazide diuretics may be needed. In fact, in patient cohorts in which optimal disease control still remains elusive, replacement therapy with recombinant parathyroid hormone analogs may be contemplated. Despite the predominantly neuromuscular symptoms of hypoparathyroidism, further effects of parathyroid hormone deficiency at the muscle cell level remain poorly understood. Thus, the aim of our study was to evaluate the effects of hypocalcemia in combination with hyperphosphatemia on muscle cells differentiation in vitro.MethodsC2C12 cells, an in vitro model of muscle cells, were differentiated for 2 or 6 days in the presence of hypocalcemia (CaCl2 0.9 mmol/l) and moderate (PO4 1.4 mmol/l) or severe (PO4 2.9 mmol/l) hyperphosphatemia, or combinations of both conditions. Cell differentiation and expression of genes linked to muscle differentiation were evaluated.ResultsThe combination of hypocalcemia with hyperphosphatemia induced a significant reduction (50%) in differentiation marker levels, such as MyoD (protein 1 for myoblast determination) and myogenin on the 1st day of differentiation, and MHC (myosin heavy chains) after 6 days of differentiation compared to control. Furthermore, this condition induced a statistically significant reduction of insulin-like growth factor-1 (IGF-1) mRNA expression and inhibition of IGF signaling and decrease in ERK phosphorylation compared to control cells.ConclusionsOur results showed that a condition of hypocalcemia with hyperphosphatemia induced an alteration of muscle cell differentiation in vitro. In particular, we observed the reduction of myogenic differentiation markers, IGF-1 signaling pathway, and ERK phosphorylation in differentiated skeletal myoblasts. These data suggest that this altered extracellular condition might contribute to the mechanisms causing persistence of symptoms in patients affected by hypoparathyroidism.

Possible involvement of somatotrophic hormones in the postpartum regulation of ovarian activity in mares

In horses, resumption of ovarian function after parturition occurs early and the majority of mares ovulate within two weeks after parturition. Previous research suggests that early postpartum increases in the peripheral concentration of prolactin and somatotrophic hormones are the major stimulus for resumption of ovarian activity and ovulation. In the present study, we investigated if placental tissue may be a source of somatotrophic hormones and thus influence ovarian activity. Ovarian follicular development and concentrations of prolactin, insulin like growth factor-1 (IGF1) and LH were determined in 14 healthy Haflinger mares (9.2±3.8 years). From D15 before until D15 after parturition (pp), transrectal ultrasonography of ovarian activity was performed and blood samples were collected for determination of hormone concentrations (prolactin, IGF1 by ELISA; LH by RIA). All foals were healthy, mature and stayed with their dams. Samples from the allantochorion and amnion were obtained immediately after delivery. The relative abundance of mRNA for IGF1 and the prolactin receptor (PRLR) was analyzed by qPCR. Formalin fixed samples were analyzed for protein expression of IGF1 and PRLR by immunohistochemistry. Statistical analysis (IBM SPSS 27.0) was done by analysis of variance (for repeated measures) comparing data from mares that ovulated(group ov; n=5) and did not ovulate until D15 pp (group non-ov, n=9). Values are mean ± SD. Mares from group ov foaled later in the year than non-ov mares (day of year 84±29 vs. 50±20, p<0.05), but no differences in gestation length, placental weight or size were detected. Already on D15 before parturition, ovarian follicles <2 cm diameter were present in all mares, but growth to preovulatory size occurred only pp with a higher growth rate in ov mares (day x group p<0.05). Prolactin and IGF1 concentrations increased from D5 before parturition, peaked at D1 pp (ov vs. non-ov: prolactin 13.0±6.3 vs. 7.9±4.4 ng/ml, p<0.05; IGF1 409±71 vs. 350±65 ng/ml, p<0.05) and declined thereafter. The LH concentration increased from D1 pp in group ov (D12 pp: 23.82±15.96 ng/ml) but not in non-ov (D12 pp: 10.82±5.26 ng/ml; day x group p<0.001). There was a high abundance of mRNA and protein for IGF1 and PRLR in placental tissue irrespective of group. In conclusion, ovarian follicles are detectable before foaling, but develop to preovulatory size only pp when prolactin and LH concentrations increase. In agreement with the situation in cattle, IGF1 stimulates ovarian activity in lactating mares. Placental tissue produces IGF1 that may be under the influence of prolactin as suggested by the presence of PRLR in both amnion and allantochorion.

Protective Activity against Oxidative Stress in Dermal Papillae with Extracted Herbal Essential Oils

Dermal papilla cells (DPCs) are a rich source of nutrients and secrete multiple growth factors that can affect hair growth. As oxidative stress leads to hair loss in humans, it is considered to be one of the factors that can impair the function of DPCs. Herb-derived phytochemicals exhibit potent antioxidant activities; therefore, this study investigated whether a set of essential oils (lavender, lemongrass, rosemary, and chamomile oils) promote the hair-growth activity of DPCs. Intracellular reactive oxygen species (ROS) increased markedly in ultraviolet B-irradiated DPCs (50 mJ/cm2) and were efficiently blocked by essential oils. Essential oils upregulated the mRNA and protein levels of phase II enzymes (detoxifying and antioxidant), including heme oxygenase-1, NAD(P)H quinone oxidoreductase-1, and glutathione S-transferase pi. They also upregulated and activated nuclear factor E2-related factor 2, an essential transcription factor for phase II enzymes. Regarding biomarkers for hair growth, essential oils significantly increased vascular endothelial cell growth factor and insulin-like growth factor-1 mRNA levels. In conclusion, phytochemicals in essential oils enhance hair growth through ROS-scavenging activity in DPCs.

Expressions of c-myc and Insulin-Like Growth Factor-1 mRNA in the Liver of Growing Rats Vary Reciprocally in Response to Changes in Dietary Protein , ,

To investigate molecular mechanisms of growth control by diets, we examined the effects of nutrition on the expression of c-myc and insulin-like growth factor-1 (IGF-1) genes in the liver of growing rats. In the first study, rats were fed either a 24% casein, 24% zein or protein-free diet, or were starved for 3 d. The levels of the two mRNAs in the tissues were then determined by Northern blot hybridization. In the liver, levels of the two mRNAs varied in a reciprocal anner in response to changes in either quantity or quality of diet. The expression of c-myc mRNA was greatly enhanced by consumption of the protein-free diet or by starvation, whereas the IGF-1 mRNA levels were reduced markedly by consumption of the zein diet or the protein-free diet or by starvation. In another study, the casein and zein diets were fed to rats that had been adapted to a 2-h meal-feeding pattern, first with nonpurifled diet for 10 d and then with the protein-free diet for 3 d before the experiment. In rats fed casein, the level of c-myc mRNA decreased 75% within 8 h after consumption of the casein diet, whereas the IGF-1 mRNA level increased 100% during that period. Consumption of the zein diet did not affect the level of either mRNA. Because quantity of food intake did not differ between the rats fed casein and those fed zein, expression of the two genes in the liver was affected by the quality of the protein consumed. These results indicate that quality and quantity of diets changed the expression of c-myc and IGF-1 genes and thus demonstrate the possibility that nutrition not only supplies material for body components but also affects the signal transduction for growth in young growing rats.

Effect of maternal periconceptional undernutrition in sheep on cortisol regulation in offspring from mid-late gestation, through to adulthood.

Maternal periconceptional undernutrition (PCUN) alters fetal hypothalamic-pituitary-adrenal axis (HPAA) function and placental glucocorticoid metabolism in sheep. The effects of PCUN on HPAA function in adult life are not known. We investigated the effects of PCUN on fetal adrenal development across gestation and on cortisol regulation in adult offspring. Ewes were undernourished from 61 days before to 30 days after conception ('PCUN') or fed ad libitum ('N'). mRNA expression in the fetal adrenal gland of ACTH receptor (ACTHR), steroidogenic acute regulatory protein (STAR), cytochrome P450 17A1 (CYP17A1), 11beta-hydroxysteroid-dehydrogenase type 2 (11βHSD2), insulin-like growth factor-2 (IGF2), and in the fetal hippocampus of 11βHSD1, 11βHSD2, mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) was determined at 50 (adrenal only), 85, 120 and 131 days of gestation (term=148 days). In adult offspring (≥ 3 years, N; 10 female, 5 male, PCUN; 10 female, 10 male) a combined arginine vasopressin (AVP, 0.1 μg/kg) and corticotropin-releasing hormone (CRH, 0.5 μg/kg) challenge and a metyrapone (40 mg/kg) challenge were undertaken. mRNA expression of ACTHR, STAR and CYP17A1 were determined in adult adrenals. Fetal adrenal STAR, CYP17A1 and IGF2 mRNA expression were not different between groups in early gestation but were higher in PCUN than N at 131 days' gestation (all p<0.01). PCUN reduced fetal hippocampal MR and GR mRNA expression by 50% at 85 day, but not in later gestation. Adult offspring plasma cortisol responses to AVP+CRH or metyrapone were not different between groups. Plasma ACTH response to AVP+CRH was lower in PCUN males but ACTH response to metyrapone was not different between groups. Adult adrenal ACTHR, STAR, and CYP17A1 mRNA expression were not affected by PCUN. We conclude that the effects of PCUN on fetal HPAA function that became apparent in late gestation, are not reflected in adrenal cortisol secretion in mid-adulthood.

Mechanism and inhibitory effects of cactus (Opuntia dillenii) extract on melanocytes and its potential application for whitening cosmetics

Penghu cactus (Opuntia dillenii [Ker.] Haw) is a cactus plant that commonly grows in Penghu Island, Taiwan, Republic of China (ROC). However, still lack of scientific study on the Opuntia dillenii [Ker.] Haw extract on skin-whitening-associated tyrosinase activity and melanin production. The activities of its extract in melanogenesis were investigated in this article. In this experiment, we used an extract from the Penghu cactus (Opuntia dillenii [Ker.] Haw) to study its tyrosinase inhibition, anti-melanin generation, UV-protection effects and wound healing capacity in B16-F10 melanocytes. Without reducing cell growth greatly or causing cell death, 20 g/L cactus extract effectively inhibited the melanin production of B16-F10 cells, and melanogenesis was induced by 3-isobutyl-1-methylxanthine. The cactus extract could also promote cell proliferation. Cactus extract treatment decreased the mRNA expression of insulin-like growth factor 1 (IGF-1) and vascular endothelial growth factor (VEGF) and increased that of transforming growth factor β (TGF-β). Thus, it could reduce cell melanin production and promote cell growth but by also reducing IGF-1 and VEGF mRNA expression, may reduce wound scarring and prevent tumor proliferation and swelling. Increasing TGF-β mRNA expression can help increase collagen to remove wrinkles and help in wound healing. Skin patch test results agreed with in vitro results with B16-F10 melanoma cells. The cactus extract significantly inhibited tyrosinase activity and reduced melanin production, showing a whitening effect on skin tests. Cactus may be a good natural candidate for inhibiting melanin production and promoting cell proliferation.

Oxidative stability of milk and expression of oxidative stress response, lipogenic-related, and IGF-1 genes in early-lactation cows supplemented with peanut and sunflower oils

Context Feeding dairy cows with plant oils rich in unsaturated fatty acids is a recommended strategy for increasing the proportion of polyunsaturated fatty acids in milk for human consumption. Aims We evaluated the effects of supplementation with peanut oil (high oleic acid content) and sunflower oil (high linoleic acid content) on the fatty acid profile and oxidative stability of milk and the expression of oxidative stress response, lipogenic-related and insulin-like growth factor 1 (IGF-1) genes in pasture-fed Jersey cows in early lactation. Methods Twenty-four cows were distributed in a randomised block design. Treatments were four dietary supplements of concentrate alone, or concentrate plus oil (59.6 g/kg dry matter): peanut oil, sunflower oil, or a 1:1 (v/v) mixture of peanut and sunflower oils. Analyses were performed at 30 and 60 days in milk. Key results Both peanut oil and mixed oil diets increased the hypocholesterolemic/hypercholesterolemic ratio (P &lt; 0.001) and health-promoting index (P &lt; 0.001) of milk. The sunflower oil diet increased the thrombogenicity index (P = 0.005) and conjugated diene production (P &lt; 0.001). mRNA abundance of fatty acid synthase (P = 0.003), acetyl-CoA carboxylase alpha (P = 0.001), and stearoyl-CoA desaturase (P &lt; 0.001) decreased with the sunflower oil diet. mRNA abundance of superoxide dismutase decreased (P = 0.01) with sunflower oil and mixed oil diets. The control diet tended (P = 0.08) to reduce IGF-1 mRNA abundance. Conclusions and implications Supplementation with peanut oil increases the health-promoting index of milk, whereas supplementation with sunflower oil increases susceptibility to lipid peroxidation. Supplementation with sunflower oil reduces the expression of lipogenic and antioxidant enzyme genes, and supplementation with unsaturated fats may reduce IGF-1 gene expression in early-lactation cows. The relatively small sample size means that more studies are needed.

Comprehensive Analysis of lncRNA and mRNA Reveals the Effect of ZBED6 on Spleen Growth in Pigs

Transcription factor zinc-finger BED domain-containing protein 6 (ZBED6) is unique to placental mammals and regulates insulin-like growth factor 2 (IGF2) expression, which lead to muscle growth. However, the effect of ZBED6 on the growth of spleen is still elusive. In this study, we explored the regulation of ZBED6 on spleen growth, and the results showed ZBED6 knockout (ZBED6 KO) pigs had heavier spleens than wild-type (WT) pigs. To analyze the mechanism of increased spleen weight in ZBED6 KO pigs, long noncoding RNAs (lncRNAs) and mRNAs in the spleen samples (WT:ZBED6 KO pigs = 3:3) were analyzed to identify differentially expressed lncRNAs (DE-lncRNAs) and genes (DEGs) based on the RNA sequencing (RNA-seq) method. Then, 142 DEGs and 82 DE-lncRNAs were obtained. The qRT-PCR results were consistent with those of the RNA-seq, indicating that the data were reliable. The heavier spleen weight of ZBED6 KO pigs coincided with the significantly upregulated IGF2 mRNA. Functional enrichment analysis of DEGs showed enrichment mainly in myofibril assembly and sarcomere. In addition, 252 cis- and 109 trans-acting target genes of 82 DE-lncRNAs were predicted. By conjoint analysis of lncRNA and mRNA revealed that IGF2, DE-lnRNAs (XLOC_113021, XLOC_078852, NONSUSG004057.1, NONSUSG014354.1, and NONSUSG009750.1), and their target gene ACTN2 may be the key candidate genes in promoting spleen growth in ZBED6 KO pigs. This study provides new directions to understand the global functions of ZBED6 and lncRNAs in spleen growth in pigs.

IL-1β/NF-κB signaling inhibits IGF-1 production via let-7f-5p in dendritic epidermal T cells.

Dendritic epidermal T cells (DETCs) are the main source of insulin-like growth factor-1 (IGF-1) in epidermal tissue, which promote re-epithelialization and wound healing. In refractory wounds, IL-1β has been shown to activate NF-κB and suppress IGF-1 expression in DETCs. Nevertheless, the underlying mechanisms remain unclear. In this study, chromatin immunoprecipitation analysis revealed that IL-1β did not inhibit NF-κB binding to IGF-1 promoter, indicating that IL-1β/NF-κB may suppress IGF-1 expression by alternative mechanisms. MiRNAs negatively regulate gene expression predominantly by base pairing to the 3' untranslation region (UTR) of target mRNAs. Let-7f-5p, miR-1a-3p, and miR-98-5p have been identified as IGF-1-specific miRNAs that can bind directly to the 3'UTR of IGF-1 mRNA and dysregulate IGF-1 mRNA and protein levels. In IL-1β-treated epidermis around wounds or DETCs in vitro, NF-κB promoted the expression of let-7f-5p, and IGF-1 expression was impeded via NF-κB/let-7f-5p pathway. As pre-let-7f-5p, let-7f-1 is located in the 3'UTR of LOC118568094, and let-7f-2 is located in the intron of HUWE1. We discovered that NF-κB p65 bound to the promoters of LOC118568094 and HUWE1 to accelerate let-7f-5p expression, but NF-κB p65 did not affect the methylation levels of LOC118568094 and HUWE1 CpG islands. Injections of Let-7f-5p antagomir into IL-1β-treated and ischemic wound margins restored IGF-1 secretion in DETCs and promoted wound healing. In conclusion, we demonstrated that NF-κB signaling pathway activated by IL-1β could increase let-7f-5p expression to inhibit IGF-1 production in DETCs and delay wound healing. And let-7f-5p antagomir utilized in wound margin could effectively promote refractory wound healing.

Genome-wide characterization of three IGFs in hybrid yellow catfish (Pseudobagrus fulvidraco ♀× Pseudobagrus vachellii ♂) and the association of IGF2 allelic variants with growth traits

“Huangyou No. 1”, a new hybrid yellow catfish (Pseudobagrus fulvidraco ♀× Pseudobagrus vachellii ♂) with heterosis in growth traits, is an economically important freshwater fish in China. Among genes related to growth traits, Insulin-like growth factors (IGFs) are recognized as key factors during embryonic development and growth period. Thus, we identified and characterized IGF1, IGF2 and IGF3 gene from hybrid yellow catfish. Phylogenetic analysis showed that the IGF genes were evolutionarily-constrained and their function was conserved. Transcriptional studies indicated that three IGF mRNA expression were most abundant in liver, despite the ubiquitous expression in nine tested tissues. Further, two opposite temporal expression patterns were observed in these three IGF genes during 16 embryogenesis and larval development stages of hybrid yellow catfish, of which IGF2 and IGF3 shared the similar profile highly expressed after gastrula. Finally, single nucleotide polymorphisms (SNPs) of IGF2 were screened and genotyped in 264 hybrid yellow catfish by Sanger sequencing. Only a non-synonymous mutation (SNP 97 T > C) obeyed Hardy-Weinberg equilibrium. Association analysis indicated SNP 97 T > C significantly associated with nine growth traits (body length, full length, head length, body height, caudal peduncle height, caudal peduncle length, body width, body weight and fatness; P < 0.05). This association was also validated in a second population with 183 individuals. Overall, results from our studies highlight the importance of IGF system in mediating growth development of hybrid yellow catfish, and provide a potential marker for “Huangyou No. 1” breeding.

Molecular characterization, expression pattern and genetic variant of insulin-like growth factor 2 mRNA-binding protein 3 gene in goose (Anser cygnoides)

ABSTRACT As a member of the insulin-like growth factor mRNA binding proteins (IGFBPs) family, IGF2BP3 plays important role in skeletal muscle development by regulating IGF2, which has been considered a key gene related to growth and reproductive traits in many agricultural animal species. However, study on the goose IGF2BP3 gene is still scarce. In this study, the full-length cDNA sequence of goose IGF2BP3 gene was cloned and characterized. The goose IGF2BP3 cDNA is 1825 bp in length, containing an open reading frame (ORF) of 1755 bp that encodes a protein of 584 amino acids, which consists of two RNA-recognition motifs (RRMs) and four K-homologous (KH) domains. RT-qPCR analysis indicates that the IGF2BP3 mRNA expression level is the highest in ovary tissue, followed in hypothalamus, spleen, leg muscle and liver, and low level in breast muscle and muscular stomach of the adult Zhedong White goose. Meanwhile, 25 single nucleotide polymorphisms (SNPs) and 5 insertions/deletions (InDels) variants were identified in about 10,764 bp of goose IGF2BP3 genomic DNA sequence. These results serve as a foundation for further investigations of the function of IGF2BP3 gene in geese.

Effects of Pregnant Fatigue on the Development of Offspring in Rats.

To explore the correlation between pregnant fatigue and intrauterine physical and neural development of offspring in rats. Sprague-Dawley pregnant rats were randomly divided into a normal control group, a mild fatigue group (stand in water for 6 hours/day), and a severe fatigue group (stand in water for 15h/day). The levels of lactic acid, 5-Hydroxytryptamine and Interleukin-6 in cardiac serum of rats were used to evaluate the fatigue. The expression of S100β in the telencephalon, Insulin-like growth factor-1 (IGF-1) in the liver and Cyclooxygenage-2 (COX-2) in the small intestine tissues of fetal rats were examined. Frozen sections were taken from the telencephalon of rat pups to observe morphological changes in the hippocampal primordium. Pregnant fatigue led to a decrease in food intake (F = 37.586, p = 0.000) and water intake (F = 23.608, p = 0.000) in rats. The IGF-1 mRNA expression of fetal rats in the severe fatigue group was lower than that in the control group (p = 0.0003). The expression of S100β mRNA (p = 0.000) and COX-2 mRNA (p = 0.0002) of fetal rats were higher in the severe fatigue group than in the control group. HE staining of the telencephalon of fetal rats in the pregnant fatigue group revealed sparse and irregular cell arrangement and increased gaps in the hippocampal primordial site. Pregnant fatigue rats had both physical fatigue and mental fatigue. Fatigue during pregnancy affects physical development and neurodevelopment of offspring. Further research should elucidate the mechanisms of pregnant fatigue and its effects on offspring.

Metabolic memory determines gene expression in liver and adipose tissue of undernourished ewes

The aim of this work was to determine the effects of undernutrition on gene expression in liver and adipose tissue, and on the hepatic concentration of insulin like growth factor 1 (IGF1) and insulin, in pregnant ewes with different initial body condition score (iBCS). Thirty-six Rasa Aragonesa ewes were divided into 2 groups with different BCS: BCS > 2.75 (n = 19; high, H) and BCS < 2.25 (n = 17; low, L), and were randomly assigned to two nutritional treatments: 1.5 (control, C) or 0.5 (undernourished, U) times the daily maintenance requirements. Thus, the final four groups were: high-iBCS control (HC, n = 9), high-iBCS undernourished (HU, n = 10), low-iBCS control (LC, n = 9) and low-iBCS undernourished (LU, n = 8). High-iBCS ewes presented greater liver IGF1 mRNA expression (P < 0.05), which is consistent with higher liver IGF1 and insulin concentrations (P = 0.10 and P < 0.05, respectively), and with greater liver growth hormone receptor (GHR) gene expression (P < 0.05). These data, and the trend for more leptin receptor type b (LEPR-b) mRNA in high-iBCS ewes (P = 0.09) suggest a better anabolic status. Furthermore, undernourished ewes presented lower hepatic insulin like growth factor binding protein 3 (IGFBP3) gene expression (P = 0.05), suggesting a shift in the availability of IGFs in plasma and therefore, in their action in target tissues. In the adipose tissue, U ewes presented lower GHR mRNA expression than C animals (P < 0.05), being greatest in the high-iBCS control group. As expected, leptin (LEP) gene expression in adipose tissue was greater in high-iBCS than in low-iBCS ewes (P < 0.05). Nevertheless, the most interesting finding was the organ-specific pattern of LEPR-b gene expression: in contrast to the liver, the greatest adipose tissue LEPR-b gene expression was observed in the low-iBCS undernourished ewes (P < 0.05). In summary, the present study demonstrates that the initial body condition score evokes different, even contrasting, responses in key metabolic organs (i.e., liver and adipose tissue) of control and undernourished ewes.

The Impact of Curcumin on Growth Performance, Growth-Related Gene Expression, Oxidative Stress, and Immunological Biomarkers in Broiler Chickens at Different Stocking Densities.

Simple SummaryThe primary goal of global poultry production is to optimize the amount of chicken produced per square meter of floor area. Consequently, stocking density (SD) and curcumin supplementation on broiler performance were investigated. Our results revealed that supplemental curcumin improved birds’ growth, behaviours, and immunity by lowering oxidative stress, enhancing humoral immune response, and modulating the suppression of growth-related gene expressions in broilers raised in high stocking density circumstances.Curcumin’s antioxidant properties reduce free radicals and may improve broiler growth. Therefore, the influence of stocking density (SD) and administration of curcumin in the diet on broiler performance was explored to clarify the impact of HSD and curcumin on the performance of growth, behavioural patterns, haematological, oxidant/antioxidant parameters, immunity markers, and the growth-related genes expression in broiler chickens. A total of 200 broiler chickens (Cobb 500, 2-weeks old) were allotted into 4 groups; SD (moderate and high) and curcumin (100 and 200 mg/kg diet)-supplemented HSD, respectively. Behavioural observations were performed. After a 28-day experimental period, tissue and blood samples were collected for analysis. Expressions of mRNA for insulin-like growth factor-1 (IGF-1), growth hormone receptor (GHR), myostatin (MSTN), and leptin in liver tissues were examined. HSD birds exhibited lower growth performance measurements, haematological parameters, circulating 3,5,3-triiodothyronine and thyroxine levels, antioxidant activities (GSH-Px, catalase, superoxide dismutase), immunoglobulins (A, G, M), and hepatic GHR and IGF-1 expression values. However, HSD birds even had an increment of serum corticosterone, malondialdehyde, pro-inflammatory cytokine (TNF-a, IL-2, IL-6) levels, hepatic leptin and MSTN expression. Moreover, HSD decreased drinking, feeding, crouching, body care, and increased standing and walking behaviour. The addition of curcumin, particularly at a 200 mg/kg diet, alleviated the effect of HSD through amending growth-related gene expression in the chickens. In conclusion, curcumin can enhance birds’ growth performance, behavioural patterns, and immunity by reducing oxidative stress and up-regulating the growth-related gene expressions of broilers under stressful conditions due to a high stocking density.