Anti-Insra+b polyclonal antibody

Glycogen content in mink uterine glandular and luminal epithelia (GE and LE) is maximal during estrus and is depleted before implantation while embryos are in diapause. Uterine glycogen synthesis in vivo is stimulated by estradiol (E-2) while its mobilization is induced by progesterone (P-4). Nevertheless, treatment of an immortalized mink uterine epithelial cell line (GMMe) with E-2 did not affect glycogen production. Interestingly, insulin alone significantly increased synthesis of the nutrient and glycogen content in response to insulin + E-2 was greater than for insulin alone. Our objectives were to determine: 1) If insulin receptor protein (INSR) is expressed by mink uterine GE and LE in vivo and if the amount differs between estrus, diapause and pregnancy; 2) if E-2, P-4 or insulin regulate insulin receptor gene (Insr) expression by GMMe cells, and 3) if E-2 and P-4 act independently to regulate glycogen metabolism by GMMe cells and/or if their effects are mediated in part through the actions of insulin. The mean (+/-S.E.) percent INSR content of uterine epithelia was greatest during diapause (GE: 15.65 +/- 0.06, LE:16.56 +/- 1.25), much less during pregnancy (GE: 2.53 +/- 0.60, LE:2.25 +/- 0.32) and barely detectable in estrus (GE: 0.03 +/- 0.01, LE:0.02 +/- 0.01). Glycogen concentrations in GMMe cells increased 10-fold in response to insulin and 20-fold with insulin + E-2 when compared to controls. Expression of Insr was increased 2-fold by insulin and insulin + E-2 when compared to controls and there was no difference between the two hormone treatments, indicating that E-2 does not increase Insr expression in insulin treated cells. To simulate E-2-priming, cells were treated with Insulin + E-2 for 24 h, followed by the same hormones + P-4 for the second 24 h (Insulin + E-2 P-4) which resulted in Insr and glycogen levels not different from controls. Similarly, cells treated with Insulin + P-4 resulted in glycogen concentrations not different from controls. We conclude that the glycogenic actions of E-2 on GMMe cells are due to increased responsiveness of the cells to insulin, but not as a result of up-regulation of the insulin receptor. Glycogen mobilization in response to P-4 was the result of decreased glycogenesis and increased glycogenolysis occurring concomitantly with reduced Insr expression. Mink uterine glycogen metabolism appears to be regulated in a reproductive cycle-dependent manner in part as a result of the actions of E-2 and P-4 on cellular responsiveness to insulin. (C) 2019 Elsevier Inc. All rights reserved.

The objective was to investigate the of effect chemical composition of dietary fat on transcription of genes involved in lipid metabolism in adipose tissue and the liver via transcriptional profiling in growing pigs. A total of 48 Genetiporc 6.0 x Genetiporc F25 (PIC, Inc., Hendersonville, TN) barrows (initial BW of 44.1 +/- 1.2 kg) were randomly allotted to 1 of 6 dietary treatments. Each experimental diet included 95% of a corn-soybean meal basal diet and 5% cornstarch (control; CNTR), animal-vegetable blend (AV), coconut oil (COCO), corn oil (COIL), fish oil (FO), or tallow (TAL). Pigs were sacrificed on d 10 (final BW of 51.2 +/- 1.7 kg) to collect tissues. Expression normalization across samples was performed by calculating a delta cycle threshold (Delta Ct) value using RPL32. Delta delta cycle threshold (Delta Delta Ct) values were expressed relative to the CNTR treatment. In adipose tissue, adding dietary fat, regardless of the source, decreased the mRNA abundance of FASN compared with the CNTR (P = 0.014). Pigs fed a COIL-based diet tended to have greater adipose tissue expression of FASN (P = 0.071) than pigs fed the other dietary fat sources tested. Abundance of PRKAG-1 mRNA was greater in adipose tissue of barrows a fed COIL-based diet than barrows fed CNTR or FO-based diets (P = 0.047). In the liver, adding dietary fat, regardless of source, increased the mRNA abundance of ACACA, ATGL, INSR, PPAR-a, PRKAG-1, and SCD (P <= 0.020) and tended to increase the abundance of HSL (P = 0.071) and SREBP-1 (P = 0.086) compared with the CNTR. Pigs fed a TAL-based diet had greater hepatic transcription of HSL than pigs fed CNTR-, COCO-, or FO-based diets (P = 0.013). Hepatic transcription of FASN tended to be greater in pigs fed COCO than in pigs fed other dietary fat sources (P = 0.074). Dietary omega-3 fatty acid content tended to negatively correlate with mRNA abundance of PRKAG-1 (P = 0.065) in adipose tissue and ATGL (P = 0.063) in the liver. Dietary fat SFA content was negatively correlated with PPAR-a in the liver (P <= 0.039). Dietary fat MUFA content tended to be positively correlated with ACACA, PPAR-alpha, and PRKAG-1 mRNA abundance in the liver (P <= 0.100). To conclude, the intake of omega-3 fatty acids suppressed the mRNA abundance of genes involved in lipolysis in both adipose tissue and the liver. Dietary SFA are greater inhibitors of lipogenesis in adipose tissue than omega-6 fatty acids. Intake of medium-chain fatty acids alters hepatic lipid metabolism differently than intake of long-chain fatty acids.