The Hyperresponsiveness of Cells Expressing Truncated Erythropoietin Receptors Is Contingent on Insulin-Like Growth Factor-1 in Fetal Calf Serum

Blood ◽  
1998 ◽  
Vol 92 (2) ◽  
pp. 425-433 ◽  
Author(s):  
Jacqueline E. Damen ◽  
Jana Krosl ◽  
Donna Morrison ◽  
Steven Pelech ◽  
Gerald Krystal
Keyword(s):  
Cell Death ◽  
Growth Factor ◽  
Neutralizing Antibodies ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Negative Regulator ◽  
Combined Effects ◽  
Insulin Like Growth Factor ◽  
Wild Type

Abstract We demonstrate herein that the well documented hyperresponsiveness to erythropoietin (Epo) of Ba/F3 cells expressing C-terminal truncated erythropoietin receptors (EpoRs) is contingent on these cells being in fetal calf serum (FCS). In the absence of FCS, their Epo-induced proliferation is far poorer than Ba/F3 cells expressing wild-type (WT) EpoRs. This hyporesponsiveness in the absence of serum is also seen in DA-3 cells expressing these truncated EpoRs. In fact, long-term proliferation studies performed in the absence of serum show that even at saturating concentrations of Epo, Ba/F3 cells expressing these truncated receptors die via apoptosis, while cells bearing WT EpoRs do not, and this programmed cell death correlates with an inability of Epo-stimulated Ba/F3 cells expressing truncated EpoRs to induce the tyrosine phosphorylation of MAPK and the activation of p70S6K. Using neutralizing antibodies to insulin-like growth factor (IGF)-1, we show that a major non-Epo factor in FCS that contributes to the hyperresponsive phenotype of Ba/F3 cells expressing truncated EpoRs is IGF-1. Our results suggest that the Epo-hypersensitivity of truncated EpoR expressing Ba/F3 cells is due to the combined effects of these EpoRs not possessing a binding site for the negative regulator, SHP-1, and the triggering of proliferation-inducing/apoptosis-inhibiting cascades, lost through EpoR truncation, by IGF-1.

The Hyperresponsiveness of Cells Expressing Truncated Erythropoietin Receptors Is Contingent on Insulin-Like Growth Factor-1 in Fetal Calf Serum

Blood ◽  
1998 ◽  
Vol 92 (2) ◽  
pp. 425-433
Author(s):  
Jacqueline E. Damen ◽  
Jana Krosl ◽  
Donna Morrison ◽  
Steven Pelech ◽  
Gerald Krystal
Keyword(s):  
Cell Death ◽  
Growth Factor ◽  
Neutralizing Antibodies ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Negative Regulator ◽  
Combined Effects ◽  
Insulin Like Growth Factor ◽  
Wild Type

We demonstrate herein that the well documented hyperresponsiveness to erythropoietin (Epo) of Ba/F3 cells expressing C-terminal truncated erythropoietin receptors (EpoRs) is contingent on these cells being in fetal calf serum (FCS). In the absence of FCS, their Epo-induced proliferation is far poorer than Ba/F3 cells expressing wild-type (WT) EpoRs. This hyporesponsiveness in the absence of serum is also seen in DA-3 cells expressing these truncated EpoRs. In fact, long-term proliferation studies performed in the absence of serum show that even at saturating concentrations of Epo, Ba/F3 cells expressing these truncated receptors die via apoptosis, while cells bearing WT EpoRs do not, and this programmed cell death correlates with an inability of Epo-stimulated Ba/F3 cells expressing truncated EpoRs to induce the tyrosine phosphorylation of MAPK and the activation of p70S6K. Using neutralizing antibodies to insulin-like growth factor (IGF)-1, we show that a major non-Epo factor in FCS that contributes to the hyperresponsive phenotype of Ba/F3 cells expressing truncated EpoRs is IGF-1. Our results suggest that the Epo-hypersensitivity of truncated EpoR expressing Ba/F3 cells is due to the combined effects of these EpoRs not possessing a binding site for the negative regulator, SHP-1, and the triggering of proliferation-inducing/apoptosis-inhibiting cascades, lost through EpoR truncation, by IGF-1.

213 IN VITRO DEVELOPMENT OF BOVINE EMBRYOS IN THE PRESENCE OF GROWTH HORMONE, INSULIN-LIKE GROWTH FACTOR-1, AND INSULIN IN OOCYTE MATURATION AND EMBRYO CULTURE MEDIA

2008 ◽  
Vol 20 (1) ◽  
pp. 186 ◽  
Author(s):  
C. B. Ponchirolli-Schneider ◽  
C. P. Freitas ◽  
F. C. Landim-Alvarenga
Keyword(s):  
Growth Hormone ◽  
Growth Factor ◽  
Production Rate ◽  
Dna Fragmentation ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Insulin Like Growth Factor ◽  
The Third ◽  
Fragmentation Rate

The addition of hormones and growth factors to bovine IVM and IVC media has been reported to affect early embryonic development by enhancing the blastocyst formation rate and quality of embryos produced. The purpose of this study was to investigate the influence of adding growth hormone (GH), insulin-like growth factor-1 (IGF-1), and insulin to IVM and IVC media. Blastocyst production rate and blastocyst quality, as verified by the number of cells with DNA fragmentation, were evaluated. Ovaries from an abattoir were transported to the laboratory and COC were selected and cultured in IVM medium 199 (Earle's salts, Sigma, St. Louis, MO, USA), 10% fetal calf serum (Sigma), 50 µg mL–1 of sodium pyruvate, 1 µg mL–1 of estradiol (Sigma), 50 µg mL–1 of hCG (Profasi hp�, 5000 IU, Serono Inc., Rockland, MA, USA), 5 µg mL–1 of FSH (Folltropin�, Vetrepharm, Ontario, Canada), and 75 µg mL–1 of gentamicin sulfate for 24 h. After IVF (18 h), zygotes were partially denuded and transferred to IVC medium HTF (HTF�, Irvine Scientific, Santa Ana, CA, USA) and BME (BME�, Sigma), in a 1:1 proportion (HTF:BME), 0.6% BSA (Sigma), 0.01% myoinositol (Sigma), and 75 µg mL–1 of gentamicin sulfate, at 38.5�C, in a humidified atmosphere of 5% CO2 in air, supplemented with 10% fetal calf serum at Day 3 of culture. Three different experiments were performed. The first and second experiments were analyzed using the chi-square test (P < 0.05). The third experiment was analyzed with the general linear model of SAS� (SAS Institute Inc., Cary, NC, USA) and the Tukey test (P < 0.1). In the first experiment, oocytes were cultured in IVM medium supplemented with GH (10 ng mL–1), IGF-1 (100 ng mL–1), insulin (1 µg mL–1), or all 3 combined. In the second experiment, IVC medium was supplemented with GH, IGF-1, insulin, or all 3 combined (same concentrations as above). In the third experiment, the quality of the embryos produced in the first 2 experiments was determined by the percentage of cells with DNA fragmentation. After 96 h of culture, embryos were stained with orange acridin (100 µg mL–1) and propidium iodide (100 µg mL–1) and slides were evaluated by fluorescence microscopy (450 to 490 nm). Rates of blastocyst production (blastocysts/oocytes) in the first experiment (29, 28, 28, 26, and 28% for control, GH, IGF-1, insulin, or all 3 combined, respectively) and in the second experiment (35, 35, 36, 35, and 31%) were not statistically different among the groups. In the third experiment, the addition of GH, IGF-1, or insulin to IVM medium did not affect the DNA fragmentation rate (11, 5, 2, 12, and 12%). However, the addition of insulin to IVC medium led to a higher DNA fragmentation rate (24%), when compared with the other groups (11, 10, 6, and 8% for control, GH, IGF-1, and all 3 combined). The addition of GH or IGF-1 to bovine IVM and IVC media did not affect the blastocyst production rate or the quality of embryos produced. The quality of embryos cultured in the presence of insulin was negatively affected.

Comparison of the effects of growth hormone, insulin-like growth factor-I and fetal calf serum on mouse molar odontogenesis in vitro

1995 ◽  
Vol 40 (9) ◽  
pp. 789-799 ◽  
Author(s):  
W.G. Young ◽  
J.V. Ruch ◽  
M.R. Stevens ◽  
C. Bègue-Kirn ◽  
C.Z. Zhang ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Growth Factor ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Insulin Like Growth Factor ◽  
Factor I ◽  
Growth Factor I

Cell survival and proliferation are modified by insulin-like growth factor 2 between days 9 and 10 of mouse gestation

Development ◽  
2001 ◽  
Vol 128 (19) ◽  
pp. 3819-3830 ◽  
Author(s):  
Jason L. Burns ◽  
A. Bassim Hassan
Keyword(s):  
Cell Death ◽  
Growth Factor ◽  
Cell Survival ◽  
Cell Number ◽  
S Phase ◽  
Paternal Allele ◽  
Insulin Like Growth Factor ◽  
Wild Type ◽  
Post Implantation ◽  
Cell Survival And Proliferation

The size of mammalian species involves the interaction of multiple genetic modifiers that control the timing and extent of growth mechanisms. Disruption of the paternal allele of the imprinted embryonic gene coding for insulin-like growth factor 2 (IGF2, Igf2+m/−p), results in viable mice that are 60% the weight of wild-type littermates. Differences in weight are first detected at embryonic day (E) 11, and the growth deficit is maintained throughout life. We report the mechanisms that account for this unusual phenotype. In order to quantify growth, we used novel methods to generate single cell suspensions of post-implantation mouse embryos. We were then able to quantify cell number, cell proliferation and cell death between E8.5 and E11.5 using flow cytometry. Determination of total embryo cell number also allowed us to time litters by a method other than by plugging. Wild-type and Igf2+m/−p embryos accumulated similar total cell numbers up to E9.25, but cell number began to diverge by around E9.5, with significant differences by E11 (75% of wild type). A relative increase in pyknotic nuclei, sub-GI cytometry counts and caspase activity, all indicative of cell death, occurred in Igf2+m/−p embryos at E9.25, reverting to wild-type levels by E9.75. This was followed at E9.75 by a significant reduction in the proportion of cells in S phase, quantified by S-phase cytometry counts and BrdU labelling. No significant differences in cell size were detected. We conclude that the majority of the cell number differences between wild-type and Igf2+m/−p mice can be accounted for by modification of cell survival and proliferation during the period (E9 to E10) of post-implantation development.

Long-term treadmill training ameliorates endothelium-dependent vasorelaxation mediated by insulin and insulin-like growth factor-1 in hypertension

2015 ◽  
Vol 119 (6) ◽  
pp. 663-669 ◽  
Author(s):  
Yi-Yuan Lin ◽  
Shin-Da Lee ◽  
Chia-Ting Su ◽  
Tsung-Lin Cheng ◽  
Ai-Lun Yang
Keyword(s):  
Growth Factor ◽  
Insulin Receptor ◽  
Treadmill Training ◽  
Control Group ◽  
Insulin Like Growth Factor ◽  
Hypertensive Rats ◽  
Protein Levels ◽  
Significant Difference ◽  
Nos Inhibitors

Dysfunction of insulin and insulin-like growth factor-1 (IGF-1) is associated with the pathophysiology of hypertension. The influence of long-term exercise on vascular dysfunction caused by hypertension remains unclear. We investigated whether long-term treadmill training improved insulin- and IGF-1-mediated vasorelaxation in hypertensive rats. Eight-week-old male spontaneously hypertensive rats (SHR) were randomly divided into sedentary and exercise (SHR-EX) groups. The SHR-EX group was trained on a treadmill for 60 min/day, 5 days/wk, for 8 wk. Wistar-Kyoto rats (WKY) were used as the normal control group. After training, aortic insulin- and IGF-1-mediated vasorelaxation was evaluated in organ baths. Additionally, the roles of phosphatidylinositol 3-kinase (PI3K), nitric oxide synthase (NOS), and aortic protein expression were examined in the three groups. Compared with sedentary SHR and WKY groups, insulin- and IGF-1-mediated vasorelaxation was significantly enhanced to a nearly normal level in the SHR-EX group. After endothelial denudation, blunted and comparable vasorelaxation was found among the three groups. Pretreatment with selective PI3K and NOS inhibitors attenuated insulin- and IGF-1-mediated vasorelaxation, and no significant difference was found among the three groups after the pretreatment. The aortic protein levels of the insulin receptor (IR), IGF-1 receptor (IGF-1R), insulin receptor substrate-1 (IRS-1), and endothelial NOS (eNOS) were also significantly increased in the SHR-EX group compared with the other two groups. These results suggested that treadmill training elicited the amelioration of endothelium-dependent insulin/IGF-1-mediated vasorelaxation partly via the increased activation of PI3K and NOS, as well as the enhancement of protein levels of IR, IGF-1R, IRS-1, and eNOS, in hypertension.

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