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  • COA 106107

    Document Type:
    Certificate of Analysis
    Product Catalog Number:
    106107

  • Significance of IGFBP-4 in the Development of Fetal Growth Restriction. 22689691

    Background: Fetal growth restriction (FGR) is a leading cause of perinatal mortality and morbidity. Animal studies suggest dysregulation of IGF-binding protein (IGFBP)-4 is significant in the development of FGR, although human data are lacking. We postulated that IGFBP-4 is expressed at the maternal fetal interface and plays a role in regulating IGF bioavailability. Thus, maternal serum levels of IGFBP-4 may be associated with complications of abnormal placental growth and development including FGR. Methods: Circulating levels of IGFBP-4 and its protease, pregnancy-associated plasma protein-A (PAPP-A), were examined in healthy pregnancies. Their expression in villi and bed as possible sources of the circulating products were examined by immunohistochemistry. From the large Ottawa and Kingston (OaK) Birth Cohort, a nested case-control study was conducted to examine circulating levels of IGBP-4, PAPP-A, IGF-I, and IGF-II by Western blot in early gestation in 36 women who went on to develop FGR and 36 controls having normal-weight babies. Results: IGFBP-4 was elevated in early pregnancy compared with nonpregnant women and women in later pregnancy, consistent with the presence of abundant extravillous trophoblasts and decidual cells that highly expressed IGFBP-4. High expression of PAPP-A was observed in extravillous trophoblasts and decidual cells in early pregnancy but hardly detectable in the circulation at this time, suggesting maternal circulating PAPP-A originates more likely from syncytiotrophoblasts. Increased IGFBP-4 in the maternal circulation in early pregnancy was associated with the development of FGR [0.48 (0.28-0.74) in control vs. 1.22 (0.66-1.65) in FGR; odds ratio = 22 (95% confidence interval = 2.7-181)]. No difference was observed in circulating PAPP-A, IGF-I and IGF-II in the FGR vs. control group. Conclusion: Our findings support the role of IGFBP-4 in regulating IGF bioavailability and provide new clues for the prevention and treatment of FGR, raising the possibility of clinical use of IGFBP-4 as an early biomarker for this condition.
    Document Type:
    Reference
    Product Catalog Number:
    06-107

  • Molecular recognition characteristics in the insulin-like growth factor (IGF)-insulin-like growth factor binding protein -3/5 (IGFBP-3/5) heparin axis. 15691886

    Insulin-like growth factor binding proteins (IGFBPs) -3 and -5 are known to interact with various components of the extracellular matrix (ECM; e.g. heparin and heparan sulphate) and this interaction is believed to affect the affinity of both IGFBP species for their cognate ligands--IGF-I and -II. There is little detail on the nature of the molecular complex formed between ECM components, IGFBPs and IGFs although the glycosaminoglycan (GAG) heparin has been reported to reduce the affinity of IGFBP-5 for IGF-I. In order to investigate this phenomenon further, we have undertaken an extensive surface plasmon resonance based biosensor study to report the affinity of IGFBP-3 and -5 for binding heparin (22 and 7 nM respectively). We have also shown that pre-complexation of IGFBP with IGF-I and -II inhibits the subsequent association of IGFBP with heparin and conversely that heparin complexation of IGFBP-3 and -5 inhibits IGFBP binding to biosensor surfaces containing immobilised IGF-I. In addition we have used both IGF-I and heparin coated biosensor surfaces in an attempt to build ternary IGF-IGFBP-heparin complexes in order to gain some insight into the nature of inhibition by heparin of IGFI-IGFBP complex formation. Our data lead us to conclude that the inhibition by heparin is partly competitive in nature, and that ternary complexes of IGF-IGFBP-heparin are either unable to form, or only form unstable transient complexes. The potential biological significance of our data is highlighted by the demonstration that IGF-I and IGF-II can displace endogenous IGFBP-5 from monolayer cultures of the mouse mammary epithelial cell line HC11.
    Document Type:
    Reference
    Product Catalog Number:
    06-107

  • Seasonal effects on the response of ovarian follicles to IGF1 in mares. 18687788

    The response of follicles to IGF1 was compared between the transition into the ovulatory season (transitional period) and the ovulatory season (ovulatory period) in eight mares using a cross-over experimental design within periods. Granulosa cells were collected from follicles 15-24 or 25-34 mm and expression of IGF1R, IGF2R, FSHR, LHCGR and PAPPA was determined by qPCR. In addition, 10 mg IGF1 or vehicle were injected into the largest follicle (transitional period) or the second largest follicle (ovulatory period) of a follicular wave before the beginning of diameter deviation between the two largest follicles (mean diameters at injection 19.2 and 20.0 mm during transitional and ovulatory periods respectively). Follicular fluid was collected 24 h after injection for determination of free IGF1, IGFBP, inhibin A and oestradiol levels. Granulosa cells from follicles 25-34 mm, but not follicles 15-24 mm, expressed higher levels of IGF1R (P=0.01), FSHR (Pless than 0.007) and LHCGR (P=0.09) during the ovulatory period than during the transitional period, whereas IGF2R expression was higher in transitional than ovulatory follicles (P=0.06). Follicular IGFBP2 levels were not different (Pgreater than 0.1) between periods and treatments, whereas IGFBP5 levels were higher (Pless than 0.05) during the ovulatory period. Finally, IGF1 injection before the beginning of deviation induced an approximately twofold increase (P=0.01) in follicular inhibin A levels during each period and did not affect oestradiol (Pgreater than 0.1). These results suggest that, as during ovulatory waves, equine follicles during transitional waves are responsive to IGF1 before the beginning of deviation and that, therefore, inadequate IGF1 responsiveness before deviation may not underlie the deficient development of dominant follicles during transition.
    Document Type:
    Reference
    Product Catalog Number:
    Multiple

  • Effects of decreased estradiol-17beta on the serum and anterior pituitary IGF-I system in pigs. 16423816

    To further delineate the role of estradiol in the IGF system an experiment was conducted to determine the dosage of the aromatase inhibitor, anastrozole, needed to decreases serum concentrations of estradiol-17beta (E2) in maturing boars. A second experiment was conducted to determine if administration of anastrozole to growing boars decreased serum concentrations of E2 and affected components of the serum and anterior pituitary gland (AP) IGF system vs untreated boars and barrows. In Experiment 1, 12 crossbred boars (292 days, 158 kg) were administered either 0, 1 or 10 mg/day anastrozole (n = 4/group) beginning on day 1. Blood samples were collected every 7-14 days. Mean serum concentrations of E2 were decreased (P less than 0.05) in the 10 mg group vs the 0 and 1 mg groups by day 36; however, no difference (P greater than 0.05) existed between the 0 and 1 mg groups. In Experiment 2, 24 crossbred boars and 12 barrows (101 days, 44 kg) were stratified by litter to one of three treatment groups (n = 12): boars administered 10 mg/day anastrozole, boars administered 0 mg/day, and barrows administered 0 mg/day. Blood samples were collected and pigs were weighed on day 0 and every 14 days thereafter, then killed on day 84 when blood and APs were collected. The 10 mg/day pigs were fed the anastrozole-amended diet beginning on day 1. Mean serum concentrations of E2 did not differ (P greater than 0.05) between the 10 mg/day pigs and 0 mg/day pigs on day 0; however, on day 15 through to 84 mean serum concentrations of E2 were greater (P less than 0.05) in 0 mg/day pigs than in the 10 mg/day pigs. Mean percentage increase in serum concentrations of IGF-I was greater (P less than 0.05) in untreated boars than anastrozole-treated boars and barrows from day 58 through to 84. Mean percentage of basal IGF-I increased (P less than 0.05) from day 29 through to 84 in untreated boars. Mean relative amounts of AP IGF-binding protein (IGFBP)-2 and -5 were less (P less than 0.01) in 10 mg/day pigs than in the 0 mg/day pigs, but each was greater (P less than 0.01) than in barrows administered 0 mg/day. These results indicate anastrozole administered at a dosage of 10 mg/day suppresses serum concentrations of E2 in pigs. Administration of anastrozole to boars reduced the percentage increase in serum concentrations of IGF-I and relative amounts of AP IGFBP-2 and -5. These data further support a role for E2 in regulating components of the IGF system in pigs.
    Document Type:
    Reference
    Product Catalog Number:
    Multiple

  • Components of the porcine anterior pituitary insulin-like growth factor system throughout the estrous cycle. 21055896

    Components of the circulating and anterior pituitary insulin-like growth factor (IGF) system vary in response to steroids in pigs. However, whether serum and anterior pituitary concentrations of the IGF system vary throughout the estrous cycle has not been determined. To further examine this relationship, estrus was synchronized in 40 gilts of similar age and weight (180 d; 120 kg) by feeding 15 mg altrenogest for 15 d to synchronize estrus. Gilts were checked twice daily for expression of estrus beginning 3 d after the end of altrenogest treatment and continuing for 7 d. The first day each gilt exhibited estrus was designated as day 1 of the estrous cycle. Blood samples were obtained by jugular venipuncture on days 1, 4, 7, 10, 13, 16, 19, and 22 of the estrous cycle. On days 7, 13, 19, and 22 of the estrous cycle 10 pigs were killed and anterior pituitary glands (AP) were collected. Serum concentrations of IGF-I and AP concentrations of IGF-I were determined by radioimmunoassay. Relative amounts of AP IGF binding protein (IGFBP) were determined by western ligand blot analysis. Relative expression of AP IGF-I, IGF-I receptor (IGF-I-R), gonadotropin-releasing hormone receptor (GnRHR), and luteinizing hormone (LH)-β subunit were determined by real-time reverse transcription polymerase chain reaction. Serum concentrations of IGF-I fluctuated throughout the estrous cycle. Mean serum concentrations of IGF-I decreased (P < 0.02) from day 1 through day 10, increased (P < 0.02) on days 13 through 16, and then decreased (P < 0.02) from days 19 through 22. Mean AP concentrations of IGF-I were greater (P < 0.03) on day 19 than on all other days, whereas no difference was detected (P > 0.05) in mean AP concentrations of IGF-I on days 7, 13, and 22. Mean relative amounts of AP IGFBP-2 and -5 were each greater (P < 0.02) in gilts on day 19 than on all other days, whereas no difference was detected (P > 0.05) in mean relative amounts of AP IGFBP-2 and -5 among pigs on days 7, 13, and 22 of the estrous cycle. Relative expression AP IGF-I was greater (P < 0.05) on days 13, 19, and 22 than on day 7 of the estrous cycle. Similarly, the relative expression of AP IGF-IR was increased (P < 0.05) in gilts on days 13, 19, and 22 compared with day 7. The relative expression of GnRHR was greater (P < 0.05) on days 13 and 22 of the estrous cycle than on day 7. The relative expression of LHβ subunit was greater (P < 0.05) on day 19 of the estrous cycle than on days 7, 13, and 22. Anterior pituitary release of LH throughout the porcine estrous cycle may be modulated by changes in the intrapituitary IGF system.
    Document Type:
    Reference
    Product Catalog Number:
    Multiple

  • Administration of estradiol-17beta increases anterior pituitary IGF-I and relative amounts of serum and anterior pituitary IGF-binding proteins in barrows. 11831520

    Two experiments were conducted to determine whether 1) administration of estradiol-173 (E2) implants to barrows elevates serum concentrations of E2 to levels similar to those of adult boars and subsequently affects the anterior pituitary gland IGF system and 2) administration of E2 to barrows increases serum concentrations of E2, serum and anterior pituitary concentrations of IGF-I, and relative amounts of serum and anterior pituitary IGF-binding proteins (IGFBP), vs boars and unimplanted barrows. In Exp. 1, 20 crossbred barrows (150 +/- 6 d, 103 +/- 8 kg) were administered varying number of E2 implants (0, 2, 3, 4; n = 5/group) on d 1. Blood samples were collected weekly by jugular venipuncture, beginning on d 1. Pigs were killed on d 36 when a blood sample and anterior pituitary were collected. Serum concentrations of E2 were increased (P 0.05) in pigs with 2,3, and 4 implants vs 0 implants, but no difference (P > 0.05) was detected in serum concentrations of E2 among pigs with 2, 3, and 4 implants. Orthogonal contrasts identified that three or four E2 implants were necessary to increase serum concentrations of E2 to that similar to boars. Serum and anterior pituitary concentrations of IGF-I were increased (P 0.05) in pigs with 2, 3, and 4 implants vs 0 implants. Relative amounts of anterior pituitary IGFBP-2 and - 5 increased (P 0.05) in response to administration of E2. In Exp. 2, three treatment groups were randomly allotted by litter; boars (n = 11), E2-implanted barrows (n = 9), and unimplanted barrows (n = 12). A blood sample was taken from all pigs on d 1 and every 14 d thereafter. Implanted pigs received four implants on d 1. Pigs were killed on d 91, when a blood sample and anterior pituitary were collected. Mean serum concentrations of E2 were greater (P 0.05) in implanted pigs vs boars. Mean serum concentrations of IGF-I (ng/mL) were greater (P 0.05) in boars (238.7 +/- 6.8) than in implanted barrows (170.2 +/- 8.9) and unimplanted (150.4 +/- 6.7) pigs and tended to be greater (P = 0.08) in implanted vs unimplanted pigs. Mean anterior pituitary concentrations of IGF-I (ng/mg tissue) were greater (P 0.05) in implanted (773.6 +/- 57.0) pigs than boars (251.9 +/- 51.6) and unimplanted (185.6 +/- 49.4) pigs. Relative amounts of serum IGFBP-2 were greater (P 0.05) in implanted pigs vs boars. Relative amounts of anterior pituitary IGFBP-2 and -5 were greater (P 0.05) in boars than in implanted and unimplanted pigs. These data suggest that E2 may influence components of the porcine IGF system in the serum and anterior pituitary. Other gonadal factors present in boars may additionally affect the serum and anterior pituitary IGF system.
    Document Type:
    Reference
    Product Catalog Number:
    Multiple