Increased Concentrations of Insulin-Like Growth Factor Binding Protein (IGFBP)-2, IGFBP-3, and IGFBP-4 Are Associated With Fetal Mortality in Pregnant Cows

Figure 1. Analysis of insulin-like growth factor-binding proteins (IGFBPs) in samples from pregnant and non-pregnant cows by Western ligand blotting. Due to protein glycosylation, IGFBP-3 and -4 can be detected as band doublets (Co: internal control; St: human recombinant IGFBP-standard).

Figure 2. Semi-quantitative pregnancy-associated glycoproteins (PAG) values given in optical density (OD) units and insulin-like growth factor 1 (IGF-1) concentrations. (A,B) Study trial I in not pregnant [np, n = 43, day of AI (0) and 18 days after AI] and pregnant (p, n = 34, day of AI, and 18, 42, 60 and 100 days after AI) pluriparous Holstein–Friesian cows as well as cows with fetal mortality (fm, n = 4, day of AI and until day 60 after AI). (C,D) Study trial II in not pregnant (np, n = 20, 10–14 (IGF-1), 24–27, 34–37 days after AI) and pregnant (p, n = 30, 24–27, 34–37 and 54–57 days after AI) Holstein–Friesian cows as well as cows with embryo mortality (em, n = 7, 24–27, 34–37 days after AI) or late embryo/fetal mortality (em/fm, n = 8, 24–27, 34–37, and 54–57 days after AI). Values are given as mean ± SEM.

Figure 3. Study trial I: insulin-like growth factor-binding protein (IGFBP) (A): -2, (B): -3 and (C): -4 concentration and (D): IGFBP-3/IGFBP-2 ratio in not-pregnant [np, n = 43, day of AI (0) and 18 days after AI] and pregnant (p, n = 34, day of AI, and 18, 42, 60 and 100 days after AI) pluriparous Holstein–Friesian cows as well as cows with fetal mortality (fm, n = 4, day of AI and until day 60 after AI). Values are given in% relative to day 0 of non-pregnant cows. Values are given as mean ± SEM.

Figure 4. Study trial II: insulin-like growth factor-binding protein (IGFBP) (A): -2. (B): -3. (C): -4 and (D): IGFBP3/IGFBO2 ratio in non pregnant (np, n = 20) and pregnant (p, n = 30) Holstein Friesian cows and cows with embryo mortality (em, n = 7) between days 24-27 and 34-37 or late embryo mortality/early fetal mortality (em/fm, n = 8) between days 24-27 and 54-57. Values are given in percentege relative to days 10–14 of non-pregnant cows. Values are given as mean ± SEM.

Figure 5. Western ligand blots with serum samples of not-pregnant and pregnant cows before (−I) and after (+I) incubation (24 h, 39°C) with Insulin-like Growth factor 4 (IGFBP-4) and insulin-like growth factor 1 (IGF-1) spiked to the serum. As positive control serum of a pregnant woman was available for which proteolytic activity IGFBP-4 is clearly proven.