The GH/IGF system is a complex regulation network strongly dependent on nutrient availability. While the effect of starvation on the GH/IGF system has been extensively studied, the time course of events leading to the restoration of GH/IGF system activity after starvation is largely unknown. We, therefore, measured the plasma levels of GH, IGF-I and IGF-II and the expression of the GH/IGF system in liver and muscle. Starvation increased the plasma GH level and 1 day of refeeding completely restored it (1$10G0$27 vs 1$12G0$28 ng/ml). Thereafter, plasma GH continued to decrease until day 7 and returned to control values from day 15. Starvation decreased plasma IGF-I and IGF-II and refeeding raised plasma IGF-I only from day 4. In contrast, the plasma IGF-II level doubled after 1 day's refeeding (26$5G1$9 vs 44$0G3$4 ng/ml; P!0$01). Starved fish exhibited higher GH receptor (GHR)1 mRNA abundance in liver and muscle than in controls, whereas GHR2 mRNA abundance was increased only in muscle. In liver, 1 day of refeeding, decreased GHR1 (twofold), but increased GHR2 mRNA abundance (twofold). Thereafter, a progressive return to normal values was observed. Liver IGFBP-4 mRNA abundance was lowered in starved fish followed by a progressive restoration during refeeding. Starvation had no effect on liver IGFBP-2 and IGFBP-6 mRNA abundance, whereas refeeding provoked a peak of IGFBP-2 and IGFBP-6 expression at day 7. In muscle, starvation led to a decrease of the IGFBP-2 mRNA level, which was restored only from day 7. IGFBP-4 mRNA abundance in starved fish was lower than in the controls and refeeding led to a transient upregulation (sevenfold) of IGFBP-4 gene at day 1. IGF-I, IGFBP-5, and IGFBP-related protein 1 (rP1) expression profiles were similar, showing a decrease of expression after starvation, a first peak of expression at day 2, a second peak at day 7, and a return to normal value from day 15. Moreover, IGF-I, IGFBP-5, and IGFBP-rP1 mRNA abundance were positively correlated (rZ0$6-0$8; P!0$0001). In conclusion, plasma IGF-I was restored later than plasma GH level, which suggests that plasma IGF-I levels cannot account for plasma GH changes. The coordinated regulation of IGF-I, IGFBP-5, and IGFBP-rP1 expression would be a signature for the resumption of myogenic activity.
In the present study we report the full coding sequence of rainbow trout IGF-binding protein-1 (IGFBP1), -2, -3, -5, and -6 and IGFBP-related protein-1 (IGFBP-rP1) mRNAs as well as the partial coding sequence of IGFBP-4 mRNA. To our knowledge, this is the first report of IGFBP4, IGFBP6, and IGFBP-rP1 in a nonmammalian species. The tissue distribution of all mRNAs was studied, and the ovarian expression profiles of IGFBP2 to -6 and IGFBP-rP1 between late vitellogenesis and oocyte maturation were characterized. In addition, in vitro hormonal regulation by the maturation-inducing steroid 17,20beta-dihydroxy-4-pregnen-3-one (17,20betaP), gonadotropin, and estradiol were studied. We observed that besides IGFBP1, which was only found in liver, IGFBP2 to -6 and IGFPB-rP1 were expressed in the preovulatory ovary. IGFBP3 was also detected in liver, trunk, kidney, skin, and gills, whereas IGFBP2 to -6 and IGFBP-rP1 exhibited a wider tissue distribution. In the preovulatory ovary, IGFBP3 was strongly down-regulated during the postvitellogenesis period, whereas IGFBP5 exhibited a limited up-regulation. In addition, IGFBP6 and IGFBP-rP1 were up-regulated during oocyte maturation. Hormonal regulation data indicated that all ovarian IGFBPs and IGFBP-rP1 transcripts are regulated under gonadotropic stimulation at a concentration that induced 100% oocyte maturation. In addition, IGFBP2 to -5 transcripts are regulated by 17,20betaP and estradiol. Together, our observations strongly suggest that during final oocyte maturation, a down-regulation of IGFBP3, -4, and -5 occurs in the oocyte in response to gonadotropic and 17,20betaP (IGFBP3 and -5) stimulation, whereas an up-regulation of IGFBP2 and -6 occurs in follicular layers or extrafollicular tissue in response to gonadotropic stimulation.
Relationships between insulin-like growth factor-I (IGF-I) and thyroxine (T4) and cortisol hormones were studied in female brood stocks of Persian sturgeon [Acipenser persicus; caught from both sea water (SW) and fresh water (FW)] during late stages of sex maturation. A number of biometrical traits were also studied that could represent the reproductive and/or growth states of brood stocks, and the possible relationship between IGF-I and growth was assessed in juvenile Persian sturgeons between 1 and 4 years of age. IGF-I, T4 and cortisol were measured in serum samples using commercially available kits. A four-parameter logistic model test was performed between the standard curves and the sample dilutions for each hormone. Parallelism, linearity and regression coefficients for the linearized standard curves and serial dilutions of samples were not significantly different (P < 0.05). Serum IGF-I levels in the juveniles were higher than those in the SW brood stocks, and cortisol levels in the former were lower than those in both brood stocks (P < 0.05). T4 levels in serum samples of juveniles were below the detectable level. IGF-I concentrations in juveniles were correlated with total weight, total length and fork length, but they did not increase significantly with increasing age from 1 to 4 years old (P < 0.05). Compared with SW brood stock, the FW brood stock was younger, had a smaller girth, smaller polarization index and higher ova diameter (P < 0.05). There were no differences between IGF-I and T4 levels in the two brood stocks, but cortisol levels were significantly higher in the FW brood stock. Percentage of fertilization was correlated with serum IGF-I in both brood stocks (P < 0.05). Our results support a role for IGF-I during the juvenile growth and reproductive physiology in female brood stocks of the Persian sturgeon.
Two levels of clove oil concentrations (0, 75, and 150 ppm) were prepared in 3 separate aquariums, each include 15 fish goldfish, Carassius auratus (average weight of 65 ± 5 g). Fish were exposed to different concentrations of clove oil and kept in aquariums at 18° C until they reach to stage 4 of anesthesia. Blood samples were taken from caudal vein at 0, 4, and 24 hours after anesthesia. Red blood cell count (RBC), hemoglobin concentration (Hb), hematocrit (PCV), white blood cell count (WBC) and the differential leukocyte count (leukogram) were determined by standard hematology method. Results showed that there were no significant differences between values of Hb, PCV and leukogram in each treatment in compare to the same control group (P> 0.05); however, WBC was significantly lower at 4 hours for 150 ppm clove oil treatment group and then returned to normal level 24 hours post anesthesia (P< 0.05). Moreover, RBC in this group was increased significantly after 24 h post anesthesia (P< 0.05). The induction time was less for 150 than 75 ppm clove oil treatment group (90 and 180 seconds respectively). Our results verified that using clove oil up to 75 ppm dose not have irreversible hematological side effect to the goldfish.
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