Domestic Animal Endocrinology

Editorial Board

2012-05-01

The impact of maternal overnutrition and obesity on hypothalamic-pituitary-adrenal axis response of offspring to stress

N.M. Long, P.W. Nathanielsz, S.P. Ford — 2012-01-24

Abstract: We evaluated the effect of maternal obesity before and throughout gestation on offspring hypothalamic-pituitary-adrenal axis function. Multiparous Rambouillet by Columbia crossbred ewes were fed either 100% of National Research Council (NRC) recommendations (control, C) or 150% of NRC recommendations (obese, OB) from 60 d before mating until lambing. Ten lambs born to OB ewes (five males and five females), and eight lambs born to C ewes (three male and five female) were studied. From delivery to weaning lambs were maintained with their mothers, who were all fed 100% NRC recommendations. After weaning, all lambs were group housed and fed the same diet to meet NRC requirements. At 19 mo of age lambs were placed in individual pens and fed a pelletized diet to meet maintenance requirements. Jugular vein catheters were placed and 2 d later lambs received an intravenous (i.v.) adrenocorticotropic hormone (ACTH) challenge followed by an i.v. corticotropin-releasing hormone (CRH)/arginine vasopressin (AVP) challenge 1 d later. Thirty d later offspring were again catheterized and placed into metabolism crates for 2 d before receiving an isolation stress test. ACTH and cortisol responses to the isolation stress test and CRH/AVP challenge and cortisol responses to ACTH challenge were determined. Cortisol was quantified via radioimmunoassay and ACTH was quantified using an Immulite 1000; both were analyzed using repeated measures using the MIXED procedure of SAS. Offspring from OB ewes had elevated basal plasma ACTH and cortisol compared with C offspring before all three challenges (P < 0.05). Offspring from OB mothers tended (P = 0.06) to have a greater ACTH response after an i.v. CRH/AVP injection than offspring from C mothers (12,340 ± 1,430 vs 8,170 ± 1,570 area under the curve, respectively). Cortisol response to the CRH/AVP and ACTH challenges was not influenced by maternal nutrition (P = 0.46) and averaged 4.77 ± 0.2 μg/dL and 1.94 ± 0.01 μg/dL, respectively. The ACTH response following the isolation stress test was also similar (P = 0.82) for OB and C offspring (147 ± 20 pg/mL), and cortisol response during the isolation stress test was similar between C and OB offspring (P = 0.64, 5.25 ± 0.3 μg/dL). These findings suggest that maternal obesity before and during gestation does not affect stress responses by the offspring, but has an impact on hypothalamic-pituitary-adrenal sensitivity. The lack of differences in cortisol release under the influence of difference concentrations of ACTH during the CRH/AVP challenge could indicate adrenal dysfunction in OB offspring.

Cysteamine improves growth performance and gastric ghrelin expression in preweaning piglets

G. Du, Z. Shi, D. Xia, X. Wei, L. Zhang, N. Parvizi, R. Zhao — 2012-01-12

Abstract: The aim of the present study was to investigate the effect of cysteamine on growth performance of preweaning piglets and gastric expression of ghrelin mRNA in vivo and in vitro. Twelve litters of newborn piglets were allocated randomly to control and treatment groups. From 15 d of age, piglets in the control group were fed basal creep diet, whereas the treatment group received basal diet supplemented with 120 mg cysteamine per kg of diet until weaning on 35 d of age. Body weight gain, creep feed consumption, and diarrhea rates were recorded, and gastric mucosal tissues were collected for quantifying mRNA expression. To evaluate the direct effect of cysteamine on gastric ghrelin expression, primary cultures of gastric mucosal cells isolated from 35-d-old piglets were exposed to cysteamine for 20 h at 0, 1, 10, and 100 μg/mL, respectively. Dietary cysteamine increased (P < 0.05) average daily creep feed consumption and BW gain in preweaning pigs, which was accompanied by reduction in diarrhea rates. At 35 d of age, piglets treated with cysteamine showed increased (P < 0.05) ghrelin and gastrin and decreased (P < 0.05) somatostatin mRNA expression in gastric mucosa. Moreover, dietary cysteamine treatment increased serum concentration of gastrin (P < 0.05). In vitro, cysteamine significantly increased ghrelin mRNA expression in gastric mucosal cells at the concentration of 10 μg/mL. In conclusion, dietary cysteamine is effective in improving the growth performance and health condition of preweaning piglets, which is associated with its stimulatory effects on gastric ghrelin mRNA expression both in vivo and in vitro.

Stromal-epithelial interactions modulate the effect of ovarian steroids on goat uterine epithelial cell interleukin-18 release

X.F. Qi, Z.C. Nan, Y.P. Jin, Y.Y. Qu, X.J. Zhao, A.H. Wang — 2012-01-09

Abstract: A primary role of epithelial-stromal interactions in mediating steroid hormone action in the uterus has been established. The present study was undertaken to determine the mode of ovarian steroid action in regulating IL-18 release by goat endometrial epithelial cells (EECs) in the presence and absence of endometrial stromal cells (ESCs). Primary and telomerase-immortalized goat EECs grown alone or cocultured with ESCs were treated with two ovarian steroids, 17β-estradiol (E2) and progesterone (P4). The IL-18 mRNA and protein expression in EECs were studied by reverse transcript (RT) PCR, ELISA, and Western blot assay. The E2 and/or P4 treatment of EECs led to a significant increase in both IL-18 mRNA and protein expression either in the primary or in the immortalized EECs compared with that in EECs without the steroid treatment. However, in the presence of ESCs, IL-18 expression by EECs treated with steroids was significantly decreased compared with cells untreated with E2 and/or P4. In addition, significantly high abundance of IL-18 mRNA and protein expression by primary and telomerase-immortalized goat EECs was observed in the presence of ESCs compared with those cells without ESCs. These findings suggest that steroids are important for the control of IL-18 expression in goat EECs. Underlying ESCs are needed to mediate the inhibitory effects of steroids on the IL-18 secretory activity of goat EECs in vitro. The IL-18 abundance expressed by goat EECs in vitro are enhanced by underlying ESCs without the treatment of E2 and/or P4.

Yellowtail insulin-like growth factor 1: molecular cloning and response to various nutritional conditions

H. Fukada, K. Murashita, T. Furutani, T. Masumoto — 2012-01-24

Abstract: Insulin-like growth factor 1 (IGF1) plays an important role in fish growth. This study investigated the IGF1 response to various nutritional conditions in yellowtail. First, we cloned 1,075 bp of yellowtail IGF1 cDNA, which codes for a protein of 185 amino acids (aa). This is composed of 44 aa for the signal peptide; 68 aa for the mature peptide comprising the B, C, A, and D domains; and 73 aa for the E domain. The mature yellowtail IGF1 showed high identity to IGF1 of other teleosts. Insulin-like growth factor 1 mRNA expression in the liver and white muscle was measured to observe the IGF1 response to various nutritional conditions, because the liver has the highest IGF1 expression and white muscle comprises the largest fraction of the fish body. Only white muscle IGF1 mRNA expression decreased significantly by 3 wk of fasting and recovered by refeeding. In subsequent feeding ratio (1%, 2%, and 3%/BW/d) experiments, significant correlations to growth were observed in white muscle IGF1 mRNA expression at 2- and 6-wk points and in hepatic IGF1 mRNA expression at 4 wk point. These data suggest that IGF1 expression both in hepatic and white muscle is important for somatic growth in yellowtail. Furthermore, white muscle IGF1 mRNA expression showed better responses to somatic growth and nutrition status in our two experiments than hepatic IGF1 mRNA expression.

Disruption of fibroblast growth factor receptor signaling in bovine cumulus-oocyte complexes during in vitro maturation reduces subsequent embryonic development

K. Zhang, A.D. Ealy — 2012-01-24

Abstract: Several fibroblast growth factors (FGF) mediate folliculogenesis and oogenesis in cattle but it is unclear whether FGFs are required during the final stages of oocyte maturation. The objectives of this work were to determine whether blocking FGF receptor (FGFR) activity during in vitro maturation (IVM) affects oocyte fertilization and embryo development; examine changes in FGFR transcript profiles in cumulus cells and oocytes during IVM; and evaluate whether gonadotropins modulate FGFR transcript abundance during IVM. In the first set of studies, bovine cumulus-oocyte complexes (COCs) were matured in the presence of one of two FGFR kinase inhibitors (SU5402 or PD173074). After maturation, COCs were washed and cultured without inhibitors. Inhibitors did not affect cleavage rates but the percentage of ≥ 8-cell embryos at d 3 and blastocysts at d 7 and d 8 postfertilization were decreased when COCs were matured with either inhibitor. Profiles of FGFR mRNA variants were examined in cumulus cells and oocytes separated either immediately before (0 h) or at 6 or 21 h after beginning IVM. In cumulus cells, increases in R1b, R2b, and R2c abundance were detected when cultured in the absence of follicle-stimulating hormone (FSH). Supplementing FSH (1 or 25 μM) increased the abundance of R1b, R1c, R2b, and R2c. In oocytes, no time- or FSH-dependent changes in FGFR transcript abundance were detected. These observations implicate FGFs as crucial components of bovine oocyte competency and indicate that FSH augments FGFR mRNA abundance in cumulus cells during the final stages of oocyte maturation.

Molecular and functional characterization of grass carp squint/nodal-related 1: a potential regulator of activin signaling in teleost pituitary cells

T. Zhao, X. Wang, H. Wei, M. Yang, F. Zeng, H. Zhou — 2012-02-15

Abstract: Nodal, a member of the transforming growth factor-β superfamily, plays important roles in embryogenesis in vertebrates, including fish. However, the functional characterization of the fish nodal-related gene in nonembryonic cells is still unclear. In teleost, three nodal-related genes, nodal-related (ndr)1/squint, ndr2/cyclops, and ndr3/southpaw have been reported. In this study, a full-length cDNA for grass carp squint (gcSqt) was cloned, and its transcript was detected in the selected organs, including pituitary, brain, heart, head kidney, kidney, spleen, and gonad. To further define its functional role, recombinant grass carp squint (rgcSQT) was produced in Escherichia coli in a homodimer form. Furthermore, we examined the effects of rgcSQT on activin and its receptor gene expression with the use of grass carp pituitary cell as a model. Results showed that rgcSQT stimulated the mRNA expression of activin βA and βB subunit, as well as activin receptor ActRIB and ActRIIB. These findings not only contribute to the understanding of nonembryonic functions of nodal gene in fish, but they also provide new insight into the regulation of activin signaling in vertebrates.

Fecal corticosterone metabolites and plasma corticosterone in Japanese quail selected for low or high plasma corticosterone responses to brief restraint

J.F. Cockrem, D.G. Satterlee, E.J. Candy, S.A. Castille — 2012-02-13

Abstract: Fecal corticosterone metabolites and plasma corticosterone in Japanese quail selected for low- or high-plasma corticosterone responses to brief mechanical restraint (low- and high-stress lines), and in a line of unselected quail, were measured in this study. No line differences were observed in baseline plasma corticosterone concentrations, but fecal corticosterone metabolite concentrations and daily fecal corticosterone metabolite production were 20% higher in quail of the high-stress line than in unselected or low-stress quail for males and females living together in group cages (P < 0.05). No differences were observed between lines in corticosterone metabolite concentrations and production for male birds in individual cages. Baseline plasma corticosterone concentrations, fecal corticosterone metabolite concentrations, and production appeared to be higher for males and females in group cages compared with males in individual cages. This difference might have been because of greater corticosterone secretion by male quail living in mixed sex groups than living individually. Correlations between baseline plasma corticosterone concentrations and fecal corticosterone metabolite concentrations in low-stress and high-stress quail, and for all birds combined, were r = 0.521 (P = 0.038), r = 0.604 (P = 0.013), and r = 0.431 (P = 0.002), respectively. The low- and high-stress lines that have been selected for low- and high-corticosterone responses differ in other characteristics, including growth and reproductive performance, and the current results are consistent with the assumption that these other differences are a consequence of greater daily corticosterone secretion in quail of the high-stress line.

Feline parathyroid hormone: validation of hormonal assays and dynamics of secretion

C. Pineda, E. Aguilera-Tejero, A.I. Raya, E. Diez, M. Rodriguez, I. Lopez — 2012-02-27

Abstract: Validated assays for quantification of intact parathyroid hormone (I-PTH) are no longer available. Moreover, the third-generation PTH assay that only detects the whole PTH molecule (W-PTH) has never been tested in cats. The work presented here is aimed to validate a commercially available assay for measurement of I-PTH and W-PTH in cats and to study the dynamics of PTH secretion in healthy cats. Our results show that both assays are reliable for the measurement of feline PTH. In healthy adult cats W-PTH concentration (15.1 ± 1.6 pg/mL) was greater (P < 0.001) than I-PTH concentration (9.1 ± 0.7 pg/mL). The dynamics of PTH secretion in response to changes in extracellular calcium (Ca2+) were investigated in 13 cats by studying PTH-Ca2+ curves. PTH-Ca2+ curves were obtained by intravenous infusion of disodium ethylenediaminetetraacetic acid and CaCl2. PTH was measured using both I-PTH and W-PTH assays. During hypocalcemia a sigmoidal curve that was similar when measured with I-PTH or W-PTH was obtained. The maximal PTH concentration in response to hypocalcemia was greater with W-PTH (179.6 ± 41.9 pg/mL) than with I-PTH (67.6 ± 10.5 pg/mL; P = 0.01). However, hypercalcemia resulted in an equivalent PTH inhibition, with both assays yielding PTH concentrations as follows: W-PTH = 4.0 ± 0.4 pg/mL and I-PTH = 4.9 ± 0.3 pg/mL (NS). Parameters of the feline PTH-Ca2+ curve are similar to what has been previously reported in dogs.