The importance of thyroid hormone from embryonic through neonatal life has been documented in both avian and mammalian species. However, the regulation of thyroid hormone production during this period is not completely understood. The objective of this study was to characterize expression of chicken TSHbeta messenger RNA (mRNA) compared with that of thyroid hormones and GH in embryonic and neonatal chickens. Total pituitary RNA was extracted on embryonic days (e-) 11, 13, 15, 17, and 19 and neonatal days (d-) 1, 3, 6, 9, and 12 and subjected to ribonuclease protection assays (RPA) for chicken TSHbeta mRNA. TSHbeta mRNA levels increased through e-19, with e-19 levels being greater than those at all other embryonic ages (P < 0.05). Levels decreased markedly on d-1, then slowly increased to d-6 and stayed elevated through d-12. RIAs were performed for T4, T3, and GH at the same ages. Serum T4 levels increased slowly from less than 1.0 ng/ml on e-11 to a peak of 6.6 ng/ml on d-1 (P < 0.05). After the peak on d-1, posthatch T4 levels stabilized between 3.5-4.5 ng/ml through d-12 (P < 0.05). T3 concentrations were less than 0.25 ng/ml on e-11, increased dramatically between e-19 and d-1 (P < 0.05), and remained high throughout the rest of the experiment, with a concentration of 3.25 ng/ml on d-6 (P < 0.05). GH levels for e-11 through e-17 were below the sensitivity of the GH RIA. On e-19, the GH level was 3 ng/ml and continued to increase through d-12 to a level of 130 ng/ml. As thyroid hormone levels were preceded by maximal TSHbeta mRNA levels on e-19, we next determined whether TSHbeta gene expression on e-19 was under TRH and T3 regulation. E-19 anterior pituitary cells were cultured in serum-free medium with either TRH (10[-8]) or T3 (10[-8]) for 20-24 h. Treatment with T3 significantly decreased levels of TSHbeta mRNA (P < 0.05). However, TRH did not produce a significant increase in TSHbeta mRNA, although TRH did increase TSHbeta mRNA by 60%, on the average, in this study. Therefore, these results indicate that an increase in pituitary TSH production probably regulates thyroid hormone levels during late embryonic development and that negative feedback inhibition of TSH production by thyroid hormones also exists at this critical developmental stage.
Incubation behavior in birds is characterized by elevated serum prolactin (PRL) levels. The objective of this study was to determine the cellular basis for increased PRL secretion. Incubation behavior of Bantam hens was induced by allowing the hens to accumulate their eggs. In experiment 1, hens were allowed to accumulate their eggs in floor pens for 0, 2, 12, or 24 days (Day 0, Day 2, Day 12, and Day 24 hens, respectively). Anterior pituitaries were dissociated into individual cells with trypsin, and the resulting cells were then subjected to reverse hemolytic plaque assays for PRL. The percentage of PRL-secreting cells and serum PRL levels were higher in Day 24 hens than in the other groups (p < 0.05; n = 3 separate trials). In experiment 2, hens were allowed to accumulate their eggs for 2, 14, 18, or 24 days (Day 2, Day 14, Day 18, and Day 24 hens, respectively). The percentage of PRL-secreting cells and levels of serum PRL were greater in Day 18 and Day 24 hens than in Day 2 hens (p < 0.05; n = 4). Results from Day 14 hens were intermediate and different (p < 0.05) from those from both Day 2 and Day 18 groups. Data indicate that an increase in the proportion of PRL secretors occurred between 14 and 18 days after hens began to accumulate their eggs. The area of plaques formed by lactotrophs from Day 14 and Day 18 hens was also increased (p < 0.05). In experiment 3, hens were allowed to accumulate their eggs for 0 and 24 days (Day 0 and Day 24 hens, respectively). Their anterior pituitaries were dissected into cephalic, middle, and caudal regions, and the percentage of PRL cells were analyzed in each region separately. The percentage of PRL secretors was increased (p < 0.05) in Day 24 compared to Day 0 hens in both the cephalic and middle regions of the anterior pituitary but not in the caudal portion. We conclude that increased PRL secretion during incubation behavior involves the recruitment of additional pituitary cells into the PRL-secreting population and an increase in the capacity of these cells to secrete hormone (p < 0.05). Furthermore, this increase in PRL-secreting cells is localized to the cephalic and middle regions of the anterior pituitary.
The importance of thyroid hormone from embryonic through neonatal life has been documented in both avian and mammalian species. However, the regulation of thyroid hormone production during this period is not completely understood. The objective of this study was to characterize expression of chicken TSHbeta messenger RNA (mRNA) compared with that of thyroid hormones and GH in embryonic and neonatal chickens. Total pituitary RNA was extracted on embryonic days (e-) 11, 13, 15, 17, and 19 and neonatal days (d-) 1, 3, 6, 9, and 12 and subjected to ribonuclease protection assays (RPA) for chicken TSHbeta mRNA. TSHbeta mRNA levels increased through e-19, with e-19 levels being greater than those at all other embryonic ages (P < 0.05). Levels decreased markedly on d-1, then slowly increased to d-6 and stayed elevated through d-12. RIAs were performed for T4, T3, and GH at the same ages. Serum T4 levels increased slowly from less than 1.0 ng/ml on e-11 to a peak of 6.6 ng/ml on d-1 (P < 0.05). After the peak on d-1, posthatch T4 levels stabilized between 3.5-4.5 ng/ml through d-12 (P < 0.05). T3 concentrations were less than 0.25 ng/ml on e-11, increased dramatically between e-19 and d-1 (P < 0.05), and remained high throughout the rest of the experiment, with a concentration of 3.25 ng/ml on d-6 (P < 0.05). GH levels for e-11 through e-17 were below the sensitivity of the GH RIA. On e-19, the GH level was 3 ng/ml and continued to increase through d-12 to a level of 130 ng/ml. As thyroid hormone levels were preceded by maximal TSHbeta mRNA levels on e-19, we next determined whether TSHbeta gene expression on e-19 was under TRH and T3 regulation. E-19 anterior pituitary cells were cultured in serum-free medium with either TRH (10[-8]) or T3 (10[-8]) for 20-24 h. Treatment with T3 significantly decreased levels of TSHbeta mRNA (P < 0.05). However, TRH did not produce a significant increase in TSHbeta mRNA, although TRH did increase TSHbeta mRNA by 60%, on the average, in this study. Therefore, these results indicate that an increase in pituitary TSH production probably regulates thyroid hormone levels during late embryonic development and that negative feedback inhibition of TSH production by thyroid hormones also exists at this critical developmental stage.
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