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ABSTRACT

Changes in neuropeptide gene expression in the hypothalami of sheep subjected to psychological stress (isolation, 1 h; n=3) or dehydration (48 h; n=3) were examined using in-situ hybridization histochemistry. Compared with non-stressed euhydrated control animals (n=3), isolation induced significant accumulation of mRNA for corticotrophin-releasing hormone, pro-enkephalin and pro-dynorphin (DYN) in the paraventricular nucleus (PVN), but no change in mRNA content within the supraoptic nucleus (SON). By contrast, dehydration significantly increased DYN mRNA in the magnocellular neurones of the PVN and SON. However, neither isolation nor dehydration altered the expression of mRNA for vasopressin (AVP) in either the PVN or the SON. These results indicate that in the ovine hypothalamus (1) stress represents a powerful stimulus to co-ordinated neuropeptide synthesis and (2) expression of DYN mRNA and AVP mRNA may be independently regulated during changes in plasma osmolality.

ABSTRACT

To examine the role of 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1) in the control of glucocorticoid actions in the ovine pituitary during development, we have sought developmental changes in the distribution and the level of 11β-HSD1 mRNA by in situ hybridization. In the pars distalis, 11β-HSD1 mRNA was present by day 60; its amount did not change significantly until term (days 145–147) when it increased dramatically. The level of 11β-HSD1 mRNA increased further during the postnatal period. In contrast, 11β-HSD1 mRNA in the pars intermedia was not detectable until day 135; it increased in amount at days 140–143, but did not change significantly thereafter through to adulthood.

We have also measured levels of both dehydrogenase and reductase activities of 11β-HSD1 in the pars distalis of fetal sheep at day 140 and term, and of postnatal sheep at 1–2 months of age, to determine whether changes in 11β-HSD1 mRNA are reflected in the levels of enzyme activities. There were progressive increases in both dehydrogenase and reductase activities from day 140 to 1–2 months postnatally, although dehydrogenase activity was consistently higher than reductase activity.

Finally, we have determined the effect of short-term intrafetal cortisol infusion (5 μg/min for 12 h) on levels of pituitary 11β-HSD1 mRNA by in situ hybridization. There was no effect of cortisol infusion on 11β-HSD1 mRNA expression.

The present results demonstrate that 11β-HSD mRNA and enzyme activity in the pars distalis of fetal sheep increase dramatically at term when plasma levels of both ACTH and cortisol are elevated. This suggests that 11β-HSD1 may contribute to the proposed resetting of cortisol negative feedback within the fetal pituitary at that time.

ABSTRACT

Ontogenic changes in pituitary pro-opiomelanocortin (POMC) mRNA and prolactin (PRL) mRNA were examined during gestation and early neonatal life using in situ hybridization histochemistry. Pituitaries were harvested from fetuses at days 60–80, 100–120, 135–140 and 142–143 of gestation and at term, and from lambs at days 1–7 and 30–60 of age and adults.

POMC mRNA, present by day 60, rose during mid- and late gestation. Concurrently there was a change in corticotroph distribution, resulting in a relatively greater quantity of POMC mRNA at the base of the pars distalis. At term, there was a significant (P<0·05) further elevation of POMC mRNA. POMC mRNA levels remained high in the newborn lamb but decreased in the adult. Cells in the pars intermedia expressed large amounts of POMC mRNA early in fetal life and this pattern persisted throughout gestation and into the neonatal period. Changes in the expression of the POMC gene correlated closely with the presence of immunoreactive (ir)ACTH in the pituitary; in fetuses the proportion of irACTH-positive cells rose to 10% of pars distalis cells by day 100 and did not change significantly thereafter. The lactotrophs contained PRL mRNA by day 60, and the quantity increased towards parturition (P<0·05). PRL mRNA subsequently decreased in the neonate, but rose as the lamb matured.

These results indicate that in the fetal pituitary: (1) the POMC gene is highly expressed during gestation in both the pars distalis and the pars intermedia, (2) changes in the amounts of POMC mRNA and PRL mRNA in the pars distalis correlate with the distribution of irACTH and irPRL respectively, and (3) POMC mRNA is distributed primarily in the inferior aspect of the pars distalis, and in this region its quantity is highest immediately prior to parturition.

Abstract

The effect of nutritional status on IGF-I mRNA expression in the liver and brain of juvenile barramundi (Lates calcarifer) was investigated. Fish were either fed a satiety ration (SAT) or starved (STV) for 6 weeks. Starved fish demonstrated significantly lower condition factor and hepatic IGF-I mRNA expression at 3 and 6 weeks, when compared with the SAT group. IGF-I mRNA expression in the brain was 10 fold lower than the liver and was not affected by ration size. These results suggest the liver is the major site of IGF-I mRNA synthesis and hepatic but not brain IGF-I mRNA expression is regulated by food availability in juvenile barramundi.