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The role of the pituitary gland and ACTH in the regulation of mRNAs encoding proteins essential for adrenal steroidogenesis in the late-gestation ovine fetus.

To further understand the relative roles of the pituitary gland and ACTH in the regulation of mRNAs encoding proteins that are essential for adrenal development, we investigated the effects of, first, an ACTH infusion and labour in intact fetuses and, secondly, the effect of an ACTH infusion to fetuses with and without a pituitary gland, on the relative abundance of the mRNA encoding for the ACTH receptor (MC2R), steroidogenic factor 1 (SF-1), cholesterol side-chain cleavage enzyme (P450(scc)), 3beta-hydroxysteroid dehydrogenase (3betaHSD) and 17alpha-hydroxylase (P450(C17)) in the fetal adrenal gland. ACTH(1-24) infusion (14.7 pmol/kg per h) to intact fetuses was without effect on the abundance of mRNA encoding MC2R and SF-1, irrespective of whether the infusion was given for 18 (115-132 days of gestation) or 32 days (115 days to term (147 days of gestation)). Hypophysectomy (HX) did not alter the expression of MC2R mRNA; however, the abundance of SF-1 mRNA fell by approximately 50% following the removal of the pituitary gland. ACTH(1-24) infusion to HX fetuses failed to restore levels of SF-1 mRNA to that seen in intact animals. P450(scc) and 3betaHSD mRNAs were increased by ACTH(1-24) infusion for 18 days in intact animals, although no effects of the infusion were seen on P450(C17) mRNA levels. For all three of these mRNAs, there was a significant increase in their abundance between 132 days of gestation and term in intact fetuses. By term, ACTH(1-24) infusion was without any additional effect on their abundance. HX decreased the expression of P450(scc), 3betaHSD and P450(C17) mRNAs, while ACTH(1-24) infusion to HX fetuses increased the expression of these mRNAs to levels seen in intact animals. There were significant correlations between the abundance of the mRNA for P450(scc), 3betaHSD and P450(C17), but not MC2R and SF-1, and premortem plasma cortisol concentrations. These results emphasise the importance of the pituitary gland and ACTH in the regulation of the enzymes involved in adrenal steroidogenesis. Factors in addition to ACTH may also play some role, as the infusion was not always effective in increasing the abundance of the mRNAs. Surprisingly, the mRNA for MC2R and SF-1 did not appear to be regulated by ACTH in the late-gestation ovine fetus, though a pituitary-dependent factor may be involved in the regulation of SF-1 mRNA abundance.

Adrenal Glands↗

Contrast enhanced area of posterior pituitary gland in early dynamic MRI exceeds hyperintense area on T1-weighted images.

Seventeen normal pituitary glands were studied by midline sagittal (and in one case axial) dynamic MRI before and immediately after injection of Gd-DTPA, using spin-echo sequences with a short repetition time. Images were obtained at 1 min intervals for 6 min at 1.5 T. A well-defined focal area with marked enhancement was seen in the posterior portion of the pituitary gland on the first image after injection of Gd-DTPA. The size of the early enhancing area in the posterior pituitary (EEPP) was compared with the high signal of the posterior pituitary (HSPP) on the precontrast image. The EEPP was larger than the HSPP in 10 cases (59%). The two were equal in size in seven cases (41%). The EEPP was anteriorly convex in 11 cases (65%), whereas HSPP was anteriorly convex in 4 cases (24%). We believe that EEPP closely represents the real extent of the posterior lobe. Our results support the hypothesis that HSPP may involve only a portion of the posterior pituitary.

Contrast Media↗

Electron-microscopic study on the anterior pituitary gland of spontaneous dwarf rats.

The spontaneous dwarf rat is a novel experimental model animal on the study of pituitary dwarfism. The fine structure of the anterior pituitary cells was studied in the immature and mature dwarf rats. Pituitary glands were removed from 5-, 10-, 20-day-old immature dwarfs, adult (45 days-16 weeks) dwarfs and normal 3-month-old rats and processed for electron-microscopic observation. In the control animals, growth hormone cells were readily identified by their ultrastructural characteristics, such as the presence of numerous electron-dense secretory granules, 300-350 nm in diameter, well developed rough endoplasmic reticulum and a prominent Golgi complex. In contrast, growth hormone cells were not found in the anterior pituitary gland of the spontaneous dwarf rat at any age examined. Other pituitary cell types, i.e., luteinizing hormone/follicle stimulating hormone, thyroid stimulating hormone, adrenocorticotropic hormone and prolactin cells, appeared similar in their fine structure to those found in the control rats. In the pituitary gland of dwarf rats, a number of polygonal cells were observed either with no or relatively few secretory granules. The rough endoplasmic reticulum was arranged in parallel cisternae and the Golgi complex was generally prominent in these cells. In addition, many were found to have abundant lysosomes. A few minute secretory granules were occasionally observed; however, the immunogold technique failed to localize growth hormone or prolactin in the granules. The nature of these cells remained obscure in this study.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Normal development of the pituitary gland: assessment with three-dimensional MR volumetry.

BACKGROUND AND PURPOSE: Currently available data on pituitary volume have been based on indirect methods of measurement and are mostly limited to adult populations. The purpose of this study was to determine the normal development of pituitary volume by means of direct measurements made on thin-section 3D MR acquisitions. METHODS: The volume of the normal pituitary gland in children and adolescents was measured by using 3D MR sequences with a section thickness of 0.6 to 0.75 mm. The clinical study group consisted of 199 pediatric patients with clinically normal pituitary function and no abnormal findings on routine MR studies. The volume of the posterior pituitary was also measured in all the subjects. RESULTS: A phantom study revealed measurement errors within 25%. In the clinical study, the measured pituitary volumes showed a growth spurt during puberty, which was more prominent in girls. Posterior pituitary volumes showed gradual growth without such a spurt. Among 5- to 9-year-olds, the posterior pituitary volumes were significantly larger for boys than for girls. CONCLUSION: Normal development of the pituitary gland and posterior pituitary was determined by means of 3D MR volumetry. With this technique, we found a gender difference in the volume of the posterior pituitary.

Adolescent↗

Isolation and biological properties of secretory granules from rat anterior pituitary glands.

A method is described for the isolation of secretory granules from rat anterior pituitary glands. The method consists of differential and isopycnic gradient centrifugations, followed by filtration of the zones containing granules on Nuclepore filters to remove mitochondria. Highly purified granules were obtained as indicated by electron microscopy. Major parts of the thyrotropin (TSH) and adrenocorticotropin (ACTH) were recovered in a single fraction of granules as were follicle-stimulating (FSH) and luteinizing (LH) hormones. The somatotropin (STH) and prolactin (LTH) were recovered in separate granule fractions. The major parts of the six different hormones were associated with their respective granule fractions as shown by bioassays specific for each of the hormones. The diameters of granules in sections of intact rat pituitary glands and in isolated pellets were measured, and the means and ranges were in close agreement. These results contribute to the identification of the cell types which produce the different pituitary hormones.

Adrenocorticotropic Hormone↗

Serotonin and dopamine receptors in the rat pituitary gland: autoradiographic identification, characterization, and localization.

S2 serotonin and D2 dopamine receptors were identified, characterized, and localized in rat pituitary gland by quantitative light microscopic autoradiography. [3H]Spiperone was used to localize S2 serotonin and D2 dopamine receptors. A high concentration of D2 dopamine receptors [1 microM 2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene (ADTN)- or sulpiride-displaceable [3H]spiperone binding] was found in the rat intermediate lobe with much lower concentrations present in the anterior and posterior lobes. Significant densities of cinanserin-displaceable [3H]spiperone binding sites (i.e. S2 serotonin receptors) were present in all three lobes of the pituitary gland. [125I]Lysergic acid ([125I]LSD) was used to characterize further and selectively visualize S2 serotonin receptors in the rat pituitary. Data analysis by densitometry showed that [125I]LSD binding the rat intermediate pituitary was saturable and of high affinity with an apparent dissociation constant (Kd) of 1.2 nM. Data from competition studies using a variety of compounds showed a S2 serotonin receptor profile at this [125I]LSD binding site in rat pituitary. The highest concentration of [125I]LSD binding sites was found in the intermediate lobe with progressively lower concentrations present in the posterior and anterior lobes, respectively. There is a uniform pattern of distribution of S2 serotonin and D2 dopamine receptors within each lobe of the rat pituitary gland. The results of the present study provide the first identification of S2 serotonin receptors in the pituitary and confirm the heterogeneous distribution of D2 dopamine receptors within the rat pituitary. These data provide further evidence for the importance of dopamine in regulating pituitary function and suggest a physiological role for serotonin in regulating pituitary hormone secretion.

Animals↗

Localisation and physiological regulation of corticotrophin-releasing factor receptor 1 mRNA in the Xenopus laevis brain and pituitary gland.

In Xenopus laevis, corticotrophin-releasing factor (CRF) and urocortin 1 are present in the brain and they both are potent stimulators of alpha-melanophore stimulating hormone (MSH) secretion by melanotroph cells in the pituitary gland. Because both CRF and urocortin 1 bind with high affinity to CRF receptor type 1 (CRF1) in mammals and Xenopus laevis, one of the purposes of the present study was to identify the sites of action of CRF and urocortin 1 in the Xenopus brain and pituitary gland. Moreover, we raised the hypothesis that the external light intensity is a physiological condition controlling CRF1 expression in the pituitary melanotroph cells. By in situ hybridisation, the presence of CRF1 mRNA is demonstrated in the olfactory bulb, amygdala, nucleus accumbens, preoptic area, ventral habenular nuclei, ventromedial thalamic area, suprachiasmatic nucleus, ventral hypothalamic area, posterior tuberculum, tectum mesencephali and cerebellum. In the pituitary gland, CRF1 mRNA occurs in the intermediate and distal lobe. The optical density of the CRF1 mRNA hybridisation signal in the intermediate lobe of the pituitary gland is 59.4% stronger in white-adapted animals than in black-adapted ones, supporting the hypothesis that the environmental light condition controls CRF1 mRNA expression in melanotroph cells of X. laevis, a mechanism likely to be responsible for CRF- and/or urocortin 1-stimulated secretion of alpha-MSH.

Adaptation, Physiological↗

Islet-1 expression and its colocalization with luteinising hormone, thyroid-stimulating hormone and oestrogen receptor alpha in the developing pituitary gland of the sheep foetus.

Islet-1 has been reported to be involved in pituitary development in the early stages of mouse embryogenesis. Oestrogen receptor (ER) and its expression may be involved in regulating pituitary development and its hormone-secreting function. Islet-1 expression and its correlations to ER in the developing pituitary gland are unknown. We therefore determined the pituitary cell specific expression of Islet-1 and its colocalization with ER alpha (ERalpha) in sheep foetus by immunohistochemistry. The results demonstrated that Islet-1-immunoreactive (ir) cells were distributed throughout the pituitary gland from day 60 of gestation until birth. The Islet-1-ir cell number was significant higher at day 90 and 120 of gestation compared to that at day 60 and at birth. All of the ERalpha-ir cells were colocalized with Islet-1 at day 60 of gestation, although a few ERalpha-ir cells were negative for Islet-1 in the later stage of gestation. The dominant cell type expressing Islet-1 is the gonadotroph, although small proportions of thyrotrophs and lactotrophs also express Islet-1. The proportion of luteinising hormone-ir gonadotrophs possessing Islet-1 kept rising from day 60 to day 120 of gestation and persisted until birth. The proportion of thyroid-stimulating hormone-ir cells expressing Islet-1 was at a high level from day 60-120 of the gestation and significantly declined at birth. The percentage of prolactin (PRL)-ir cells expressing Islet-1 was about 20% at day 60 of gestation. Very few PRL-ir cells contained Islet-1 in later stages of gestation. These data suggest that the development and functional establishment of pituitary gonadotrophs, thyrotrophs and lactotrophs might be regulated by the expressions of Islet-1 and ERalpha and by their interactions, although any mechanisms need to be elucidated further.

Animals↗

The stability of historical control data for common neoplasms in laboratory rats: adrenal gland (medulla), mammary gland, liver, endocrine pancreas, and pituitary gland.

Time-related changes in the incidences of spontaneous neoplasms in adrenals (medulla), mamma, liver, pituitary, and (endocrine) pancreas were assessed statistically in Wistar, Sprague-Dawley, and F344 rats employed by BASF, Germany and major European contract research organizations over the last 20 years. Negative trends (7 of 80 cases) were observed for pituitary pars distalis adenomas in Sprague-Dawley males and females, for pancreas islet cell adenomas in BASF Wistar males and females, for benign adrenal pheochromocytomas in Sprague-Dawley males, for malignant pheochromocytomas in F344 males, and for mammary gland fibroadenomas in BASF Wistar females. Positive trends (13 of 80 cases) were observed for benign pheochromocytomas, mammary gland adenocarcinomas, and pancreas islet cell carcinomas in HanWistar females, for malignant pheochromocytomas and islet cell carcinomas in BASF Wistar males, for benign pheochromocytomas and islet cell adenomas in F344 males, for mammary gland fibroadenomas in Sprague-Dawley females, and for benign hepatocellular tumors in HanWistar males and females, and in BASF Wistar and Sprague-Dawley females. In 60 of 80 cases there were no statistically significant trends. These results indicate that the majority of tumor types showed no time trends and that, in each rat strain, certain tumor types are susceptible to slight positive or negative time trends. Accordingly, the validity and use of historical control data should be based on an organ- and strain-specific statistical analysis of tumor incidence over time.

Adrenal Gland Neoplasms↗

Angiogenesis in pituitary adenomas and the normal pituitary gland.

Angiogenesis is essential for tumor growth beyond a few millimeters in diameter, and the intratumoral microvessel count that represents a measure of angiogenesis has been correlated with tumor behavior in a variety of different tumor types. To date no systematic study has assessed pituitary tumors of different secretory types, correlating vascular count with tumor size. The vascular densities of pituitary tumors and normal anterior pituitary were therefore assessed by counting vessels labeled using the vascular markers CD31 and ulex europaeus agglutinin I. One hundred and twelve surgically removed pituitary adenomas (30 GH-secreting, 25 prolactinomas, 15 ACTH-secreting, and 42 nonfunctioning tumors) were compared with 13 specimens of normal anterior pituitary gland. The vascular counts in the normal anterior pituitary gland were significantly higher (P < 0.05) than those in the tumors using both CD31 and ulex europaeus agglutinin I. In addition, microprolactinomas were significantly less vascular (P < 0.05) than macroprolactinomas, although there was no such difference between vascular densities of microadenomas and macroadenomas producing GH. ACTH-secreting tumors were, like microprolactinomas, of much lower vascular density than the normal pituitary and other secreting and nonsecreting tumor types. In marked contrast to other tumors, pituitary adenomas are less vascular than the normal pituitary gland, suggesting that there may be inhibitors of angiogenesis that play an important role in the behavior of these tumors.

Adenoma↗

Effects of season and sex on the distribution of cytosolic estrogen receptors within the brain and the anterior pituitary gland of sheep.

This study was undertaken to evaluate the effects of season and sex on the distribution of cytosolic estradiol-17 beta receptors (E2-receptors) within multiple regions of the ovine brain and the anterior pituitary gland. Our data confirm the existence of a saturable, high-affinity receptor specific for estradiol-17 beta (E2) in the hypothalamus and anterior pituitary gland. The concentrations of E2-receptors in tissues from castrated ewes and rams killed in December (breeding season) and May ( nonbreeding season) were determined by incubating cytosols with a saturating concentration of [3H] E2. For both sexes, the anterior pituitary gland contained the highest concentration of E2-receptor. Within the brain, the highest concentrations of E2-receptor were in the hypothalamus and preoptic area (POA), and within the hypothalamus, the median eminence-infundibular stalk (ME) contained a higher concentration of E2-receptor than the anterior and posterior hypothalamus. The numbers of E2-receptors in the anterior hypothalamus and ME of ewes were 2- 3X those for rams. In ewes, there was little seasonal difference in the numbers of E2-receptors in the POA, anterior hypothalamus and ME. In rams, there was little seasonal difference in the numbers of E2-receptors in the POA and anterior hypothalamus. For both sexes, there were fewer E2-receptors in the anterior pituitary gland and posterior hypothalamus in May than in December. Thus, during the nonbreeding season, the major neuroendocrine site of action of E2 probably is within the brain, specifically in the POA, anterior hypothalamus and ME of the ewe, and in the POA and the anterior hypothalamus of the ram. These data do not support a mechanism whereby the increased sensitivity to sex steroids during the nonbreeding season is due to enhanced numbers of E2-receptors.

Animals↗

Effects of corticotrophin-releasing hormone on corticotrophs in anterior pituitary gland allografts in hypophysectomized, orchidectomized hamsters.

We investigated the effects of corticotrophin-releasing hormone (CRH) on the percentage of anterior pituitary gland (APG) cells which are corticotrophs as well as the size and shape of corticotrophs. Pituitary glands were removed from 7-week-old male hamsters and placed beneath the renal capsules of hamsters that had been hypophysectomized and orchidectomized 3 weeks previously. Beginning 6 days after each host had received a single allograft, each was injected subcutaneously twice daily with 4 micrograms CRH or vehicle for 16 days. Six hosts in each group were decapitated 16 h after the last injection. Sections of anterior pituitary tissue were stained for ACTH and with hematoxylin. The percentage of corticotrophs among APG cells was greater in allografts exposed to exogenous CRH (approximately 20%) than in allografts exposed to vehicle (approximately 15%). Exposure to exogenous CRH increased the cross-sectional area of corticotroph cells in allografts to values greater than those measured for corticotrophs in allografts exposed to vehicle, without altering the shape of cells. Results of subsequent studies suggested that hamsters with allografts injected with vehicle do not release ACTH and that exogenous CRH causes an abrupt release of ACTH from allografts. These results indicate that CRH releases ACTH from ectopic corticotrophs and that administration of CRH can increase corticotroph size and the percentage of APG cells that are corticotrophs.

Adrenocorticotropic Hormone↗

Genetic defects in the development and function of the anterior pituitary gland.

Genetic defects affecting the hypothalamic-pituitary-target organ axes can cause a variety of diseases involving restricted or broad disruptions of human development and physiology. At the level of the anterior pituitary gland, mutations in the genes encoding key transcription factors, hypothalamic releasing and inhibiting hormone receptors, and the pituitary hormones themselves, can all result in the loss of action of one or more of the specialized hormone-secreting cell types. This article focuses on the effects of inherited and sporadic mutations on the development and function of the anterior pituitary. Mutations in the genes encoding the HESX1, PITX2, LHX3, LHX4, PROP1, PIT1, SF1, and TPIT developmental transcription factors are associated with combined pituitary hormone deficiency diseases. By contrast, deleterious alterations in the genes that encode hypothalamic releasing hormone receptors or pituitary hormones, such as the growth hormone releasing hormone receptor or growth hormone genes, usually result in phenotypes that reflect specific defects in the hormone-secreting capacities of individual anterior pituitary cell types.

Basic Helix-Loop-Helix Proteins↗

Actin is unevenly distributed in the pituitary gland.

In view of the suggestion that actin-like proteins might be involved in the final steps leading to hormone secretion, the actin content of pituitary glands of adult rats was determined by sodium dodecylsulfate-polyacrylamide gel electrophoresis (for total actin), by the DNAse method (which measures predominantly monomeric actin) and by immunocytochemistry. The amount of actin present in the neural lobe, expressed per mg total protein, was found to be comparable to that of other neural tissues. In contrast, in the anterior lobe, the ratio was significantly lower. The intensity of immunofluorescent staining with anti-actin antibodies was higher in the neural lobe than in either anterior or intermediate lobes. The intensity and distribution of tubulin immunofluorescent staining with anti-tubulin antibodies resembled that of anti-actin antibodies. Thus, three independent methods point to an uneven distribution of actin in the subdivisions of the pituitary gland, although all these subdivisions are believed to secrete their hormones by exocytosis. These data suggest that the bulk of actin present in pituitary cells is unlikely to be involved only in exocytosis, but may be implicated also in the intracellular translocation of secretory products.

Actins↗

Localization of fertility factor SP22 to specific cell types within the anterior pituitary gland.

Sperm protein 22 (SP22) was recently identified in the anterior pituitary gland (AP) of male Golden Syrian hamsters using ion trap mass spectrometry. SP22 has been implicated in apoptosis, androgen receptor function, fertility, and ontogeny of early-onset Parkinson's disease. However, the role of SP22 in the pituitary has not been investigated. We cloned the cDNA for full-length SP22 from AP and posterior lobe (posterior pituitary and intermediate lobe) of the pituitary gland in adult male rats and Golden Syrian hamsters, confirming the presence of SP22 mRNA in the AP and posterior lobe. Because gonadal steroids are important regulators of AP function, and SP22 is associated with androgen receptor function, we used Western blots to compare SP22 in the AP of intact and orchidectomized male rats given placebo or a low or high dose of testosterone. SP22 did not differ with treatment, indicating that AP SP22 concentration was not regulated by testosterone. To localize SP22 to specific cells of the AP, mirror-image paraffin sections were labeled against SP22 and either luteinizing hormone (LH)beta, thyroid-stimulating hormone (TSH)beta, prolactin, adrenocorticotropic hormone (ACTH), or growth hormone (GH) using peroxidase-conjugated secondary antibody. Additional sections were colabeled with SP22 and one of the AP hormones using fluorescent secondary antibodies. SP22 colocalized in somatotropes and thyrotropes in rat and hamster. We identified SP22 in a small percentage of corticotropes, gonadotropes, and lactotropes. This is the first report that SP22 mRNA is present specifically in the AP, and SP22 is localized primarily in somatotropes and thyrotropes. SP22 may help regulate AP function and be particularly important for the control of GH and TSH secretion.

Animals↗