Immunoelectron microscopy for growth hormone and prolactin in pituitary adenomas

Growth hormone (GH)- and prolactin (Prl)-producing pituitary adenomas were studied with immunoelectron microscopy using protein A-gold complex, and were compared with the normal pituitary gland. GH-producing cells were readily identifiable by numerous, uniformly dense, round secretory granules in both adenomas and normal pituitary gland. In contrast, Prl secretory granules in normal pituitary gland were much larger in size than the scarce, smaller, secretory granules of Prl-producing adenomas. Thus immunoelectron microscopic identification of Prl is more valuable for prolactinoma. With more specific antigens available as tumor markers, immunoelectron microscopy appears to be a powerful tool for tumor diagnosis.     

The pituitary in cirrhosis: ultrastructure, growth hormone, and prolactin concentrations.

Micronodular cirrhosis was induced in male SUAH substrain Wistar rats by combined phenobarbitone and carbon tetrachloride treatment. Both pituitary and serum concentrations of growth hormone were significantly reduced in cirrhotic rats compared with age-related untreated rats or those treated only with phenobarbitone. Ultrastructurally growth hormone-secreting cells (somatotrophs) of pituitaries of cirrhotic rats appeared relatively inactive, having few hormone-containing granules, sparse rough endoplasmic reticulum, and small nuclei with areas of condensed chromatin. The cells themselves were smaller than similar cells of untreated rats with a reduced cytoplasmic area. In addition immunocytochemistry of pituitaries at light microscope level, using sheep anti-rat growth hormone antibody, showed that somatotrophs of cirrhotic rats were more heteromorphic and disorganized than those in controls. There was marked development of the folliculo-stellate cell system in pituitaries of cirrhotic rats, the cells were enlarged with distinct golgi, and numerous microvilli were projecting into dilated follicular lumena.     

Mixed growth hormone cell- prolactin cell pituitary adenoma with acromegaly: α-subunit most growth hormone cells

A 51 -year-old woman with mixed growth hormone (GH) cell-prolactin (PRL) cell pituitary adenoma is presented. She had clinical signs due to hypersecretion of GH and PRL. Resected tissue was studied immunohistochemically and morphologically. The serial sections revealed that GH and α-subunit were co-localized in most cells, while GH and PRL were localized in different cells.     

Human growth hormone and prolactin secreting pituitary adenomas analyzed by in situ hybridization

Acidophilic pituitary adenomas commonly produce growth hormone (GH) or prolactin (PRL), according to studies employing immunohistochemical and ultrastructural methods. To examine this question, in situ hybridization with oligonucleotide probes was done on routinely processed tissues received in the pathology laboratory to analyze for the presence of GH and PRL messenger RNA (mRNA) in 4 normal pituitaries, 10 prolactinomas, and 16 GH-secreting adenomas. Most acidophilic cells in normal pituitaries expressed either GH or PRL hormone and the respective mRNAs, but GH mRNA and PRL hormone were also detected in some of the same cells. Patients with a clinical diagnosis of prolactinoma had cells with only PRL mRNA in their tumors, while most (14 of 16) patients with a clinical diagnosis of acromegaly or gigantism had both GH and PRL mRNAs in their tumors. The GH adenomas varied in these studies. In situ hybridization was helpful in characterizing the adenoma from a patient with acromegaly who had immunoreactive PRL, but no immunoreactive GH in the resected tumor; in situ hybridization analysis revealed mRNAs for both GH and PRL in the same tumor cells. Our findings indicate that pituitary adenomas from patients with acromegaly commonly express PRL mRNA. It is concluded that in situ hybridization provides new information about the clinical biology and the histopathologic classification of pituitary adenomas.     

Pergolide for the treatment of pituitary tumors secreting prolactin or growth hormone

We gave pergolide mesylate, a new long-acting ergot derivative with dopaminergic properties, to 47 patients with hypersecretion of prolactin or growth hormone. Single doses produced long-lasting reductions of serum prolactin levels; after 24 hours, the values remained depressed at a mean of 28.8 per cent of the base-line value. Among 41 patients (22 women and 19 men) with hyperprolactinemia who took pergolide for three months or more, prolactin levels fell to normal in 37 and remained slightly elevated in 2. In the two patients in whom the levels fell to only 38 to 52 per cent of base line, treatment was regarded as a failure. The level of growth hormone fell to a mean of 52.8 per cent of base line in patients with acromegaly who were taking 100 micrograms of pergolide per day. Among patients for whom adequate CT scans were available, definite tumor shrinkage occurred in 10 of 13 with macroadenomas and definite or probable shrinkage in 5 of 9 with microadenomas. Menses returned in 76 per cent of treated women and testosterone levels rose in 10 of 14 men. We conclude that pergolide reduces hypersecretion and shrinks most prolactin-secreting macroadenomas. In some patients long-term pergolide therapy may be superior to surgery and x-ray treatment.     

In vitro pituitary prolactin, growth hormone and follicle stimulating hormone secretion during sexual maturation of female rats primed with melatonin

The influence of in vivo melatonin administration on in vitro pituitary follicle stimulating hormone (FSH), growth hormone (GH) and prolactin secretion, as well as the possible influence of dopamine (DA) were evaluated in prepubertal (31-day-old), pubertal (33-day-old) and adult female rats at diestrus phase of the sexual cycle. The in vitro pituitary hormone secretions were evaluated at basal rate for the first hour of incubation only, in Krebs Ringer phosphate (KRP) (I1) and after a second hour of incubation with KRP (I2) or with KRP+DA (I2 plus DA). I1PRL secretion was significantly higher in 33-day-old control and melatonin treated (MEL) rats as compared to I2 periods. However, in 31-day-old rats I1 secretion was higher than in the I2 or I2+DA periods, in MEL rats. In vitro GH secretion was significantly higher at I1 than during I2 periods in the control 31- and 33-day-old groups, but not in MEL rats. The only significant effect of DA was the elevation of GH in prepubertal MEL rats. In vitro FSH release was increased by melatonin in 31- and 33-day-old female rats. No differences in PRL, GH and FSH secretion were found in adult rats. In conclusion, the results show that melatonin effects upon in vitro pituitary gland activity are reproductive-stage-dependent modifying the secretory capacity of the lactotrop, gonadotrop and somatotrop during prepubertal and pubertal ages but not in adult rats studied at a quiescent phase of the sexual cycle.     

Effects of propylthiouracil on growth hormone and prolactin messenger ribonucleic acids in the rat pituitary

The effects of hypothyroidism induced by propylthiouracil (PTU) treatment on growth hormone (GH) and prolactin (PRL) messenger ribonucleic acid (mRNA) levels were analyzed in adult female rat adenohypophyses by in situ hybridization histochemistry and Northern hybridization analyses. Twenty-eight days of PTU treatment produced a significant decrease in GH mRNA levels and a smaller decrease in PRL mRNA determined by both in situ hybridization histochemistry and Northern hybridization analyses. A combined procedure of in situ hybridization histochemistry followed by immunochemistry on the same sections revealed mammosomatotropic cells expressing GH mRNA and PRL protein in the same pituitary cells from all treatment groups. Cells expressing GH mRNA and thyroid-stimulating hormone protein were not detected by this method. Immunochemical staining revealed a decrease in GH cells and an increase in thyroid-stimulating hormone cells in hypothyroid rats. Cells expressing both GH and thyroid-stimulating hormone protein were not detected by immunostaining. These results indicate that hypothyroidism produces significant decreases in GH mRNA and also decreases PRL mRNA and that mammosomatotropic cells can be detected in pituitaries from normal and hypothyroid rats.     

Effect of acrylonitrile on the rat pituitary: enlargement of Golgi region in prolactin cells, crinophagy in prolactin cells and growth hormone cells

Since it has been shown that acrylonitrile prevents the appearance of spontaneous pituitary adenomas, we have investigated its effect in acute experiments on rat pituitaries by histology, immunocytochemistry, electron microscopy and morphometry; in addition, serum prolactin and growth hormone levels were measured by radioimmunoassay. Electron microscopy of prolactin cells revealed hypertrophy of the Golgi region without significant change in volume densities and diameters of forming and storage granules. In the 24 h group, crinophagy was observed in prolactin cells and growth hormone cells. Corticotrophs, thyrotrophs and gonadotrophs were unaltered. Dilation, congestion and rupture of capillaries, as well as pericapillary and intercellular oedema were evident in the 24 h group. One hour after intravenous acrylonitrile injection, serum prolactin levels were within the normal range, whereas at 24 h, hyperprolactinemia was noted. Serum growth hormone concentrations were unchanged. It can be concluded that acrylonitrile has a complex effect on prolactin cells. Hypertrophy of Golgi complex and hyperprolactinemia may reflect increased prolactin synthesis and release. Since volume densities and diameters of secretory granules in prolactin cells remained unchanged, it appears that newly synthesized prolactin was preferentially released and not the prolactin stored in secretory granules. Crinophagy may be the morphological manifestation of a discrepancy between hormone synthesis and release suggesting increased degradation of unused hormone by lysosomes.     

The effect of age on the number of pituitary cells immunoreactive to growth hormone and prolactin

The results of a combined immunocytochemical, morphometric, and clinicopathologic analysis of growth-hormone-producing and prolactin-producing pituitary cells in 28 subjects ranging in age from 16 to 90 are reported. There was a significant age-related decline in the number and size of growth-hormone-producing cells, which was most marked in the transition from youth to middle age. There was also a significant age-related decline in the number of pituitary parenchymal cells but not in pituitary weight. Prolactin cells did not show a significant decline in number with age.     

Melatonin reduces the production and secretion of prolactin and growth hormone from rat pituitary cells in culture

A culture of clonal tumour cells from rat pituitary gland that secrete both prolactin and growth hormone were used to investigate whether the pineal hormone melatonin can act directly on the pituitary gland to control prolactin production. Melatonin inhibition of prolactin and growth hormone production was significant but mild. Concentrations of between 10(-8) M and 10(-6) M reduced both prolactin and growth hormone production and prolactin secretion by 10-50%. 17 beta-oestradiol (OE) and thyrotrophin-releasing hormone (TRH) stimulated prolactin production but had no significant effect on growth hormone production. Melatonin reduced the effects of both of these compounds. Both TRH and vasoactive intestinal peptide (VIP) stimulated secretion of prolactin, and TRH also of growth hormone. Melatonin reduced these effects significantly. TRH and VIP increased cAMP production two- and 12-fold, respectively. Melatonin had no effect on basal or stimulated cAMP production. The melatonin-induced changes in prolactin and growth hormone production and secretion seen here do not approach the magnitude of the fluctuations seen in plasma in vivo. It is concluded that while melatonin does have a direct effect on the lactotroph in the regulation of prolactin production, its main physiological target must be elsewhere.     

Effect of acrylonitrile on the rat pituitary: enlargement of Golgi region in prolactin cells,crinophagy in prolactin cells and growth hormone cells.

Since it has been shown that acrylonitrile prevents the appearance of spontaneous pituitary adenomas, we have investigated its effect in acute experiments on rat pituitaries by histology, immunocytochemistry, electron microscopy and morphometry; in addition, serum prolactin and growth hormone levels were measured by radioimmunoassay. Electron microscopy of prolactin cells revealed hypertrophy of the Golgi region without significant change in volume densities and diameters of forming and storage granules. In the 24 h group, crinophagy was observed in prolactin cells and growth hormone cells. Corticotrophs, thyrotrophs and gonadotrophs were unaltered. Dilation, congestion and rupture of capillaries, as well as pericapillary and intercellular oedema were evident in the 24 h group. One hour after intravenous acrylonitrile injection, serum prolactin levels were within the normal range, whereas at 24 h, hyperprolactinemia was noted. Serum growth hormone concentrations were unchanged. It can be concluded that acrylonitrile has a complex effect on prolactin cells. Hypertrophy of Golgi complex and hyperprolactinemia may reflect increased prolactin synthesis and release. Since volume densities and diameters of secretory granules in prolactin cells remained unchanged, it appears that newly synthesized prolactin was preferentially released and not the prolactin stored in secretory granules. Crinophagy may be the morphological manifestation of a discrepancy between hormone synthesis and release suggesting increased degradation of unused hormone by lysosomes.     

The effects of estrogens on tumor growth and on prolactin and growth hormone mRNA expression in rat pituitary tissues.

Estrogens inhibit tumor growth and modify PRL and GH expression in the MtT/W15 transplantable rat pituitary tumor. The effects of estradiol (E2) and diethylstilbestrol (DES) on PRL and GH mRNA levels were investigated. Estrogens increased GH mRNA levels and decreased PRL mRNA levels as detected by in situ hybridization and Northern blot hybridization with oligonucleotide probes, while inhibiting tumor growth. Similar changes in immunoreactive GH and PRL were seen in the tumor cells. The pituitary glands of tumor-bearing rats treated with estrogen for 3 weeks were increased in weight with a concurrent increase in pituitary PRL mRNA when analyzed by dot blot hybridization. These results indicate that estrogens have an inhibitory effect on the growth of the MtT/W15 tumor and increase GH protein and mRNA levels, while causing PRL protein and mRNA levels to decrease. The pituitaries of tumor-bearing rats concomitantly undergo PRL cell hyperplasia with an increase in PRL mRNA. These results also demonstrate a paradoxical effect of estrogens on different pituitary tissues.     

Ultrastructure,immunohistochemistry and hormone release of pituitary adenomas in relation to prolactin production

Summary Fifteen cases of pituitary adenoma, 14 of which were associated with hyperprolactinemia, were studied by observation and granule morphometry of electron micrographs, immunohistochemistry and sequential observation of in vitro release with regard to hormone production, storage and secretion. Adenoma cells of 6 cases with marked elevation of plasma prolactin were sparsely granulated, showed characteristic ultrastrucures including the presence of small secretory granules, well developed Golgi and rough membranes, misplaced exocytosis, and positive or negative immunostaining for prolactin. These adenomas also showed vigorous release of the hormone into the circulation and/or culture medium. In vitro studies showed that negative immunostaining of adenoma cells did not preclude the production and secretion of the hormone. One densely granulated adenoma containing cells with numerous lactotroph type granules showed moderate release of prolactin into the circulation. In an acromegalic case associated with both high plasma growth hormone and prolactin, some cells were shown by immunohistochemistry to store both hormones. There were 4 adenomas which could not be shown to produce, store and secrete prolactin by any method available.Abbreviations Used in this Paper ACTH adrenocorticotropic hormone - -MSH -melanocyte stimulating hormone - hGH human growth hormone - hPRL human prolactin - LH luteinizing hormone - FSH follicle stimulating hormone - TSH thyroid stimulating hormone - TRH Thyrotropin-releasing hormone This work was supported in part by Grants-in-Aid for Cancer Research (No. 50-14) and for Specific Diseases (Disorder of Hypothalamic and Pituitary System) from the Ministry of Health and Welfare, and for Cancer Research (No. 401034) from the Ministry of Education, Science and Culture, Japan     

Humoral and cell-mediated immunity in mice with genetic deficiencies of prolactin, growth hormone, insulin-like growth factor-I, and thyroid hormone

Prolactin (PRL), growth hormone (GH), insulin-like growth factor-I (IGF-I), and thyroid hormones have been proposed as critical immunoregulatory mediators, and their clinical use is being considered. The precise role played by each of these hormones in the generation of humoral and cell-mediated immune responses was assessed in a panel of mice with mutations that result in a selective reduction of PRL, GH, IGF-I, and/or thyroid hormone production. A surprising result, in view of previous studies indicating an immunoregulatory role for these hormones, was that all mice generated normal humoral and cell-mediated immune responses following challenge with T-independent and T-dependent antigens and with Listeria monocytogenes. A review of these findings in the context of previous data has resulted in the formulation of a working hypothesis proposing that these hormones act as anabolic and/or stress modulating mediators with effects on most cells, including those of the immune system. When considered in this context, it is possible to reconcile the contradictory data.     

Ontogeny of immunoreactive somatolactin, prolactin and growth hormone secretory cells in the developing pituitary gland of Cichlasoma dimerus (Teleostei, Perciformes)

Prolactin, growth hormone and somatolactin constitute a hormone family because they are structurally related and are secreted by acidophilic cells of different regions of the adenohypohyisis. In this work, we report the ontogeny of ir-prolactin, ir-growth hormone and ir-somatolactin cells in the developing pituitary gland of the cichlid fish Cichlasoma dimerus (Teleostei; Perciformes). Antisera raised against fish pituitary hormones were used. In this species hatching occurs 54 hs after fertilization and the three different cell types were recognized two days later. The neurohypophysis was recognized on day 14 after hatching and in later stages it began to show the characteristic deep interdigitations of the adults. On day 42 (juvenile stage) the distribution of ir-PRL, ir-GH and ir-SL showed the pattern described for adults of this species. The ir-SL cells were not PAS-positive in larvae as they are in adults. This would suggest the presence of a nonglycosilated form of somatolactin in early stages of development which may coexist in adults with a glycosilated form. The appearence of these hormones so early in development suggest their importance in the survival of fish larvae but further studies focused on the ontogeny of hypothalamic factors that regulate their synthesis and secretion must be performed.