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Isolation and characterization of alpha-endorphin and gamma-endorphin from single human pituitary glands.

alpha-Endorphin and gamma-endorphin, two closely related peptides of the pro-opiomelanocortin family with characteristic biological activities, were purified to homogeneity from single human pituitary glands and chemically identified. Isolation of the peptides was based on size fractionation by Sephadex G-75 chromatography followed by two HPLC steps using reverse-phase and paired-ion reverse-phase systems and was monitored by radioimmunoassay. During the isolation procedure alpha- and gamma-endorphin-sized material behaved chromatographically and immunologically indistinguishably from synthetic alpha- and gamma-endorphin. The amino acid composition and NH2-terminus of isolated peptides demonstrated their identity as authentic alpha-endorphin and gamma-endorphin. Acetylated forms were absent. In addition, evidence is provided that large forms with alpha- and gamma-endorphin immunoreactivity detected during gel filtration are human lipotropin-(1-74) and -(1-75), respectively. The data substantiate that alpha-endorphin and gamma-endorphin exist as endogenous peptides in the human pituitary gland.

Amino Acids↗

Displacement of the normal pituitary gland by sellar and juxtasellar tumours: surgical-MRI correlation and use in differential diagnosis.

We compared the position of the normal pituitary gland as estimated by gadolinium (Gd)-DTPA-enhanced MRI, with its position at surgery in 40 patients with intra- and juxtasellar tumours: 22 pituitary adenomas, 4 craniopharyngiomas, 7 meningiomas, 2 germinomas, and 5 Rathke cleft cysts. In 37 of these, the normal gland showed more intense contrast enhancement than the adjacent tumour, from which it could be differentiated by Gd-DTPA-enhanced MRI, especially in the sagittal plane. The direction of displacement of the normal pituitary gland correlated well with tumour type, so that its position proved helpful in the differential diagnosis. The normal gland was typically displaced superiorly by pituitary adenomas, inferiorly by craniopharyngiomas, and anteriorly by germinomas. It showed variable displacement by Rathke cleft cysts, and was not usually displaced by meningiomas.

Adenoma↗

Distribution of cadmium in ovaries, adrenals and pituitary gland after chronic administration in rats.

Wistar rats were given 0.25, 0.5 or 1.0 mg/kg/week CdCl2 for 14, 18 or 22 weeks and the body weight, Cd content of ovaries, adrenals, pituitary gland, furthermore the progesterone and oestradiol-17 beta secretion of ovary were checked. Cd treatment caused a slight decrease in the body weight, but failed to alter the weight of endocrine organs. CdCl2 in a dose of 0.25 mg/kg/week resulted in almost the same Cd content in all the three organs. Rising the amount of Cd administered the pituitary gland accumulated more Cd than the adrenals, and the lowest levels were found in the ovary. CdCl2 even in the dose of 1.0 mg/kg/week failed to alter the ovarian cycle, progesterone and oestradiol-17 beta production of ovary. The data point also to a developing tolerance to Cd as the cumulative dose of CdCl2 lies close to the LD50 levels.

Adrenal Glands↗

GnRH molecular variants in the brain and pituitary gland of pejerrey, Odontesthes bonariensis (Atheriniformes). Immunological and chromatographic evidence for the presence of a novel molecular variant.

Gonadotropin-releasing hormone (GnRH) molecular variants in the brain and pituitary gland of pejerrey, Odontesthes bonariensis (Atheriniformes), were characterized by gradient reverse phase high performance liquid chromatography (RP-HPLC). Eluted fractions were tested in radioimmunoassays with different antisera. The results show that the brain extract contains three forms of GnRH: one is immunologically and chromatographically similar to cIIGnRH (chicken II), and another is similar to sGnRH (salmon). A third GnRH appears to be chromatographic and immunologically different from the nine other known forms of the vertebrate hormone. This is the only variant present in the pituitary gland.

Animals↗

Hyperprolactinemia reduces vasoactive intestinal peptide in the anterior pituitary glands of rats.

The concentration of vasoactive intestinal polypeptide (VIP) in the anterior pituitary gland of female rats was significantly reduced by pituitary implants, prolactin-secreting tumours or dopamine blockade-induced hyperprolactinemia. Oestradiol implants increased plasma prolactin (PRL) to 12 times and increased pituitary VIP to 3.5 times that of controls after one week. The dopamine agonist bromocriptine significantly reduced pituitary VIP at doses within the clinically used range. These results provide evidence for a suppressive effect of plasma PRL on pituitary VIP, which may be effected by a dopaminergic mechanism, but is over-ridden by oestrogen.

Animals↗

Relationship between unbound oestrogen receptors in the uterus and anterior pituitary gland after implantation of oestradiol-17 beta into rats.

This paper describes the effect of oestradiol-17 beta implants on unbound cytosolic and nuclear oestrogen receptors in the uterus and anterior pituitary gland of ovariectomized adult rats. Rats were ovariectomized and implanted 1 week later with oil or oestradiol-17 beta/oil solution in silicone elastomer capsules. In the latter animals the capsules either remained in situ until decapitation (subgroup 1) or were removed 48 h after implantation (subgroup 2). Implants of oestradiol-17 beta caused a significant depletion in both forms of oestrogen receptor in the uterus and anterior pituitary gland of rats in subgroup 1. In subgroup 2, however, there was an almost complete return to control concentrations of uterine cytosolic receptors and anterior pituitary cytosolic and nuclear receptors. Concentrations of uterine nuclear oestrogen receptors showed only a partial recovery. These data suggest that both forms of oestrogen receptor constitute an integrated population of oestrogen receptors, without dependence on intracellular location, and that in the presence of oestradiol these receptors are bound more quickly than they are synthesized. The results also indicate the existence of a dynamic equilibrium of unbound receptors between the cytosolic and nuclear compartments for both target organs, but show that in the absence of oestradiol this equilibrium is restored in the anterior pituitary sooner than in the uterus.

Animals↗

Localization and identification of nuclear radioactivity in the pituitary gland and genital tract after administering 3H-testosterone, 3H-dihydrotestosterone, or 3H-estradiol to male rhesus monkeys.

Target cells for testosterone, dihydrotestosterone, and estradiol in the pituitary gland and genital tract of the male primate were localized by thaw-mount autoradiography, and high performance liquid chromatography was used to identify the metabolites of these steroids in cell nuclei. Castrated rhesus monkeys were injected with 3H-testosterone, 3H-dihydrotestosterone, or 3H-estradiol and killed 60 min later. In the anterior pituitary gland, fewer cells were labeled and less radioactivity was taken up by cell nuclei following the administration of either 3H-testosterone (4% of pars distalis cells and 5 dpm/micrograms DNA) or 3H-dihydrotestosterone (5% of cells and 13 dpm/micrograms DNA) than following the administration of 3H-estradiol (43% of cells and 214 dpm/micrograms DNA). Most of the radioactivity in nuclei was in the form of the unmetabolized parent compound (78-94%). In prostate, seminal vesicles, and penis, 3H-dihydrotestosterone was the predominant form of nuclear radioactivity following both 3H-testosterone (67-90%) and 3H-dihydrostestosterone (94-97%) administration, and both androgens labeled epithelial and smooth muscle cells. In contrast, 3H-estradiol was taken up in unchanged form, by cell nuclei of the genital tract and it labeled connective tissue fibroblasts, but not epithelial cells. Thus, the distributions of target cells for androgens and estrogens were clearly different in all these tissues, and the uptake of testosterone resembled that of its androgenic rather than that of its estrogenic metabolite.

Animals↗

Cloning, expression and regulation of chicken ovalbumin upstream promoter transcription factors (COUP-TFII and EAR-2) in the rat anterior pituitary gland.

Chicken ovalbumin upstream promoter transcription factors (COUP-TF)-II (NR2F2) and EAR-2 (NR2F6) are structurally related orphan members of the nuclear receptors superfamily. There are growing evidences that these factors play important roles during processes of differentiation and proliferation of several tissues. To better understand their role in the differentiated adult rat pituitary gland, we cloned COUP-TFII and EAR-2 cDNAs from an anterior pituitary cDNA library. Subsequently, we raised and characterized specific antibodies to the N-terminal domain of both nuclear receptors. We next examined their cellular and subcellular distribution in the pituitary gland and determined their regulation during pregnancy. COUP-TFII and EAR-2 pituitary genes display, respectively, 90 and 100% homologies with their human and mouse homologues. Cellular expression of both nuclear receptors was mainly detected in the lactotropes of male and female rats, with a prominent distribution in the nuclear compartment for EAR-2, and interestingly both proteins were significantly upregulated in pituitaries of pregnant vs. cycling female rats. Thus, our results have characterized cloning of rat pituitary COUP-TFII and EAR-2 genes, demonstrated that they are both specifically expressed in lactotropes, and strongly suggested that they may play an important role in modulating prolactin (PRL) gene expression during pregnancy.

Amino Acid Sequence↗

Nasal pyriform aperture stenosis and absence of the anterior pituitary gland: report of two cases.

We describe two female infants with congenital nasal pyriform aperture stenosis and severe pituitary insufficiency. The anterior pituitary gland was undetectable with magnetic resonance imaging. Consanguinity of parents in both cases suggests autosomal recessive inheritance of this disorder. An early fetal developmental defect may explain this syndrome, which affects midline craniofacial structures. In patients with congenital pyriform aperture stenosis, magnetic resonance imaging of the brain and endocrine investigations should be performed for rapid diagnosis and treatment of the latter to avoid major neurologic complications.

Abnormalities, Multiple↗

Changes in the processing of beta-endorphin in the hypothalamus and pituitary gland of female rats during sexual maturation.

Puberty in the female rat is accompanied by a marked attenuation of the opioid inhibition of luteinizing hormone secretion. One factor which may contribute to this altered role is a change in the metabolism of opioid peptides during sexual maturation. beta-Endorphin undergoes a considerable degree of metabolism through both C-terminal proteolysis and N-acetylation, and these metabolites do not possess opioid activity. The processing of beta-endorphin in the hypothalamus and in the anterior and neurointermediate lobes of the pituitary gland in prepubertal and adult female rats was studied using gel filtration and high performance liquid chromatography coupled with radioimmunoassay. In the anterior lobe, high molecular weight precursors of beta-endorphin (pro-opiomelanocortin and beta-lipotropin) were present in prepubertal (28 days old) rats, but little authentic beta-endorphin was detected. In contrast, only beta-lipotropin and beta-endorphin were present in mature (70 days old) animals. Only beta-endorphin-sized peptides were present in the neurointermediate lobes of both prepubertal and adult rats. However, the proportion of N-acetylated metabolites was higher in sexually mature animals. In the hypothalamus, only beta-endorphin-sized peptides were present in both juvenile and adult animals. However, C-terminal proteolysis increased with age (no acetylated metabolites were detectable in this tissue). The proportion of the total beta-endorphin-like immunoreactivity attributable to beta-endorphin was lower in young adult (first dioestrus after vaginal opening) (55%) and mature (dioestrus, 61-64 days old) rats (56%) compared to prepubertal (30 days old) animals (75%) and the proportions of non-acetylated metabolites [beta-endorphin-(1-27) in young adults and beta-endorphin-(1-26) in adults] were increased concomitantly. These changes were correlated with a reduced luteinizing hormone response to the opiate antagonist naloxone in adult compared to prepubertal rats. beta-Endorphin is processed differently in the two lobes of the pituitary gland and in the hypothalamus and the degree of metabolism increases as the rat reaches sexual maturity. The increased metabolism of beta-endorphin in the hypothalamus, the site most likely to be involved in the control of luteinizing hormone secretion, results in a reduction in the relative proportion of the opioid active parent peptide. Thus, increased inactivation of beta-endorphin may contribute to the attenuation of the opioid inhibition of luteinizing hormone secretion observed during puberty.

Animals↗

Hypertrophic cranial pachymeningitis involving the pituitary gland: a case report.

BACKGROUND: As the use of magnetic resonance imaging (MRI) has become widespread, the reported cases of pachymeningitis have increased. Various inflammations are known to occur in the pituitary gland. However, an association between pachymeningitis and pituitary inflammation has not been recognized previously. CASE DESCRIPTION: This 56-year-old woman complained of incessant headache and deafness. MRI showed thickened dura at the right convexity and a dumbbell shaped pituitary enlargement, which was isointense to the gray matter (T1-weighted images) and homogeneously enhanced. Microscopic examination of the dura mater showed a marked increase of collagen fibers and various inflammatory cells. The anterior pituitary lobe was infiltrated with lymphocytes, and normal tissues were displaced by increased collagen fibers. CONCLUSION: Hypertrophic cranial pachymeningitis, a rare clinical entity, is a progressive lesion prone to include the dura mater at the skull base and can be fatal. We report a case of hypertrophic cranial pachymeningitis involving the pituitary gland, and discuss the possible etiology of the association.

Diagnosis, Differential↗

Nitric oxide synthase and cGMP in the anterior pituitary gland: effect of a GnRH antagonist and nitric oxide donors.

Using immunohistochemistry and in situ hybridization, it has been previously shown that gonadotropes and folliculo-stellate cells in the rat anterior pituitary gland express nitric oxide (NO) synthase (NOS), and that NOS expression is increased by gonadectomy. Using the indirect immunofluorescence technique in conjunction with antibodies raised to conjugated cGMP, we have attempted to establish the target cells for NO in the anterior pituitary and to define the mediator of NO regulation. After incubation of pituitary slices with several NO donors, numerous endocrine cells, but no folliculo-stellate cells, expressed cGMP. Most of these cells stained for LH, that is they were gonadotropes. However, there were apparently cGMP-positive, LH-negative and LH-positive, cGMP-negative endocrine cells. The increase in cGMP could be virtually completely blocked by a guanylyl cyclase inhibitor. cGMP was not expressed in corticotropes, but cGMP-positive cells often contained NOS-like immunostaining. Incubation with GnRH did not result in detectable levels of cGMP. However, when castrated rats were pretreated with a potent longlasting GnRH antagonist, antide, the castration-induced increase in NOS was completely blocked. This suggests that GnRH is involved in the in vivo upregulation of NOS after castration, but that GnRH cannot induce cGMP accumulation in normal pituitary slices in vitro. Taken together, the present results give further evidence for a role of NO in the control of, in particular, LH secretion from the anterior pituitary gland in the rat.

Animals↗

NGF content and expression in the rat pituitary gland and regulation by thyroid hormone.

The involvement of nerve growth factor (NGF) in neuroendocrine regulation is supported by several lines of evidence. In this paper, we investigated the NGF content and expression in the pituitary gland and other endocrine organs during dysendocrine states (thyroidectomized, adrenalectomized and gonadectomized male rats). We found an increase of NGF-IR in the pituitary gland and testis of hypothyroid rats whereas no differences were found in the adrenal gland and blood. Also, NGF mRNA expression had increased in the anterior pituitary of hypothyroid rats whereas it had not changed after adrenalectomy and gonadectomy. Moreover, other neurotrophins and neurotrophin high-affinity receptors were unchanged in the anterior pituitary of hypothyroid rats. These data indicate that pituitary NGF is selectively modulated by thyroid status of the animal, further supporting a close link between NGF and thyroid hormone.

Adrenal Glands↗

Immunocytochemical study of the GH cells in the anterior pituitary gland of human fetus II. Anencephalic fetus.

In order to elucidate the effects of hypothalamic regulation on the morphology of GH cells, light and electron microscopic immunocytochemical examinations were carried out comparing GH cells in the anterior pituitary gland of anencephalic fetus with those of normal fetuses. Three types of GH cells were identified in the anterior pituitary gland of anencephalic fetus as well as in the normal fetus. Type-I is a small, round cell containing a few small secretory granules. Type-III is a large, polygonal cell with numerous large secretory granules. Type-II is a polygonal cell with medium-sized secretory granules. The Type-II GH cell was predominant in both anencephalic and normal fetuses. The most striking difference between anencephalic and normal fetuses was the presence of atypical forms of the Type II cell. These were polygonal cells containing secretory granules, which were either immunopositive or immunonegative to anti-human GH (anti-hGH) serum. Furthermore, two other types of GH cells were identified. The somatomammotroph (SM cell) contained GH and PRL in different granules within the same cell. Also, a different type of the GH cell was noted containing two varieties of secretory granules; one was immunolabeled only with anti-hGH and the other was not immunolabeled to either anti-hGH or anti-human PRL (anti-hPRL). From these results, we suggest that an absence of hypothalamic regulation in the anencehpalic does not seriously modify GH cell morphology but induces an altered GH storage pattern in some of the cells.

Anencephaly↗