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Pituitary gland and axial skeletal malformations in human fetuses with spina bifida.

The purpose of the present study was to describe the pituitary gland and axial skeleton, including the sella turcica, in human fetuses with spina bifida. Ten fetuses with gestational ages (GA) 15 1/2-28 weeks were investigated radiographically (Faxitron X-ray apparatus) and immunohistochemically. Four of the fetuses have been described previously. The study showed that nine fetuses had minor or no skeletal abnormalities in the vertebral bodies of the spine, and one fetus had severely malformed vertebral bodies. In all cases the sella turcica and the pituitary gland were malformed. Adenopituitary tissue was in all cases located in both the sella turcica and the pharyngeal submucosa. The most severe sella turcica/pituitary gland malformation was seen in the fetus with the malformed spine. The connection between the prenatally registered sella turcica/pituitary gland malformation and the endocrinological status of children with spina bifida should be emphasized in future studies.

Fetus↗

Abundance of endothelin-3 in rat intestine, pituitary gland and brain.

We established a highly sensitive and specific sandwich-enzyme immunoassay (EIA) for endothelin-3 (ET-3), which showed no crossreactivity with endothelin-1 (ET-1), endothelin-2 (ET-2) and big-endothelin-1 (big-ET-1). We had previously established a sensitive sandwich-EIA for ET-1, which fully crossreacted with ET-2, but not with ET-3 or big-ET-1. These EIAs were used to examine the tissue distribution of immunoreactive (ir-) ET-3 and compare them with those of ir-ET-1 (including ir-ET-2) in Sprague-Dawley rats. High concentrations of ir-ET-3 were found in the intestine, lung, pituitary gland and brain (greater than 100 pg/g wet tissue), ir-ET-1(ET-2) showed widespread distribution, with large amounts in the lung and colon (greater than 1000 pg/g wet tissue). The pituitary gland was the only organ containing higher amounts of ir-ET-3 than ir-ET-1 (ET-2). In reverse phase-high performance liquid chromatography coupled with EIAs, the ir-ET-3 was exclusively eluted at the position of synthetic ET-3, indicating that the ir-ET-3 was identical to ET-3. The abundance of ET-3 in the intestine, pituitary gland and brain indicates that ET-3 is a new brain-gut peptide which may have a physiological function in nervous and endocrine systems.

Animals↗

The dopaminergic innervation of the intermediate lobe and of the neural lobe of the pituitary gland.

The intermediate and the neural lobe of the pituitary gland are innervated by two, virtually independent, groups of dopaminergic neurons which, until recently, were considered as a uniform system and referred to as the "tubero-hypophyseal dopamine system". Some aspects of the separate physiological functions of these neurons in the intermediate and in the neural lobe, of their microanatomy and biochemistry as well as of dopamine release from their terminals are discussed in this review.

3,4-Dihydroxyphenylacetic Acid↗

Cloning and partial sequencing of the porcine growth hormone (pGH) gene from pituitary gland.

High molecular weight DNA was isolated from porcine pituitary gland, and a genomic library was constructed using lambda EMBL3 as cloning vector. Five positive clones were identified by in-situ hybridization with the full-length pGH cDNA as the probe. Dot-blot hybridization, restriction analysis, and Southern hybridization showed that one of the positive clones contained the entire pGH gene. A subclone that contained the upstream sequence from the SmaI site of pGH gene was identified, and part of it was sequenced. We also compared the sequence of our pGH gene with the corresponding sequence of the published pGH gene.

Amino Acid Sequence↗

[Isolation and characterization of prolactin messenger RNA from bovine anterior pituitary glands].

The messenger RNA was extracted from bovine anterior pituitary glands and was purified by oligo(dT)-cellulose chromatography. The length of the mRNA was 1,200 nucleotides measured by agarose gel electrophoresis containing methylmercury hydroxide. The bovine prolactin (PRL) mRNA was confirmed by Northern blot analysis of the mRNA with gamma-32P labelled synthetic oligonucleotide probes based on the partial amino acid sequences of bovine PRL and could stimulate the incorporation of 35S-methionine into protein in the rabbit reticulocyte cell-free system. The translation product of bovine PRL mRNA was immunoprecipitable by rabbit anti-ovine PRL anti-antibodies. The molecular weight of the translation product corresponding to the bovine PRL precursor was estimated to be approximately 25,000 by SDS polyacrylamide gel electrophoresis and autoradiograph.

Amino Acid Sequence↗

The distribution of immunoreactive FMRF-amide, neurotensin, and galanin in the brain and pituitary gland of three species of Xiphophorus from birth to sexual maturity.

This report seeks to extend the existing information on the relationship of regulatory neuropeptides to neuroendocrine and pituitary function through a longitudinal study of the distribution of FMRF-amide, galanin (GAL), and neurotensin (NT) in the brain and pituitary gland of three species of Xiphophorus from birth to sexual maturity. In the pituitary gland, immunoreactive (ir)-NT and -GAL were localized in the three regions of the adenohypophysis; ir-FMRF-amide was found in the neurohypophsis and in cells of the rostral pars distalis, but the immune reaction to this antisera in the pituitary was of very low intensity. Ir-GAL was found to colocalize with growth hormone, prolactin, and somatolactin in pituitary cells. In the brain, ir-GAL was found in the posterior nucleus lateralis tuberis (NLT), nucleus preopticus (NPO), and in the nucleus preopticus periventricularis (NPP). Ir-NT was localized in the anterior NLT. Ir-FMRF-amide was localized in the nucleus olfactoretinalis (NOR) where it was colocalized with gonadotropin releasing hormone, as well as in tracts that appeared to extend from the NOR, through the NPO and NPP, to the NLT. The NLT and the NOR have been previously implicated in the pituitary regulation of reproductive function. The above-cited localizations suggest that these peptides are involved in the neuroendocrine regulation of growth and reproduction in this genus.

Animals↗

Intraoperative US of the pituitary gland. Work in progress.

PURPOSE: To evaluate the use of ultrasound (US) of the pituitary gland during transsphenoidal surgery as a means of detecting microadenomas in patients with Cushing disease. MATERIALS AND METHODS: Thirteen patients with Cushing disease and one with acromegaly underwent US during transsphenoidal surgery. Mechanically oscillating transducers (10 MHz [n = 8] or 15 MHz [n = 6]) mounted on a 15-cm-long probe were used. RESULTS: Seven adenomas were definitely visualized, two were poorly seen, and four were overlooked. Small tumors (< 5 mm in diameter) tended to be hypoechoic; larger ones, hyperechoic. All four overlooked adenomas were imaged with the 15-MHz transducer. The cavernous sinuses and internal carotid arteries could be seen, but minimal invasion of the medial wall of the cavernous sinus could not. CONCLUSION: Pituitary microadenomas can be detected with transsphenoidal US; however, evaluation of larger series with instrumentation developed specifically for transsphenoidal application will be necessary to determine the ultimate value of this technique.

Acromegaly↗

Epidermal tissue homeostasis. II. Cell pool size, cell birth rate and cell loss in toads deprived of the pars distalis of the pituitary gland.

Following removal of the pars distalis of the pituitary gland in toads, epidermal efflux from the stratum corneum recruitment cell pool (i.e. production of corneal layers) is greatly increased. In this investigation the cell birth rate is studied by means of the metaphase arrest technique, as a function of time after pars distalis ablation. The method allows assessment of the total cell production over 14 days after the operation, to be compared with the total efflux and changes in the epidermal cell pool size. Whereas in intact toads the rate of cell production exceeds that of cell loss by moulting by a factor of 2.7, the 'surplus' of cells neither being used for formation of corneal layers nor permanently accommodated within the living epidermis, a 'balance sheet' of efflux and influx indicates that following pars distalis ablation all cells produced are also used for the (excessive) formation of corneal cell layers. The observations lend further support to the hypothesis that controlled cell deletion is a tissue homeostatic mechanism complementary to controlled cell divisions.

Animals↗

Prolonged sensitisation of pituitary glands in vitro to repeated administration of luteinising hormone-releasing hormone: effects of pulse frequency, ovariectomy, estradiol and progesterone.

Anterior pituitary gland fragments were obtained from female Wistar-derived rats on dioestrus or pro-oestrus and perifused in Biogel columns in vitro. They were subjected at the beginning of each of 5 h of perifusion to a volley of 6 1-min pulses of luteinising hormone-releasing hormone (LHRH, 10 nM), given 4 min apart. Luteinising hormone (LH) was measured by radioimmunoassay in sequential 2-min fractions of the perifusate. Pituitary glands removed at 14.00 h on dioestrus showed a characteristic pattern of sensitisation followed by desensitisation to the repeated volleys of LHRH, whereas tissues removed at 10.00 or 14.00 h on pro-oestrus showed no evidence of desensitisation over the 5-hour period, the response to each LHRH volley being greater than the preceding one. Estradiol (E, 3-100 pg/ml) added to the medium from the start of perifusion had no significant effect on the pattern of response from tissues removed on pro-oestrus, but the highest concentration significantly enhanced the response of dioestrus pituitaries to all but the last of the LHRH volleys. Progesterone (P, 1-50 ng/ml) added to the medium produced a dose-related inhibition of the response of pro-oestrous tissues to the LHRH volleys. Groups of animals were ovariectomised (OVX) on dioestrus and used for experiment the next morning. OVX at 10.00 h on dioestrus produced the pattern of response characteristic of dioestrus the next morning, but with much higher levels of LH release, which were unaltered by the addition of E to the medium. OVX at 17.00 h on dioestrus produced an entirely different pattern of response the next day, with high basal and moderate LHRH-induced LH release, and no evidence of changes in sensitivity of the tissue.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Lack of autoreceptor mediated regulation of the spontaneous dopamine turnover in the isolated neurointermediate lobe of the rat pituitary gland in vitro.

Isolated neurointermediate lobes of the rat pituitary gland were incubated in Krebs-HEPES solution and the spontaneous outflow of endogenous dopamine and its metabolites (DOPAC, HVA and MOPET) was determined by HPLC with electrochemical detection. The spontaneous outflow of dopamine metabolites (about 1500 fmol/10 min) largely exceeded that of dopamine (about 60 fmol/10 min). Apomorphine concentration-dependently (IC50, 205 nmol/l) reduced the spontaneous outflow of the dopamine metabolites. The effect of apomorphine developed slowly and was progressive over an observation period of 70 min. After 1 h of exposure to a maximall effective concentration of apomorphine (10 mumol/l), the outflow of metabolites was inhibited by 43%. The effect of apomorphine was not affected by the dopamine D2 receptor antagonist (-)-sulpiride nor by the dopamine D1 receptor antagonist SCH 23390. Neither quinpirole nor fenoldopam significantly affected the spontaneous outflow of dopamine metabolites. It was previously shown that the high rate of spontaneous outflow of dopamine metabolites from the dopaminergic nerves in the neurointermediate lobe reflects largely the immediate catabolism of newly synthesized dopamine. This high rate of spontaneous dopamine synthesis in the neurointermediate lobe is not controlled by dopamine autoreceptors. Apomorphine appears to inhibit the spontaneous dopamine turnover by an inhibition not mediated by dopamine receptors.

3,4-Dihydroxyphenylacetic Acid↗

ACTH and corticosterone secretion in rats following removal of the neurointermediate lobe of the pituitary gland.

Surgical removal of the neurointermediate lobe of the pituitary gland (NIL-X) in the rat resulted in two abnormalities in ACTH secretion: (1) plasma ACTH and corticosterone levels were elevated in the morning and over a 24-hour period compared with levels in control (NIL-C) rats, and (2) although NIL-X and NIL-C rats had acute increases in plasma ACTH and corticosterone of equal magnitude after interoceptive stimuli (hemorrhage, surgery), NIL-X rats demonstrated markedly smaller elevations in plasma levels of these hormones after neurotropic stimuli (noise, novel environment). This subnormal adrenocortical response of NIL-X rats was not due to an impairment in perception of a neurotropic stimulus; these rats had normal latencies to paw licking and to jumping off a heated surface, yet smaller increases in plasma corticosterone after the stimulus. The impairment in ACTH response was not related to stimulus intensity, as NIL-X and NIL-C rats had equal ACTH and corticosterone secretion during both low and high levels of insulin-induced hypoglycemia. NIL-X rats demonstrated a significant elevation in daily water intake, although hematocrit, plasma Na+, K+, osmolality and protein were normal. Significant diurnal rhythms in plasma corticosterone levels and in water intake were maintained as well. The elevated morning plasma ACTH levels, the blunted hormone increases after noise, and the increase in water intake persisted in NIL-X rats 2 months after surgery. These data indicate that removal of the NIL results in (1) chronic elevations in basal ACTH and corticosterone secretion, and (2) chronic impairment in adrenocortical responses to neurotropic stimuli, but not to interoceptive stimuli. The deficit is not due to impaired perceptual capacity nor to the intensity of the stimulus.

Adrenocorticotropic Hormone↗

Magnetization transfer imaging of the pituitary gland.

Magnetization transfer (MT) techniques provide tissue contrast which depends mainly on the concentration of macromolecules. The magnetization transfer phenomenon is determined by the restricted macromolecular protons and is quantified by the magnetization transfer ratio (MTR). Since many macromolecular structures are implicated in the secretory activity of the pituitary gland, this imaging technique has recently been used for the study of the normal adenohypophysis and the pre-operative and post-operative evaluation of pituitary adenomas. Significant changes of the MTR with age occur in normal subjects. In patients with hyperprolactinemia, the MTR of prolactin-secreting tumors is higher and that of the non secreting adenomas is lower compared to the MTR values of the normal pituitary gland of age and sex matched controls. In some patients operated on for growth hormone adenomas, although classic MR images were negative for residual tumor, increased MTR values were highly suggestive of persistence of adenoma tissue, in agreement with biochemical findings of persisting secretory activity. These data suggest that MT techniques might be an additional useful tool in the assessment of pituitary gland disorders.

Journal Article↗

Effect of mating on the metabolic activity of the brain and pituitary gland assessed by [14C]2-deoxyglucose in a reflex ovulator, the vole (Microtus agrestis).

The neural pathways involved in reflex ovulation in the vole (Microtus agrestis) have been investigated with the [14C]2-deoxyglucose (2-DG) method. Female voles were injected i.p. with 2-DG and either not mated, sham-mated (mounted by males by vagina was taped) or mated for a period of 45 min after the injection, after which the animals were decapitated. The brain was processed for autoradiography and the relative metabolic activities (rma) of selected areas of the brain and pituitary gland were determined. The plasma separated from trunk blood was assayed for the concentration of luteinizing hormone (LH). The lordosis quotients (LQ) were (mean +/- S.E.M.) 81 +/- 8 (n = 5) in mated compared with 47 +/- 8 (n = 5) in sham-mated voles. The rma of the midbrain central grey (CG) and reticular formation (RF) were significantly greater in mated and sham-mated voles compared with the values in unmated voles. There were no other between-group differences in the rma of the other 28 areas of the brain or pituitary gland studied in spite of the fact the plasma LH concentration in mated voles was 36.9 +/- 9.6 ng NIH-LH-S18/ml compared with undetectable (less than 2.5 ng/ml) values in all the sham-mated and unmated animals. These results show that in the vole increased metabolic activity of the CG and RF is associated with lordosis, but that the reflex release of LH is not accompanied by any significant change in the metabolic activity of the brain or the pituitary gland.

Animals↗

Cell proliferation and death in the irradiated pituitary gland and its modification by growth stimulants.

PURPOSE: This study was undertaken to show whether the rate of expression of radiation injury in the rat pituitary gland could be accelerated by the use of growth stimulants. METHODS AND MATERIALS: Rat pituitary glands were irradiated in situ with a range of single doses up to 20 Gy. The rats were then given subcutaneous slow-release implants containing 17beta-estradiol (E2) and sulpiride (S) to stimulate lactotroph proliferation. Two sequential cycles were used, each consisting of stimulation (3 weeks) and withdrawal (2 weeks). Measurements were made of gland weight; BrdU-labeled, giant, and apoptotic cells; lactotrophs; as well as pituitary prolactin content, in response to exogenous thyroid-releasing hormone (TRH). RESULTS: The two cycles of stimulation/withdrawal resulted in marked changes in gland weight, BrdU-labeling index, and serum prolactin (PRL) levels in unirradiated rats. The proportion of immunopositive growth-hormone-producing (GH) cells increased after irradiation. Radiation inhibited the hypertrophic response to E2 + S and also inhibited increases in BrdU-labeling index and serum PRL levels. Also, giant lactotrophs were observed in the irradiated pituitaries. However, they were not seen in the unirradiated rats or in the irradiated rats treated with E2 + S. TRH promoted PRL secretion in the unirradiated rat. In contrast, TRH inhibited PRL secretion in the irradiated rat and in all treatment groups receiving E2 + S. Apoptosis was induced by irradiation and was substantially increased in lactotrophs and in other cell types by withdrawal of the E2 and S stimulus, although the highest observed incidence was only 7 per 10,000 cells. CONCLUSION: Both irradiation and E2 + S treatment removed the hypothalamic control of PRL secretion, which reveals this important inhibitory action of TRH upon PRL secretion. This suggests that it is not suitable as a dynamic test of pituitary PRL reserves in such abnormal situations, where there may also be damage to the hypothalamic-pituitary vasculature. The increasing proportion of GH cells after irradiation indicates that lactotrophs respond more rapidly to irradiation. The stimulation by E2 + S somehow prevented the radiation-damaged lactotrophs from becoming giant cells. Also, the ratio of apoptotic cells to BrdU-labeled cells was increased by the E2 + S treatment, indicating that the E2 + S did enhance radiation-induced cell death relative to cell renewal. However, overall, the E2 + S stimulus protocol did not promote a dramatic increase in cell death (apoptosis) nor a marked decrease in residual gland weight after irradiation. Hence, its use would probably not be beneficial in the treatment of slow-responding prolactinomas, if malignant lactotrophs respond similarly to the normal pituitary lactotrophs. However, the observation of induced apoptosis after hormone and drug withdrawal suggests that agents which promote tumor shrinkage may be effective by causing rapid apoptosis of tumor cells in vivo.

Animals↗

Microcalcifications in the anterior pituitary gland of the fetus and the newborn: a histochemical and immunohistochemical study.

Calcified concretions are a normal and constant finding in the anterior pituitary gland of fetuses and newborns. Their light and electron microscopic characteristics have been recently reported by the authors. In this study, undecalcified and decalcified sections from 20 neonatal and 60 fetal anterior pituitary glands were studied by histochemical and immunohistochemical methods to further clarify their nature and mechanism of formation. All the glands revealed homogeneous and/or laminar calcifications located either within the interstitium or follicular structures. They were composed of a diastase-resistant periodic acid-Schiff-positive carbohydrate-rich matrix. The Feulgen method for DNA was negative. Their core frequently reacted to Alcian blue and epithelial membrane antigen (EMA). EMA also stained the apical membranes of adjacent epithelial cells. Other immunostains (vimentin, keratin, and pituitary hormones) were negative. The positive staining for Alcian blue and EMA and the negative staining with the Feulgen method for DNA suggest that the core of the calcifications consists of acidic mucosubstances and EMA-positive proteinaceous material previously secreted by viable pituitary cells. The EMA-negative periphery of the concretions probably develops from further extracellular peripheral mineralization that leads to larger, sometimes laminated psammoma bodies. The occurrence of pituitary calcifications in states of adult physiological and pathological hyperprolactinemia suggests that the marked proliferation of lactotrophs occurring during the fetal life play an important role in the pathogenesis of the fetal and neonatal concretions.

Biomarkers↗

Occurrence of colloid-containing follicles in the pars distalis of pituitary glands from aging guinea pigs.

Colloid-containing follicles in the pars distalis of pituitary glands from guinea pigs at various ages ranging from 5 days to 36 months were examined by the periodic acid-Schiff (PAS) reaction, immunohistochemistry, and electron microscopy. The follicles storing PAS-positive colloid were first detected in 6-month-old animals, in which only a few follicles were present and mean diameters of colloid deposits were small: 4.3 +/- 1.0 microns in males and 4.1 +/- 0.4 microns in females. Thereafter, the follicles gradually increased in number and size with age. The largest number of follicles was observed in the senile groups: 410.5 +/- 92.3 in males, 454.7 +/- 84.7 in females. Mean diameters of colloid masses in the senile groups were more than 2 times larger than those in 6-month-old animals: 10.0 +/- 0.1 microns in males, 9.7 +/- 0.1 microns in females. These findings suggest that the formation of colloid-containing follicles in the guinea-pig pars distalis is an aging phenomenon. The follicular lumina were mainly surrounded by thin cytoplasmic processes or cell bodies of folliculo-stellate cells immunoreactive for S-100 protein. The lining folliculo-stellate cells showed aggregations of intermediate-sized filaments, numerous lysosomes and colloid-like inclusions. Granulated cells in contact with colloid were occasionally encountered. Intracellular cavities storing colloid-like and fibrous materials were detected in the syncytial formation of GH cells.

Aging↗