Release of Pro-Opiomelanocortin-Derived Peptides from the Pars intermedia and Pars distalis of the Rat Pituitary: Effect of Corticotrophin-Releasing Factor and Somatostatin

1985 ◽  
Vol 41 (5) ◽  
pp. 363-373 ◽  
Author(s):  
Jacob Kraicer ◽  
Timothy C. Gajewski ◽  
Bruce C. Moor
Keyword(s):  
Pars Intermedia ◽  
Pars Distalis ◽  
Corticotrophin Releasing Factor ◽  
Rat Pituitary

ADRENOCORTICOTROPHIC HORMONE IN THE ANTERIOR AND NEUROINTERMEDIATE LOBES OF THE FETAL RAT PITUITARY GLAND

1981 ◽  
Vol 89 (2) ◽  
pp. 181-186 ◽  
Author(s):  
ALAIN CHATELAIN ◽  
J. P. DUPOUY
Keyword(s):  
Pituitary Gland ◽  
Functional Differentiation ◽  
Pars Intermedia ◽  
Adrenocorticotrophic Hormone ◽  
Pars Distalis ◽  
Adult Rats ◽  
Adrenal Cells ◽  
Specific Radioimmunoassay ◽  
Fetal Rat ◽  
Rat Pituitary

The concentration of ACTH in the pars distalis and pars intermedia of the fetal rat hypophysis from days 17–21 of pregnancy was measured with a specific radioimmunoassay and a bioassay using isolated adrenal cells from adult rats. In both lobes of the pituitary gland, a significant correlation was observed between immunoreactive and bioreactive values, expressed as pg equivalents synthetic human 1–39 ACTH per μg protein. In the pars distalis, ACTH concentrations increased steadily from days 17–20 and then remained unchanged to term. At this time they were tenfold higher than on day 17. In the neurointermediate lobe, ACTH was detected only from day 18; the concentration of ACTH increasing between days 18 and 19. At each of the stages of pregnancy examined, the concentration of ACTH in the pars distalis was greater than that in the pars intermedia. These data have demonstrated that ACTH is present in both anterior and neurointermediate lobes of the fetal rat hypophysis, that the functional differentiation of the pars distalis takes place earlier than that of the pars intermedia, and that the concentrations of corticotrophin in the pars distalis and in the pars intermedia have different patterns of development as gestation progresses.

Changes in glucocorticoid receptor mRNA in the developing ovine pituitary and the effects of exogenous cortisol

1995 ◽  
Vol 144 (3) ◽  
pp. 483-490 ◽  
Author(s):  
S G Matthews ◽  
K Yang ◽  
J R G Challis
Keyword(s):  
Glucocorticoid Receptor ◽  
Late Gestation ◽  
Developmental Changes ◽  
Pars Intermedia ◽  
Pars Distalis ◽  
Short Term ◽  
Neonatal Life ◽  
Inferior Aspect ◽  
Glucocorticoid Receptor Mrna

Abstract Developmental changes in pituitary glucocorticoid receptor (GR) mRNA were examined during gestation and early neonatal life using in situ hybridization. Pituitaries were harvested from sheep fetuses at days 60–80, 100–120, 130–135, 140–142 and term, and from lambs of days 0–7 and 30–60, and adults. GR mRNA was present in the pars distalis by day 60, levels increased through gestation, and there was a redistribution of GR mRNA, resulting in a relatively greater abundance at the base of the pars distalis. At term, there was a significant (P<0·05 compared with the day 140–142 fetuses) elevation of GR mRNA, which was maintained in the newborn lamb, reaching highest levels at days 30–60 of neonatal life. GR mRNA was undetectable in the pars intermedia until day 120, but subsequently increased to high levels at term. Interestingly, the expression of GR mRNA in the pars intermedia dropped precipitously in the newborn (P<0·05 compared with term), though levels recovered in the older lambs and adults. The regional and cellular distribution of GR mRNA correlated closely with the presence of immuno-reactive GR (irGR) in the pituitary; the majority of irGR was present in the nuclei. Intrafetal infusion of cortisol (12 h; 5 μg/min) in late gestation (day 135) had no effect on GR mRNA expression in either the pars distalis or pars intermedia. These results indicated that, in the fetal pituitary, (1) the GR gene is expressed in both the pars distalis and pars intermedia, (2) levels of GR mRNA in the fetal pituitary correlated with the distribution of nuclear irGR, (3) GR mRNA is present at higher levels in the inferior aspect of the pars distalis, its abundance increases immediately prior to parturition and is maintained in the newborn, and (4) cortisol infusion for 12 h does not affect GR mRNA in either region of the pituitary, suggesting that, in the short term, glucocorticoids do not directly regulate GR synthesis. Journal of Endocrinology (1995) 144, 483–490

Production and utilization of monoclonal antibodies to human/rat corticotrophin-releasing factor-41

1990 ◽  
Vol 5 (2) ◽  
pp. 159-166 ◽  
Author(s):  
N. G. N. Milton ◽  
E. W. Hillhouse ◽  
S. A. Nicholson ◽  
C. H. Self ◽  
A. M. McGregor
Keyword(s):  
Monoclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dependent Manner ◽  
Corticotrophin Releasing Factor ◽  
Tissue Extracts ◽  
Rat Pituitary ◽  
Hypothalamic Tissue ◽  
Dose Dependent

ABSTRACT Murine monoclonal antibodies against human/rat corticotrophin-releasing factor-41 (CRF-41) were produced and characterized for use in the immunological and biological characterization of CRF-41. Spleen cells from BALB/c mice immunized with CRF-41 conjugated to bovine γ-globulin were fused with a BALB/c-derived non-secretor X-63 myeloma line. Hybridomas were selected for CRF antibody production by enzyme-linked immunosorbent assay, and positive hybridomas cloned twice. Three monoclonal antibodies were obtained (KCHMB001, KCHMB002 and KCHMB003) and characterized as IgG1, IgG1 and IgG2a isotypes respectively, with affinity constants for rat CRF-41 of 30, 53 and 34 nmol/l respectively. All three monoclonal antibodies recognize an epitope contained between residues 34 and 41 of the human/rat sequence. The antibodies were able to neutralize the ACTH-releasing activity of rat CRF-41, applied to rat pituitary fragments in vitro, in a dose-dependent manner. Isoelectric focusing showed that KCHMB 003 detected bands of synthetic rat CRF-41 and rat [Met(O)21,38]-CRF-41 at pH 7·1 and 6·8 respectively. Use of KCHMB003 in a two-site enzyme-amplified immunoassay showed that this antibody recognizes both synthetic rat CRF-41 and immunoreactive CRF-41 in rat hypothalamic tissue extracts.

scholarly journals In situ hybridization analysis of anterior pituitary hormone gene expression during fetal mouse development.

1994 ◽  
Vol 42 (8) ◽  
pp. 1117-1125 ◽  
Author(s):  
M A Japón ◽  
M Rubinstein ◽  
M J Low
Keyword(s):  
Gene Expression ◽  
Anterior Pituitary ◽  
Beta Subunit ◽  
Pars Intermedia ◽  
Pituitary Hormone ◽  
Pars Distalis ◽  
Mouse Development ◽  
Glycoprotein Hormone ◽  
Anterior Pituitary Hormone ◽  
Stimulating Hormone

We used 35S-labeled oligonucleotides and cRNAs (riboprobes) to detect the temporal order and spatial pattern of anterior pituitary hormone gene expression in (B6CBF1 x B6CBF1)F2 fetal mice from embryonic Day 9.5 (E9.5) to postnatal Day 1 (P1). Pro-opiomelanocortin (POMC) mRNA was expressed in the basal diencephalon on Day E10.5, in the ventromedial zone of the pars distalis on Day E12.5, and in the pars intermedia on Day E14.5. The common alpha-glycoprotein subunit (alpha-GSU) mRNA first appeared in the anterior wall of Rathke's pouch on Day E11.5 and extended to the pars tuberalis and ventromedial zone of the pars distalis on Day E12.5. Thyroid-stimulating hormone-beta (TSH beta) subunit mRNA was expressed initially in both the pas tuberalis and ventromedial pars distalis on Day E14.5, with an identical spatial distribution to alpha-GSU at the time. In contrast, luteinizing hormone-beta (LH beta) subunit and follicle-stimulating hormone beta (FSH beta) subunit mRNAs were detected initially only in the ventromedial pars distalis on Days E16.5 and E17.5, respectively, in an identical distribution to each other. POMC-, alpha-GSU-, TSH beta, LH beta-, and FSH beta-positive cells within the pars distalis all increased in number and autoradiographic signal with differing degrees of spatial expansion posteriorly, laterally, and dorsally up to Day P1. POMC expression was typically the most intense and extended circumferentially to include the entire lateral and dorsal surfaces of the pars distalis. The expression of both growth hormone (GH) and prolactin (PRL) started coincidentally on Day E15.5. However PRL cells localized in the ventromedial area similarly to POMC and the glycoprotein hormone subunits, whereas GH cells were found initially in a more lateral and central distribution within the lobes of the pars distalis. Somatotrophs increased dramatically in number and autoradiographic signal, extending throughout the pars distalis except for the most peripheral layer of cells on Day E17.5. Mammotrophs also increased in number but less abundantly than somatotrophs, and PRL expression remained more confined to central-medial and ventrolateral areas of the pars distalis up to Day P1. These data demonstrate distinctive patterns of expression for each of the major anterior pituitary hormone genes during development of the mouse pituitary gland and suggest that different groups of committed cells are the immediate precursors to the terminally differentiated hormone-secreting cell types.

Vasopressin-binding sites in the pig pituitary gland: competition by novel vasopressin antagonists suggests the existence of an unusual receptor subtype in the anterior lobe

1994 ◽  
Vol 141 (3) ◽  
pp. 383-391 ◽  
Author(s):  
Y Arsenijevic ◽  
M Dubois-Dauphin ◽  
E Tribollet ◽  
M Manning ◽  
W H Sawyer ◽  
...  
Keyword(s):  
Pituitary Gland ◽  
Rat Liver ◽  
Arginine Vasopressin ◽  
Binding Sites ◽  
Anterior Lobe ◽  
Receptor Subtype ◽  
Lower Affinity ◽  
Corticotrophin Releasing Factor ◽  
Rat Pituitary ◽  
Acth Release

Abstract Arginine vasopressin (AVP) acts in the pituitary gland, in synergy with corticotrophin-releasing factor, to induce ACTH release in response to stressful stimuli. Pituitary AVP receptors in the rat are coupled to phospholipase C, as are the so-called V1-type AVP receptors. The present study examined [3H]AVP binding in membranes prepared from the anterior lobe of the pituitary gland of the pig. [3H]AVP, alone or in competition with analogues, bound to sites in the pig anterior lobe which are pharmacologically similar to those described previously by others in the rat pituitary gland. For comparison, the same competition studies were performed on membrane preparations from the rat liver which contain the classic V1-type AVP receptor. Pituitary and liver AVP-binding sites were dissimilar; both cyclic and linear V1 antagonists had, in general, a much lower affinity for pituitary AVP-binding sites than for those in the liver. Thus, Phaa-d-Tyr(Et)-Phe-Gln-Asn-Lys-Pro-Arg-NH2 (Phaa=phenylacetyl) has a 2500-fold greater affinity for the latter (negative logarithm of inhibition constant (pKi)=9·64) than for the former (pKi=6·22). One linear antagonist, Pa-d-Tyr-Phe-Val-Asn-Arg-Pro-Arg-Arg-NH2 (Pa=propionyl) had about equal affinities for liver and pituitary membranes (pKi=6·39 and 6·53 respectively). Another compound, Phaa-d-Tyr-Phe-Val-Asn-Arg-Pro-Arg-Arg-NH2 had the highest affinity found to date for binding to AVP sites in the pituitary (pKi=7·43). These findings suggest some ideas for the design of more potent and/or selective AVP analogues acting in the pituitary gland. Journal of Endocrinology (1994) 141, 383–391

SULPHUR METABOLISM IN THE PITUITARY AND HYPOTHALAMUS OF THE RAT: A STUDY OF RADIOISOTOPE-UPTAKE AFTER THE INJECTION OF 35S dl-CYSTEINE, METHIONINE, AND SODIUM SULPHATE

1960 ◽  
Vol 20 (1) ◽  
pp. 9-23 ◽  
Author(s):  
J. C. SLOPER ◽  
D. J. ARNOTT ◽  
BARBARA C. KING
Keyword(s):  
Sodium Sulphate ◽  
Pars Intermedia ◽  
Posterior Pituitary ◽  
Pars Distalis ◽  
Active Protein ◽  
Lateral Border ◽  
Nuclear Regions ◽  
Radioisotope Uptake ◽  
Different Parts ◽  
Supraoptic Nuclei

SUMMARY 1. The relative radioactivity of different parts of the pituitary and hypothalamus has been assessed with a flow counter, and, more satisfactorily, both visually and by granule-counts in autoradiographs. These experiments have involved 103 rats killed between 15 sec and 97 hr after the administration of 35S dl-cysteine, dl-methionine and sodium sulphate. 2. Subarachnoid injections proved more satisfactory than intraperitoneal or intracarotid ones. They were followed by the rapid localization of radioisotope in the adenohypophysis as well as in nervous tissue. 3. The early and marked uptake of radioisotope shown by the cell bodies of neurones in various nuclear regions, and in particular in the supraoptic nuclei, has been interpreted as evidence of active protein synthesis; this pattern of uptake was observed after the injection of labelled cysteine and methionine, but not sodium sulphate. 4. A similar, early, but less marked uptake of radioisotope was noted in the pars distalis after the injection of both cysteine and methionine. Only after the injection of methionine was there a marked uptake in the pars intermedia, and this was confined to its lateral border. 5. Uptake by the infundibular process of the neurohypophysis became greater than that in pars distalis or in the superjacent hypothalamus 9½ hr and longer after injection of labelled cysteine, but not methionine. This pattern of uptake was confirmed by granule counts in twenty-five animals. 6. It is suggested that the late neurohypophysial uptake of radioisotope reflects the storage in the nerve-terminals of the gland of slowly metabolizing proteins or polypeptides synthesized in the supraoptic and paraventricular nuclear regions. These substances probably include the posterior pituitary principles, since the latter are rich in cystine, but lack methionine.

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