The functional significance of glycosylation of proopiomelanocortin in melanotrophs of the mouse pituitary gland

1985 ◽  
Vol 107 (3) ◽  
pp. 365-374 ◽  
Author(s):  
B. G. Jenks ◽  
A. G. H. Ederveen ◽  
J. H. M. Feyen ◽  
A. P. van Overbeeke
Keyword(s):  
Pituitary Gland ◽  
Functional Significance ◽  
Peptide Hormones ◽  
Terminal Region ◽  
Pars Intermedia ◽  
Apparent Effect ◽  
Glycoprotein Precursor ◽  
Tunicamycin Treatment ◽  
Mouse Pituitary

ABSTRACT Pro-opiomelanocortin (POMC) is a glycoprotein precursor for a number of neuropeptides and peptide hormones. The functional significance of the glycosylation of POMC has never been established. Using the antibiotic tunicamycin to block glycosylation of the prohormone in the mouse pars intermedia, we have compared processing of non-glycosylated prohormone with that of glycosylated prohormone in pulse-chase experiments. The peptides produced from non-glycosylated prohormone were shown to be correct cleavage products. Therefore it was concluded that, with the possible exception of peptides from the N-terminal region of the prohormone, the carbohydrate on POMC plays no role in directing cleavage or in protecting the prohormone from random proteolysis. Tunicamycin treatment retarded N-terminal acetylation of melanotrophin but had no apparent effect on acetylation of β-endorphin. The mouse pars intermedia synthesizes two forms of POMC which differ in their degree of glycosylation. Our results indicated that, during secretion, the melanotrophs make no distinction between peptides derived from the two prohormones. J. Endocr. (1985) 107, 365–374

SEPARATION OF A LABELLED FRAGMENT FROM PROTEIN SYNTHESIZED IN VITRO BY THE PARS INTERMEDIA OF THE MOUSE PITUITARY GLAND

1977 ◽  
Vol 75 (1) ◽  
pp. 181-182
Author(s):  
H. M. ISHERWOOD ◽  
V. F. THORNTON
Keyword(s):  
Molecular Weight ◽  
Pituitary Gland ◽  
Pars Intermedia ◽  
Biosynthetic Activity ◽  
Cellulose Acetate Electrophoresis ◽  
King's College ◽  
Large Molecular Weight ◽  
Pituitary Glands ◽  
Mouse Pituitary

Department of Anatomy, King's College, London, WC2R 2LS (Received 12 May 1977) The biosynthetic activity of the pars intermedia of the mouse pituitary gland was investigated by studying the fate of labelled amino acids incorporated into relatively large-molecular-weight products (Thornton & Isherwood, 1975). It was reported that label was actively incorporated in vitro into a protein (referred to as 'labelled P1') with a molecular weight of about 75 000 (Isherwood & Thornton, 1977). However, more recent observations suggest that 'labelled P1' is a complex between a protein and a labelled peptide. The purpose of this communication is to give a brief account of some of the findings which led to this conclusion. Full details of the incubation and cellulose acetate electrophoresis procedures are given in Isherwood & Thornton (1977). Posterior pituitary glands from male, Schofield mice were incubated in media containing [3H]leucine and [3H]phenylalanine (50 μCi/ml). In one experiment, posterior

Improved immunolabelling of ultrathin cryosections using antibody conjugated with 1nm gold particles

1990 ◽  
Vol 48 (3) ◽  
pp. 928-929
Author(s):  
Morten H. Nielsen ◽  
Lone Bastholm
Keyword(s):  
Pituitary Gland ◽  
Liquid Nitrogen ◽  
Electron Density ◽  
Small Diameter ◽  
Cytochemical Staining ◽  
Gold Particles ◽  
Labelling Density ◽  
Ultrathin Cryosections ◽  
Mouse Pituitary ◽  
Electron Microscopical

During the last 5 years the diameter of the gold probes used for immuno-cytochemical staining at the electron microscopical (EM) level has been decreased. The advantage of small diameter gold probes is an overall increased labelling density. The disadvantage is a lower detectability due to the low electron density of smaller gold particles consequently an inconvenient high primary magnification needed for EM examination. Since 1 nm gold particles are barely visible by conventional EM examination the need for enlargement by silverenhancement of the gold particles has increased.In the present study of ultrathin cryosectioned material the results of immunostaining using 5 nm gold conjugated antibody and 1 nm gold conjugated antibodies are compared after silverenhancement of the 1 nm gold particles.Slices of freshly isolated mouse pituitary gland were immersion fixed for 20 min in 2 % glutaraldehyde /2 % paraformaldehyde. Blocks cryoprotected with 2.3 M sucrose were frozen in liquid nitrogen and ultra-cryosectioned on a RMC cryoultra-microtome.

scholarly journals IGF-I regulates pro-opiomelanocortin and GH gene expression in the mouse pituitary gland

2003 ◽  
Vol 178 (1) ◽  
pp. 71-82 ◽  
Author(s):  
J Honda ◽  
Y Manabe ◽  
R Matsumura ◽  
S Takeuchi ◽  
S Takahashi
Keyword(s):  
Pituitary Gland ◽  
Anterior Pituitary ◽  
Northern Blotting ◽  
Pituitary Cells ◽  
Mrna Levels ◽  
Gh Treatment ◽  
Gh Receptor ◽  
Pomc Mrna ◽  
Igf I ◽  
Mouse Pituitary

IGF-I is expressed in somatotrophs, and IGF-I receptors are expressed in most somatotrophs and some corticotrophs in the mouse pituitary gland. Our recent study demonstrated that IGF-I stimulates the proliferation of corticotrophs in the mouse pituitary. These results suggested that somatotrophs regulate corticotrophic functions as well as somatotrophic functions by the mediation of IGF-I molecules. The present study aimed to clarify factors regulating pituitary IGF-I expression and also the roles exerted by IGF-I within the mouse anterior pituitary gland. Mouse anterior pituitary cells were isolated and cultured under serum-free conditions. GH (0.5 or 1 microg/ml), ACTH (10(-8) or 10(-7) M), GH-releasing hormone (GHRH; 10(-8) or 10(-7) M), dexamethasone (DEX; 10(-8) or 10(-7) M) and estradiol-17beta (e2; 10(-11) or 10(-9) M) were given for 24 h. IGF-I mRNA levels were measured using competitive RT-PCR, and GH and pro-opiomelanocortin (POMC) mRNA levels were measured using Northern blotting analysis. GH treatment significantly increased IGF-I mRNA levels (1.5- or 2.1-fold). ACTH treatment did not alter GH and IGF-I mRNA levels. IGF-I treatment decreased GH mRNA levels (0.7- or 0.5-fold), but increased POMC mRNA levels (1.8-fold). GH treatment (4 or 8 microg/ml) for 4 days increased POMC mRNA levels. GHRH treatment increased GH mRNA levels (1.3-fold), but not IGF-I mRNA levels. DEX treatment significantly decreased IGF-I mRNA levels (0.8-fold). e2 treatment did not affect IGF-I mRNA levels. GH receptor mRNA, probably with GH-binding protein mRNA, was detected in somatotrophs, and some mammotrophs and gonadotrophs by in situ hybridization using GH receptor cDNA as a probe. These results suggested that IGF-I expression in somatotrophs is regulated by pituitary GH, and that IGF-I suppresses GH expression and stimulates POMC expression at the transcription level. Pituitary IGF-I produced in somatotrophs is probably involved in the regulation of somatotroph and corticotroph functions.

Distribution of Met5-enkephalin-Arg6-Gly7-Leu8-immunoreactivity in the rat and mouse pituitary gland

1991 ◽  
Vol 36 (2) ◽  
pp. 271-281 ◽  
Author(s):  
S. Soinila ◽  
N. Bäck ◽  
G.J. Mpitsos
Keyword(s):  
Pituitary Gland ◽  
Mouse Pituitary

scholarly journals Reporter Expression, Induced by a Growth Hormone Promoter-Driven Cre Recombinase (rGHp-Cre) Transgene, Questions the Developmental Relationship between Somatotropes and Lactotropes in the Adult Mouse Pituitary Gland

Endocrinology ◽  
2007 ◽  
Vol 148 (5) ◽  
pp. 1946-1953 ◽  
Author(s):  
Raul M. Luque ◽  
Geraldine Amargo ◽  
Shinya Ishii ◽  
Corrinne Lobe ◽  
Roberta Franks ◽  
...  
Keyword(s):  
Pituitary Gland ◽  
Transgenic Mouse ◽  
Anterior Pituitary ◽  
Adult Mouse ◽  
Small Subset ◽  
Parental Line ◽  
Pituitary Cells ◽  
Placental Alkaline Phosphatase ◽  
Developmental Studies ◽  
Mouse Pituitary

This report describes the development and validation of the rGHp-Cre transgenic mouse that allows for selective Cre-mediated recombination of loxP-modified alleles in the GH-producing cells of the anterior pituitary. Initial screening of the rGHp-Cre parental line showed Cre mRNA was specifically expressed in the anterior pituitary gland of adult Cre+/− mice and cephalic extracts of e17 Cre+/− fetuses. Heterozygote rGHp-Cre transgenic mice were crossbred with Z/AP reporter mice to generate Cre+/−,Z/AP+/− offspring. In this model system, the GH promoter-driven, Cre-mediated recombination of the Z/AP reporter leads to human placental alkaline phosphatase (hPLAP) expression that serves to mark cells that currently produce GH, in addition to cells that would have differentiated from GH cells but currently do not express the GH gene. Double immunocytochemistry of adult male and female Cre+/−,Z/AP+/− pituitary cells revealed the majority (∼99%) of GH-producing cells of the anterior pituitary also expressed hPLAP, whereas ACTH-, TSH-, and LH-producing cells were negative for hPLAP, confirming previous reports that corticotropes, thyrotropes, and gonadotropes develop independently of the somatotrope lineage. A small subset (∼10%) of the prolactin-producing cells was positive for hPLAP, consistent with previous reports showing lactotropes can arise from somatotropes during pituitary development. However, the fact that 90% of prolactin-producing cells were negative for hPLAP suggests that the majority of lactotropes in the adult mouse pituitary gland develop independently of the somatotrope lineage. In addition to developmental studies, the rGHp-Cre transgenic mouse will provide a versatile tool to study the role of a variety of genes in somatotrope function and neoplastic transformation.

PERFUSION OF INTERCELLULAR SPACES IN THE PARS DISTALIS AND THEIR VARIATION WITH TIME OF DAY

1975 ◽  
Vol 66 (3) ◽  
pp. 385-NP
Author(s):  
W. B. QUAY
Keyword(s):  
Pituitary Gland ◽  
Functional Significance ◽  
Time Of Day ◽  
Male Rats ◽  
Pars Distalis ◽  
Intercellular Spaces ◽  
India Ink ◽  
Structural Regularity ◽  
Rat Pituitary ◽  
Light Darkness

SUMMARY Microperfusion of the tissue parenchyma of regions of the rat pituitary gland with 1:1 dilutions of India ink revealed a network of fine intercellular spaces or canaliculi throughout the pars distalis. They were demonstrable in animals of all ages from 1·5 months to over 1 year, in both sexes, and at various times during the 24 h light–darkness cycle. However, 24 h rhythms were suggested in the radial perfusion distance (P < 0·05) and tissue density (P < 0·01) of the perfused canaliculi in adult male rats. The amplitude of the 24 h rhythmicity in these pituitary canaliculi was lower than that shown recently in pineal canaliculi of the same species. Structural regularity and evidence of rhythmic and physiologically correlated changes in the pituitary canaliculi suggest that they relate to natural features rather than being only artifactual, and that analysis of their possible functional significance is appropriate.

scholarly journals Epithelial cell integrin β1 is required for developmental angiogenesis in the pituitary gland

2016 ◽  
Vol 113 (47) ◽  
pp. 13408-13413 ◽  
Author(s):  
Kathleen M. Scully ◽  
Dorota Skowronska-Krawczyk ◽  
Michal Krawczyk ◽  
Daria Merkurjev ◽  
Havilah Taylor ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Pituitary Gland ◽  
Molecular Mechanisms ◽  
Cell Types ◽  
Endocrine Function ◽  
Vascular Density ◽  
Integrin Β1 ◽  
Coordinated Development ◽  
Mouse Pituitary ◽  
Insight Into

As a key component of the vertebrate neuroendocrine system, the pituitary gland relies on the progressive and coordinated development of distinct hormone-producing cell types and an invading vascular network. The molecular mechanisms that drive formation of the pituitary vasculature, which is necessary for regulated synthesis and secretion of hormones that maintain homeostasis, metabolism, and endocrine function, remain poorly understood. Here, we report that expression of integrin β1 in embryonic pituitary epithelial cells is required for angiogenesis in the developing mouse pituitary gland. Deletion of pituitary epithelial integrin β1 before the onset of angiogenesis resulted in failure of invading endothelial cells to recruit pericytes efficiently, whereas deletion later in embryogenesis led to decreased vascular density and lumen formation. In both cases, lack of epithelial integrin β1 was associated with a complete absence of vasculature in the pituitary gland at birth. Within pituitary epithelial cells, integrin β1 directs a large transcriptional program that includes components of the extracellular matrix and associated signaling factors that are linked to the observed non–cell-autonomous effects on angiogenesis. We conclude that epithelial integrin β1 functions as a critical and canonical regulator of developmental angiogenesis in the pituitary gland, thus providing insight into the long-standing systems biology conundrum of how vascular invasion is coordinated with tissue development.

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