Expression of mRNA for somatostatin receptor (sstr) types 2 and 5 in individual rat pituitary cells. A double labeling in situ hybridization analysis.

CircRNAs have been identified to be expressed differently and stably in numerous species and tissues, but their functions in growth hormone (GH) secretion are still largely unknown. In summary, we have revealed a circRNA-miRNA-mRNA network that may play a biological role in the rat pituitary gland. First, we verified the chromosome location information of circAgtpbp1 according to sequencing analysis. The circAgtpbp1 characteristics were authenticated through PCR, qRT–PCR, treating with RNase and fluorescent in situ hybridization (FISH). Second, we detected the expression pattern of circAgtpbp1 in the rat anterior pituitary by qRT–PCR. We also designed circAgtpbp1 siRNA and constructed overexpression plasmid to evaluate the effect of circAgtpbp1 function on GH secretion by qRT–PCR, ELISA and Western blot. CircAgtpbp1 is a stable, truly circular molecule. We found that circAgtpbp1 interacted with miR-543-5p and can regulate GH secretion in pituitary cells through a circAgtpbp1-miR-543-5p-GH axis. Overall, the evidence generated by our study suggests that circAgtpbp1 can act as a sponge of miR-543-5p to reduce the inhibitory effect of miR-543-5p on Gh1 and further promote GH secretion. These findings expand our existing knowledge on the mechanisms of hormone regulation in the pituitary gland.

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Effects of triiodothyronine, dexamethasone and estradiol-17β on GH mRNA in rat pituitary cells in culture as revealed by in situ hybridization

Acta Endocrinologica ◽

10.1530/acta.0.1240083 ◽

1991 ◽

Vol 124 (1) ◽

pp. 83-90 ◽

Cited By ~ 10

Author(s):

M. G. Martinoli ◽

R. Veilleux ◽

G. Pelletier

Keyword(s):

In Situ Hybridization ◽

Calf Serum ◽

Female Rats ◽

Pituitary Cells ◽

Mrna Levels ◽

Male And Female ◽

Male And Female Rats ◽

Rat Pituitary ◽

Estradiol 17Β

Abstract. The GH lines of rat pituitary tumour cells have been largely used to study the regulation of GH mRNA. In order to investigate the role of T3, dexamethasone and estradiol-17β on GH expression in non-tumoural pituitary cells, we have used in situ hybridization techniques performed on rat anterior pituitary cells in monolayer culture. The amounts of mRNA encoding for GH, as evaluated by counting the number of grains per somatotrope, were markedly reduced after 4 days of culture in a steroid-free medium supplemented with an hypothyroid calf serum. Addition of T3 or dexamethasone for 3 days increased GH mRNA levels. The concomitant administration of the two hormones produced a synergistic effect on GH mRNA levels which became higher than those observed after T3 or dexamethasone administration alone. However, this effect did not restore GH mRNA levels to those measured in monolayer pituitary cells grown in medium containing 10% fetal calf serum. Moreover GH mRNA levels appeared higher in male than in female pituitary cells. The administration of E2 to pituitary cell cultures from both male and female rats produced an increase by 15, and 12.8% in GH mRNA levels in male and female, respectively. This stimulatory effect of E2 in cell culture was competitively blocked by simultaneous incubation with the antiestrogen LY156758 (Keoxifene). These results demonstrate that T3, dexamethasone as well as E2 act directly on somatotropic cells to regulate GH gene expression.

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Plasma clearance, tissue uptake and expression of pituitary peptide 23/pancreatitis-associated protein in the rat

Journal of Endocrinology ◽

10.1677/joe.0.1450461 ◽

1995 ◽

Vol 145 (3) ◽

pp. 461-469 ◽

Cited By ~ 9

Author(s):

C Chakraborty ◽

S Sharma ◽

N Katsumata ◽

L J Murphy ◽

I C Schroedter ◽

...

Keyword(s):

Male Rats ◽

Pituitary Cells ◽

Tissue Uptake ◽

Recombinant Peptide ◽

Tissue Extracts ◽

Rat Pituitary ◽

Rat Pituitary Cells ◽

Plasma Half Life ◽

Pancreatitis Associated Protein

Abstract The secretion of peptide 23 by rat pituitary cells is stimulated by growth hormone-releasing hormone and inhibited by somatostatin. Recent cloning of the cognate cDNA for peptide 23 revealed that it is identical to pancreatitis-associated protein (PAP). In the present study, the clearance and tissue uptake of recombinant peptide 23/PAP in normal adult male rats was assessed. The plasma half-life of recombinant peptide 23/PAP was 4·8 ±1·4 (s.d.) min. Maximal accumulation of radiolabelled peptide 23/PAP was observed in the kidney, stomach, small intestine and pancreas whereas negligible uptake was seen in the liver, lung or heart. Peptide 23/PAP was detected in a variety of tissue extracts using a radioimmunoassay. Extracts of ileum contained the highest concentrations of peptide 23/PAP. In situ hybridization analysis showed that peptide 23/PAP mRNA was highly expressed in the columnar epithelial cells of ileum, jejunum and duodenum. These observations demonstrate that peptide 23/PAP, a protein previously thought to be of pituitary origin, is widely expressed in the gastrointestinal tract and that it is rapidly removed from the circulation by the kidney and by tissues which express peptide 23/PAP. Journal of Endocrinology (1995) 145, 461–469

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Expression of IL-6 mRNA in normal rat and human pituitaries and in human pituitary adenomas.

Journal of Histochemistry & Cytochemistry ◽

10.1177/42.1.8263325 ◽

1994 ◽

Vol 42 (1) ◽

pp. 67-76 ◽

Cited By ~ 24

Author(s):

B Velkeniers ◽

P Vergani ◽

J Trouillas ◽

J D'Haens ◽

R J Hooghe ◽

...

Keyword(s):

In Situ Hybridization ◽

Fibrous Tissue ◽

Anterior Lobe ◽

Pituitary Cells ◽

Human Pituitary ◽

Receptor Mrna ◽

Rat Pituitary ◽

Normal Rat ◽

Normal Human

Cells expressing IL-6 mRNA were detected by in situ hybridization in normal pituitaries. In normal untreated rat pituitary the expression was very low. Within hours after IP administration of liposaccharide, IL-6 mRNA accumulated in the anterior lobe of the pituitary. Production of IL-6 was monitored after dissociation and culture of pituitary cells. High levels (8000 pg/ml) were recovered after 72 hr in culture. In normal human pituitaries, less than 1% of cells expressed IL-6 mRNA or IL-6 receptor mRNA (IL-6-R mRNA). In gonadotropinomas, prolactinomas, and non-functioning adenomas, only rare, scattered positive cells were found for either IL-6 or IL-6-R. In contrast, both genes were highly expressed in ACTH- and GH-secreting tumors at the junction of adenoma and infiltrating fibrous tissue and around blood vessels. The combined expression of IL-6 and IL-6-R suggests that IL-6 acts in an autocrine or in a paracrine way in ACTH and GH adenomas.

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Fluorescence in situ hybridization analysis of human oocytes: Advantages of a double-labeling procedure

Fertility and Sterility ◽

10.1016/j.fertnstert.2004.03.050 ◽

2004 ◽

Vol 82 (4) ◽

pp. 919-922 ◽

Cited By ~ 5

Author(s):

F PELLESTOR ◽

T ANAHORY ◽

B ANDREO ◽

G REGNIERVIGOUROUX ◽

J SOULIE ◽

...

Keyword(s):

In Situ Hybridization ◽

Fluorescence In Situ Hybridization ◽

Hybridization Analysis ◽

Double Labeling ◽

Human Oocytes

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Combination of immunocytochemistry and in situ hybridization in the same tissue section of rat pituitary.

Journal of Histochemistry & Cytochemistry ◽

10.1177/34.1.3510246 ◽

1986 ◽

Vol 34 (1) ◽

pp. 39-43 ◽

Cited By ~ 89

Author(s):

B D Shivers ◽

R E Harlan ◽

D W Pfaff ◽

B S Schachter

Keyword(s):

In Situ Hybridization ◽

Enzymatic Degradation ◽

Cryostat Section ◽

Double Labeling ◽

Rnase Inhibitor ◽

Rat Pituitary ◽

A Cell ◽

Combination Of Methods ◽

Synthetic Capacity

A procedure is described for combining avidin-biotinylated horseradish peroxidase immunocytochemistry, for localizing peptides or proteins, with in situ hybridization for localizing mRNA autoradiographically in the same cryostat section of paraformaldehyde-fixed rat pituitary. Protection against enzymatic degradation of target mRNAs during the immunocytochemical step was necessary and was accomplished by including an RNase inhibitor, 0.04% diethylpyrocarbonate, in primary and secondary antisera. This combination of methods may be useful in other tissues, as well, for (a) determining the relation of protein content to the concentration of its encoding mRNA, (b) proving the synthetic capacity of a cell in which a protein has been localized, (c) determining immunological or nucleic acid probe specificity, or (d) as an alternative to double-labeling immunocytochemical methods.

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