Improved sensitivity to adrenocorticotrophin after purification and pre-incubation of isolated rat adrenal cells

Goverde, H.J.M.; Pesman, G. J.; Benraad, Th. J.

doi:10.1530/acta.0.0940221

Cited by 29 publications

(11 citation statements)

References 18 publications

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“…The tissue was minced into small pieces and dissociated with collagenase (type I; Sigma Chemical Co., St. Louis, MO, USA), as described previously 21 . The tissue was minced into small pieces and dissociated with collagenase (type I; Sigma Chemical Co., St. Louis, MO, USA), as described previously 21 .…”

Section: Cell Dispersion and Incubationmentioning

confidence: 99%

An Adolescent with Food-Dependent Cushing's Syndrome Secondary to Ectopic Expression of GIP Receptor in Unilateral Adrenal Adenoma

Noordam¹,

Hermus²,

Pesman³

et al. 2002

Journal of Pediatric Endocrinology and Metabolism

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ACTH-independent Cushing's syndrome may be secondary to the expression of ectopic hormone receptors in adrenal tissue. In food-dependent Cushing's syndrome the adrenals aberrantly express receptors for gastric inhibitory polypeptide (GIP). We present the first case of food- and GIP-dependent adrenal adenoma in an adolescent. A 15 year-old girl presented with signs of hypercortisolism. Urinary cortisol excretion was clearly elevated. Upon the finding of very low fasting plasma cortisol levels with a rise during the morning, food-dependent cortisol secretion was suspected. Hourly measurements of plasma cortisol and GIP levels during a day with and a day without meals showed meal- and GIP-related cortisol secretion. Plasma ACTH levels were undetectable. Abdominal computed tomography showed a tumor of 2.5 x 2.5 x 2 cm in the left adrenal. Unilateral adrenalectomy was performed and microscopic examination of the tumor showed an adrenocortical adenoma. Incubation of adenomatous cells prepared from this tumor with GIP resulted in increased cortisol secretion. Using RT-PCR amplification and cDNA hybridization, the GIP receptor was found to be overexpressed in the adenoma tissue but not in the adjacent adrenal tissue. Twelve months after the operation, hourly measurements of plasma cortisol, GIP and ACTH levels on a day with and a day without meals no longer showed GIP-related cortisol production. The present report shows that in an adolescent with corticotrophin-independent Cushing's syndrome, food-dependent hypercortisolism is a possible diagnosis.

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Section: Cell Dispersion and Incubationmentioning

confidence: 99%

An Adolescent with Food-Dependent Cushing's Syndrome Secondary to Ectopic Expression of GIP Receptor in Unilateral Adrenal Adenoma

Noordam¹,

Hermus²,

Pesman³

et al. 2002

Journal of Pediatric Endocrinology and Metabolism

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“…The assay in [22] was not widely used because of either low responsiveness of the cells (high A 5o), which would require large volumes of serum, or preparing adrenocortical cell suspension from hypothysectomized rats to lower A 5o [22].…”

Section: Discussionmentioning

confidence: 99%

Preparation of adrenocortical cell suspension highly responsive to acth or dibutylryl‐cyclic AMP

Trzeciak

Mathé

1981

FEBS Letters

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“…The fractions collected from the columns described above were assayed for inhibition of ACTH-stimulated corticosterone production using a Sayers' type rat adrenal cell preparation (7) to which synthetic human ACTH (hACTH) was added (150 pg/ml). The modifications to the Sayers bioassay systems have been described (8)(9)(10). The buffers were prewarmed to 37°C on a shaking water bath and gassed with 95% 02/5% CO2 prior to use.…”

Section: Methodsmentioning

confidence: 99%

Isolation and structure of corticostatin peptides from rabbit fetal and adult lung.

Zhu

Mulay

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

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A 34-amino acid peptide and three other structurally related peptides were isolated from rabbit fetal and adult lung. These cationic arginine-and cysteine-rich peptides inhibit corticotropin (ACTH)-stimulated rat adrenal cell corticosterone production. The peptide was called corticostatin (CSI). CSI was purified by reverse-phase HPLC and was shown to be homogenous from its amino acid analysis. Its sequence was determined on a gas-phase sequenator. The structure of CSI is Gly-Ile-Cys-Ala-Cys-Arg-Arg-Arg-PheCys-Pro-Asn-Ser-Glu-Arg-Phe-Ser-Gly-Tyr-Cys-Arg-ValAsn-Gly-Ala-Arg-Tyr-Val-Arg-Cys-Cys-Ser-Arg-Arg. CSI was found to markedly inhibit ACTH-stimulated corticosterone production by rat adrenal cells in vitro but did not affect basal levels. CSI did not affect the stimulation of aldosterone synthesis by angiotensin I in rat zona glomerulosa cells but it did suppress ACTH-stimulated aldosterone synthesis in whole adrenal cells, demonstrating that CSI is a specific inhibitor of ACTH-stimulated corticosteroid synthesis. The minimum effective concentration of CSI inhibiting ACTH-stimulated (33 pM) corticosterone production was 5 nM (20 ng/ml), the ED50 (50% effective dose) was 25 nM and steroidogenesis was completely inhibited at concentrations >500 nM (2 jpg/ml).Glucocorticoids are known to stimulate fetal lung development in humans, rabbits (1)(2)(3)(4), and several other species. Secretory products from the lung are not known to affect adrenal function. In the course of trying to isolate and sequence corticotropin (ACTH) from rabbit fetal lung, we found a peak of immunoreactive material with the elution position of the peptide hormone after purification of the extract on HPLC. It was, however, present in such small amounts as to preclude chemical identification. The ACTHlike peak material stimulated corticosterone production in rat adrenal cells in vitro, but adjacent to it was material that suppressed corticosterone production. In this communication, we describe the isolation of peptides from adult and fetal rabbit lung that inhibit ACTH-stimulated corticosterone production by rat adrenal cell suspensions.MATERIAL AND METHODS Lung Extraction and Purification. A total of 100 pairs of rabbit fetal lungs obtained during late gestation (Pel-Freez) were homogenized at 4°C in 100 ml of an acidic medium consisting of 1 M HCI/5% formic acid/1% NaCl (wt/vol)/ 1% trifluoroacetic acid. After centrifugation (3300 x g for 15 min), the pellet was reextracted twice with 100 ml of medium and the supernatants were combined and divided into three equal parts. Each was passed through a set of five ODSsilica cartridges (C18 Sep-Pak, Waters Associates) connected in series. Prior to use, the ODS-silica was wetted with 20 ml of 80% acetonitrile containing 0.1% trifluoroacetic acid and equilibrated with 30 ml of 0.1% trifluoroacetic acid. After loading the eluates, the cartridges were washed with 50 ml of 0.1% trifluoroacetic acid and eluted with 10 ml of 80% acetonitrile in 0.1% trifluoroacetic acid. The eluates were then concen...

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Improved sensitivity to adrenocorticotrophin after purification and pre-incubation of isolated rat adrenal cells

Cited by 29 publications

References 18 publications

An Adolescent with Food-Dependent Cushing's Syndrome Secondary to Ectopic Expression of GIP Receptor in Unilateral Adrenal Adenoma

An Adolescent with Food-Dependent Cushing's Syndrome Secondary to Ectopic Expression of GIP Receptor in Unilateral Adrenal Adenoma

Preparation of adrenocortical cell suspension highly responsive to acth or dibutylryl‐cyclic AMP

Isolation and structure of corticostatin peptides from rabbit fetal and adult lung.

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