TISSUE CULTURE STUDIES ON HUMAN PITUITARY TUMOURS: LONG TERM RELEASE OF ANTERIOR PITUITARY HORMONES INTO THE CULTURE MEDIUM

ABSTRACT A study was undertaken to determine the length of time that human pituitary tumours are capable of releasing anterior pituitary polypeptide hormones in vitro under basal conditions and to study the spectrum of hormone release by functioning and "non-functioning" pituitary neoplasms. Fragments from the pituitary tumours of 10 patients in the following categories: 1 Cushing's disease, 2 with amenorrhoea-galactorrhoea, 3 with acromegaly, and 4 with "non-functioning" pituitary tumours and from 2 normal human anterior pituitary glands were placed in primary culture immediately after surgery. The in vitro release of human growth hormone (hGH), prolactin (Prl), thyrotrophin (TSH), adrenocorticotrophin (ACTH), luteinizing hormone (LH), and follicle stimulating hormone (FSH) was measured by specific radioimmunoassays at the end of each week in culture. Hormone release was surveyed from 6 weeks to 6 months depending upon the survival of the culture. Hormone release patterns were compared with clinical and pathological data. In the initial week of the study, all 6 anterior pituitary polypeptides were detected in the media from the 2 control pituitaries and from 4 of the tumours (1 amenorrhoea-galactorrhoea and 3 acromegaly) in concentrations up to 100 ng/ml of medium while 5 of the 6 hormones were readily detectable in the media from 2 additional tumour samples (Cushing's disease and 1 "non-functioning" pituitary tumour). The media of the remaining 4 tumours contained at least 3 of the 6 hormones (1 amenorrhoea-galactorrhoea and 3 "non-functioning" pituitary tumours). After 6 months in culture, the 6 hormones were readily detectable in at least 1 of the 5 surviving cultures and hGH (up to 800 ng/ml) and LH were each detectable in the media from 2 cultures. Although most of the hormone concentrations in the media decreased with length of time in culture, there were 2 exceptions. First, in the media from 5 of the 12 cultures from both controls and tumours, Prl concentrations increased after 50 to 80 days culture. This increase usually lasted for several weeks before Prl levels again began to decline. The second unusual finding occurred in a tumour from a patient with acromegaly in the media of which hGH levels rose from 60 ng/ml to 800 ng/ml between days 125 and 174. These findings of prolonged hormone release in vitro give promise of future usefulness of tissue culture methods for study of polypeptide hormone releasing mechanisms and long-term production of human anterior pituitary hormones for use in research and possible therapy.

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TISSUE CULTURE STUDIES ON HUMAN PITUITARY TUMOURS: RADIOIMMUNOASSAYABLE ANTERIOR PITUITARY HORMONES IN THE CULTURE MEDIUM

Acta Endocrinologica ◽

10.1530/acta.0.0880239 ◽

1978 ◽

Vol 88 (2) ◽

pp. 239-249 ◽

Cited By ~ 16

Author(s):

Loren G. Lipson ◽

Inese Z. Beitins ◽

Paul D. Kornblith ◽

Janet W. Mc Arthur ◽

Henry G. Friesen ◽

...

Keyword(s):

Tissue Culture ◽

Cushing’S Disease ◽

Anterior Pituitary ◽

Pituitary Hormones ◽

Cushing's Disease ◽

Hormone Release ◽

Pituitary Tumours ◽

Human Pituitary ◽

Anterior Pituitary Hormones

ABSTRACT A tissue culture study was undertaken to determine if human non-functioning pituitary tumours secrete polypeptide anterior pituitary hormones in vitro and to study the spectrum of hormone release by functioning pituitary neoplasms. Fragments from 48 human pituitary tumours (from patients - 2 with Cushing's disease, 1 with Nelson's syndrome, 5 with amenorrhoea-galactorrhoea, 10 with acromegaly and 30 with non-functioning pituitary tumours) and three normal human anterior pituitary glands (controls) were placed in tissue culture immediately after surgery. The in vitro release of human growth hormone (HGH), prolactin (Prl), thyrotrophin (TSH), adrenocorticotrophin (ACTH), luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured by radioimmunoassays at the end of one week in culture. Clinical and pathological data were compared to hormone release patterns. In the culture media from control pituitaries the concentrations of the six hormones tested were 100 to 10 000 times greater than in peripheral blood. The medium surrounding the fragments from functioning pituitary tumours contained the following: a) Acromegaly - high levels of HGH and variable concentrations of the other hormones. b) Cushing's disease - ACTH and Prl predominantly. c) Amenorrhcea-galactorrhoea syndrome - prolactin in 4 out of 5 patients, all six polypeptides in one patient. In the media from the 30 patients diagnosed as having non-functioning pituitary tumours, 60 % of the samples contained at least one hormone at a concentration similar to that of the controls and 100 % of the samples contained detectable quantities of at least one hormone.

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STUDIES ON THE GROWTH AND HORMONE PRODUCTION OF RAT HYPOPHYSIS IN TISSUE CULTURE

Acta Endocrinologica ◽

10.1530/acta.0.0430147 ◽

1963 ◽

Vol 43 (1) ◽

pp. 147-154 ◽

Cited By ~ 2

Author(s):

Stig Kullander

Keyword(s):

Tissue Culture ◽

Pituitary Gland ◽

Anterior Pituitary ◽

Anterior Lobe ◽

Anterior Pituitary Gland ◽

The Other ◽

Pituitary Tumours ◽

Hormone Production ◽

Other Hand

ABSTRACT The anterior lobe of the pituitary gland of the rat was studied in tissue culture. Oestrone, progesterone and androsterone did not have any effect on the growth. On the other hand, oestrogen-induced pituitary tumours in tissue culture grew more quickly in medium containing oestrone or androsterone. The anterior pituitary gland produced prolactin in vitro.

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THE EFFECT OF ANTERIOR PITUITARY HORMONES ON CARBOHYDRATE METABOLISM IN VITRO IN THE PREPUBERTAL RAT OVARY

Journal of Endocrinology ◽

10.1677/joe.0.0400073 ◽

1968 ◽

Vol 40 (1) ◽

pp. 73-80 ◽

Cited By ~ 2

Author(s):

PATRICIA W. MAJOR ◽

D. T. ARMSTRONG

Keyword(s):

Carbohydrate Metabolism ◽

Anterior Pituitary ◽

Lactic Acid Production ◽

Pituitary Hormones ◽

Thyroid Stimulating Hormone ◽

Anterior Pituitary Hormones ◽

The Media ◽

Rat Ovary ◽

Low Levels

SUMMARY Small doses of luteinizing hormone (LH) have been shown to exert a stimulating effect on the in vitro carbohydrate metabolism of the prepubertal rat ovary. In order to determine the specificity of the response, studies were undertaken comparing the effects caused by LH with those of other anterior pituitary hormones. Preparations of thyroid stimulating hormone (TSH) caused responses similar to those caused by LH, when added to the media in which prepubertal rat ovaries were incubated. A detailed study of the effects of TSH preparations (prepared by different procedures with known low levels of LH contamination) on lactic acid production, has been undertaken. All TSH preparations tested caused considerably greater stimulation of glycolysis than could be accounted for on the basis of their contamination with LH when the latter was appraised by the ovarian ascorbic acid depletion assay.

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Early glucocorticoid feedback in anterior pituitary corticotrophs: differential inhibition of hormone release induced by vasopressin and corticotrophin-releasing factor in vitro

Journal of Endocrinology ◽

10.1677/joe.0.1290261 ◽

1991 ◽

Vol 129 (2) ◽

pp. 261-268 ◽

Cited By ~ 14

Author(s):

M. J. Shipston ◽

F. A. Antoni

Keyword(s):

Anterior Pituitary ◽

Actinomycin D ◽

Feedback Inhibition ◽

Female Rats ◽

Hormone Release ◽

Type Ii ◽

Acth Secretion ◽

Corticotrophin Releasing Factor ◽

Acth Release

ABSTRACT Vasopressin and 41-residue corticotrophin-releasing factor (CRF-41) are physiological mediators of the hypothalamic control of pituitary ACTH secretion, whilst adrenocortical glucocorticoids are the major inhibitory factors regulating ACTH output. In the present study it was investigated in vitro whether the characteristics of early glucocorticoid inhibition of stimulated ACTH secretion would differ depending on the nature of the stimulus and the temporal relationship between secretagogue and steroid. The experiments were carried out using perifused segments of rat adenohypophysis obtained from randomly cycling female rats. Repeated pulses (5 min) of CRF-41 or vasopressin were given at 1-h intervals for up to 7 h. The net release of ACTH became stable after the second secretagogue pulse. Administration of 0·1 μmol corticosterone/l 30 min before and during a 5-min pulse of 10 nmol CRF-41/l inhibited CRF-41-stimulated ACTH release to 60% of control. Stimulated hormone release remained suppressed at 90 min after the start of the corticosterone infusion and returned to control levels by 150 min. If corticosterone treatment (35 min total exposure) was started simultaneously with the CRF-41 pulse, no inhibitory effect of the steroid was observed at any subsequent time-point examined (60,90,120 and 150 min). In contrast, vasopressin-stimulated ACTH release was inhibited by approximately 50% when corticosterone was applied before, or simultaneously with, a 5-min pulse of 10 nmol vasopressin/l. The synthetic glucocorticoid type II receptor agonist RU28362, administered 30 min before and during a 5-min pulse of 10 nmol CRF-41/l, reduced CRF-41-stimulated ACTH release to 50% of control up to 2·5 h after the start of RU28362 application (although inhibition after 35 min exposure was not statistically significant). Inhibition of ACTH release stimulated by 10 nmol vasopressin/l was observed within 35 min of steroid application and was maintained up to 2·5 h after the initial application of RU28362. The action of RU28362 on CRF-41-stimulated ACTH release was blocked by inhibitors of transcription (actinomycin D) and translation (puromycin); notably these drugs did not modify the ACTH response to CRF-41. In contrast, actinomycin D as well as puromycin reduced vasopressin-stimulated ACTH release. The data suggest that: (1) the timing of steroid application is important in determining the early glucocorticoid inhibition of CRF-41- but not vasopressin-stimulated ACTH secretion; (2) CRF-41 and vasopressin mobilize different pools of ACTH from the anterior pituitary gland; (3) type II glucocorticoid receptors and synthesis of new protein(s) are involved in the early inhibitory action of glucocorticoids; (4) depending on the timing and nature of the incident secretagogue, differential negative feedback inhibition of ACTH secretion may occur at the pituitary level in vivo. Journal of Endocrinology (1991) 129, 261–268

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Vasopressin as a neuroendocrine regulator of anterior pituitary hormone secretion

Acta Endocrinologica ◽

10.1530/acta.0.1290489 ◽

1993 ◽

Vol 129 (6) ◽

pp. 489-496 ◽

Cited By ~ 33

Author(s):

Andreas Kjær

Keyword(s):

Arginine Vasopressin ◽

Mode Of Action ◽

Anterior Pituitary ◽

Hormone Secretion ◽

Pituitary Hormones ◽

Pituitary Hormone ◽

Anterior Pituitary Hormones ◽

Anterior Pituitary Hormone

Secretion of the anterior pituitary hormones adrenocorticotropin (ACTH), β-endorphin and prolactin (PRL) is complex and involves a variety of factors. This review focuses on the involvement of arginine-vasopressin (AVP) in neuroendocrine regulation of these anterior pituitary hormones with special reference to receptor involvement, mode of action and origin of AVP. Arginine-vasopressin may act via at least two types of receptors: V1− and V2−receptors, where the pituitary V1−receptor is designated V1b. The mode of action of AVP may be mediating, i.e. anterior pituitary hormone secretion is transmitted via release of AVP, or the mode of action may be permissive, i.e. the presence of AVP at a low and constant level is required for anterior pituitary hormones to be stimulated. Under in vivo conditions, the AVP-induced release of ACTH and β-endorphin is mainly mediated via activation of hypothalamic V1− receptors, which subsequently leads to the release of corticotropin-releasing hormone. Under in vitro conditions, the AVP-stimulated release of ACTH and β-endorphin is mediated via pituitary V1b− receptors. The mode of action of AVP in the ACTH and β-endorphin response to stress and to histamine, which is involved in stress-induced secretion of anterior pituitary hormones, is mediating (utilizing V1− receptors) as well as permissive (utilizing mainly V1− but also V2−receptors). The AVP-induced release of PRL under in vivo conditions is conveyed mainly via activation of V1−receptors but V2−receptors and probably additional receptor(s) may also play a role. In stress- and histamine induced PRL secretion the role of AVP is both mediating (utilizing V1 −receptors) and permissive (utilizing both V1− and V2− receptors). Arginine-vasopressin may be a candidate for the PRL-releasing factor recently identified in the posterior pituitary gland. Arginine-vasopressin of both magno- and parvocellular origin may be involved in the regulation of anterior pituitary hormone secretion and may reach the corticotrophs and the lactotrophs via three main routes: the peripheral circulation, the long pituitary portal vessels or the short pituitary portal vessels.

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A STUDY OF PROLACTIN-LIKE ACTIVITY IN INDIVIDUAL HUMAN PITUITARY GLANDS

Journal of Endocrinology ◽

10.1677/joe.0.0480639 ◽

1970 ◽

Vol 48 (4) ◽

pp. 639-647 ◽

Cited By ~ 3

Author(s):

PATRICIA M. NICHOLSON

Keyword(s):

Growth Hormone ◽

Gel Electrophoresis ◽

Anterior Pituitary ◽

Post Partum ◽

Human Pituitary ◽

Lactating Women ◽

Pituitary Glands ◽

Human Anterior Pituitary ◽

Pituitary Prolactin ◽

Individual Human

SUMMARY Polyacrylamide disc gel electrophoresis of aqueous extracts of individual human anterior pituitary glands failed to identify a protein with lactogenic activity which was characteristic of pregnancy and the post-partum period. Lactogenic activity, determined by a semi-quantitative rabbit mammary gland organ culture assay, was largely associated with the growth hormone fraction. The total prolactin activity of individual anterior pituitary glands was determined by a 'local' intradermal pigeon crop sac method. The glands from pregnant and parturient women did not contain a higher concentration of prolactin than those of men or non-pregnant non-lactating women. These results do not provide any evidence for the existence of a human pituitary prolactin distinct from growth hormone. Reasons for this are discussed.

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Long-term maintenance of reaggregated hypothalamic cultures developed from embryonic rat hypothalamus: prostaglandin release during synaptogenesis in vitro

Journal of Cell Science ◽

10.1242/jcs.34.1.159 ◽

1978 ◽

Vol 34 (1) ◽

pp. 159-171

Author(s):

A. Gyevai ◽

P.J. Chapple ◽

W.H. Douglas

Keyword(s):

Adult Animal ◽

Morphological Differentiation ◽

Adult Rat ◽

Monolayer Cultures ◽

Prostaglandin Release ◽

Pge2 Concentration ◽

The Media ◽

Term Maintenance

Hypothalamic aggregate cultures were developed from hypothalami taken from rat embryos at 17–19 days gestation. The aggregate cultures exhibited a prominent morphological differentiation during 3–4 weeks in culture. The fine structure of the synapses formed in the aggregates resembled synapses in tha adult animal. During synaptogenesis the aggregates spontaneously release prostaglandin E2 (PGE2). The amount of PGE2 released in the media was reversed upon the morphological differentiation of the hypothalamic cultures. Media containing a higher PGE2 concentration increased the extracellular prolactin accumulation in monolayer cultures developed from adult rat hypophysis.

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Phloroglucinol Mediated Plant Regeneration of Ornithogalum dubium as the Sole “Hormone-Like Supplement” in Plant Tissue Culture Long-Term Experiments

Plants ◽

10.3390/plants9080929 ◽

2020 ◽

Vol 9 (8) ◽

pp. 929

Author(s):

Carloalberto Petti

Keyword(s):

Tissue Culture ◽

Plant Regeneration ◽

Callus Formation ◽

Total Phenolic ◽

Naturally Occurring ◽

Long Term Experiments ◽

Direct Embryogenesis ◽

The Media ◽

Synthetic Hormones

Tissue culture is an essential requirement in plant science to preserve genetic resources and to expand naturally occurring germplasm. A variety of naturally occurring and synthetic hormones are available to induce the processes of dedifferentiation and redifferentiation. Not all plant material is susceptible to tissue culture, and often complex media and hormone requirements are needed to achieve successful plant propagations. The availability of new hormones or chemicals acting as hormones are critical to the expansion of tissue culture potentials. Phloroglucinol has been shown to have certain hormone-like properties in a variety of studies. Ornithogalum dubium, an important geophyte species, was used to characterise the potential of phloroglucinol as the sole plant-like hormone in a tissue culture experiment. Tissue culture, plant regeneration, total phenolic and genetic variability were established by applying a variety of methods throughout long-term experiments. Phloroglucinol did induce callus formation and plant regeneration when used as the sole supplement in the media at a rate of 37%, thus demonstrating auxin/cytokines-like properties. Callus formation was of 3 types, friable and cellular, hard and compact, and a mixture of the two. The important finding was that direct somatogenesis did occur albeit more frequently on younger tissue, whereby rates of induction were up to 52%. It is concluded that phloroglucinol acts as a “hormone-like” molecule and can trigger direct embryogenesis without callus formation.

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Characterization of the somatostatin-like immunoreactivity extracted from an adrenal medullary tumour

Bioscience Reports ◽

10.1007/bf01116377 ◽

1982 ◽

Vol 2 (3) ◽

pp. 147-154 ◽

Cited By ~ 6

Author(s):

R. Corder ◽

J. E. C. Sykes ◽

P. J. Lowry

Keyword(s):

Growth Hormone ◽

Anterior Pituitary ◽

Gel Filtration ◽

Hormone Release ◽

Growth Hormone Release ◽

Identical Composition ◽

Salt Fractionation ◽

Somatostatin 14

Significant amounts of somatostatin-like immunor reactivity (SLI) were detected in the extract of a human catecholamine-secreting adrenal medullary tumour. After salt fractionation and reconstitution the major portion of SLI was purified by gel filtration and two HPLC steps; in all three systems it eluted in the position of somatostatin-14. The purified somatostatin-like peptide inhibited, in a dose-related manner, growth hormone release from stimulated perfused rat anterior pituitary ceils in vitro. Amino acid analysis showed the purified peptide to have an identical composition to somatostatin found in other species.

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