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C Denef

Publications and source records attributed to C Denef.

At least 37 records · Page 2Linked to original sources
PubMed1994

Autoradiographic demonstration of 125I-Tyr4-bombesin binding sites on rat anterior pituitary cells.

Because several direct effects of bombesin related peptides on pituitary hormone release have been demonstrated, we chose to study the presence of bombesin binding sites in the adult male rat anterior pituitary in aggregate cell culture by autoradiographic localization of 125I-tyrosine4-bombesin (125I-Tyr4-BBN) binding and immunocytochemical localization of the anterior pituitary hormones. When aggregates were cultured in medium without hormonal supplements, the number of cells with detectable 125I-Tyr4-BBN binding was below 1%. In cell aggregates cultured in the presence of 1 nM estradiol (E2) 125I-Tyr4-BBN binding was detected on 5.4 +/- 0.8% of the cells after redispersion and on 5.8 +/- 1.1% of the cells in sections of paraffin embedded aggregates. The binding cell types were mainly lactotrophs and somatotrophs. The binding of 125I-Tyr4-BBN (3 or 5 nM) was specific because it was inhibited by the addition of an excess of unlabelled Tyr4-BBN or the bombesin receptor antagonist L686,095. In aggregates cultured in the presence of 1 nM E2 and 4 nM dex, the percentage of cells with detectable 125I-Tyr4-BBN binding was significantly lower than that in aggregates cultured in the presence of 1 nM E2 alone. Binding on somatotrophes almost completely disappeared. The present data show that specific 125I-Tyr4-BBN binding sites are present on anterior pituitary cells, are detectable mainly on a small subpopulation of lactotrophs and somatotrophs, and are affected by hormonal conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Isolation of two peptides from rat gonadotroph-conditioned medium displaying an amino acid sequence identical to fragments of secretogranin II.

Pituitary cells from 14-day-old rats were separated by unit gravity velocity sedimentation, and a highly enriched population of gonadotrophs was established in reaggregate cell culture in serum-free and serum albumin-free defined culture media. The medium conditioned by these aggregates was concentrated, ultrafiltrated, and the concentrated substances consecutively separated by two reversed-phase HPLC steps, a gel filtration step and an additional reversed-phase purification step. Two peptides could be isolated that displayed an N-terminal amino acid sequence identical to fragments of rat presecretogranin II: one was cleaved at the dibasic amino acid residues K182-R183 and the other at the dibasic amino acid residues K569-R570. The latter peptide was completely purified and separated into three variants with the same N-terminal amino acid sequence. From an analysis by electrospray ionization mass spectrometry, it was deduced that the three forms extended up to amino acid residue 612 (A), which is, in presecretogranin II, also flanked by two basic amino acids (K613R614). Two variant forms had a molecular mass that was 17 Da higher. The present data support the hypothesis that secretogranin II is a precursor of putative regulatory peptides.

Amino Acid Sequence↗
PubMed1994

Bioactive peptides in anterior pituitary cells.

The anterior pituitary (AP) has been shown to contain a wide variety of bioactive peptides: brain-gut peptides, growth factors, hypothalamic releasing factors, posterior lobe peptides, opioids, and various other peptides. The localization of most of these peptides was first established by immunocytochemical methods and some of the peptides were localized in identified cell types. Although intracellular localization of a peptide may be the consequence of internalization from the plasma compartment, there is evidence for local synthesis of most of these peptides in the AP based on the identification of their messenger-RNA (mRNA). In several cases the release of the peptide from the AP cell has been shown and regulation of synthesis, storage and release have also been described. Because the amount of most of the AP peptides is very low (except for POMC peptides and galanin), endocrine functions are not expected. There is more evidence for paracrine, autocrine, or intracrine roles in growth, differentiation, and regeneration, or in the control of hormone release. To demonstrate such functions, in vitro AP experiments have been designed to avoid the interference of hypothalamic or peripheral hormones. The strategy is first to show a direct effect of the peptide after adding it to the in vitro system and, secondly, to explore if the endogenous AP peptide has a similar action by using blockers of peptide receptors or antisera immunoneutralizing the peptide.

Animals↗
PubMed1994

In vitro evidence that an 11-kilodalton N-terminal fragment of proopiomelanocortin is a growth factor specifically stimulating the development of lactotrophs in rat pituitary during postnatal life.

We have previously shown that in reaggregate cell cultures of 14-day-old female rat pituitary, LHRH is capable of stimulating lactotrophs to enter the S-phase of the cell cycle, and that this effect is mediated by paracrine growth factors secreted from gonadotrophs. In the present report we describe the isolation, purification, and chemical characterization of one of these growth factors. Gonadotroph-rich aggregates from 14-day-old female rats were cultured for 6-7 weeks in serum-free and serum albumin-free defined medium in the presence of 0.01 nM LHRH. A total volume of 3.2 liters containing material secreted from 2.5 x 10(8) cells was batchwise concentrated on Sep-Pak C18/125 A cartridges (10 g) and retained molecules ultrafiltrated on Centricon 3 membrane filters [mol wt (M(r)) cut-off, 3 kilodaltons (kDa)]. Material capable of increasing the number of [3H]thymidine ([3H]T)-incorporating lactotrophs in pituitary cell aggregates of 2-week-old rats was isolated by chromatography on two reverse phase HPLC columns, one HPLC gel filtration column, and a final reverse phase HPLC column. A substance was obtained which, on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions, ran as a single band with an apparent M(r) of 16 kDa. On gel filtration, the apparent M(r) was 11 kDa. N-Terminal amino acid sequence analysis revealed that the substance was a peptide; a sequence of 10 residues was obtained, which was identical to that in the N-terminal part of rat POMC. By electrospray ionization mass spectrometry, six different compounds with slightly different M(r), ranging from 10,796-11,108 daltons, were detected. The latter data suggest that the peptide extends C-terminally at least to residue 74, which in the POMC sequence is flanked by an Arg-Arg dibasic residue, a posttranslational cleavage site. The substance increased the number of [3H]T-incorporating lactotrophs in pituitary cell aggregates without affecting [3H]T labeling of other pituitary cell types. Authentic human POMC-(1-76), at a concentration of 10 nM provoked a similar stimulation of the [3H]T-labeled lactotroph number without affecting other cell types. It is concluded that POMC-(1-74) is a growth factor that specifically targets lactotrophs during postnatal development of the rat anterior pituitary.

Animals↗
PubMed1994

In vitro evidence that LHRH stimulates the recruitment of prolactin mRNA-expressing cells during the postnatal period in the rat.

We have studied the effect of LHRH and neuropeptide Y (NPY) on prolactin (PRL) mRNA levels in pituitary reaggregate cell cultures from 14-day-old female rats, by means of in situ hybridization and Northern blot analysis. As estimated by computer-image analysis, addition of LHRH on day 5 in culture for 40 h resulted in a 37% increase in the total cytoplasmic areas of cells containing PRL mRNA, visualized using a digoxigenin-labelled PRL cRNA. The size of individual PRL-expressing cells was not influenced, nor was the content of PRL mRNA per cell. A similar effect of LHRH was found by dot blot hybridization of extracted RNA. PRL mRNA levels were not affected by NPY. LHRH induced a 29% increase in the number of PRL mRNA-expressing cells processing through the S phase of the cell cycle, visualized by the incorporation of [3H]thymidine ([3H]T) into DNA over 16 h. The fraction of [3H]T-labelled cells was 10-12% of the total cell population. NPY did not influence the number of [3H]T-positive cells expressing PRL mRNA, but completely blocked the effect of LHRH on the latter population. The present data suggest that LHRH, probably via a paracrine action of gonadotrophs, stimulates the recruitment of new lactotrophs, an action which is negatively modulated by NPY. Since the magnitude of this effect was the same in the total pituitary cell population as in cells processing through the S phase of the cell cycle and presumably mitosis, recruitment of lactotrophs seems to be based on differentiation of progenitor or immature cells into PRL-expressing cells, rather than on a mitogenic action on pre-existing lactotrophs alone.

Analysis of Variance↗
PubMed1993

Expression of the genes encoding bombesin-related peptides and their receptors in anterior pituitary tissue.

The bombesin-related peptides gastrin-releasing peptide (GRP) and neuromedin B (NMB) have been demonstrated in the anterior pituitary (AP) on an immunological basis. We studied the presence of mRNAs for these peptides and for their receptors by RNAse protection assay using fresh adult male rat AP, AP cell reaggregates cultured in the presence of estradiol and the rat AP derived GH3 cell line. In total RNA from fresh AP we detected high amounts of NMB mRNA and much smaller amounts of GRP mRNA, while finding a weak signal for GRP-receptor (GRP-R) and NMB-receptor (NMB-R) mRNAs. In total RNA from the reaggregate cell cultures we detected high levels of NMB mRNA as well as a strong signal for GRP-R mRNA. Finally, in GH3 cells, high levels of NMB mRNA and GRP-R mRNA were found, while GRP mRNA and NMB-R mRNA remained undetectable even in high amounts (200 micrograms) of total RNA. We conclude that mRNAs encoding both bombesin-related peptides and each of the mRNAs encoding their receptors are expressed in rat AP tissue. NMB mRNA is more prominent than GRP mRNA in all AP-like tissues examined (fresh AP, estradiol-treated reaggregate AP cell cultures and GH3 cells). NMB-R mRNA and GRP-R mRNA are both present in low levels in fresh AP whereas the GRP-R mRNA is predominant in GH3 cells and estradiol treated AP reaggregate cell cultures. Compared to fresh AP tissue, NMB mRNA and GRP-R mRNA expression is enhanced in estradiol-treated reaggregate cell cultures.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Immunocytochemical evidence that S-100-positive cells of the mouse anterior pituitary contain interleukin-6 immunoreactivity.

We have previously shown that bioactive interleukin-6 (IL-6) is produced by rat and mouse (anterior) pituitary cells in vitro. Since the amount produced correlated with the presence of S-100-containing folliculostellate (FS) cells, these cells were suggested to be a source of IL-6 in the anterior pituitary (AP) lobe. In the present study we used immunocytochemical techniques to confirm this presumption. Freshly isolated mouse pituitary cells were subjected to immunocytochemical procedures whereby two different (neutralizing) monoclonal antibodies (MAb) against mouse IL-6 (6B4 and 20F3) and a polyclonal antiserum raised against bovine S-100 were used as primary antibodies. Single immunostaining revealed a small portion of mouse pituitary cells (about 6.5%) to be positive for IL-6 immunoreactivity with both antibodies. Importantly, the same proportion of cells was found to be IL-6 positive if only the AP was used as the cell source. About 7.5% of the pituitary cells stained for the presence of S-100 immunoreactivity. Positive staining for IL-6 was also found in pituitary cell samples from 2-day-old monolayer cultures and from redispersed 9-day-old histotypic aggregates, which both secreted bioassayable IL-6. In contrast, no IL-6 staining was found in AtT-20 cells, an established ACTH-secreting tumor cell line of the mouse pituitary which did not secrete bioactive IL-6. The specificity of the IL-6 immunostaining was demonstrated by a total loss of staining when MAb 6B4 was omitted or replaced by irrelevant rat IgG. Conclusively, pre-adsorption of the anti-IL-6 MAb (6B4) with recombinant mouse IL-6 totally abolished staining of pituitary cells. Double immunostaining for IL-6 and S-100 revealed that most if not all of the IL-6-containing pituitary cells were positive for S-100. Few of the S-100-containing cells did not stain for IL-6. These results confirm our previous hypothesis that FS cells, characterized by immunostaining of S-100 protein, contain bioactive and immunoreactive IL-6 and therefore are very likely producers of IL-6 in the AP. Furthermore, our results suggest that IL-6 is implicated in the local regulatory role ascribed to FS cells in the pituitary gland.

Animals↗
PubMed1992

Isolation of cleaved prolactin variants that stimulate DNA synthesis in specific cell types in rat pituitary cell aggregates in culture.

Using an affinity column of monoclonal antibodies against rat prolactin (PRL), PRL immunoreactive material secreted by rat pituitary aggregate cell cultures was purified in milligram amounts from the culture medium. Further purification of the PRL immunoreactive molecules was achieved by anion-exchange and reversed-phase h.p.l.c. At least four variants could be distinguished which showed PRL-like bioactivity in the Nb2 lymphoma bioassay. On SDS/PAGE two variants migrated as single bands under both reducing and non-reducing conditions. The two other variants migrated as single bands under non-reducing conditions, but yielded under reducing conditions two fragments, the larger of which had molecular masses of approx. 16 and 17 kDa respectively. These variants were therefore considered to be cleaved PRL (clPRL) variants, and were designated clPRL-1 and clPRL-2 respectively. These variants were not artefactually produced at low pH during the h.p.l.c. purification step. Each of the clPRLs represented about 0.6-1.0% of total PRL. The cleavage site of clPRL-1, determined by N-terminal and C-terminal amino acid sequence analysis, was in the large disulphide loop between Tyr-145 and Leu-146. Purified clPRL-1 added to pituitary aggregate cell cultures from 14-day-old female rats in the presence of the dopamine agonist bromocriptine increased [3H]thymidine incorporation into DNA in gonadotrophs and thyrotrophs, but not in other pituitary cell types. Under the same experimental conditions the main forms of PRL and clPRL-2 were inactive. These data demonstrate that PRL is cleaved within the pituitary gland between specific amino acid residues, and that PRL cleaved between Tyr-145 and Leu-146 may have a specific biological role as a paracrine growth regulator in this tissue.

Amino Acid Sequence↗
PubMed1992

Negative regulation by dexamethasone of the potentiation of neuromedin C-induced growth hormone and prolactin release by estradiol in anterior pituitary cell aggregates.

The bombesin-like peptide neuromedin C (NMC) stimulated the release of growth hormone (GH) and prolactin (PRL) from adult male rat anterior pituitary cell aggregates cultured in serum-free medium. This effect, particularly on GH release, was significantly increased when the cells were cultured in the presence of estradiol (E2). The GH response to NMC was not affected by 4 nM dexamethasone (Dex) but was enhanced in the presence of 80 nM Dex. However, the strong stimulatory action of E2 on the GH response to NMC was completely abolished by 4 nM Dex. In contrast, the PRL response to NMC was reduced by 4 nM Dex and this to a proportionally similar extent in the presence or absence of E2. The present data show that glucocorticoids can be both positive and negative regulators of the same response system.

Animals↗
PubMed1992

Stimulation of prolactin secretion from rat pituitary by luteinizing hormone-releasing hormone: evidence against mediation by angiotensin II acting through a (Sar1-Ala8)-angiotensin II-sensitive receptor.

In aggregate cell cultures of 15- to 20-day-old rat pituitary maintained in serum-free medium, luteinizing hormone-releasing hormone (LHRH) (10 nM) stimulated prolactin (PRL) release, confirming our previous results and those of others with serum-supplemented medium. Since angiotensin II (AII) stimulates PRL release and a renin-angiotensin system is expressed in gonadotrophs, LHRH stimulation of PRL release might be mediated by AII. To evaluate this hypothesis, the influence of (Sar1,Ala8)AII and (Sar1,Ile8)AII two peptide AII receptor antagonists, of DUP753, a nonpeptide and stable AII receptor antagonist, of a converting enzyme inhibitor, and of angiotensinogen on LHRH-induced PRL release was tested in various in vitro conditions of 15- to 20-day-old female rat pituitary. In aggregates maintained in serum-free medium with or without dexamethasone (DEX) and triiodothyronine (T3), or maintained in serum-supplemented medium, the effect of LHRH on PRL release was not affected by (Sar1Ala8)AII (0.1 microM), (Sar1,Ile8)AII (10 microM) or DUP753 (10 microM). Only a high dose (10 microM) of (Sar1,Ala8)AII attenuated the LHRH-induced PRL release. The latter attenuation was seen only with aggregates cultured in the DEX/T3 medium and not with aggregates cultured in the presence of serum. A dose of 1 or 10 nM (Sar1,Ala8)AII also failed to block the effect of LHRH used at 1 nM. In contrast, (Sar1,Ala8)AII dose dependently as well as DUP753 (10 microM) abolished the AII-induced PRL release. (Sar1,Ala8)AII also failed to affect the LHRH-induced PRL release in pituitary cell aggregates from 6-week-old male rats. However, in aggregates from both immature and 6-week-old rats, (Sar1,Ala8)AII provoked a small and statistically significant attenuation of the LHRH-induced PRL release when a 100 nM dose of LHRH was used. In freshly isolated hemipituitaries from 5-day-old rats, (Sar1,Ala8)AII (1 or 10 microM) did not affect the LHRH- (10 nM) induced PRL release. In single cells obtained by redispersion of aggregates and mounted in a Biogel P2 column, LHRH still stimulated PRL release. Again this effect could not be blocked by DUP753. Treatment of aggregate cell cultures with the angiotensin-converting enzyme inhibitor captopril or with angiotensinogen did not alter the LHRH-induced PRL release. It is concluded that AII is not the paracrine factor mediating the effect of LHRH at low nanomolar doses on PRL release, at least not through the classical AII receptor. The involvement of AII acting on a non-(Sar1,Ala8)AII-sensitive receptor cannot be excluded and warrants further investigation.

1-Sarcosine-8-Isoleucine Angiotensin II↗
PubMed1992

Angiotensin II is retained in gonadotrophs of pituitary cell aggregates cultured in serum-free medium but does not mimic the effects of exogenous angiotensins and luteinizing-hormone-releasing hormone on growth hormone release.

Angiotensin II (AII)-like immunoreactivity (LIR) was detected by immunostaining in 7.5 +/- 1.1% of cells obtained by redispersion of pituitary cell aggregates from 15- to 20-day-old female rats, cultured for 5-7 days in serum-free medium supplemented with thyroid hormone and dexamethasone. Also, renin-LIR was retained in these cultures. As shown by double immunostaining of paraffin-embedded sections of the aggregates, this AII-LIR was localized only in gonadotrophs. AII-LIR was detected at least up to 5 weeks in culture. On reversed-phase, high-performance liquid chromatography (HPLC), this AII-LIR co-migrated with authentic AII. In perifused aggregate cell cultures of 15- to 20-day-old female rat pituitary maintained in serum-free medium supplemented with dexamethasone (DEX) and triiodothyronine (T3), AII stimulated GH release. AI and AIII had a similar effect. To evaluate the possible involvement of endogenous AII in the local regulation of GH release, gonadotrophs were stimulated with luteinizing hormone-releasing hormone (LHRH). LHRH displayed a transient inhibitory effect on GH release, which was followed by a rebound of GH release after withdrawal of the peptide. Treatment of aggregates with pertussis toxin reversed this inhibitory effect into a significant stimulation of GH release. In aggregates cultured in serum-supplemented medium, LHRH provoked a significant stimulation of GH release which was still followed by a post-stimulus rebound release. In hemipituitaries from 5-day-old rats, a significant stimulatory effect of LHRH on GH release was found without rebound secretion. To evaluate the possible involvement of endogenous AII in the effects of LHRH on GH release, the influence of (Sar1,Ala8)AII, a peptide AII receptor antagonist, and of DUP753, a non-peptide AII receptor blocker was tested in various in vitro conditions. The effect of LHRH on GH release in aggregates cultured either in serum-free medium supplemented with DEX and T3 or in serum-supplemented medium was not affected by (Sar1,Ala8)AII, not even after enhancing the LHRH-induced GH release by treatment of the aggregates with pertussis toxin. A hundred times lower concentration of (Sar1,Ala8)AII, however, abolished the AII-induced changes in GH release. Also DUP753 (10 microM) failed to block LHRH-induced GH release in aggregates. (Sar1,Ala8)AII also failed to block the effect of LHRH on GH release from hemipituitaries. It is concluded that LHRH has inhibitory and stimulatory effects on GH release in cultured pituitary cell aggregates.(ABSTRACT TRUNCATED AT 400 WORDS)

Angiotensin II↗
PubMed1992

Luteinizing hormone-releasing hormone and neuropeptide Y influence deoxyribonucleic acid replication in three anterior pituitary cell types. Evidence for mediation by growth factors released from gonadotrophs.

Treatment of anterior pituitary reaggregate cell cultures from 14-day-old female rats with physiological doses of the gonadotropin-releasing hormone LHRH or neuropeptide Y (NPY) for 40 h dose-dependently increased [3H]thymidine ([3H]T) incorporation into DNA of cells expressing PRL immunoreactivity (PRL-ir) and of those expressing ACTH-ir, whereas these peptides decreased the number of [3H]T-labeled cells expressing GH-ir. The effects of NPY were of the same magnitude as those of LHRH. The effects of LHRH were not seen in a gonadotroph-deprived cell population obtained by sequential velocity and buoyant density gradient sedimentation. When the latter cell population was coaggregated with purified gonadotrophs from 14-day-old rats, LHRH did enhance [3H]T labeling of lactotrophs and decreased that of somatotrophs. Gonadotroph-conditioned medium obtained by continuous perifusion of gonadotroph-rich reaggregates contained four different high molecular weight substances mimicking the effects of LHRH and NPY on [3H]T incorporation in the respective pituitary cell types. These substances were partially purified and separated from each other by concentration on a Bond-elut C18-reversed phase cartridge, ultrafiltration, and C18-reversed phase HPLC. One factor stimulated [3H]T labeling of lactotrophs, another that of corticotrophs, and two others inhibited [3H]T labeling of somatotrophs. The present data suggest that the development of PRL-, GH-, and ACTH-containing cells in the pituitary is modulated by LHRH and/or NPY and that the action of LHRH and probably also of NPY is mediated by specific paracrine growth factors released from gonadotrophs.

Adrenocorticotropic Hormone↗
PubMed1992

Evidence that folliculo-stellate cells mediate the inhibitory effect of interferon-gamma on hormone secretion in rat anterior pituitary cell cultures.

Interferon-gamma (IFN-gamma) is known to inhibit the release of ACTH, PRL, and GH by rat anterior pituitary (AP) cells, stimulated by appropriate hypothalamic releasing factors. In the present study we examined the mechanisms underlying this inhibition. Dose-response studies, revealing a maximal inhibitory effect with an IFN-gamma dose as small as 10 U/ml, suggested the existence of a limiting intermediate step. In addition, in perifusion experiments with aggregates of established hormone-secreting tumor cell lines (AtT-20 and GH3), IFN-gamma had no inhibitory effect, suggesting that an accessory cell type was involved. Studies on differentially enriched cell populations of normal rat AP, obtained by velocity and buoyant density sedimentation, indicated that the inhibitory effect of IFN-gamma on stimulated ACTH and GH release was absent in those populations that contained only few folliculo-stellate (FS) cells. The presence of a minimal proportion of FS cells was found to be necessary for the inhibition to be manifest. This was seen in monolayers, but also in cultures of AP cell aggregates, which are well known to closely mimic the behavior of the AP gland in vivo. Definitive evidence for the role of FS cells was obtained by reconstitutive coculture experiments; FS cell-poor populations, which by themselves resisted the inhibitory effect of IFN-gamma, became sensitive when cocultured with an FS cell-rich population. Basal ACTH and GH release were not influenced by preincubation with IFN-gamma in either original or fractionated AP cell populations. In contrast, basal PRL release was inhibited in both systems. In cultures of AP cell populations, separated by velocity sedimentation, this inhibition showed a pattern similar to that observed for stimulated release of ACTH and GH, i.e. more inhibition in fractions with a higher proportion of FS cells. However, in FS cell-poor cultures, inhibition of basal PRL release did occur, although to a lesser degree than in FS cell-rich cultures. Our results indicate that IFN-gamma affects AP hormone secretion through the FS cell. In addition, they suggest that IFN-gamma and the FS cell constitute a system through which the pituitary gland perceives changes in the activation state of the immune system.

Adrenocorticotropic Hormone↗
PubMed1991

Selectivity of juxtaposition between cup-shaped lactotrophs and gonadotrophs from rat anterior pituitary in culture.

Semi-thin sections of three-dimensional reaggregates from adult female rat pituitary, cultured in serum-free defined medium, were stained for prolactin, gonadotropin, thyrotropin, growth hormone and S-100, using the double immunolabelling technique. The frequency of juxtaposition between lactotrophs and gonadotrophs was enumerated and compared with the expected frequency at random distribution of polygonal cell profiles in a hexagonal configuration. The proportions of lactotrophs and gonadotrophs in the aggregate sections were determined using stereometrical analysis. The observed frequency of juxtaposition did not differ significantly from the expected frequency. Hence, no reason was found to assume a selective adhesion between lactotrophs and gonadotrophs in adult female rat pituitary reaggregates. A constant proportion of lactotrophs was found to meet the criteria of a cup-shaped morphology, and 70% +/- 9% (mean +/- S.D.) of these so-called cup-shaped lactotrophs were found to be juxtaposed at their concave side to gonadotrophs. Administration of 0.01 nM 17 beta-oestradiol to the culture medium resulted in a significant reduction of the proportion of cup-shaped lactotrophs but did not affect the selectivity of juxtaposition to gonadotrophs. The selectivity of juxtaposition between cup-shaped lactotrophs and gonadotrophs may be the morphological correlate of the functional relationship between these cells, which are known to be involved in an intra-pituitary paracrine communication system.

Animals↗
PubMed1991

Release of interleukin-6 from anterior pituitary cell aggregates: developmental pattern and modulation by glucocorticoids and forskolin.

The release of the immunologically active cytokine interleukin-6 (IL-6) by cultured anterior pituitary cell aggregates was found to increase with the age of the donor rats. The glucocorticoid hormone dexamethasone (DEX) dose-dependently inhibited the release of IL-6, the IC50 being 2.43 +/- 0.93 nM. The sex steroids dihydrotestosterone and estradiol were without detectable effect. The adenylate cyclase activator forskolin increased IL-6 release in aggregate cultures in the presence of DEX. It is concluded that IL-6 release by anterior pituitary cells is subject to developmental changes and under regulatory control of biologically active substances.

Adenylyl Cyclases↗
PubMed1991

Evidence for the presence of gastrin-releasing peptide immunoreactivity in rat anterior pituitary corticotrophs and lactotrophs, AtT20 cells, and GH3 cells: failure to demonstrate participation in local control of hormone release.

Using antisera raised against the N-terminal (1-16) or the C-terminal part of the bombesin (BBN)-like peptide gastrin-releasing peptide (GRP) and against the C-terminus of pro-GRP, GRP- and pro-GRP-like immunoreactivity (IR) was detected by immunostaining in freshly dispersed rat anterior pituitary (AP) cells and in reaggregate AP cells cultured in serum-free medium. Depending on the antiserum used, 6.4 +/- 0.4% to 12.4 +/- 2.7% of freshly dispersed cells were immunoreactive. Solid phase preabsorption of the antisera with the respective antigens abolished the staining. GRP-IR was detectable by double immunostaining in a subpopulation of PRL cells and ACTH-containing cells. In contrast, only very few somatotrophs, gonadotrophs, and thyrotrophs were immunoreactive, and they were much smaller than the typical mature forms of these cell types. GRP- and pro-GRP-IR were also detected in the ACTH-secreting AtT20 cell line and in the PRL- and GH-secreting GH3 cell line. GRP- and pro-GRP-IR were present in AP cell aggregates maintained in culture for 4 weeks. The finding of both GRP- and pro-GRP-IR in a subpopulation of lactotrophs and corticotrophs and their persistence in culture under serum-free conditions strongly suggest that GRP-like peptides are produced in rat AP. Although the potent GRP receptor antagonist L 686,095-001C002 effectively blocks stimulation of GH and PRL release by exogenous BBN-like peptides, it failed to affect basal GH and PRL release from perifused reaggregate AP cell cultures as well as GH and/or PRL release stimulated by epinephrine, vasoactive intestinal peptide, TRH, GH-releasing factor, and angiotensin-II or PRL release inhibited by dopamine. Thus, the latter data do not support the hypothesis often suggested that peptides endogenously present in AP cells are involved in the paracrine regulation of AP hormone secretion.

Adrenocorticotropic Hormone↗
PubMed1990

New perspectives in the function of pituitary folliculo-stellate cells.

Classical morphological studies of the folliculo-stellate (FS) cells of the anterior pituitary have suggested that these cells play roles as supporting cells, in metabolism and in macromolecular transport. Over the last 10 years the details of their activity in both trophic and catabolic processes has been clarified, and recent work has demonstrated several transport systems in these cells. Various novel peptides with growth factor or cytokine activity have been identified in FS cells and/or FS cell conditioned media. These recent functional experiments confirm and extend previous morphological and experimental studies, and in addition open new perspectives on the physiological roles of FS cells.

Animals↗