Biomedical subjects

M Szabo

Publications and source records attributed to M Szabo.

At least 19 recordsLinked to original sources

PubMed2005

Mutation scanning of the p53 tumor suppressor gene in renal and liver transplant patients in Hungary.

INTRODUCTION: The increased incidence of malignancies among transplanted patients is well known. Abnormal function of the p53 tumor suppressor gene has been reported in more than half of all tumors. The aim of our study was to detect point mutations of p53 gene in transplanted patients because the presence of mutations may be a predictive factor for tumor development. An earlier diagnosis can help to develop new strategies for immunosuppressive therapies. METHODS: Three point mutations were chosen based on the literature: exon5-codon175, exon7-codon248, exon8-codon273. Genomic DNA from the plasma of 60 liver, 362 renal transplants, and 45 nontransplanted patients with different tumors and 20 suspected healthy patients were analyzed with a real-time PCR method using the Roche LightCycler. The mutations were evaluated by melting curve analysis. RESULTS: We elaborated a special protocol for scanning the above mentioned p53 point mutations, which were proved by sequencing as well. Among 487 patients, 486 showed a wild-type genotype. The only patient carrying a mutation at codon 273 (heterozygous) was a liver transplant patient, who developed pancreas carcinoma and had already died. CONCLUSION: Our data suggest that mutations of the targeted codons in leukocyte DNA seem to be rare, but a mutation could be lethal. The evaluated three point mutations of p53 gene were not predictive for tumor development.

Base Sequence↗

PubMed2004

Measurement of NF-kappaB in normal and reconstructed human skin in vitro.

The survival of grafted donor skin for the treatment of burn injuries depends on several factors including wound bed vascularisation and the intensity of acute inflammation shortly after injury. However, acceptance rates approximate 50% at best and therefore a clinical need exists for improvement. The aim of the study was to develop a method for assessing the inflammatory response of cells in skin tissue based on activation of the NF-kappa B (NF-kappaB) transcription factor complex, thereby providing a basis for analysing the inflammatory component and anti-inflammatory strategies for tissue-engineered treatments. We have extended a standard method of measuring NF-kappaB in monolayer cultures that relies on determining translocation of the p65 subunit from the cytoplasm to the nucleus. Normal human skin and tissue engineered skin was analysed using an immunofluorescence microscopy technique, that revealed base line NF-kappaB activation in the epidermis and dermis were different. It was possible to determine the activation of NF-kappaB in skin tissue, enabling correlation that NF-kappaB measurement is a sensitive indicator of cellular responses in 3-D tissue. The approach will provide a basis for early responses of skin cells in determining the efficacy of anti-inflammatory delivery via tissue-engineered scaffolds for burn injuries.

Dermis↗

PubMed2003

Anti-inflammatory and anti-invasive effects of alpha-melanocyte-stimulating hormone in human melanoma cells.

Alpha-melanocyte stimulating hormone (alpha-MSH) is known to have pleiotrophic functions including pigmentary, anti-inflammatory, antipyretic and immunoregulatory roles in the mammalian body. It is also reported to influence melanoma invasion with levels of alpha-, beta- and gamma-MSH correlated clinically with malignant melanoma development, but other studies suggest alpha-MSH acts to retard invasion. In the present study, we investigated the action of alpha-MSH on three human melanoma cell lines (HBL, A375-SM and C8161) differing in metastatic potential. alpha-melanocyte-simulating hormone reduced invasion through fibronectin and also through a human reconstructed skin composite model for the HBL line, and inhibited proinflammatory cytokine-stimulated activation of the NF-kappaB transcription factor. However, A375-SM and C8161 cells did not respond to alpha-MSH. Immunofluorescent microscopy and Western blotting identified melanocortin-1 receptor (MC-1R) expression for all three lines and MC-2R on HBL and A375-SM lines. Receptor binding identified a similar affinity for alpha-MSH for all three lines with the highest number of binding sites on HBL cells. Only the HBL melanoma line demonstrated a detectable cyclic adenosine monophosphate (cAMP) response to alpha-MSH, although all three lines responded to acute alpha-MSH addition (+(-)-N(6)-(2-phenylisopropyl)-adenosine (PIA)) with an elevation in intracellular calcium. The nonresponsive lines displayed MC-1R polymorphisms (C8161, Arg (wt) 151/Cys 151; A375-SM, homozygous Cys 151), whereas the HBL line was wild type. Stable transfection of the C8161 line with wild-type MC-1R produced cells whose invasion was significantly inhibited by alpha-MSH. From this data, we conclude that alpha-MSH can reduce melanoma cell invasion and protect cells against proinflammatory cytokine attack in cells with the wild-type receptor (HBL).

Cytokines↗

PubMed2003

Development of a stable chemically defined surface for the culture of human keratinocytes under serum-free conditions for clinical use.

Within the field of tissue engineering there is a need to develop new approaches to achieve effective wound closure in patients with extensive skin loss or chronic ulcers. This article exploits the well-known interdependency of epithelial keratinocytes and stromal fibroblasts in conjunction with plasma surface technology. The aim was to produce a chemically defined surface, which with the aid of a feeder layer of lethally irradiated dermal fibroblasts would improve the attachment and proliferation of the keratinocyte cell from which subconfluent cells can be transferred to wound bed models. Plasma copolymers of acrylic acid/octa-1,7-diene have been prepared and characterized by X-ray photoelectron spectroscopy. The fibroblasts and keratinocytes were cultured on plasma polymer-coated 24-well plates. Cell attachment and proliferation were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide-eluted stain assay (MTT-ESTA) and DNA assay. Attachment and proliferation of both cell types on plasma polymer surfaces were compared with tissue culture plastic and collagen I, plus a negative control of a pure hydrocarbon layer. A pure acrylic acid surface, fabricated at a power of 10 W and containing 9.2% carboxylate groups, was found to promote both fibroblast and keratinocyte attachment and proliferation and permit the serum-free coculture of keratinocytes and irradiated fibroblasts.

Acrylates↗

PubMed2003

Pathologic features of suicidal deaths caused by explosives.

Suicidal explosions that lack a terrorist background are only rarely encountered in the field of forensic pathology. The investigation of explosion-related fatalities can be a substantial challenge in medicolegal casework. Determining whether the manner of death is suicide, homicide, or accident in such cases can present an especially difficult task to the forensic pathologist. This study considers the pathologic features of suicidal deaths caused by explosives without a terrorist background. The modus operandi of the decedents reflected familiarity and proficiency, or at least a degree of specialized knowledge, with the construction and use of explosive devices. All explosions were set off in confined spaces. The injury patterns consisted of a combination of primary blast injuries (e.g. decapitation, traumatic amputation of limbs, gross lacerations of the body surface, blast injuries of gas-containing and hollow organs), secondary blast injuries (e.g. splinter-induced penetrating trauma), tertiary blast injuries (e.g. abrasions and contusions), and burn injuries (mostly of the flash type). The previously described symmetric distribution pattern of injuries in suicidal explosions was apparent only to a certain degree in the present series. Our observation of superficially sharp-edged wound margins with bridging in the depths of the lesion in blast-induced lacerations of the skin should deserve further attention in forthcoming cases of explosion-related fatalities because this finding is a diagnostic possibility that may support the theory of an explosion-related fatality under special circumstances, e.g. when the body has been dumped away from the place of death. Because a terrorist attack may be initially suspected in each case of suicide involving explosives, the importance of a joint inquiry based on expertise from police investigators, bomb experts, and forensic pathologists is evident.

Adult↗

PubMed2003

Prognostic value of FDG-PET in malignant lymphoma.

Lymphomas have represented an indication for nuclear medicine investigations for 30 years. Gallium-67 scintigraphy has been shown to be a valuable complementary method in Hodgkin's disease and non-Hodgkin lymphoma for detecting viable residual lesions after chemotherapy and for diagnosis of a relapse. Thallium-201 is of interest in differentiating cerebral lymphomas from infectious lesions in AIDS patients but less useful in extra-cerebral lymphomas. PET with fluorine-18-FDG is more accurate than 67Ga in lymphoma. In patients with a positive PET scan after chemotherapy an early relapse occurs in up to 100%, while more than 80% of patients with a negative PET will have a long-term remission. Most studies show that FDG-PET is significantly correlated with patient outcome whereas there is much weaker or even no correlation for CT. The main reason is that PET is not bound to morphological criteria like lymph node size while CT is often not able to differentiate between residual tumour and post-therapeutic fibrosis. Therefore, based on a considerable number of clinical studies, FDG-PET gains increasing significance for staging, restaging and therapy monitoring in malignant lymphomas.

Citrates↗

PubMed2001

Progesterone receptors as neuroendocrine integrators.

Intracellular progesterone receptors (PRs) are ligand-inducible transcription factors that mediate the majority of the effects of progesterone (P) on neuroendocrine functions. During the past decade, evidence has accumulated which suggest that PRs can also be activated independently of P, by signals propagated through membrane-bound receptors to the interior of cells. The activation of PRs by this type of "cross-talk" mechanism has been implicated in the physiological regulation of several important neuroendocrine processes, including estrous behavior and periovulatory hormone secretions. We review evidence that both ligand-dependent and ligand-independent activation of PRs occurs in central neurons and in anterior pituitary cells and that the convergence and summation of these signals at the PR serves to integrate neural and endocrine signals which direct several critically important neuroendocrine processes. An integrative function for PRs is reviewed in several physiological contexts, including the display of lordosis behavior in female rodents, the neurosecretion of gonadotropin-releasing hormone surges, secretion of preovulatory gonadotropin surges, and release of periovulatory follicle stimulating hormone surges. The weight of evidence indicates that cross talk at the intracellular PR is an essential component of the integrative mechanisms that direct each of these neuroendocrine events. The recurrence of PR's integrative actions in several different physiological contexts suggests that other intracellular steroid receptors similarly function as integrators of neural and endocrine signals in other neuroendocrine processes.

Animals↗

PubMed2001

[Death in an unusual body position].

Death in head-down position is rare and uncommon. The mechanism of death in such cases is rather complicated. The authors present a case of unknown man who was found dead in a wood, hanging in head-down position fixed at the branchfork of a tree by his right foot. The authors analyse the possible mechanisms of death in so-called postural asphyxia.

Accidents↗

PubMed2000

Progesterone receptor A and B messenger ribonucleic acid levels in the anterior pituitary of rats are regulated by estrogen.

In target tissues of most mammalian and avian species, progesterone receptors (PR) are expressed as structurally related, but functionally distinct, isoforms A and B, and they are regulated by estrogen (E) as well as by their cognate ligand, progesterone (P(4)). The objectives of the present work were to identify mRNA expression for the A and B isoforms of PR in the anterior pituitary of the rat, to examine its regulation by gonadal steroids, and to compare this regulation with that in the primary target organ, the uterus. Messenger RNAs for the PR isoforms, determined by two separate reverse transcription-polymerase chain reaction protocols, one that detects PR A and PR B equally and the other specific for PR B, were identified in anterior pituitary of female and male rats. In anterior pituitary of cycling female rats, steady-state mRNA levels for both PR A+B and PR B were highest at 0900 h on proestrus, declined rapidly to nadir values at 0900 h on metestrus (PR A+B) or 0900 h on estrus (PR B), and remained below proestrous values through 2100 h on diestrus. Administration of E to intact proestrous female rats caused significant increases in mRNA for both PR A+B and PR B on estrus and metestrus. Blockade of P(4) action by administration of the antiprogestins RU-486 and ZK-98299 on proestrus had no effect on PR mRNA levels on the morning of estrus. Ovariectomy two and ten days after surgery markedly reduced mRNA levels for both PR A+B and PR B. Whereas treatment of 10-day-ovariectomized rats with E led to marked induction of mRNA for PR A+B and PR B two days later, treatment with P(4) one day after treatment had no effect on basal or E-stimulated PR mRNA. Regulation of PR mRNA expression in the pituitary differed from that in the uterus, in which P(4) treatment of ovariectomized rats antagonized the E-induced rise in mRNA for PR B, and antiprogestins increased mRNA for both isoforms. In addition to induction of PR mRNA in the pituitary of female rats by E in vivo, we also demonstrated induction by E in primary culture of anterior pituitary cells in vitro. We conclude that in the anterior pituitary of female rats, both the A and B isoforms of PR are expressed and regulated by E.

Animals↗

PubMed2000

Follistatin suppresses steroid-enhanced follicle-stimulating hormone release in vitro in rats.

Previous in vitro and in vivo studies from our laboratory showed that progesterone (P(4)), corticosterone (B), and testosterone (T) increase intracellular content and release of FSH in the anterior pituitary. Activin (Act) and inhibin (Inh) are structurally related proteins with antagonistic actions, as Act stimulates and Inh inhibits FSH secretion from the anterior pituitary. Together with follistatin (FS), a protein that bioneutralizes Act, they form an autocrine-paracrine loop in the anterior pituitary that tightly regulates FSH secretion. The objective of the present study was to test the hypothesis that P(4), B, and T modulate this autocrine-paracrine loop to favor increased FSH secretion. If Act were to mediate steroid-induced FSH release, FS would be expected to block these effects. To test this interaction, cell cultures were prepared from anterior pituitaries of male and female rats, and treated with Act, B, P(4), or T in the absence or presence of FS. Act, B, P(4), and T increased FSH release; FS suppressed both basal and Act- and steroid-stimulated FSH release to approximately 50% below basal levels. Cell cultures from anterior pituitary of female rats were used to compare the interaction of incremental concentrations of FS on dose-related Act- and P(4)-stimulated FSH release. With increasing concentrations of Act, the FS-induced suppression of FSH release was attenuated and eventually abolished; in contrast, maximally stimulatory concentrations of P(4) did not fully overcome the FS-induced suppression of FSH release. The effects of P(4), B, and Act in the presence and absence of estradiol on steady-state mRNA levels of FSHbeta, Actbeta(B), and FS were determined in primary pituitary cell cultures from metestrous female rats by reverse transcription-polymerase chain reaction. Whereas Act, P(4), B increased FSHbeta mRNA levels, only Act raised the level of FS mRNA, and neither steroid increased Actbeta(B) mRNA. The results support the hypothesis that endogenous Act is a common mediator of the action of P(4), B, and T in the rat primary anterior pituitary cell culture. We conclude that the stimulation of FSH release and intracellular content in the gonadotroph by P(4), B, and T is mediated, in part, by Act and involves modulation of a tightly regulated Act/FS autocrine-paracrine loop.

Activins↗

PubMed2000

Development of chick axial mesoderm: specification of prechordal mesoderm by anterior endoderm-derived TGFbeta family signalling.

Two populations of axial mesoderm cells can be recognised in the chick embryo, posterior notochord and anterior prechordal mesoderm. We have examined the cellular and molecular events that govern the specification of prechordal mesoderm. We report that notochord and prechordal mesoderm cells are intermingled and share expression of many markers as they initially extend out of Hensen's node. In vitro culture studies, together with in vivo grafting experiments, reveal that early extending axial mesoderm cells are labile and that their character may be defined subsequently through signals that derive from anterior endodermal tissues. Anterior endoderm elicits aspects of prechordal mesoderm identity in extending axial mesoderm by repressing notochord characteristics, briefly maintaining gsc expression and inducing BMP7 expression. Together these experiments suggest that, in vivo, signalling by anterior endoderm may determine the extent of prechordal mesoderm. The transforming growth factor (beta) (TGFbeta) superfamily members BMP2, BMP4, BMP7 and activin, all of which are transiently expressed in anterior endoderm mimic distinct aspects of its patterning actions. Together our results suggest that anterior endoderm-derived TGFbetas may specify prechordal mesoderm character in chick axial mesoderm.

Activins↗

PubMed1998

125I-labeled fasciculin 2: a new tool for quantitation of acetylcholinesterase densities at synaptic sites by EM-autoradiography.

Radio-iodinated fasciculin 2 (Fas2), a polypeptide anticholinesterase toxin from Mamba venom, was used as a new probe for localizing and quantifying acetylcholinesterase (AChE) at mouse neuromuscular junctions (NMJs) by quantitative electron microscope autoradiography. We demonstrate that 125I-Fas2 binds very specifically to the NMJs of mouse sternomastoid muscles, with very little binding to other regions in the muscles. Junctional AChE-site densities obtained from the autoradiograms were similar to those previously obtained for the same muscles using 3H-DFP. The use of 125I-Fas2 with EM-autoradiography is simpler and provides higher resolution and sensitivity, as well as considerably lower non-specific binding than previously attainable with 3H-DFP. The advantages and limitations of this procedure are discussed.

Acetylcholinesterase↗

PubMed1998

Hypothalamic-pituitary-gonadal axis in the mutant weaver mouse.

The weaver (wv) mutant mouse manifests severe locomotor defects, a deficiency in granule cells of the cerebellum, and cellular deficits in the midbrain dopaminergic system. The wv phenotype is associated with a missense mutation in the pore region of the G-protein-gated inwardly rectifying potassium channel, GIRK2. The homozygous male wv mouse is essentially infertile due to an inadequate level of sperm production. Females are fertile although they also manifest the neurological phenotype. Homozygotes of both sexes have reduced body weight. We have evaluated the hypothalamic-pituitary-gonadal axis in heterozygote and homozygote male and female wv mutants in comparison with wild-type controls. Testicular weight was significantly reduced in the homozygous males, due to degenerative changes of seminiferous epithelium. Serum and pituitary content of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin were normal in all groups, and the normal sex differences were noted (FSH and LH higher in males, prolactin higher in females). Pituitary growth hormone (GH) concentration was normal, with control and mutant males showing higher GH than females. Serum testosterone levels were normal in the mutants, as was testicular testosterone. Testicular alpha-inhibin content was mildly reduced, but high in proportion to testicular weight. The defect in spermatogenesis appeared predominantly in the postmeiotic stages. In situ hybridization was consistent with expression of some GIRK2 mRNA isoforms in seminiferous epithelium. There were no significant differences between genotypes in the levels of dopamine, dihydroxyphenylacetic acid, serotonin and 5-hydroxyindoleacetic acid in the mediobasal and preoptic hypothalamic regions. Homovanillic acid levels in these two areas were, however, reduced in wv homozygotes compared to wild-type animals. In the light of normal pituitary hormone levels, normal hypothalamic monoamine concentrations and normal sex differences in gonadotropins, we conclude that the infertility in the male homozygote wv mouse lies within the tubule and is probably a primary defect in the germ cells. The hormonal data suggest that Leydig cell function, and at least some aspects of Sertoli cell function, are normal in the mutant mice.

Animals↗

PubMed1998

Antiprogestins suppress basal and activin-stimulated follicle-stimulating hormone secretion in an estrogen-dependent manner.

Previous studies from our laboratory, demonstrating that suppression of serum FSH by RU486 requires a high estrogen (E) background, suggested that E-inducible progesterone receptors play a role in the regulation of FSH secretion. We demonstrated further that the type II antiprogestin RU486 and the type I antiprogestin ZK98299 both suppressed the elevated serum FSH and FSHbeta messenger RNA levels similarly on the evening of proestrus, but had divergent effects on the morning of estrus, when only RU486, but not ZK98299, lowered the elevated serum FSH level (secondary FSH surge). In the present work we used primary anterior pituitary cell culture to examine whether RU486 caused direct, E-dependent suppression of basal and recombinant human activin A (activin)-induced FSH secretion in the gonadotrope and to compare this direct effect, if any, with that of ZK98299. Primary cell cultures were prepared from anterior pituitaries collected from cycling female rats either on metestrous or proestrous morning and cultured in DMEM, supplemented with charcoal-stripped serum without or with 10 nM estradiol (E2) for 96 h; exposure to test agents occurred during the last 48 h of culture. FSH released into the medium and intracellular FSH content were determined by RIA. In cells from the anterior pituitary of metestrous rats cultured in E2-free medium, neither antiprogestin (10 nM) affected FSH release; in contrast, when cells were cultured in medium to which E2 had been added, both antiprogestins caused profound suppression of both basal and activin (10 ng/ml)-stimulated FSH release. In cell cultures from proestrous rats, both antiprogestins caused a slight, but significant, suppression of basal FSH release even in the absence of added E2; activin-stimulated FSH release, however, was not affected. Upon exposure of the cells from proestrous rats to E2, the antiprogestins potently suppressed both basal and activin-stimulated FSH secretion. Because the foregoing incubations were performed in culture medium devoid of progesterone (P4), the actions of the antiprogestins on FSH secretion were independent of the natural ligand. Addition of P4 (10 nM) to the cell cultures stimulated basal and activin-induced FSH release more in the presence than in the absence of E2. The FSH response to P4 was completely blocked by both antiprogestins in both the absence and presence of E2. Finally, both RU486 and ZK98299 blocked the stimulatory effect of corticosterone (1 microM) on FSH secretion. The observed effects of P4 and antiprogestins were specific for FSH secretion; LH secretion was not similarly suppressed by either antiprogestin, but was, in fact, stimulated by ZK98299 in E2-treated cells. We conclude that 1) E2-inducible progesterone receptors interact with activin-mediated signal transduction to regulate FSH secretion, and 2) unlike on the morning of estrus in vivo, RU486 and ZK98299 affect FSH secretion similarly in the gonadotrope in vitro.

Activins↗

PubMed1997

The antiprogestins RU486 and ZK98299 affect follicle-stimulating hormone secretion differentially on estrus, but not on proestrus.

Previous in vivo studies from our laboratory indicated that administration of the antiprogestin RU486 on proestrus suppresses both the preovulatory gonadotropin surges and the secondary FSH surge, suggesting a role for the progesterone receptor (PR) in the generation of these surges. The present study was designed to test the effects of another antiprogestin, ZK98299, which has been reported to block the PR through a mechanism different from that of RU486, on gonadotropin secretion in vivo. RU486 and ZK98299 (2 and 6 mg/kg) were administered s.c. at 1230 h on proestrus; uterine intraluminal fluid content, serum gonadotropins, and gonadotropin subunit messenger RNAs (mRNAs) were determined at 1830 h on proestrus and at 0900 h on estrus. At 1830 h on proestrus, both RU486 and ZK98299 at both doses caused equal suppression of the preovulatory FSH surge and FSHbeta mRNA. Both antiprogestins also equally attenuated the preovulatory LH surge at this time, with the higher doses causing greater suppression. In contrast, at 0900 h on estrus, the antiprogestins affected serum FSH differentially; only RU486 suppressed the secondary FSH surge despite the fact that both drugs prevented the release of uterine intraluminal fluid, confirming blockade of progesterone action at the level of the uterus. Neither drug had a significant effect on FSHbeta mRNA at 0900 h on estrus. ZK98299 at the higher dose caused a small, but significant, increase in serum LH. In a subsequent experiment, we compared the effects of RU486 and ZK98299 (6 mg/kg, s.c.), administered at 1230 h on proestrus, on serum FSH raised above the natural secondary FSH surge on the morning of estrus by passive immunization with an antiserum to inhibin-alpha (anti-I) at 1700 h on proestrus. Consistent with the results of the first experiment, both antiprogestins blocked the release of uterine intraluminal fluid, but only RU486 lowered serum FSH in both the normal sheep serum-treated controls and anti-I-treated rats; in contrast, ZK98299 actually increased serum FSH in the normal sheep serum-treated control animals. ZK98299 also increased FSHbeta mRNA in the control group; RU486, on the other hand, reduced FSHbeta mRNA only in the anti-I group. The results demonstrate unequivocally that whereas the effects of the two antiprogestins on serum FSH and FSHbeta mRNA are similar on proestrus, they are divergent on estrus. The data suggest that the functional state of the PR/transcriptional activation complex in the gonadotrope on the morning of estrus is different from that on the evening of proestrus.

Animals↗

PubMed1997

[Anesthesiologic characteristics of isolated hyperthermic liver perfusion with mitomycin C].

The isolated hyperthermic liver perfusion with mitomycin C presents a new technique of regional therapy for irresectable liver tumours. The advantage is a high local concentration of the antitumour agent with reduced systemic side-effects. Isolated hyperthermic liver perfusion is an extensive surgical procedure requiring a veno-venous bypass and a heart-lung machine. Disturbances affecting the base-acid hemostasis, the coagulation system and the cardiocirculatory function can occur. To date, there has been little experience with this technique. The intraoperative changes during the isolated hyperthermic liver perfusion in our series were similar to those seen during orthotopic liver transplantation. In contrast to orthotopic liver transplantation, heparin is given during the anhepatic phase. The reperfusion after isolated hyperthermic liver perfusion was not complicated by severe cardiocirculatory changes. A decrease in body temperature was not observed probably due to the absence of cold, potassium-rich perfusate flowing into the systemic circulation. Two patients developed signs of a reperfusion syndrome within the first hour after reperfusion (decrease in peripheral systemic resistance).

Adult↗

PubMed1996

Differential desensitization response of the neonatal and adult rat somatotroph to growth hormone-releasing hormone and phorbol ester.

Elevated levels of circulating growth hormone (GH) in the perinatal animal may be caused in part by relative resistance to the desensitizing effects of GH secretagogues. We compared the effects of 4-day exposure of primary pituitary cell cultures from adult male and 2-day-old rats to GH-releasing hormone (GHRH; 10 nM) or 12-O-tetradecanoyl-phorbol-13-acetate (TPA; 1 microM) on subsequent acute GH response to these secretagogues. Prolonged exposure to GHRH reduced subsequent GHRH-induced GH release from pituitary cells of both age groups, but the reduction in GH response was significantly less in neonates than adults. In addition, GH secretion from neonatal pituitaries rose progressively during each day of GHRH exposure, to reach levels almost 7 times basal; by contrast, GH secretion from adult pituitaries increased only transiently and then declined. Prolonged exposure to TPA reduced the subsequent GH response to TPA equally in neonates and adults, but differentially affected the GH response to GHRH; TPA exposure reduced the GH response to GHRH in neonates, but not in adults. These data suggest a fundamental difference between the GH regulatory processes of neonatal and adult pituitaries. The ability of the somatotroph to exhibit attenuated GH response on exposure to secretagogue is developmentally regulated, and relative resistance of the immature somatotroph to homologous desensitization by GHRH may contribute to elevated serum GH levels during the perinatal period.

Animals↗

PubMed1996

Mechanism of the inhibitory action of RU486 on the secondary follicle-stimulating hormone surge.

Recent evidence utilizing RU486 has implicated progesterone (P) and glucocorticoids, in addition to a drop in serum inhibin, in the development of the secondary FSH surge on the morning of estrus. To assess the role of these steroids, we treated proestrous female rats with the antiprogestin/antiglucocorticoid RU486 (6 mg/kg sc) at 1230 h, and with dexamethasone (dex; 8.4 or 16.2 mg/kg sc), or with the steroid biosynthesis inhibitor aminoglutethimide (AG; 150 mg/kg ip) at 1030 h, alone or in combination with RU486. The effects of these treatments on uterine ballooning and intraluminal fluid content (an index of P action), ovulation, and serum levels of P, corticosterone (B), FSH, LH, and inhibin-alpha at 1830 h proestrus and 0900 h estrus were examined. In accord with previous work from our laboratory, RU 486 caused uterine intraluminal fluid retention on the morning of estrus and significantly suppressed the preovulatory surges of both FSH and LH, and the secondary surge of FSH without affecting the fall in inhibin-alpha. Treatment with dex alone raised serum FSH at both 1830 h proestrus and 0900 h estrus, coincident with suppression of serum inhibin-alpha. When administered in combination with RU486, dex partially reversed the increased uterine intraluminal fluid retention at 0900 h estrus, but did not modify the inhibitory effect of RU486 on the primary gonadotropin surges or the secondary surge of FSH. AG alone significantly suppressed serum P, B, and gonadotropins (LH to a greater extent than FSH) at 1830 h proestrus and blocked ovulation and uterine intraluminal fluid release at 0900 h estrus; it did not, however, suppress the secondary FSH surge or prevent the fall in serum inhibin-alpha. When administered 2 h before RU486, AG did not prevent the RU486-induced inhibition of the primary gonadotropin surges or the secondary FSH surge. We conclude from these results that development of the secondary FSH surge does not require P or glucocorticoid action and that RU486 suppression of the secondary FSH surge does not involve blockade of binding of these steroids to their receptors. Our data are compatible with ligand-independent activation of the P receptor, susceptible to blockade by RU486, as the mechanism underlying the enhanced secretion of FSH from the gonadotrope on the morning of estrus.

Aminoglutethimide↗