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Biomedical subjects

Peter J Fuller

Publications and source records attributed to Peter J Fuller.

At least 19 recordsLinked to original sources
PubMed2007

A direct effect of aldosterone on endothelin-1 gene expression in vivo.

Aldosterone regulates sodium reabsorption in epithelial tissues such as the kidney and colon, via a pathway involving the activation of intracellular mineralocorticoid receptors (MR), induction of specific target genes, and a subsequent increase in sodium channel activity. Characterized aldosterone target genes in epithelia include the serum and glucocorticoid-regulated kinase 1 and the corticosteroid hormone-induced factor. Endothelin-1 (ET-1) is a potent vasoconstrictor that alters both sodium transport and hydrogen ion secretion in the kidney. Recent studies in a mouse medullary collecting duct cell line and rat A-10 smooth muscle cells have demonstrated an acute response of ET-1 gene expression to aldosterone. In the present study, we have investigated the ET-1 gene in vivo as a potential direct aldosterone-regulated target gene in the kidney and colon. Adrenalectomized rats given a single dose of aldosterone were found to have a 2-fold increase in ET-1 mRNA levels in the kidney and colon after 1 h. No significant changes in mRNA levels were detected for the related isoforms ET-2 or ET-3. Cotreatment with aldosterone and potassium canrenoate, a MR antagonist, blocked induction of ET-1 mRNA, suggesting that induction was mediated via the MR. In a time course study, ET-1 mRNA levels were induced rapidly by aldosterone, with levels of ET-1 mRNA maximally increased 2- and 2.5-fold after 1 h in the kidney and colon, respectively. These results suggest that ET-1 is a direct aldosterone gene target in the kidney and colon and may play an important role in aldosterone-regulated ion homeostasis.

Aldosterone↗
PubMed2006

Gene expression in the adapting small bowel after massive small bowel resection.

BACKGROUND: Intestinal adaptation occurs in the residual bowel following the loss or resection of a proportion of the small bowel. The purpose of the adaptive response is to return absorptive and digestive properties to near normal levels. This study employed a rat model of massive small bowel resection (MSBR) to study the adaptive response in the residual terminal ileum and the jejunum. The time points were chosen to reflect changes in gene expression early on in the response, because these are the genes that alter to initiate and maximize the response observed during adaptation. METHODS: Sprague Dawley rats underwent an 80% resection. Differential display polymerase chain reaction (DD-PCR) analysis was performed on mRNA extracted from the remnant ileum and jejunum 0, 1, 2, 4, and 7 days post-MSBR. RESULTS: DD-PCR identified 11 genes that were possibly regulated following MSBR. Genes confirmed to be regulated were 16S ribosomal RNA, lymphocyte antigen 6 (LY6)-like molecule, Krüppel-like factor-3 (KLF-3), G-protein-binding protein (CRFG), system A transporter 2 (SAT2), and an intestine-specific gene (similar to mKIAA0493). CONCLUSIONS: DD-PCR analysis showed regulation of a number of genes not previously known to be involved in adaptation after MSBR or previously characterized in the intestine. These genes may be important in bringing about the complement of changes seen during the adaptive response.

Adaptation, Physiological↗
PubMed2006

The role of the glucocorticoid receptor in mineralocorticoid/salt-mediated cardiac fibrosis.

The pathophysiological consequences of excess mineralocorticoid for salt status include hypertension, vascular inflammation, and cardiac fibrosis. Mineralocorticoid receptor (MR) blockade can both prevent and reverse established inflammation and fibrosis due to exogenous mineralocorticoids or endogenous glucocorticoid activation of the MR. Glucocorticoids also exert potent antiinflammatory effects via glucocorticoid receptors (GR) in the vascular wall. We propose that GR signaling may ameliorate mineralocorticoid/salt-induced vascular inflammation and fibrosis in the mineralocorticoid/salt model. In the present study, the role of GR in the mineralocorticoid/salt model was explored in uninephrectomized rats that were maintained on 0.9% saline solution to drink and treated as follows: control (CON), no further treatment; deoxycorticosterone (DOC; 20 mg/wk) for 4 wk (DOC4); DOC for 8 wk (DOC8); DOC for 8 wk plus the GR antagonist RU486 (2 mg/d) wk 5-8 (DOC8/RU486); and DOC for 8 wk plus RU486 and the MR antagonist eplerenone (EPL; 50 mg/kg.d) for wk 5-8 (DOC8/RU486+EPL). DOC treatment significantly increased systolic blood pressure, cardiac fibrosis, inflammation (ED-1-positive macrophages and osteopontin), and mRNA for markers of oxidative stress (p22phox, gp91phox, and NAD(P)H-4). GR blockade reduced the DOC-mediated increase in systolic blood pressure and the number of infiltrating ED-1-positive macrophages but had no effect on fibrosis, oxidative stress, or osteopontin mRNA levels. EPL reversed DOC-induced pathology in the absence or presence of GR blockade. Thus, blocking agonist activity at the GR neither enhances nor attenuates the fibrotic response, although it may modulate systolic blood pressure and macrophage recruitment in the mineralocorticoid/salt model.

Animals↗
PubMed2006

Mineralocorticoid receptor blockade but not steroid withdrawal reverses renal fibrosis in deoxycorticosterone/salt rats.

The pathophysiologic effects of nonepithelial mineralocorticoid receptor (MR) activation include vascular inflammation followed by renal and cardiac remodeling in experimental animals. We have recently shown that MR blockade can reverse established cardiac fibrosis and vascular inflammation; the present study explores whether a similar protection is seen in renal tissue. Rats were uninephrectomized and maintained on 0.9% NaCl solution to drink and treated as follows: control, vehicle; deoxycorticosterone (DOC), 20 mg/wk sc for 4 wk and then killed; DOC for 8 wk; DOC for 4 wk and no steroid for wk 5-8; DOC for 8 wk and eplerenone 100 mg/kg.d in the food for wk 5-8. DOC increased renal collagen at 4 and 8 wk; rats given DOC for 4 wk and killed at 8 wk showed levels of fibrosis identical with those killed at 4 wk, whereas rats given DOC for 8 wk plus eplerenone for wk 5-8 were indistinguishable from control. The inflammatory markers ED-1, osteopontin, and cyclooxygenase-2 remained significantly elevated despite the withdrawal of DOC (DOC404), whereas eplerenone restored cyclooxygenase-2 expression (but not that of ED-1 or osteopontin) to control levels. In addition, markers of oxidative stress and TGFbeta were determined. We hypothesize that continuing tubular inflammation and fibrosis despite DOC withdrawal indicates that the renal tissue may reflect MR activation in the context of tissue damage.

Animals↗
PubMed2006

Mammalian K-ras2 is a corticosteroid-induced gene in vivo.

Aldosterone acts via the mineralocorticoid receptor to regulate gene expression. A number of aldosterone-induced genes have been characterized in the distal colon and/or the distal nephron. Using the Xenopus kidney-derived A6 cell line, the K-ras transcript of the K-ras gene was identified as aldosterone induced, with a role in epithelial sodium transport. This study sought to establish whether K-ras expression is also increased in mammalian epithelia in vivo in response to aldosterone. RNA was extracted from the kidney and distal colon of rats treated with aldosterone or dexamethasone. Northern blot analysis and real-time RT-PCR were performed using probes and primers specific for the K-rasA isoform and for total K-ras. The expression of both total K-ras and of the A isoform is induced in the distal colon by aldosterone and by dexamethasone. Given the relative abundances of the two isoforms, this would appear to indicate induction of both isoforms. The time course of the response is consistent with a primary transcriptional response. In contrast to the documented up-regulation in the amphibian kidney, we did not observe regulation by corticosteroids in the kidney. However, regulation in a subpopulation of cells cannot be excluded.

Adrenal Cortex Hormones↗
PubMed2005

Mechanisms of mineralocorticoid action.

Sodium transport in epithelial tissues is regulated by the physiological mineralocorticoid aldosterone. The response to aldosterone is mediated by the mineralocorticoid receptor (MR), for which the crystal structure of the ligand-binding domain has recently been established. The classical mode of action for this receptor involves the regulation of gene transcription. Several genes have now been shown to be regulated by aldosterone in epithelial tissues. Of these, the best characterized is serum- and glucocorticoid-regulated kinase, which increases sodium influx through the epithelial sodium channel. Turnover of these channels in the cell membrane is mediated by Nedd4-2, a ubiquitin protein ligase; serum- and glucocorticoid-regulated kinase interacts with and phosphorylates Nedd4-2, thereby rendering it unable to bind the sodium channels. In nonepithelial tissues, particularly the cardiovascular system, aldosterone also has direct effects, activating an inflammatory cascade, leading to cardiac fibrosis. A critical role for the MR in cardiovascular disease has now been demonstrated by the beneficial response to MR blockade in 2 large clinical trials in patients with cardiac failure. It is these nonepithelial actions of MR activation that need to be exploited for the development of antagonists that target the cardiovascular system while avoiding the undesirable side effects of renal MR blockade.

Aldosterone↗
PubMed2005

Expression of steroid receptor coactivators in cultured cells from paired myometrial and fibroid tissues.

OBJECTIVES: Fibroid tumor growth in the myometrium appears to be regulated by estrogens but the role of estrogen receptor (ER) coregulators, such as the steroid receptor coactivator (SRC) family members, in fibroid growth is currently unknown. The aims of this study were to compare the expression of the SRC-1, SRC-2, and SRC-3 coactivators between fibroids and normal myometrium in pure populations of cultured smooth muscle cells (SMC) and microvascular endothelial cells (MEC), and also between both cell types, and to identify any relationship between the SRC expression profiles and the known ER status of the SMC and MEC samples examined in this study. METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR) coupled with Southern blot analysis was used to derive a semiquantitative estimate of the relative levels of SRC-1, SRC-2, and SRC-3 expression in pure populations of SMC (>98% alpha-smooth muscle actin [SMA](+)) and MEC (>99% CD31(+)) isolated and cultured from eight samples of paired human myometrial and fibroid tissue. RESULTS: The mean levels of SRC-1, SRC-2, and SRC-3 were each similar in normal myometrium compared to fibroids for SMC and also for MEC. However, SRC-1, SRC-2, and SRC-3 levels were each significantly higher in SMC compared to MEC from both myometrial and fibroid samples, although for SRC-3 there was a trend for higher levels in myometrial samples that did not reach significance. While all SMC samples expressed ERalpha and high coactivator levels, there does not appear to be a relationship between coactivator expression levels and the presence or absence of ERalpha in MEC samples. CONCLUSION: Coactivators may be more important in ERalpha-mediated growth of SMC than for MEC. Although the SRC family members are likely to play a role in the response of fibroid SMC to estrogen, via ERalpha, changes in their levels do not appear to contribute to the increased sensitivity of fibroid SMC to estrogen.

Acetyltransferases↗
PubMed2005

Seladin-1/DHCR24 expression in normal ovary, ovarian epithelial and granulosa tumours.

OBJECTIVE: The human DIMINUTO/DWARF1 homolog seladin-1/DHCR24 has been recently reported to be up-regulated in adrenocortical adenomas. Seladin-1 expression has been reported in the normal ovary. Granulosa cell tumours of the ovary (GCT) as with adrenocortical adenomas arise from a steroidogenic tissue, respond to pituitary hormone stimulation and synthesize steroid hormones. DESIGN: To test the hypothesis that seladin-1 may also have a role in the pathogenesis of GCT, we determined the expression of seladin-1 in a cohort of GCT and in mucinous and serous cystadenocarcinomas and in normal ovary. MEASUREMENTS: Expression was determined by RT-PCR using gene specific primers and probes combined with Southern blot analysis of the PCR products. RESULTS: Seladin-1 expression was identified in the normal ovary, mucinous cystadenocarcinomas and serous cystadenocarcinomas of the ovary whereas no expression was observed in the GCT. CONCLUSIONS: Based on our results, seladin-1 is not expressed in the granulosa cells or at least not in those that give rise to GCT.

Adolescent↗
PubMed2004

Transrepression of estrogen receptor beta signaling by nuclear factor-kappab in ovarian granulosa cells.

Estrogen receptor (ER) beta is the predominant ER in granulosa cells of the ovary. ERbeta is expressed at high levels in granulosa cell tumors (GCT) and in the human GCT-derived cell lines, COV434 and KGN. To gain insight into ERbeta function in granulosa cells and in GCT, we have used the COV434 and KGN cell lines. Although the cells bind estradiol (E2), transcriptional activation of a transfected estrogen-responsive reporter vector construct (ERE2-luc) by E2 was not observed. Transactivation was also not observed with cotransfected ERalpha or beta. This transcriptional resistance is specific to steroid receptor transactivation; reporter plasmids that are activated by the transcription factors activator protein 1 (AP-1) and nuclear factor kappaB (NF-kappaB) demonstrate both constitutive and inducible transactivation. AP-1 and NF-kappaB are known to cause transrepression of both ERalpha- and glucocorticoid receptor-mediated transcription. We therefore examined the possibility that activation of these pathways was responsible for the lack of a response to estrogen by using inhibitors of AP-1 or NF-kappaB. The AP-1 inhibitors alone had no effect, whereas inhibition of NF-kappaB signaling allowed a 3- to 4-fold E2-mediated induction of ERE2-luc. This response was both ligand and ER dependent. Repression of ERbeta signaling by NF-kappaB has not previously been reported. Recent evidence suggests that ERbeta may function to promote differentiation. The inhibition of ERbeta in combination with the antiapoptotic properties of NF-kappaB may therefore contribute to pathogenesis of GCT.

Animals↗
PubMed2004

Mineralocorticoid receptor binding, structure and function.

The isolation of aldosterone 50 years ago was a critical first step in elucidating the mechanism by which corticosteroids regulate electrolyte homeostasis. The broad principles of this mechanism involving an intracellular receptor acting on specific genes to induce the expression/repression of aldosterone-induced proteins (AIP) were established 30 years ago. The cloning of the mineralocorticoid receptor (MR) has enabled studies of the subcellular mechanisms of aldosterone action, including the molecular dissection of structure-function relationships in the receptor. We have exploited the close structural and functional similarity of the MR with the glucocorticoid receptor to identify the regions in the MR that confer ligand-binding specificity. The critical region is located, not as might be expected in the ligand-binding pocket but rather on the surface of the molecule. These studies have been extended to an analysis of the interactions between the N-terminal and ligand-binding domains of the MR. In the last decade, AIP have been identified; the regulation of the genes encoding these AIP are discussed.

Aldosterone↗
PubMed2004

Signalling pathways in the molecular pathogenesis of ovarian granulosa cell tumours.

Granulosa cell tumours represent approximately 5% of all malignant ovarian tumours. They exhibit many morphological, biochemical and hormonal features of proliferating normal preovulatory granulosa cells, including both estrogen and inhibin biosynthesis. The aetiology of these tumours remains unclear, although many recent studies and several animal models have focused on signalling pathways that are known to be important in the regulation of granulosa cell proliferation. Here, we discuss the information currently available about the possible molecular changes that might lead to neoplastic transformation in human granulosa cells. The identification of these changes might lead not only to improved diagnosis, management and treatment of granulosa cell tumours, but could also offer new insights into normal granulosa cell biology.

Animals↗
PubMed2004

Cortisol resistance in the New World revisited.

Insights into the molecular basis of glucocorticoid action have been obtained from the analysis of cortisol resistance. The glucocorticoid receptor (GR) in both New World primates and guinea pigs has a decreased affinity, in vivo, for cortisol; this is achieved by two distinct mechanisms. In the New World primates recent studies have identified a key role for co-chaperones. The amino acids responsible for cortisol resistance in the guinea pig GR lie not in the ligand-binding pocket but on the surface of the receptor. We hypothesize that this region might be the site of interaction between the co-chaperones and the GR, and hence that the resistance occurs through the same mechanism, albeit from opposite sides.

Animals↗
PubMed2004

Expression status and mutational analysis of the ras and B-raf genes in ovarian granulosa cell and epithelial tumors.

OBJECTIVES: The molecular pathogenesis of granulosa cell tumors of the ovary (GCT) is not understood. Activating mutations in the K-, N-, and H-ras protooncogenes have been identified in a wide range of human cancers, including ovarian epithelial tumors. Furthermore, an apparent association has recently been reported between the presence of ras and B-raf mutations in the same cancer types. Activation of the ras/raf pathway would be predicted to be tumorigenic in granulosa cells. METHODS: Gene expression patterns of the three ras and B-raf genes were determined in a panel of GCT and ovarian epithelial tumors, and in normal premenopausal ovaries. Expression was determined by RT-PCR using gene-specific primers combined with Southern blot analysis of the PCR products using gene-specific (32)P-labeled probes. Direct sequence analysis was used to screen for known activating mutations. RESULTS: Widespread expression of the four genes was observed in all tumor types examined. Compared to the normal ovaries, none of the genes was expressed at significantly higher levels in any of the tumor types examined. A heterozygous point mutation in codon 12 of the K-ras gene was found in five of the 10 mucinous tumors. No B-raf mutations were detected in the mucinous tumors. No mutations were detected in any of the genes in the cohort of GCT. CONCLUSION: These results suggest that neither overexpression nor activating mutations of the ras or B-raf genes are associated with the development of GCT.

Adult↗
PubMed2004

Plasma GLP-2 levels and intestinal markers in the juvenile pig during intestinal adaptation: effects of different diet regimens.

Adaptation of the residual small bowel following resection is dependent on luminal and humoral factors. We aimed to establish if circulating levels of glucagon-like peptide (GLP-2) change under different dietary regimens following resection and to determine if there is a relationship between plasma GLP-2 levels and markers of intestinal adaptation. Four-week-old piglets underwent a 75% proximal small bowel resection (n = 31) or transection (n = 14). Postoperatively they received either pig chow (n = 14), nonpolymeric (elemental) infant formula (n = 7), or polymeric infant formula alone (n = 8) or supplemented either with fiber (n = 6) or with bovine colostrum protein concentrate (CPC; n = 10) for 8 weeks until sacrifice. Plasma GLP-2 levels were measured at weeks 0, 2, 4, and 8 postoperatively. In addition, end-stage parameters were studied at week 8 including weight gain, ileal villus height, crypt depth, and disaccharidase levels. Plasma GLP-2 levels were higher in resected animals compared to transected animals fed the same diet. Plasma GLP-2 levels were significantly increased in the colostrum protein isolate-supplemented animals following resection compared to all other diet groups. The increase in plasma GLP-2 (pM) was greatest in the first 2 weeks postresection (week 0, 15.5; week 2, 30.9), followed by a plateau at weeks 2 to 4 and a decrease in GLP-2 levels from week 4 to week 8. At week 8, no relationships were found between the plasma GLP-2 levels and the measurements of weight gain, villus height, lactase, sucrase, maltase, crypt depth, or villus/crypt ratio. Plasma GLP-2 levels increase in the first weeks following massive small intestinal resection. The increase in plasma GLP-2 levels was enhanced by supplementation of the diet with CPC. The changes in GLP-2 levels observed in this study may suggest that GLP-2 plays a role in the adaptive response in the intestine following resection in this preclinical model.

Adaptation, Physiological↗
PubMed2004

Colostrum protein concentrate enhances intestinal adaptation after massive small bowel resection in juvenile pigs.

OBJECTIVES: Short bowel syndrome (SBS) usually results from the surgical removal of a large segment of small intestine. Patient outcome depends on the extent of intestinal resection and adaptation of the remaining intestine. We evaluated the impact of colostrum protein concentrate (CPC) on intestinal adaptation after massive small bowel resection in a porcine model of infant SBS. METHODS: Four-week-old piglets underwent an approximate 75% small bowel resection (R, n = 23) or a control transection operation (C, n = 14). Postoperatively, animals from both groups received either pig chow (R = 6, C = 5), polymeric infant formula (R = 6, C = 3) or polymeric infant formula supplemented with CPC (R = 11, C = 6) for 8 weeks until sacrifice. Clinical outcome measures included weight gain and stool consistency. Morphologic measures were intestinal villus height and crypt depth. Functional outcome measure was mucosal disaccharidase activity. RESULTS: Resected animals fed polymeric infant formula alone had reduced weight gain compared with controls fed the same diet (P < 0.005). Despite massive small bowel resection, animals fed pig chow or polymeric infant formula supplemented with CPC grew at an equivalent rate to controls fed polymeric infant formula alone. Resected animals supplemented with CPC had increased villus length and crypt depth in the jejunum (P < 0.001) and ileum (P < 0.001) compared with resected animals fed either pig chow or polymeric infant formula alone. CONCLUSION: In an animal model of SBS, CPC supplementation of polymeric infant formula resulted in normal weight gain and features of enhanced morphologic adaptation.

Adaptation, Physiological↗
PubMed2004

Differences in the determinants of eplerenone, spironolactone and aldosterone binding to the mineralocorticoid receptor.

The importance of mineralocorticoid receptor (MR) antagonists in the treatment of cardiovascular disease has been emphasised by two recent clinical trials, one using spironolactone and the other using a new selective MR antagonist, namely eplerenone. Eplerenone has a very low affinity for the glucocorticoid receptor (GR). Determinants of binding specificity of eplerenone to the MR were investigated using chimeras created between the ligand-binding domains (LBD) of the MR and the GR. These chimeras had been used previously to investigate aldosterone and spironolactone binding specificity to the MR. Eplerenone competed strongly for [(3)H]-dexamethasone binding to a MR/GR chimera containing amino acids 804-874 of the MR and weakly to a chimera containing amino acids 672-803 of the MR. Within the 804-874 region, eplerenone competed for [(3)H]-dexamethasone binding to a chimera containing amino acids 820-844 of the MR, although the calculated affinity was approximately 10-fold lower than for binding to the full-length MR LBD. Similar results were obtained using another MR antagonist, namely spironolactone. Modelling of eplerenone binding to the MR LBD, based on the GR LBD crystal structure, suggests that amino acids 820-844 affect the overall shape of the ligand-binding pocket and that eplerenone acts as an MR antagonist because it fails to stabilize the active conformation of the receptor. In contrast with results with the MR antagonists eplerenone and spironolactone, amino acids 820-844 are sufficient in themselves to confer high-affinity aldosterone binding to the MR, suggesting that the binding determinants of the two antagonists are similar to each other but differ from those of aldosterone.

Aldosterone↗
PubMed2003

Endocrine disorders of sodium regulation. Role of adrenal steroids in genetic defects causing sodium loss or sodium retention.

Salt and water homoeostasis is tightly regulated by a variety of control mechanisms with the adrenal steroid hormone aldosterone playing a central role. Defects or disturbances in these systems lead to either salt loss, which is life threatening in the neonatal period, or sodium retention causing hypertension. Rapid and accurate diagnosis is required to avoid severe complications. During the last few years molecular genetic advances have been identified as the basic genetic defects for a number of clinical syndromes. This knowledge has considerably increased our understanding of the basic pathways involved in sodium and water homoeostasis and of the pathophysiology of these syndromes, particularly the hypertension. In this review we have summarized the biochemical, physiological and genetic basis for clinical syndromes presenting with salt loss and failure to thrive as well as the rare but important genetic syndromes causing sodium retention and hypertension. Early diagnosis and identification will help to prevent severe complications, but it has to be emphasized that the complicated cascade of aldosterone action is still relatively poorly understood. Further syndromes may exist which once identified will help to better understand the basic physiology of aldosterone action.

Adrenal Cortex Hormones↗