Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 308
Filtrar
1.
Am J Physiol Endocrinol Metab ; 315(4): E435-E445, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-29763375

RESUMO

The human (h) placental lactogenic hormone chorionic somatomammotropin (CS) is highly produced during pregnancy and acts as a metabolic adaptor in response to maternal insulin resistance. Maternal obesity can exacerbate this "resistance", and a >75% decrease in CS RNA levels was observed in term placentas from obese vs. lean women. The genes coding for hCS ( hCS-A and hCS-B) and placental growth hormone ( hGH-V) as well as the hCS-L pseudogene and pituitary growth hormone (GH) gene ( hGH-N) are located at a single locus on chromosome 17. Three remote hypersensitive sites (HS III-V) located >28 kb upstream of hGH-N as well as local hCS gene promoter and enhancer regions are implicated in hCS gene expression. A placenta-specific chromosomal architecture, including interaction between HS III-V and hCS but not hGH gene promoters, was detected in placentas from lean women (BMI <25 kg/m2) by using the chromosome conformation capture assay. This architecture was disrupted by pre-pregnancy maternal obesity (BMI >35 kg/m2), resulting in a predominant interaction between HS III and the hGH-N promoter, which was also observed in nonplacental tissues. This was accompanied by a decrease in hCS levels, which was consistent with reduced RNA polymerase II and CCAAT/enhancer-binding protein-ß association with individual hCS promoter and enhancer sequences, respectively. Thus, pre-pregnancy maternal obesity disrupts the placental hGH/CS gene locus chromosomal architecture. However, based on data from obese women who develop GDM, insulin treatment partially recapitulates the chromosomal architecture seen in lean women and positively affects hCS production, presumably facilitating prolactin receptor-related signaling by hCS.


Assuntos
Cromossomos Humanos/genética , Hormônio do Crescimento/genética , Hormônio do Crescimento Humano/genética , Obesidade/genética , Placenta/metabolismo , Hormônios Placentários/genética , Lactogênio Placentário/genética , Complicações na Gravidez/genética , Índice de Massa Corporal , Imunoprecipitação da Cromatina , Cromossomos Humanos/metabolismo , Feminino , Expressão Gênica , Hormônio do Crescimento/metabolismo , Hormônio do Crescimento Humano/metabolismo , Humanos , Immunoblotting , Resistência à Insulina , Obesidade/metabolismo , Hormônios Placentários/metabolismo , Lactogênio Placentário/metabolismo , Gravidez , Complicações na Gravidez/metabolismo , Regiões Promotoras Genéticas , Pseudogenes , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa
2.
Reprod Fertil Dev ; 29(3): 458-467, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28442058

RESUMO

Abnormal placental development is frequent in nuclear transfer (NT) pregnancies and is likely to be associated with altered epigenetic reprogramming. In the present study, fetal and placental measurements were taken on Day 60 of gestation in cows with pregnancies produced by AI, IVF and NT. Placentas were collected and subjected to histological evaluation, the expression of genes important in trophoblast differentiation and expression of the placental imprinted gene pleckstrin homology-like domain, family A, member 2 (PHLDA2), as well as chromatin immunoprecipitation (ChIP) for histone marks within the promoter of PHLDA2. Fewer binucleated cells were observed in NT cotyledons, followed by IVF and AI cotyledons (P<0.05). Expression of heart and neural crest derivatives expressed 1 (HAND1), placental lactogen (PL), pregnancy-associated glycoprotein 9 (PAG-9) and PHLDA2 was elevated in NT cotyledons compared with AI cotyledons. Expression of PHLDA2 was higher in IVF than AI samples (P<0.05). ChIP revealed an increase in the permissive mark dimethylation of lysine 4 on histone H3 (H3K4me2), surprisingly associated with the silent allele of PHLDA2, and a decrease in the inhibitory mark H3K9me2 in NT samples. Thus, genes critical for placental development were altered in NT placentas, including an imprinted gene. Allele-specific changes in the permissive histone mark in the PHLDA2 promoter indicate misregulation of imprinting in clones. Abnormal trophoblast differentiation could have resulted in lower numbers of binucleated cells following NT. These results suggest that the altered expression of imprinted genes associated with NT are also caused by changes in histone modifications.


Assuntos
Expressão Gênica , Código das Histonas , Histonas/metabolismo , Proteínas Nucleares/metabolismo , Técnicas de Transferência Nuclear/veterinária , Placenta/metabolismo , Alelos , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Bovinos , Feminino , Histonas/genética , Proteínas Nucleares/genética , Lactogênio Placentário/genética , Lactogênio Placentário/metabolismo , Placentação/fisiologia , Gravidez , Proteínas da Gravidez/genética , Proteínas da Gravidez/metabolismo , Trofoblastos/metabolismo
3.
Domest Anim Endocrinol ; 58: 84-89, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27743526

RESUMO

Leptin is involved in various reproductive processes in humans and rodents, including placental development and function. The specific ways that leptin influences placental development and function in cattle are poorly understood. This work was completed to explore how leptin regulates hormone, cytokine and metalloprotease transcript abundance, and cell proliferation in cultured bovine trophoblast cells. In the first set of studies, cells were cultured in the presence of graded recombinant bovine leptin concentrations (0, 10, 50, 250 ng/mL) for 6 or 24 h. Transcript profiles were examined from extracted RNA. Leptin supplementation did not affect abundance of the maternal recognition of pregnancy factor, interferon-tau (IFNT), but leptin increased (P < 0.05) abundance of chorionic somatomammotropin hormone 2 (CSH2; ie, placental lactogen) at both 6 and 24 h at each concentration tested. At 24 h, the greatest CSH2 abundance (P < 0.05) was detected in cells supplemented with 50 ng/mL leptin. Transcript abundance of the remodeling factor, metalloprotease 2 (MMP2), was greater (P < 0.05) in leptin-treated cells at 24 h but not at 6 h. The 24 h MMP2 response was greatest (P < 0.05) at 250 ng/mL. Transcript abundance for MMP9 was not altered by leptin treatment. In a separate set of studies, cell proliferation assays were completed. Leptin supplementation did not affect bovine trophoblast cell line proliferation at any dose tested. In conclusion, leptin supplementation did not affect bovine trophoblast cell proliferation or IFNT expression, but leptin increases CSH2 and MMP2 transcript abundance. Both of these factors are involved with peri-implantation and postimplantation placental development and function, and this implicates leptin as a potential mediator of early placental development and function in cattle.


Assuntos
Bovinos , Leptina/fisiologia , Trofoblastos/fisiologia , Animais , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/fisiologia , Células Cultivadas , Feminino , Expressão Gênica/efeitos dos fármacos , Interferon Tipo I , Leptina/administração & dosagem , Metaloproteinase 2 da Matriz/genética , Placenta/fisiologia , Lactogênio Placentário/genética , Gravidez , Proteínas da Gravidez , RNA Mensageiro/análise , Proteínas Recombinantes/administração & dosagem , Trofoblastos/citologia , Trofoblastos/efeitos dos fármacos
4.
Genesis ; 54(7): 389-97, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-27124574

RESUMO

Spermatogenesis is a complex and highly regulated process by which spermatogonial stem cells differentiate into spermatozoa. To better understand the molecular mechanisms of the process, the Cre/loxP system has been widely utilized for conditional gene knockout in mice. In this study, we generated a transgenic mouse line that expresses Cre recombinase under the control of the 2.5 kbp of the Prolactin family 3, subfamily b, member 1 (Prl3b1) gene promoter (Prl3b1-cre). Prl3b1 was initially reported to code for placental lactogen 2 (PL-2) protein in placenta along with increased expression toward the end of pregnancy. PL-2 was found to be expressed in germ cells in the testis, especially in spermatocytes. To analyze the specificity and efficiency of Cre recombinase activity in Prl3b1-cre mice, the mice were mated with reporter R26GRR mice, which express GFP ubiquitously before and tdsRed exclusively after Cre recombination. The systemic examination of Prl3b1-cre;R26GRR mice revealed that tdsRed-positive cells were detected only in the testis and epididymis. Fluorescence imaging of Prl3b1-cre;R26GRR testes suggested that Cre-mediated recombination took place in the germ cells with approximately 74% efficiency determined by in vitro fertilization. In conclusion, our results suggest that the Prl3b1-cre mice line provides a unique resource to understand testicular germ-cell development. genesis 54:389-397, 2016. © 2016 Wiley Periodicals, Inc.


Assuntos
Diferenciação Celular/genética , Proteínas Imediatamente Precoces/biossíntese , Proteínas Tirosina Fosfatases/biossíntese , Espermatogênese/genética , Espermatozoides/metabolismo , Animais , Regulação da Expressão Gênica no Desenvolvimento , Técnicas de Inativação de Genes , Células Germinativas/crescimento & desenvolvimento , Células Germinativas/metabolismo , Proteínas Imediatamente Precoces/genética , Masculino , Camundongos , Lactogênio Placentário/genética , Proteínas Tirosina Fosfatases/genética , Espermatozoides/crescimento & desenvolvimento , Células-Tronco/metabolismo , Testículo/crescimento & desenvolvimento , Testículo/metabolismo
5.
Neuro Endocrinol Lett ; 36(2): 136-42, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26071582

RESUMO

OBJECTIVES: Somatotroph adenomas secrete supraphysiological amounts of GH, causing acromegaly. We have previously shown epithelial splicing regulator 1 (ESRP1) to play a role in epithelial mesenchymal transition (EMT) progression in these adenomas and account for poor treatment response. We evaluated if the mRNA levels of the GH/CSH gene cluster in somatotroph adenomas are associated with an epithelial phenotype and response to SA treatment. METHODS: We investigated the associations between ESRP1 and the growth hormone/chorionic somatomammotropin (GH/CSH) gene cluster by RNA sequencing (RNAseq). CSH2 isoform 3 mRNA was further evaluated in 65 somatotroph adenomas and associations with disease severity and treatment response. RESULTS: mRNA for all genes in the GH/CSH cluster were expressed, however, only chorionic somatomammotropin 2/placental lactogen 2 (CSH2) displayed an alternative splicing pattern. CSH2 isoform 3 was associated with a dense granulation pattern and an epithelial phenotype with high levels of ESRP1 and E-cadherin expression. Further, CSH2 isoform 3 was associated with reduced serum GH and IGF-I levels after somatostatin analog treatment. CONCLUSIONS: Attenuated CSH2 isoform 3 was associated with mesenchymal phenotype and a blunted clinical response to somatostatin analog treatment in patients with acromegaly.


Assuntos
Processamento Alternativo/genética , Adenoma Hipofisário Secretor de Hormônio do Crescimento/genética , Neoplasias Hipofisárias/genética , Lactogênio Placentário/genética , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
6.
Pathol Res Pract ; 211(3): 226-34, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25499719

RESUMO

The objective of the present study was to evaluate the effect of the thyroid hormones in the gene transcription and immunohistochemical expression of hormonal and angiogenic factors in the placenta of rats. Seventy-two adult female rats were divided equally into propylthiouracil (PTU)-treated, thyroxine (T4)-treated, and control groups. The animals were sacrificed at 10, 14, and 19 days of gestation. We evaluated the immunohistochemical expression of VEGF and its receptor Flk-1. The gene transcription of VEGF, Flk-1, PGF, sFlt1, PL-1, and rPlf was evaluated in placental discs by real-time RT-PCR. The data were analyzed using a Student-Newman-Keuls (SNK) test. At day 10, T4-treated rats presented increased VEGF and PGF gene expression, while PTU-treated rats showed increased rPlf gene expression. Both groups showed reduced Flk-1 and PL-1 gene expression at day 10. At day 14, PTU-treated rats showed reduced VEGF, PGF, and rPlf gene expression. PTU-treated group showed reduced VEGF immunostaining in the placental labyrinth at 14 and 19 days of gestation but it showed increased VEGF immunostaining in the spongiotrophoblast layer at day 14. PTU-treated rats showed increased Flk-1 expression at 14 days of gestation. At days 14 and 19, T4-treated group showed increased PL-1 gene expression and reduced VEGF immunostaining. T4-treated rats also showed reduced Flk-1 and sFlt-1 expression at day 19. Both groups showed increased rPlf gene expression at day 19. In conclusion, rats treated with PTU and T4 have differential effects on the expression of factors involved in placental angiogenic and hormonal activity, and these effects are dependent on the gestational period.


Assuntos
Antitireóideos/farmacologia , Placenta/efeitos dos fármacos , Propiltiouracila/farmacologia , Tiroxina/farmacologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Animais , Feminino , Placenta/metabolismo , Fator de Crescimento Placentário , Lactogênio Placentário/genética , Lactogênio Placentário/metabolismo , Gravidez , Proteínas da Gravidez/genética , Proteínas da Gravidez/metabolismo , Ratos , Fator A de Crescimento do Endotélio Vascular/genética , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/genética , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genética
7.
Placenta ; 36(1): 97-9, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25468542

RESUMO

The objective of the present study was to evaluate the effects of different doses of T3 (10(-4) M, 10(-7) M, 10(-9) M) on the in vitro gene expression of Tpbp, Prl3b1, VEGF, PGF, PL-1, and INFy in mouse trophoblast cells by real-time RT-PCR. Doses of 10(-7) and 10(-9) M T3 increased the mRNA levels of Tpbp, Pl3b1, VEGF, PGF, INFy and PL-1. In contrast, the dose of 10(-4) M reduced the gene expression of PL-1 and VEGF. T3 affected the gene expression of differentiation, hormonal, immune and angiogenic factors in mouse trophoblast cells.


Assuntos
Expressão Gênica/efeitos dos fármacos , Tri-Iodotironina/farmacologia , Trofoblastos/efeitos dos fármacos , Animais , Relação Dose-Resposta a Droga , Interferon gama/genética , Interferon gama/metabolismo , Camundongos , Fator de Crescimento Placentário , Lactogênio Placentário/genética , Lactogênio Placentário/metabolismo , Proteínas da Gravidez/genética , Proteínas da Gravidez/metabolismo , Proteína B de Ligação a Transferrina/genética , Proteína B de Ligação a Transferrina/metabolismo , Trofoblastos/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo
8.
Nucleic Acids Res ; 42(8): 4906-21, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24561805

RESUMO

The robust and tissue-specific activation of the human growth hormone (hGH) gene cluster in the pituitary and placenta constitutes an informative model for analysis of gene regulation. The five-gene hGH cluster is regulated by two partially overlapping sets of DNase I hypersensitive sites (HSs) that constitute the pituitary (HSI, II, III and V) and placental (HSIII, IV, and V) locus control regions (LCRs). The single placenta-specific LCR component, HSIV, is located at -30 kb to the cluster. Here we generate a series of hGH/BAC transgenes specifically modified to identify structural features of the hGH locus required for its appropriate placental expression. We find that placental specificity is dependent on the overall multigene configuration of the cluster whereas the distance between the cluster and its LCR impacts the level of placental expression. We further observe that a major function of the placental hGH LCR is to insulate the transgene locus from site-of-integration effects. This insulation activity is linked to placenta-specific occupancy of the chromatin architectural protein, CTCF, at HSIV. These data reveal a remarkable combination of structural configurations and regulatory determinants that must work in concert to insure robust and tightly controlled expression from a complex multigene locus.


Assuntos
Hormônio do Crescimento Humano/genética , Elementos Isolantes , Família Multigênica , Lactogênio Placentário/genética , Proteínas Repressoras/metabolismo , Animais , Fator de Ligação a CCCTC , Desoxirribonuclease I , Feminino , Regulação da Expressão Gênica , Loci Gênicos , Humanos , Região de Controle de Locus Gênico , Camundongos , Camundongos Transgênicos , Especificidade de Órgãos , Placenta/metabolismo , Gravidez , Transcrição Genética
9.
PLoS One ; 9(1): e87325, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24475273

RESUMO

INTRODUCTION: Several studies reported that the pregnancy-specific hormone placental lactogen (hPL) is expressed at both mRNA and protein levels in breast cancer. The overall objective was to establish hPL, the product of the CSH1 and CSH2 genes, as a biomarker for breast cancer. METHODS: CSH expression was determined at the mRNA level in breast cancer cell lines (BCC) and primary carcinomas by real-time and conventional PCR and the products verified as CSH1 by sequencing. Expression of hPL protein was examined by western blots and immuno-histochemistry, using commercial and custom-made polyclonal and monoclonal antibodies. RESULTS: Variable levels of CSH mRNA were detected in several BCC, and in some primary tumors. We detected a protein, slightly larger than recombinant hPL by western blotting using several antibodies, leading us to postulate that it represents an hPL variant ('hPL'). Furthermore, some monoclonal antibodies detected 'hPL' by immunohistochemistry in breast carcinomas but not in normal breast. However, further examination revealed that these antibodies were non-specific, as efficient suppression of CSH mRNA by shRNA did not abolish the 'hPL' band. Custom-made monoclonal antibodies against recombinant hPL detected hPL of the correct size in placental lysate and hPL-overexpressing BCC, but not in unmodified cells or primary carcinomas. hPL protein was detected only when mRNA was increased several thousand fold. CONCLUSIONS: We call into question previous reports of hPL expression in breast cancer which relied on mRNA levels as surrogates for protein and/or used improperly validated antibodies to measure hPL protein levels. Our data suggests that an inhibitory mechanism(s) prevents translation of CSH mRNA in breast cancer when not highly expressed. The mechanism by which translation of CSH mRNA is inhibited is intriguing and should be further investigated.


Assuntos
Artefatos , Neoplasias da Mama/genética , Carcinoma/genética , Regulação Neoplásica da Expressão Gênica , Lactogênio Placentário/genética , RNA Mensageiro/genética , Anticorpos Monoclonais/química , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Carcinoma/diagnóstico , Carcinoma/metabolismo , Carcinoma/patologia , Linhagem Celular Tumoral , Feminino , Humanos , Placenta/química , Placenta/metabolismo , Lactogênio Placentário/deficiência , Gravidez , Biossíntese de Proteínas , RNA Mensageiro/metabolismo , Transdução de Sinais
10.
Prenat Diagn ; 34(4): 345-9, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24375464

RESUMO

OBJECTIVE: This study aimed to identify a set of predominantly placental (PP) mRNAs, which are associated with later-developing twin-to-twin transfusion syndrome (TTTS). METHOD: First, out of 50 PP mRNAs we previously reported, we select target mRNAs that are ordinarily detectable in maternal plasma. Plasma concentrations of these PP mRNAs were measured in monochorionic diamniotic twin (MCDA-T) pregnancies complicated by TTTS later (n = 11) and in uncomplicated MCDA-T pregnancies (n = 17). Finally, the diagnostic values of the PP mRNAs in plasma were evaluated. RESULTS: From 50 PP mRNAs, nine [human placental lactogen (hPL); pregnancy-specific glycoproteins 2 (PSG2); human pregnancy-specific glycoproteins 3 (PSG3); syncytin; syncytin 2; retinoic acid-induced 14; A disintegrin and metalloproteinase domain-containing protein 12 (ADAM12); chorionic glycoprotein hormones, alpha polypeptide; and chorionic glycoprotein hormones, and beta polypeptide] were selected as target mRNAs. Changes in six PP mRNAs [increased hPL, PSG2, and PSG3 and decreased syncytin, syncytin2, and ADAM12] in maternal plasma were detected in MCDA-T pregnant women who subsequently developed TTTS. Finally, mRNA signatures gave elevated AUCs (hPL/PSG2: 0.8717; hPL/PSG3: 0.8449; hPL/ADAM12: 0.8396) compared with single hPL mRNA. CONCLUSION: Quantitative aberration of plural cell-free PP mRNAs in maternal plasma precedes the appearance of clinically apparent TTTS. This suggests that pathophysiological changes in the placenta are associated with morbid conditions of TTTS.


Assuntos
Transfusão Feto-Fetal/genética , Placenta/metabolismo , RNA Mensageiro/genética , Proteínas ADAM/genética , Proteína ADAM12 , Adulto , Área Sob a Curva , Feminino , Transfusão Feto-Fetal/sangue , Transfusão Feto-Fetal/diagnóstico , Perfilação da Expressão Gênica , Produtos do Gene env/genética , Humanos , Proteínas de Membrana/genética , Peptídeos/genética , Lactogênio Placentário/genética , Gravidez , Proteínas da Gravidez/genética , Gravidez de Gêmeos , RNA Mensageiro/sangue , Gêmeos Monozigóticos , Adulto Jovem
11.
Artigo em Inglês | MEDLINE | ID: mdl-23502141

RESUMO

Growth is effected via a complex interaction of genetic, nutritional, environmental and growth factors. Hormonal factors such as the growth hormone (GH) and insulin-like growth factor (IGF) signaling system, the human placental lactogen, and insulin play an integral role in early growth. Genetic factors affecting the GH-IGF system and insulin secretion and actions, and epigenetic mechanisms including DNA methylation have been further implicated as contributory factors. These hormonal systems, on a background of genetic susceptibility, together with other factors including maternal nutrition, placental and environmental factors, regulate not only early growth but also development. These interactions may impact on later health consequences in adult life. Accumulating data in the last few decades on developmental programming and later life metabolic disorders has provided a novel perspective on the possible pathogenesis of metabolic dysregulation. Despite postulations put forward to elucidate the mechanism underlying the association between early growth and later life metabolic disorders, it remains unclear what the dominant factor(s) would be, how any underlying mechanisms interact, or whether these mechanisms are truly causal.


Assuntos
Hormônio do Crescimento Humano/genética , Doenças Metabólicas/genética , Epigênese Genética , Feminino , Desenvolvimento Humano/fisiologia , Hormônio do Crescimento Humano/metabolismo , Humanos , Placenta/metabolismo , Lactogênio Placentário/genética , Lactogênio Placentário/metabolismo , Gravidez
12.
J Clin Endocrinol Metab ; 98(3): E429-36, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23337725

RESUMO

CONTEXT: Fetal growth restriction (FGR) is a leading cause of perinatal mortality, yet no reliable screening test exists. Placental specific mRNA in the maternal circulation may reflect changes in the placental transcriptome in FGR and could be a novel biomarker for FGR. OBJECTIVE: The aim of the study was to identify placental specific RNA detectable in the maternal circulation and examine whether they are differentially expressed in severe preterm FGR. DESIGN: In silico screening was used to identify placental specific RNAs. Their expression in cases of severe FGR vs controls was examined in both maternal blood and placenta by microarray, RT-PCR, and in situ hybridization. RESULTS: Via in silico analysis, we identified 137 genes very highly expressed in the placenta relative to other tissues. Using microarray, we found that they were detectable in the maternal blood and were globally dysregulated with preterm FGR; 75 genes (55%) had a ≥1.5-fold differential expression compared to controls. Eight genes (ERVWE-1, PSG1, PLAC4, TAC3, PLAC3, CRH, CSH1, and KISS1) were validated by RT-PCR to be significantly increased in both maternal blood and placenta in a larger cohort of severe FGR compared to controls. In situ hybridization confirmed PAPPA2 and ERVWE-1 localized to the syncytiotrophoblast. CONCLUSION: There is global differential expression of placental specific mRNA in the maternal blood in pregnancies complicated by severe preterm FGR. Placental specific mRNA in maternal blood may represent a new class of biomarkers for preterm FGR.


Assuntos
Retardo do Crescimento Fetal/genética , Placenta/fisiologia , Proteínas da Gravidez/genética , RNA Mensageiro/genética , Transcriptoma , Adulto , Hormônio Liberador da Corticotropina/genética , Feminino , Retardo do Crescimento Fetal/metabolismo , Produtos do Gene env/genética , Humanos , Recém-Nascido , Recém-Nascido Prematuro/sangue , Kisspeptinas/genética , Lactogênio Placentário/genética , Gravidez , Proteína Plasmática A Associada à Gravidez/genética , Glicoproteínas beta 1 Específicas da Gravidez/genética , RNA Mensageiro/sangue , Receptores de Taquicininas/genética
13.
Mol Cell Endocrinol ; 355(1): 180-7, 2012 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-22387044

RESUMO

The human GH/CSH cluster consisting of one pituitary-expressed (GH1) and four placenta-expressed loci has been implicated in maternal metabolic adaptation to pregnancy, regulation of intrauterine and postnatal growth. We investigated how the mRNA expression profile of placental GH2, CSH1 and CSH2 genes and their alternative transcripts correlates with maternal pre-eclampsia (PE) and/or gestational diabetes mellitus (GD). The expression of studied genes in PE placentas (n=17) compared to controls (n=17) exhibited a trend for reduced transcript levels. The alternative transcripts retaining intron 4, GH2-2 and CSH1-2 showed significantly reduced expression in PE cases without growth restriction (P=0.007, P=0.008, respectively). In maternal GD (n=23), a tendency of differential expression was detected only for the GH2 gene and in pregnancies with large-for-gestational-age newborns. Our results, together with those reported by others, are consistent with a pleiotropic effect of placental hGH/CSH genes at the maternal-fetal interface relating to the regulation of fetal growth and the risk of affected maternal metabolism.


Assuntos
Diabetes Gestacional/genética , Expressão Gênica , Hormônio do Crescimento/metabolismo , Hormônios Placentários/metabolismo , Lactogênio Placentário/metabolismo , Pré-Eclâmpsia/genética , Adulto , Processamento Alternativo , Estudos de Casos e Controles , Diabetes Gestacional/metabolismo , Diabetes Gestacional/fisiopatologia , Feminino , Perfilação da Expressão Gênica , Hormônio do Crescimento/genética , Humanos , Recém-Nascido , Íntrons , Hipófise/metabolismo , Hipófise/fisiopatologia , Placenta/metabolismo , Placenta/fisiopatologia , Hormônios Placentários/genética , Lactogênio Placentário/genética , Pré-Eclâmpsia/metabolismo , Pré-Eclâmpsia/fisiopatologia , Gravidez , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , RNA Mensageiro/biossíntese
14.
Int J Gynaecol Obstet ; 117(2): 131-3, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22342059

RESUMO

OBJECTIVE: To measure and compare placental mRNA expression in the maternal circulation among women with intrauterine and ectopic pregnancies. METHODS: Plasma was collected from patients in early pregnancy at risk of ectopic pregnancy. Clinical information was prospectively collected and entered into a dedicated database. mRNA was isolated from maternal plasma and quantitative RT-PCR was performed to measure mRNA for human gonadotropin (hCG) and human placental lactogen (hPL). GAPDH mRNA expression was used as an internal control. RESULTS: Twelve women with ectopic pregnancy and 13 women with intrauterine pregnancy were enrolled. Patients with ectopic pregnancy were 6 times more likely to have undetectable levels of hPL mRNA (relative risk [RR] 6.36; 95% confidence interval [CI], 1.70-23.20; P<0.01). They were also 8 times more likely to have undetectable levels of hCG mRNA (RR 8.64, 95% CI, 1.30-57.10; P<0.01). mRNA copy numbers for hPL and hCG (normalized by GAPDH) were significantly lower in the ectopic group than in the intrauterine group. CONCLUSION: Placental mRNA is present in the maternal circulation in significantly lower copies in cases of ectopic pregnancy compared with cases of intrauterine pregnancy. Measurement of placental mRNA in the maternal circulation may help to distinguish between intrauterine and ectopic pregnancies.


Assuntos
Placenta/metabolismo , Gravidez Ectópica/diagnóstico , RNA Mensageiro/metabolismo , Adulto , Gonadotropina Coriônica/sangue , Gonadotropina Coriônica/genética , Feminino , Gliceraldeído-3-Fosfato Desidrogenases/sangue , Gliceraldeído-3-Fosfato Desidrogenases/genética , Humanos , Lactogênio Placentário/sangue , Lactogênio Placentário/genética , Gravidez , Gravidez Ectópica/sangue , Estudos Prospectivos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Adulto Jovem
15.
J Mol Endocrinol ; 47(2): 179-93, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21737519

RESUMO

The human chorionic somatomammotropin (CS) A and B genes (listed as CSH1 and CSH2 in the HUGO database) are highly expressed in placenta. A 241 bp potent enhancer, nucleotides (nts) 1-241, located at the 3' end of the CS-B gene (CS-Benh) stimulates promoter activity specifically in placental trophoblast cells in vitro. Strong activity is exerted by a 23 bp element within the CS-Benh (nts 117-139), shown to interact with transcription enhancer factor (TEF) members of the transcription enhancer activator (TEA) DNA-binding domain-containing family. An identical TEF element is present in the homologous (97.5%) CS-Aenh; however, a few nucleotide differences suppress its activity. Previously, we identified regulatory sequences distinct from the TEF element within an 80 bp modulatory domain (nts 1-80) in the CS-Benh. Using structural and functional assays we now show that CCAAT/enhancer-binding protein (C/EBP) binding sites exist in the 80 bp modulatory domains of both enhancers, and an Elk-1 binding site exists in the modulatory domain of the CS-Aenh. C/EBPα or C/EBPß strongly repressed CSp.CAT activity but stimulated CSp.CAT.CS-Benh activity. In contrast, the equivalent CS-A enhancer sequences were unable to relieve promoter repression. Elk-1 overexpression also resulted in differential effects on the CS-Aenh versus CS-Benh. Finally, we provide evidence for the association of C/EBPß with the CS-A and CS-B genes in human placental chromatin, including differential involvement of C/EBPß with the CS-Aenh versus the CS-Benh, and therefore consistent with the notion that these are regions of regulatory significance in vivo. We conclude that members of the C/EBP and Ets families can differentially modulate CS-Benh and CS-Aenh activity.


Assuntos
Proteínas Estimuladoras de Ligação a CCAAT/metabolismo , Elementos Facilitadores Genéticos/genética , Lactogênio Placentário/genética , Proteínas Proto-Oncogênicas c-ets/metabolismo , Sítios de Ligação , Linhagem Celular Tumoral , Imunoprecipitação da Cromatina , Ensaio de Desvio de Mobilidade Eletroforética , Humanos , Regiões Promotoras Genéticas/genética , Ligação Proteica/genética
16.
Mol Cell Endocrinol ; 337(1-2): 7-15, 2011 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-21241770

RESUMO

Perfluorooctanoic acid (PFOA) is a persistent pollutant worldwide and even found in human cord blood and breast milk. Some animal studies have reported that PFOA causes developmental toxicity such as fetal weight loss, but the mechanism is still unclear. This study focused on developmental toxicity of PFOA, particularly impacts of PFOA on placental endocrine function such as placental prolactin (PRL)-family hormone gene expression and fetal growth in mouse. Time-mated CD-1 mice were dosed by gavage with 0, 2, 10 and 25 mg/kg B.W/day of PFOA (n-10) dissolved with de-ionized water from gestational day (GD) 11-16. During treatment, body weight of each pregnant mouse was measured daily. On day 16, caesarean sections were performed and developmental data were observed. Three placentas from three different pregnant mice were assigned to each of the following experiments. The mRNA levels of mouse placental lactogen (mPL)-II, prolactin like protein (mPLP)-E, -F and Pit-1α and ß isotype mRNAs, a transacting factor of mPLs and mPLPs genes, were analyzed using northern blot, in situ hybridization and RT-PCR, respectively. Maternal body weight gain was significantly declined from GD 13 in the PFOA treated groups compared to control. Developmental data such as fetal and placental weights were significantly decreased in accordance with PFOA dosage. Number of dead fetuses and post-implantation losses were significantly increased in the PFOA-exposed groups. In addition, placental efficiency (fetal weight/placental weight) was significantly reduced in PFOA treated groups in accordance with PFOA dosage. Histopathologic changes were observed in placenta. Dose dependent necrotic changes were observed in both 10 mg and 25 mg PFOA treated groups. Cell frequency of glycogen trophoblast cell and parietal trophoblast giant cell were decreased dose dependently in the junctional zone. In the labyrinth zone, sinusoidal trophoblast giant cell frequency was decreased in the 25 mg PFOA treated group. Also, morphological change such as crushed nuclear (atrophy) of trophoblast cells was observed in 25 mg PFOA treated group. Finally, mRNA levels of the mPL-II, mPLP-E, -F and Pit-1α and ß were significantly reduced in the PFOA treated groups dose dependently. In addition, the changing pattern between mPL-II, mPLP-E, -F mRNA levels and fetal body weight showed positive relationship. In conclusion, the inhibitory effects of PFOA on the placental prolactin-family hormone genes expression may be secondary effects to insufficient trophoblast cell type differentiation and/or increased trophoblast cell necrosis. The impacts of PFOA on placental development and endocrine function reduced the placental efficiency and partly contributed to the fetal growth retardation in the mouse.


Assuntos
Caprilatos/efeitos adversos , Retardo do Crescimento Fetal/induzido quimicamente , Fluorcarbonetos/efeitos adversos , Hormônios Placentários/antagonistas & inibidores , Animais , Peso Corporal/efeitos dos fármacos , Contagem de Células , Citocinas/genética , Citocinas/metabolismo , Feminino , Morte Fetal/induzido quimicamente , Peso Fetal/efeitos dos fármacos , Camundongos , Placenta/efeitos dos fármacos , Placenta/patologia , Lactogênio Placentário/genética , Lactogênio Placentário/metabolismo , Gravidez , Proteínas da Gravidez/genética , Proteínas da Gravidez/metabolismo , Fator de Transcrição Pit-1/genética , Fator de Transcrição Pit-1/metabolismo , Transcrição Genética/efeitos dos fármacos , Trofoblastos/citologia , Trofoblastos/efeitos dos fármacos
17.
Placenta ; 31(11): 969-75, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-20832857

RESUMO

To understand the tissue-specific expression of the rat placental lactogen-I variant (rPL-Iv) gene, we investigated the methylation pattern of the 5'-flanking region of this gene in various rat tissues. We report that the 5'-flanking region of the rPL-Iv gene was hypomethylated in placenta that expressed the gene and hypermethylated in those tissues that did not express the gene. Moreover, the intron region of the rPL-Iv gene was hypomethylated in the placenta, but hypermethylated in the liver, kidney and pituitary. Although there are 5 CpG sites and the density of CpG dinucleotide is lower within 2 kb of the rPL-Iv 5'-flanking region, the methylated promoter reporter gene produced strong repression in the transcriptional activity of the gene. In addition, the 5'-flanking and intron regions of the rPL-Iv gene were hypomethylated on day 12 of gestation, and the methylation pattern in the placenta remained unchanged from mid-pregnancy until term. The entire genomic region of the rPL-Iv gene might be hypermethylated in tissues other than the placenta, within which its methylated status repress expression of the placenta-specific rPL-Iv gene. Interestingly, the methylation status of the intron region of the rPL-Iv in proliferating Rcho-1 cells was changed to the unmethylated status on day 8 and 12 of differentiation of Rcho-1 cells. These results demonstrate that demethylation in the rPL-Iv upstream region was induced at an early stage of placental development, and once the 5'-flanking region of the rPL-Iv had been demethylated, its status on the rPL-Iv genomic region was continued during pregnancy. Taken together, these results suggest that DNA methylation is responsible for the silencing of tissue-specific genes in non-expressing cells, while defined combinations of trophoblast factors dictate the expression of unmethylated rPL-Iv gene in placenta trophoblast cells.


Assuntos
Metilação de DNA , Regulação da Expressão Gênica no Desenvolvimento , Placenta/metabolismo , Lactogênio Placentário/metabolismo , Região 5'-Flanqueadora , Animais , Diferenciação Celular , Linhagem Celular , Feminino , Inativação Gênica , Genes Reporter , Íntrons , Especificidade de Órgãos , Lactogênio Placentário/genética , Placentação , Gravidez , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Trofoblastos/citologia , Trofoblastos/metabolismo
18.
Prenat Diagn ; 30(8): 764-70, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20661890

RESUMO

OBJECTIVE: The discovery of placental transcripts in peripheral blood of pregnant women prompted us to investigate which was the most appropriate biological specimen, between plasma and serum, to easily detect them and to exploit hPL (human placental lactogen), betahCG (human chorionic gonadotrophin beta-subunit), LOC90625, and TFPI2 (tissue factor pathway inhibitor 2) levels in order to establish whether an abnormal variation degree of presence of these placental transcripts are likely to be associated to specific fetal trisomies. METHOD: RNA was extracted from plasma and serum samples of 255 pregnant women bearing euploid fetuses, 17 bearing fetuses affected by trisomy 21 and 10 with fetuses affected by trisomy 18. Placental transcript analysis was performed by real time RT-PCR using relative quantification. RESULTS: Results obtained from euploid samples showed that fetal transcripts were more abundant in plasma than in serum samples. Euploid samples had a placental transcript abundance distinguishable from those with trisomy 21 but not from those with trisomy 18. In particular, high betahCG abundance and advanced maternal age were significantly associated with trisomy 21 pregnancy. CONCLUSION: Plasma was the most suitable tool to be employed in the detection and dosage of placental transcripts. betahCG transcript together with maternal age could be a potential marker for noninvasive prenatal screening of fetal trisomy 21.


Assuntos
Gonadotropina Coriônica Humana Subunidade beta/genética , Cromossomos Humanos Par 18 , Síndrome de Down/genética , Lipoproteínas/genética , Lactogênio Placentário/genética , Diagnóstico Pré-Natal/métodos , RNA Mensageiro/sangue , Trissomia/genética , Gonadotropina Coriônica Humana Subunidade beta/sangue , Síndrome de Down/sangue , Síndrome de Down/diagnóstico , Feminino , Marcadores Genéticos , Humanos , Lipoproteínas/sangue , Modelos Logísticos , Lactogênio Placentário/sangue , Gravidez , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Trissomia/diagnóstico
19.
J Biol Chem ; 285(26): 20022-30, 2010 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-20427283

RESUMO

Class 1 cytokines bind two receptors to create an active heterotrimeric complex. It has been argued that ligand binding to their receptors is an ordered process, but a structural mechanism describing this process has not been determined. We have previously described an obligate ordered binding mechanism for the human prolactin/prolactin receptor heterotrimeric complex. In this work we expand this conceptual understanding of ordered binding to include three human lactogenic hormones: prolactin, growth hormone, and placental lactogen. We independently blocked either of the two receptor binding sites of each hormone and used surface plasmon resonance to measure human prolactin receptor binding kinetics and stoichiometries to the remaining binding surface. When site 1 of any of the three hormones was blocked, site 2 could not bind the receptor. But blocking site 2 did not affect receptor binding at site 1, indicating a requirement for receptor binding to site 1 before site 2 binding. In addition we noted variable responses to the presence of zinc in hormone-receptor interaction. Finally, we performed Förster resonance energy transfer (FRET) analyses where receptor binding at subsaturating stoichiometries induced changes in FRET signaling, indicative of binding-induced changes in hormone conformation, whereas at receptor:hormone ratios in excess of 2:1 no additional changes in FRET signaling were observed. These results strongly support a conformationally mediated obligate-ordered receptor binding for each of the three lactogenic hormones.


Assuntos
Hormônio do Crescimento/metabolismo , Lactogênio Placentário/metabolismo , Prolactina/metabolismo , Receptores da Prolactina/metabolismo , Sítios de Ligação/genética , Ligação Competitiva/efeitos dos fármacos , Citocinas/química , Citocinas/genética , Citocinas/metabolismo , Feminino , Transferência Ressonante de Energia de Fluorescência , Hormônio do Crescimento/química , Hormônio do Crescimento/genética , Humanos , Cinética , Mutação , Lactogênio Placentário/química , Lactogênio Placentário/genética , Gravidez , Prolactina/química , Prolactina/genética , Ligação Proteica/efeitos dos fármacos , Conformação Proteica , Receptores da Prolactina/química , Receptores da Prolactina/genética , Ressonância de Plasmônio de Superfície , Zinco/metabolismo , Zinco/farmacologia
20.
J Clin Endocrinol Metab ; 95(5): 2433-42, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20233782

RESUMO

CONTEXT: The human growth hormone/chorionic somatomammotropin (hGH/CSH) locus at 17q22-24, consisting of one pituitary-expressed postnatal (GH1) and four placenta-expressed genes (GH2, CSH1, CSH2, and CSHL1), is implicated in regulation of postnatal and intrauterine growth. A positive correlation has been reported between the offspring's birth weight and serum placental GH (coded by GH2) and placental lactogen (coded by CSH1, CSH2) levels in pregnant women. OBJECTIVE: The objective of the study was the investigation of the hypothesis that the mRNA expression profile of placental hGH/CSH genes contributes to the determination of birth weight. DESIGN AND SUBJECTS: We developed a sensitive, fluorescent-labeled semiquantitative RT-PCR assay coupled with gene-specific restriction analysis, capable of distinguishing alternative splice-products of individual placental hGH/CSH genes and quantification of their relative expression levels. The detailed profile of alternative transcripts of GH2, CSH1, CSH2, and CSHL1 genes in placenta from uncomplicated term pregnancies of the REPROMETA sample collection was addressed in association with the birth weight of newborns, grouped as appropriate for gestational age (AGA; n = 23), small for gestational age (SGA; n = 15), and large for gestational age (LGA; n = 34). RESULTS: The majority of pregnancies with SGA newborn showed down-regulation of the entire hGH/CSH cluster in placenta, whereas in the case of LGA, the expression of CSH1-1, CSH2-1, and CSHL1-4 mRNA transcripts in placenta was significantly increased compared with AGA newborns (P < 0.0001, P = 0.009, P = 0.002, respectively). CONCLUSION: The expression profile of placental hGH/CSH genes in placenta is altered in pregnancies accompanied by SGA and LGA compared with AGA newborns, and thus, it may directly affect the circulating fetal and maternal placental GH and placental lactogen levels.


Assuntos
Cromossomos Humanos Par 17 , Perfilação da Expressão Gênica , Hormônio do Crescimento Humano/genética , Recém-Nascido Pequeno para a Idade Gestacional , Placenta/fisiologia , Lactogênio Placentário/genética , Processamento Alternativo , Peso ao Nascer , Primers do DNA/genética , Éxons/genética , Feminino , Humanos , Recém-Nascido , Gravidez , Mapeamento por Restrição , Transcrição Genética
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA