RESUMO
Our research aimed to compare the epigenetic alterations between placentae of in vitro fertilization (IVF) patients and spontaneous pregnancies. Additionally, the expression levels of proliferation markers (PCNA, Ki67) and glucose transporter proteins (GLUT1, GLUT3) were assessed in control and IVF placentae to examine the possible consequences of epigenetic alterations on placental development. Control group placentae were obtained from spontaneous pregnancies of healthy women (n = 16). IVF placentae were obtained from fresh (n = 16) and frozen (n = 16) embryo transfer pregnancies. A group of maternal and paternal imprint genes H19, IGF2, IGF2, IGF2R, PHLDA2, PLAGL1, MASH2, GRB10, PEG1, PEG3, and PEG10 were detected by Real-Time PCR. Additionally, PCNA, Ki67, GLUT1, and GLUT3 protein levels were assessed by immunohistochemistry and western blot. In the fresh embryo transfer placenta group (fETP), gene expression of paternal PEG1 and PEG10 was upregulated compared with the control group. Increased gene expression in paternal PEG1 and maternal IGFR2 genes was detected in the frozen embryo transfer placenta group (FET) compared with the control group. Conversely, expression levels of H19 and IGF2 genes were downregulated in the FET group. On the other hand, GLUT3 and PCNA expression was increased in FET group placentae. IVF techniques affect placental imprinted gene expressions which are important for proper placental development. Imprinted genes are differently expressed in fresh ET placentae and frozen ET placentae. In conclusion, these data indicate that altered imprinted gene expression may affect glucose transport and cell proliferation, therefore play an important role in placental development.
Assuntos
Transferência Embrionária , Expressão Gênica/fisiologia , Fator de Crescimento Insulin-Like II/metabolismo , Placenta/metabolismo , Adulto , Transferência Embrionária/métodos , Feminino , Fertilização in vitro/métodos , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Humanos , Masculino , Gravidez , Receptor IGF Tipo 2/metabolismoRESUMO
PURPOSE: Almost every female classic galactosemia patient develops primary ovarian insufficiency (POI). The unique pathophysiology of classic galactosemia, with a severely reduced follicle pool at an early age, requires a new therapeutic approach. This study evaluated the effect of dehydroepiandrosterone (DHEA) on ovarian tissue in a galactose-induced POI rat model. METHODS: Pregnant rats were fed with either a normal or a 35% galactose-containing diet from day 3 of conception continuing through weaning of the litters. Galactose-exposed female offspring were further divided into 5 groups on PND21. The first group received no application. Treatment groups were fed orally by gavage once daily with sesame oil (group 2), or DHEA at doses of 0.1 mg/kg (group 3), 1 mg/kg (group 4) or 10 mg/kg (group 5) until PND70. Fertility rates of mothers with galactosemia, body weights (BWs), and ovarian weights of the litters from PND21 to PND70 were recorded. Ovarian follicle count, immunohistochemistry for proliferation and apoptosis marker expressions and TUNEL for cell death assessment were performed in offspring ovaries. RESULTS: Decreased fertility, ovarian/body weights were observed under galactosemic conditions, together with decreased follicle number and increased atresia. Improved postnatal development, primordial follicle recruitment and follicular growth were observed after DHEA treatment. After DHEA treatment, the expression of Ki67 protein was found to be increased; elevated expression of cleaved-caspase-3 under galactosemia was found to be reduced. CONCLUSIONS: Our data suggests that DHEA treatment may be a potentially useful clinical therapy to improve ovarian ageing in women with POI-induced by galactosemia.
Assuntos
Envelhecimento/efeitos dos fármacos , Desidroepiandrosterona/farmacologia , Galactosemias/dietoterapia , Insuficiência Ovariana Primária/dietoterapia , Envelhecimento/genética , Animais , Suplementos Nutricionais , Modelos Animais de Doenças , Feminino , Galactose/toxicidade , Galactosemias/induzido quimicamente , Galactosemias/complicações , Galactosemias/patologia , Humanos , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/patologia , Gravidez , Insuficiência Ovariana Primária/induzido quimicamente , Insuficiência Ovariana Primária/patologia , RatosRESUMO
BACKGROUND: Little is known about the genetic contribution to Müllerian aplasia, better known to patients as Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome. Mutations in two genes (WNT4 and HNF1B) account for a small number of patients, but heterozygous copy number variants (CNVs) have been described. However, the significance of these CNVs in the pathogenesis of MRKH is unknown, but suggests possible autosomal dominant inheritance. We are not aware of CNV studies in consanguineous patients, which could pinpoint genes important in autosomal recessive MRKH. We therefore utilized SNP/CGH microarrays to identify CNVs and define regions of homozygosity (ROH) in Anatolian Turkish MRKH patients. RESULTS: Five different CNVs were detected in 4/19 patients (21%), one of which is a previously reported 16p11.2 deletion containing 32 genes, while four involved smaller regions each containing only one gene. Fourteen of 19 (74%) of patients had parents that were third degree relatives or closer. There were 42 regions of homozygosity shared by at least two MRKH patients which was spread throughout most chromosomes. Of interest, eight candidate genes suggested by human or animal studies (RBM8A, CMTM7, CCR4, TRIM71, CNOT10, TP63, EMX2, and CFTR) reside within these ROH. CONCLUSIONS: CNVs were found in about 20% of Turkish MRKH patients, and as in other studies, proof of causation is lacking. The 16p11.2 deletion seen in mixed populations is also identified in Turkish MRKH patients. Turkish MRKH patients have a higher likelihood of being consanguineous than the general Anatolian Turkish population. Although identified single gene mutations and heterozygous CNVs suggest autosomal dominant inheritance for MRKH in much of the western world, regions of homozygosity, which could contain shared mutant alleles, make it more likely that autosomal recessively inherited causes will be manifested in Turkish women with MRKH.
RESUMO
OBJECTIVE: To study the genetic cause of Mayer-Rokitansky-Kuster-Hauser syndrome (MRKH). Although a few candidate genes and genomic domains for have been reported for MRKH, the genetic underpinnings remain largely unknown. Some of the top candidate genes are WNT4, HNF1B, and LHX1. The goals of this study were to: 1) determine the prevalence of WNT4, HNF1B, and LHX1 point mutations, as well as new copy number variants (CNVs) in people with MRKH; and 2) identify and characterize MRKH cohorts. DESIGN: Laboratory- and community-based study. SETTING: Academic medical centers. PATIENT(S): A total of 147 MRKH probands and available family members. INTERVENTIONS(S): DNA sequencing of WNT4, HNF1B, and LHX1 in 100 MRKH patients, chromosomal microarray analysis in 31 North American MRKH patients, and characterization and sample collection of 147 North American and Turkish MRKH probands and their families. MAIN OUTCOME MEASURE(S): DNA sequence variants and CNVs; pedigree structural analysis. RESULT(S): We report finding CNVs in 6/31 people (â¼19%) with MRKH, but no point mutations or small indels in WNT4, HNF1B, or LHX1 in 100 MRKH patients. Our MRKH families included 43 quads, 26 trios, and 30 duos. Of our MRKH probands, 87/147 (59%) had MRKH type 1 and 60/147 (41%) had type 2 with additional anomalies. CONCLUSION(S): Although the prevalence of WNT4, HNF1B, and LHX1 point mutations is low in people with MRKH, the prevalence of CNVs was â¼19%. Further analysis of our large familial cohort of patients will facilitate gene discovery to better understand the complex etiology of MRKH.
Assuntos
Transtornos 46, XX do Desenvolvimento Sexual/epidemiologia , Transtornos 46, XX do Desenvolvimento Sexual/genética , Anormalidades Congênitas/epidemiologia , Anormalidades Congênitas/genética , Fator 1-beta Nuclear de Hepatócito/genética , Proteínas com Homeodomínio LIM/genética , Ductos Paramesonéfricos/anormalidades , Polimorfismo de Nucleotídeo Único/genética , Fatores de Transcrição/genética , Proteína Wnt4/genética , Adulto , Estudos de Coortes , Família , Marcadores Genéticos/genética , Predisposição Genética para Doença/epidemiologia , Predisposição Genética para Doença/genética , Humanos , Internacionalidade , Prevalência , Fatores de Risco , Adulto JovemRESUMO
PURPOSE: To evaluate the effect of the birth weight and the postconceptional age on the tear production of preterm and term newborn infants and to evaluate the changes in tear production during the first two months of life. SUBJECTS--METHODS: Both eyes of medically stable term and preterm infants were included in the study. Based on postconceptional age and birth weight, we divided preterm infants into three groups. Then, we measured the basal and reflex tear secretions of both eyes by Schirmer tests before and after instillation of topical anesthetic agent. We performed initial tear measurements on the second day of life (between first 24 hours to first 48 hours after birth): and at 2 weeks, 4 weeks and 8 weeks after birth. RESULTS: We tested a total of 138 infants (63 preterm and 75 term), 72 males and 66 females. Schirmer-1-test of preterm and terms on the 2nd day of life revealed a mean basal tear secretion (BTS) of 4.8 +/- 4.1 and 8.8 +/- 3.2 mm, respectively (p < 0.0001). The mean reflex tear secretion (RTS) within 48 hours of life was 6.1 +/- 3.9 mm and 10.3 +/- 4.4 mm in preterm and term infants, respectively (p < 0.0001). The mean values of both basal and reflex tear secretion were significantly lower in the preterm than the term infants during the first two months of the life. In preterm infants, both basal and reflex tear secretions were found to be highly correlated with postconceptional age (r: 0.79 and 0.74, respectively, both p values: 0.001); however, there was a weak correlation with birth weight (r: 0.16 and 0.19, respectively, both p values: 0.01). Statistically significant differences for both BTS and RTS existed among the three postconceptional age groups: there were no such difference among the three different birth weight groups. We found no gender or laterality dependent (side of the eyes) differences in the tear production (both p values >0.05). CONCLUSIONS: Preterm infants have significantly reduced tear secretion compared with term infants. Postconceptional age, rather than birth weight, seems to be more correlated with the tear secretion. Sex and laterality does not appear to have an effect on tear production in infants. Tear production of preterm infants is significantly reduced than that of term infants during the first two months of life. Term infants increased their tear production significantly in each examination during the neonatal period while the preterms increase tear production significantly only at mean postconceptional age of eight and a half (8.5) months.