Jerome F Strauss

Virginia Commonwealth University, Ричмонд, Virginia, United States

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Publications (376)1978.43 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: RC/BTB2 is a binding partner of sperm associated antigen 16S (SPAG16S), which is regulator of spermiogenesis in mice, a process during which sperm flagella are formed. The expression of Rc/btb2 is also regulated by multicilin, a protein that controls ciliogenesis. Given that mouse Rc/btb2 mRNA is not only expressed in tissues bearing motile cilia, but also in tissues without motile cilia, we investigated whether RC/BTB2 plays a role in the general process of ciliogenesis by studying two somatic cells lines that have primary cilia, NIH3T3 and IMCD3. We discovered that the subcellular localization of RT/BTB2 in the NIH3T3 and IMCD3 cells encompasses the pathway for ciliogenesis. RC/BTB2 was found in the Golgi bodies and centrosomes, two key structures essential for normal ciliogenesis. Knockdown of Rc/btb2 gene expression in these cell lines disrupted ciliogenesis. The percentage of cells with primary cilia was significantly reduced in stable cell lines transduced with specific Rc/btb2 shRNA viruses compared to the control cells. When cilia were formed in the knockdown cells, they were significantly shorter than those in the control cells. Knockdown of Rc/btb2 expression did not affect cell proliferation and the cell cycle. Exogenous expression of RC/BTB2 in these stable knockdown cells restored ciliogenesis. These findings suggest that RC/BTB2 is a necessary component of the process of formation of primary cilia in somatic cells, perhaps through the transportation of cargos from Golgi bodies to centrosomes for cilia assembling. This article is protected by copyright. All rights reserved. © 2015 Wiley Periodicals, Inc.
    Cytoskeleton 03/2015; DOI:10.1002/cm.21214 · 3.01 Impact Factor
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    ABSTRACT: A key event in the process of spermiogenesis is the formation of the flagella, which enables sperm to reach eggs for fertilization. Yeast two-hybrid studies revealed that meiosis-expressed gene 1 (MEIG1) and Parkin co-regulated gene (PACRG) interact, and that sperm-associated antigen 16, which encodes an axoneme central apparatus protein, is also a binding partner of MEIG1. In spermatocytes of wild-type mice, MEIG1 is expressed in the whole germ cell bodies, but the protein migrates to the manchette, a unique structure at the base of elongating spermatid that directs formation of the flagella. In the elongating spermatids of wild-type mice, PACRG colocalizes with α-tubulin, a marker for the manchette, whereas this localization was not changed in the few remaining elongating spermatids of Meig1-deficient mice. In addition, MEIG1 no longer localizes to the manchette in the remaining elongating spermatids of Pacrg-deficient mice, indicating that PACRG recruits MEIG1 to the manchette. PACRG is not stable in mammalian cells, but can be stabilized by MEIG1 or by inhibition of proteasome function. SPAG16L is present in the spermatocyte cytoplasm of wild-type mice, and in the manchette of elongating spermatids, but in the Meig1 or Pacrg-deficient mice, SPAG16L no longer localizes to the manchette. By contrast, MEIG1 and PACRG are still present in the manchette of Spag16L-deficient mice, indicating that SPAG16L is a downstream partner of these two proteins. Together, our studies demonstrate that MEIG1/PACRG forms a complex in the manchette and that this complex is necessary to transport cargos, such as SPAG16L, to build the sperm flagella. © 2015. Published by The Company of Biologists Ltd.
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    ABSTRACT: Microbial invasion of the amniotic cavity is associated with spontaneous preterm labor and adverse pregnancy outcome and Mycoplasma hominis is often present. However, the pathogenic process by which M. hominis invades the amniotic cavity and gestational tissues, often resulting in chorioamnionitis and preterm birth, remains unknown. We hypothesized that strains of M. hominis vary genetically with regards to their potential to invade and colonize the amniotic cavity and placenta. We sequenced the entire genomes of two amniotic fluid isolates and a placental isolate of M. hominis from pregnancies that resulted in preterm births, and compared them to the previously sequenced genome of the Type strain PG21. We identified genes specific to the amniotic fluid/placental isolates. We then determined the microbial burden and the presence of these genes in another set of subjects, from whom samples of amniotic fluid had been collected and were positive for M. hominis. We identified two genes encoding surface-located membrane proteins (Lmp1 and Lmp-like) in the sequenced amniotic fluid/placental isolates that were severely truncated in PG21. We also identified, for the first time, a microbial gene of unknown function referred to in this study as gene of interest C (goiC), that was significantly associated with bacterial burden in amniotic fluid and the risk of preterm delivery in patients with preterm labor. A gene in M. hominis was identified that is significantly associated with colonization and/or infection of the upper reproductive tract during pregnancy and with preterm birth. Copyright © 2015 Elsevier Inc. All rights reserved.
    American Journal of Obstetrics and Gynecology 01/2015; DOI:10.1016/j.ajog.2015.01.032 · 3.97 Impact Factor
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    ABSTRACT: Proper regulation of human leukocyte antigen-C (HLA-C) expression on trophoblast cells is essential for the immunological privilege of these cells. Endoplasmic reticulum aminopeptidase 2 (ERAP2) is known to trim antigenic peptide precursors for loading onto HLA. Altered expression of the ERAP2 could play a critical role in shaping HLA-C expression and antigen presentation, but their relationships have not been defined. We examined the genotype and expression levels of ERAP2 and HLA-C in several choriocarcinoma cell lines (BeWo, JAr, and JEG-3) by allelic discrimination qPCR, Western blotting and immunocytochemistry. Additionally, expression profiling of untreated versus gamma-interferon (IFN-γ, 20ng/ml)-treated cells was performed. Lastly, to determine if there is a relationship between ERAP2 and HLA-C expression levels, transfection of major and minor allele ERAP2 variants was performed. JAr cells were homozygous for the rs2248374 minor A allele, and express ERAP2 protein. BeWo and JEG-3 cells are homozygous for ERAP2 genetic variant (major G allele of SNP rs2248374) that causes non-sense mediated RNA decay, and thus lack ERAP2 protein expression. Interestingly, BeWo and JEG-3 cells express HLA-C, whereas, JAr cells do not. IFN-γ treatment of BeWo and JEG-3 cells increased HLA-C expression by 2-fold (p=0.0037) and 3-fold (p=0.0446) above control levels, respectively, but ERAP2 remained undetectable. In contrast, JAr cells treated with IFN-γ had increased ERAP2 protein expression by 2-fold (p=0.003), but had no effect on HLA-C expression. Transfection of JEG-3 cells (ERAP2 null) with ERAP2 plasmids with the SNP rs2549782, encoding an amino acid substitution (N392K) that alters substrate specificities and antigen presentation, revealed that the different ERAP2 isoforms do not affect the expression level of HLA-C. Our observations suggest that the expression patterns of ERAP2 and HLA-C allow choriocarcinoma cells to escape immune surveillance. E.D. Lee: None. S. Brockett: None. D. Hilliard: None. M.E. Teves: None. R. Ramus: None. J.F. Strauss: None. Copyright © 2014.
    01/2015; 5(1):88. DOI:10.1016/j.preghy.2014.10.178
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    ABSTRACT: Mammalian SPAG6 protein is localized to the axoneme central apparatus, and it is required for normal flagella and cilia motility. Recent studies demonstrated that the protein also regulates ciliogenesis and cilia polarity in the epithelial cells of brain ventricles and trachea. Motile cilia are also present in the epithelial cells of the middle ear and Eustachian tubes, where the ciliary system participates in the movement of serous fluid and mucus in the middle ear. Cilia defects are associated with otitis media (OM), presumably due to an inability to efficiently transport fluid, mucus and particles including microorganisms. We investigated the potential role of SPAG6 in the middle ear and Eustachian tubes by studying mice with a targeted mutation in the Spag6 gene. SPAG6 is expressed in the ciliated cells of middle ear epithelial cells. The orientation of the ciliary basal feet was random in the middle ear epithelial cells of Spag6-deficient mice, and there was an associated disrupted localization of the planar cell polarity (PCP) protein, FZD6. These features are associated with disordered cilia orientation, confirmed by scanning electron microscopy, which leads to uncoordinated cilia beating. The Spag6 mutant mice were also prone to develop OM. However, there were no significant differences in bacterial populations, epithelial goblet cell density, mucin expression and Eustachian tube angle between the mutant and wild-type mice, suggesting that OM was due to accumulation of fluid and mucus secondary to the ciliary dysfunction. Our studies demonstrate a role for Spag6 in the pathogenesis of OM in mice, possibly through its role in the regulation of cilia/basal body polarity through the PCP-dependent mechanisms in the middle ear and Eustachian tubes.
    PLoS ONE 11/2014; 9(11):e112879. DOI:10.1371/journal.pone.0112879 · 3.53 Impact Factor
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    ABSTRACT: Objective To investigate the association between cigarette use during pregnancy and pregnancy-induced hypertension/preeclampsia/eclampsia (PIH) by maternal race/ethnicity and age. Methods This retrospective cohort study was based on the U.S. 2010 natality data. Our study sample included U.S. women who delivered singleton pregnancies between 20 and 44 weeks of gestation without major fetal anomalies in 2010 (n = 3,113,164). Multivariate logistic regression models were fit to estimate crude and adjusted odds ratios and the corresponding 95% confidence intervals. Results We observed that the association between maternal smoking and PIH varied by maternal race/ethnicity and age. Compared with non-smokers, reduced odds of PIH among pregnant smokers was only evident for non-Hispanic white and non-Hispanic American Indian women aged less than 35 years. Non-Hispanic Asian/Pacific Islander women who smoked during pregnancy had increased odds of PIH regardless of maternal age. Non-Hispanic white and non-Hispanic black women 35 years or older who smoked during pregnancy also had increased odds of PIH. Conclusion Our study findings suggest important differences by maternal race/ethnicity and age in the association between cigarette use during pregnancy and PIH. More research is needed to establish the biologic and social mechanisms that might explain the variations with maternal age and race/ethnicity that were observed in our study.
    PLoS ONE 10/2014; 9(10):e106446. DOI:10.1371/journal.pone.0106446 · 3.53 Impact Factor
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    ABSTRACT: Humans are colonized by thousands of bacterial species, but it is difficult to assess the metabolic and pathogenic potential of the majority of these because they have yet to be cultured. Here, we characterize an uncultivated vaginal mycoplasma tightly associated with trichomoniasis that was previously known by its 16S rRNA sequence as "Mnola." In this study, the mycoplasma was found almost exclusively in women infected with the sexually transmitted pathogen Trichomonas vaginalis, but rarely observed in women with no diagnosed disease. The genomes of four strains of this species were reconstructed using metagenome sequencing and assembly of DNA from four discrete mid-vaginal samples, one of which was obtained from a pregnant woman with trichomoniasis who delivered prematurely. These bacteria harbor several putative virulence factors and display unique metabolic strategies. Genes encoding proteins with high similarity to potential virulence factors include two collagenases, a hemolysin, an O-sialoglycoprotein endopeptidase and a feoB-type ferrous iron transport system. We propose the name "Candidatus Mycoplasma girerdii" for this potential new pathogen.
    PLoS ONE 10/2014; 9(10):e110943. DOI:10.1371/journal.pone.0110943 · 3.53 Impact Factor
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    ABSTRACT: SPAG6, an axoneme central apparatus protein, is essential for function of ependymal cell cilia and sperm flagella. A significant number of Spag6-deficient mice die with hydrocephalus, and surviving males are sterile because of sperm motility defects. In further exploring the ciliary dysfunction in Spag6-null mice, we discovered that cilia beat frequency was significantly reduced in tracheal epithelial cells, and that the beat was not synchronized. There was also a significant reduction in cilia density in both brain ependymal and trachea epithelial cells, and cilia arrays were disorganized. The orientation of basal feet, which determines the direction of axoneme orientation, was apparently random in Spag6-deficient mice, and there were reduced numbers of basal feet, consistent with reduced cilia density. The polarized epithelial cell morphology and distribution of intracellular mucin, α-tubulin, and the planar cell polarity protein, Vangl2, were lost in Spag6-deficient tracheal epithelial cells. Polarized epithelial cell morphology and polarized distribution of α-tubulin in tracheal epithelial cells was observed in one-week old wild-type mice, but not in the Spag6-deficient mice of the same age. Thus, the cilia and polarity defects appear prior to 7 days post-partum. These findings suggest that SPAG6 not only regulates cilia/flagellar motility, but that in its absence, ciliogenesis, axoneme orientation, and tracheal epithelial cell polarity are altered.
    PLoS ONE 10/2014; 9(10):e107271. DOI:10.1371/journal.pone.0107271 · 3.53 Impact Factor
  • Placenta 09/2014; 35(9):A59. DOI:10.1016/j.placenta.2014.06.192 · 3.29 Impact Factor
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    ABSTRACT: Women of European ancestry are more likely to harbor a Lactobacillus-dominated microbiome, whereas African American women are more likely to exhibit a diverse microbial profile. African American women are also twice as likely to be diagnosed with bacterial vaginosis and are twice as likely to experience preterm birth. The objective of this study was to further characterize and contrast the vaginal microbial profiles in African American versus European ancestry women. Through the Vaginal Human Microbiome Project at Virginia Commonwealth University, we performed 16S rRNA gene survey to compare the microbiomes of vaginal samples from 1,268 African American women and 416 women of European ancestry. Our results confirmed significant differences in the vaginal microbiomes of the two groups and identified several taxa relevant to these differences. We found major community types dominated by Gardnerella vaginalis and the uncultivated bacterium, bacterial vaginosis-associated bacterium-1 (BVAB1)(Fredricks et al., 2005) that were common among African Americans. Moreover, prevalence of multiple bacterial taxa that are associated with microbial invasion of the amniotic cavity and preterm birth, including Mycoplasma, Gardnerella, Prevotella, and Sneathia, differed between the two ethnic groups. We investigated the contributions of intrinsic and extrinsic factors, including pregnancy, body mass index, diet, smoking and alcohol use, number of sexual partners, and household income to vaginal community composition. Ethnicity, pregnancy, and alcohol use correlated significantly with the relative abundance of bacterial vaginosis-associated species. Trends between microbial profiles and smoking and number of sexual partners were observed; however, these associations were not statistically significant. These results support and extend previous findings that there are significant differences in the vaginal microbiome related to ethnicity and demonstrate that these differences are pronounced even in healthy women.
    Microbiology 07/2014; DOI:10.1099/mic.0.081034-0 · 2.84 Impact Factor
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    ABSTRACT: Polycystic ovary syndrome (PCOS), characterized by increased ovarian androgen biosynthesis, anovulation, and infertility, affects 5-7% of reproductive-age women. Genome-wide association studies identified PCOS candidate loci that were replicated in subsequent reports, including DENND1A, which encodes a protein associated with clathrin-coated pits where cell-surface receptors reside. However, these studies provided no information about functional roles for DENND1A in the pathogenesis of PCOS. DENND1A protein was located in the cytoplasm as well as nuclei of theca cells, suggesting a possible role in gene regulation. DENND1A immunostaining was more intense in the theca of PCOS ovaries. Using theca cells isolated and propagated from normal cycling and PCOS women, we found that DENND1A variant 2 (DENND1A.V2) protein and mRNA levels are increased in PCOS theca cells. Exosomal DENND1A.V2 RNA was significantly elevated in urine from PCOS women compared with normal cycling women. Forced overexpression of DENND1A.V2 in normal theca cells resulted in a PCOS phenotype of augmented CYP17A1 and CYP11A1 gene transcription, mRNA abundance, and androgen biosynthesis. Knock-down of DENND1A.V2 in PCOS theca cells reduced androgen biosynthesis and CYP17A1 and CYP11A1 gene transcription. An IgG specific to DENND1A.V2 also reduced androgen biosynthesis and CYP17 and CYP11A1 mRNA when added to the medium of cultured PCOS theca cells. We conclude that the PCOS candidate gene, DENND1A, plays a key role in the hyperandrogenemia associated with PCOS. These observations have both diagnostic and therapeutic implications for this common disorder.
    Proceedings of the National Academy of Sciences 03/2014; 111(15). DOI:10.1073/pnas.1400574111 · 9.81 Impact Factor
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    ABSTRACT: To quantitate 2-methoxyestradiol (2-ME) in human corpus luteum (CL) of different ages and to determine the expression of cytochrome-P450-1A1 (CYP1A1) and catechol-O-methyl transferase (COMT) in CL and the action of 2-ME on P, vascular endothelial growth factor (VEGF) secretion, and luteal angiogenesis. Experimental study. University division of reproductive endocrinology. Twenty-four women of reproductive age. CL was collected from 15 women during the minilaparotomy for tubal sterilization. Granulosa lutein cells were harvested 36 hours after hCG administration in patients undergoing IVF. Levels of 2-ME were determined by high-performance liquid chromatography in CL. CYP1A1 and COMT were assessed by immunohistochemistry and Western blot. P and VEGF were measured by radioimmunoassay and ELISA. The angiogenic potential was analyzed using EA.hy926 cells. Plasma levels of E2 decreased in the late luteal phase in association with an increase in luteal tissue of 2-ME concentrations. Concomitantly, there was a significant reduction of angiogenic activity in late CL. There was no significant variation in CYP1A1 and COMT expression in all CL. In physiological doses, 2-ME inhibited basal VEGF by granulosa lutein cells and diminished the angiogenic activity in conditioned media but did not prevent P and VEGF production stimulated by hCG. These data suggest the participation of 2-ME in physiological luteolysis by reducing angiogenesis. However, 2-ME did not prevent in vitro hCG stimulation of P biosynthesis, providing a mechanism for CL rescue in the cycle of conception.
    Fertility and sterility 08/2013; DOI:10.1016/j.fertnstert.2013.07.1980 · 4.30 Impact Factor
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    ABSTRACT: Single nucleotide polymorphisms (SNPs) in the endoplasmic reticulum aminopeptidase 2 (ERAP2) gene are associated with preeclampsia (PE) in different populations. rs2549782, a coding variant (N392K) that significantly affects substrate specificity, is in linkage disequilibrium (LD) with rs2248374, a marker SNP associated with ERAP2 protein expression in previously studied populations. As a result of non-sense mediated RNA decay, ERAP2 protein is not expressed from the rs2248374 G allele. We previously reported that the fetal rs2549782 minor G allele is associated with PE in African-Americans, but not Chileans. In this study, we found that rs2549782 was in LD with rs2248374 in African-Americans, but not in Chileans. The unexpected lack of strong LD in Chileans raised the possibility that rs2248374 could be associated with PE in the absence of an association with rs2549782. However, we found no significant association for this allele with PE in Chileans. Chileans homozygous for the rs2248374 G allele did not express 110 kDa ERAP2 protein, consistent with non-sense mediated RNA decay, and carriers of the rs2248374 A allele did. We conclude that the Chilean ERAP2 haplotype structure allows for the expression of the major T allele of rs2549782 encoding 392N, which could impact peptide trimming and antigen presentation. Our discovery of racial differences in genetic structure and association with PE reveal here-to-fore unrecognized complexity of the ERAP2 locus.
    07/2013; 1(2):98-107. DOI:10.1002/mgg3.13
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    ABSTRACT: Although there is increasing evidence that genetic factors influence gestational age, it is unclear to what extent this is due to fetal and/or maternal genes. In this study, we apply a novel analytical model to estimate genetic and environmental contributions to pregnancy history records obtained from 165,952 Swedish families consisting of offspring of twins, full siblings, and half-siblings (1987-2008). Results indicated that fetal genetic factors explained 13.1% (95% confidence interval (CI): 6.8, 19.4) of the variation in gestational age at delivery, while maternal genetic factors accounted for 20.6% (95% CI: 18.1, 23.2). The largest contribution to differences in the timing of birth were environmental factors, of which 10.1% (95% CI: 7.0, 13.2) was due to factors shared by births of the same mother, and 56.2% (95% CI: 53.0, 59.4) was pregnancy specific. Similar models fit to the same data dichotomized at clinically meaningful thresholds (e.g., preterm birth) resulted in less stable parameter estimates, but the collective results supported a model of homogeneous genetic and environmental effects across the range of gestational age. Since environmental factors explained most differences in the timing of birth, genetic studies may benefit from understanding the specific effect of fetal and maternal genes in the context of these yet-unidentified factors.
    American journal of epidemiology 04/2013; 178(4). DOI:10.1093/aje/kwt005 · 4.98 Impact Factor
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    ABSTRACT: Primary ciliary dyskinesia (PCD), resulting from defects in cilia assembly or motility, is caused by mutations in a number of genes encoding axonemal proteins. PCD phenotypes are variable, and include recurrent respiratory tract infections, bronchiectasis, hydrocephaly, situs inversus and male infertility. We generated knockout mice for the Spag17 gene, which encodes a central pair (CP) protein present in the axonemes of cells with "9+2" motile cilia or flagella. Targeting of Spag17 resulted in a severe phenotype characterized by immotile nasal and tracheal cilia, reduced clearance of nasal mucus, profound respiratory distress associated with lung fluid accumulation and disruption of the alveolar epithelium, cerebral ventricular expansion consistent with emerging hydrocephalus, failure to suckle, and neonatal demise within 12 h of birth. Ultrastructural analysis revealed loss of one CP microtubule in approximately one quarter of tracheal cilia axonemes, absence of a C1 microtubule projection, as well as less frequent CP structural abnormalities. SPAG6 and SPAG16, CP proteins that interact with SPAG17, were increased in tracheal tissue from SPAG17-deficient mice. We conclude that Spag17 plays a critical role in the function and structure of motile cilia, and that the neonatal lethality is likely explained by impaired airway mucociliary clearance.
    American Journal of Respiratory Cell and Molecular Biology 02/2013; DOI:10.1165/rcmb.2012-0362OC · 4.11 Impact Factor
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    ABSTRACT: Meiosis expressed gene 1 (Meig1) was originally identified in a search for mammalian genes potentially involved in meiosis. Seven mouse Meig1 transcripts with the same coding region, but different 5'-UTRs, have been identified. These transcripts have different tissue distributions, two are only present in the testis. In the testis, Meig1 is present in germ cells and Sertoli cells. A Meig1 conditional knockout model has been generated. When Meig1 was inactivated globally by crossing with Cmv-Cre transgenic mice, the Meig1-deficient males were sterile due to severe spermiogenic defects, and had no obvious defects in meiosis. To further study its role in individual cell types in the testis, the Meig1(flox) mice were crossed with Hsp2a-Cre, Prm-Cre, and Amh-Cre mice, in which the Cre recombinase is driven by the heat shock protein 2 (Hsp2a) gene promoter (expressed in spermatocytes), the protamine 1 gene promoter (expressed in post-meiotic spermatids) and the anti-Mullerian hormone (Amh) gene promoter (expressed in Sertoli cells) respectively. Both Meig1 mRNA and protein were undetectable in testis of the Hsp2a-Cre; Meig1(flox/flox) mice and all the mutant adult males tested were sterile. This phenotype mirrors that of the Cmv-Cre; Meig1(flox/flox) mice. Even though the total testicular Meig1 mRNA and protein expression levels were dramatically reduced in testis of the Prm-Cre; Meig1(flox/flox) males, all the mice tested were fertile, and there was no significant difference in sperm count and sperm motility compared with age-matched Meig1(flox/flox) male mice. Disruption of Meig1 in the Sertoli cells did not affect the MEIG1 protein expression. Amh-Cre; Meig1(flox/flox) males were fertile, and produced the same amount of spermatozoa as age-matched Meig1(flox/flox) mice. The testicular histology was also normal. Our results indicate that MEIG1 regulates spermiogenesis through effects in germ cells alone, and that the Meig1 gene must be active during a discrete period in spermatogenesis after which it is dispensable.
    Andrology 01/2013; 1(1):37-46. DOI:10.1111/j.2047-2927.2012.00001.x · 3.37 Impact Factor
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    ABSTRACT: Hyperandrogenism is characteristic of women with polycystic ovary syndrome (PCOS). Ovarian theca cells isolated from PCOS follicles and maintained in long-term culture produce elevated levels of progestins and androgens compared to normal theca cells. Augmented steroid production in PCOS theca cells is associated with changes in the expression of genes for several steroidogenic enzymes, including CYP11A1, which encodes cytochrome P450 cholesterol side-chain cleavage. Here, we further examined CYP11A1 gene expression, at both the transcriptional and post-transcriptional level in normal and PCOS theca cells propagated in long-term culture utilizing quantitative RT-PCR, functional promoter analyses, and mRNA degradation studies. The minimal element(s) that conferred increased basal and cAMP-dependent CYP11A1 promoter function were determined. CYP11A1 mRNA half-life in normal and PCOS theca cells was compared. Results of these cumulative studies showed that basal and forskolin stimulated steady state CYP11A1 mRNA abundance and CYP11A1 promoter activity were increased in PCOS theca cells. Deletion analysis of the CYP11A1 promoter demonstrated that augmented promoter function in PCOS theca cells results from increased basal regulation conferred by a minimal sequence between -160 and -90 bp of the transcriptional start site. The transcription factor, nuclear factor 1C2, was observed to regulate basal activity of this minimal CYP11A1 element. Examination of mRNA stability in normal and PCOS theca cells demonstrated that CYP11A1 mRNA half-life increased >2-fold, from approximately 9.22+/-1.62 h in normal cells, to 22.38+/-0.92 h in PCOS cells. Forskolin treatment did not prolong CYP11A1 mRNA stability in either normal or PCOS theca cells. The 5'-UTR of CYP11A1 mRNA confers increased basal mRNA stability in PCOS cells. In conclusion, these studies show that elevated steady state CYP11A1 mRNA abundance in PCOS cells results from increased transactivation of the CYP11A1 promoter and increased CYP11A1 mRNA stability.
    PLoS ONE 11/2012; 7(11):e48963. DOI:10.1371/journal.pone.0048963 · 3.53 Impact Factor
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    ABSTRACT: Background SPAG16 is a critical structural component of motile cilia and flagella. In the eukaryotic unicellular algae Chlamydomonas, loss of gene function causes flagellar paralysis and prevents assembly of the “9 + 2” axoneme central pair. In mice, we have previously shown that loss of Spag16 gene function causes male infertility and severe sperm motility defects. We have also reported that a heterozygous mutation of the human SPAG16 gene reduces stability of the sperm axonemal central apparatus. Methods In the present study, we analyzed DNA samples from 60 infertile male volunteers of Western European (Italian) origin, to search for novel SPAG16 gene mutations, and to determine whether increased prevalence of SPAG16 single nucleotide polymorphisms (SNPs) was associated with infertility phenotypes. Semen parameters were evaluated by light microscopy and sperm morphology was comprehensively analyzed by transmission electron microscopy (TEM). Results For gene analysis, sequences were generated covering exons encoding the conserved WD40 repeat region of the SPAG16 protein and the flanking splice junctions. No novel mutations were found, and the four SNPs in the assessed gene region were present at expected frequencies. The minor alleles were not associated with any assessed sperm parameter in the sample population. Conclusions Analysis of the SPAG16 regions encoding the conserved WD repeats revealed no evidence for association of mutations or genetic variation with sperm motility and ultrastructural sperm characteristics in a cohort of Italian infertile males.
    BMC Urology 09/2012; 12(1):27. DOI:10.1186/1471-2490-12-27 · 1.94 Impact Factor
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    ABSTRACT: We analyzed 27 578 CpG sites that map to 14 495 genes in omental arteries of normal pregnant and preeclamptic women for DNA methylation status using the Illumina platform. We found 1685 genes with a significant difference in DNA methylation at a false discovery rate of <10% with many inflammatory genes having reduced methylation. Unsupervised hierarchical clustering revealed natural clustering by diagnosis and methylation status. Of the genes with significant methylation differences, 236 were significant at a false discovery rate of <5%. When data were analyzed more stringently to a false discovery rate of <5% and difference in methylation of >0.10, 65 genes were identified, all of which showed reduced methylation in preeclampsia. When these genes were mapped to gene ontology for molecular functions and biological processes, 75 molecular functions and 149 biological processes were overrepresented in the preeclamptic vessels. These included smooth muscle contraction, thrombosis, inflammation, redox homeostasis, sugar metabolism, and amino acid metabolism. We speculate that reduced methylation may contribute to the pathogenesis of preeclampsia and that alterations in DNA methylation resulting from preeclampsia may increase maternal risk of cardiovascular disease later in life.
    Reproductive sciences (Thousand Oaks, Calif.) 08/2012; 19(12). DOI:10.1177/1933719112450336 · 2.18 Impact Factor
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    ABSTRACT: Context: Polycystic ovary syndrome (PCOS) is a heterogeneous, genetically complex, endocrine disorder of uncertain etiology in women. Objective: Our aim was to compare the gene expression profiles in stimulated granulosa cells of PCOS women with and without insulin resistance vs. matched controls. Research Design and Methods: This study included 12 normal ovulatory women (controls), 12 women with PCOS without evidence for insulin resistance (PCOS non-IR), and 16 women with insulin resistance (PCOS-IR) undergoing in vitro fertilization. Granulosa cell gene expression profiling was accomplished using Affymetrix Human Genome-U133 arrays. Differentially expressed genes were classified according to gene ontology using ingenuity pathway analysis tools. Microarray results for selected genes were confirmed by real-time quantitative PCR. Results: A total of 211 genes were differentially expressed in PCOS non-IR and PCOS-IR granulosa cells (fold change ≥ 1.5; P ≤ 0.001) vs. matched controls. Diabetes mellitus and inflammation genes were significantly increased in PCOS-IR patients. Real-time quantitative PCR confirmed higher expression of NCF2 (2.13-fold), TCF7L2 (1.92-fold), and SERPINA1 (5.35-fold). Increased expression of inflammation genes ITGAX (3.68-fold) and TAB2 (1.86-fold) was confirmed in PCOS non-IR. Different cardiometabolic disease genes were differentially expressed in the two groups. Decreased expression of CAV1 (-3.58-fold) in PCOS non-IR and SPARC (-1.88-fold) in PCOS-IR was confirmed. Differential expression of genes involved in TGF-β signaling (IGF2R, increased; and HAS2, decreased), and oxidative stress (TXNIP, increased) was confirmed in both groups. Conclusions: Microarray analysis demonstrated differential expression of genes linked to diabetes mellitus, inflammation, cardiovascular diseases, and infertility in the granulosa cells of PCOS women with and without insulin resistance. Because these dysregulated genes are also involved in oxidative stress, lipid metabolism, and insulin signaling, we hypothesize that these genes may be involved in follicular growth arrest and metabolic disorders associated with the different phenotypes of PCOS.
    The Journal of Clinical Endocrinology and Metabolism 08/2012; 97(10):E2016-21. DOI:10.1210/jc.2011-3441 · 6.31 Impact Factor

Publication Stats

19k Citations
1,978.43 Total Impact Points

Institutions

  • 2006–2015
    • Virginia Commonwealth University
      • • Department of Obstetrics and Gynecology
      • • Department of Biochemistry and Molecular Biology
      Ричмонд, Virginia, United States
  • 2003–2013
    • Hospital Clinico San Borja Arriaran
      CiudadSantiago, Santiago, Chile
  • 2012
    • Johns Hopkins Medicine
      Baltimore, Maryland, United States
  • 2007
    • University of Nebraska at Lincoln
      • Department of Animal Science
      Lincoln, Nebraska, United States
    • University of Arkansas at Little Rock
      Little Rock, Arkansas, United States
  • 2000–2007
    • University of Santiago, Chile
      CiudadSantiago, Santiago Metropolitan, Chile
  • 1982–2007
    • University of Pennsylvania
      • • Department of Medicine
      • • Center for Research on Reproduction and Women's Health
      • • Department of Genetics
      • • Department of Obstetrics and Gynecology
      • • Division of Cardiothoracic Surgery
      Philadelphia, Pennsylvania, United States
  • 2005–2006
    • Northwestern University
      • Division of Endocrinology, Metabolism and Molecular Medicine
      Evanston, Illinois, United States
  • 1983–2006
    • Hospital of the University of Pennsylvania
      • • Department of Ophthalmology
      • • Department of Obstetrics and Gynecology
      • • Department of Pathology and Laboratory Medicine
      Philadelphia, Pennsylvania, United States
  • 1998–2005
    • Penn State Hershey Medical Center and Penn State College of Medicine
      • Cellular and Molecular Physiology
      Hershey, Pennsylvania, United States
  • 1983–2005
    • William Penn University
      Filadelfia, Pennsylvania, United States
  • 2004
    • Albert Einstein College of Medicine
      New York, New York, United States
  • 2002
    • National Institutes of Health
      • Chemical Biology Laboratory
      Bethesda, MD, United States
  • 2001
    • University of Chile
      • Departamento de Obstetricia y Ginecología
      CiudadSantiago, Santiago, Chile
  • 1999
    • Harvard Medical School
      Boston, Massachusetts, United States
    • Harvard University
      Cambridge, Massachusetts, United States
  • 1988–1999
    • University of California, San Francisco
      • Department of Pediatrics
      San Francisco, California, United States
    • Kimron Veterinary Institute
      Beit Dajan, Central District, Israel
  • 1997
    • Universidad Nacional Autónoma de México
      • Department of Biochemistry
      Ciudad de México, Mexico City, Mexico
  • 1996
    • Instituto Nacional de Perinatología
      Ciudad de México, The Federal District, Mexico
  • 1995
    • Montefiore Medical Center
      • Department of Pediatrics
      New York, New York, United States
  • 1983–1995
    • Temple University
      Filadelfia, Pennsylvania, United States
  • 1994
    • University of California, Los Angeles
      • Department of Medicine
      Los Ángeles, California, United States
  • 1991
    • University of Texas at San Antonio
      San Antonio, Texas, United States
    • University of California, San Diego
      • Department of Medicine
      San Diego, CA, United States
  • 1987
    • University of San Francisco
      Philadelphia, Pennsylvania, United States