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1.
RNA Biol ; 20(1): 186-197, 2023 01.
Artículo en Inglés | MEDLINE | ID: mdl-37095747

RESUMEN

Here, we provide an in-depth analysis of the usefulness of single-sample metabolite/RNA extraction for multi-'omics readout. Using pulverized frozen livers of mice injected with lymphocytic choriomeningitis virus (LCMV) or vehicle (Veh), we isolated RNA prior (RNA) or following metabolite extraction (MetRNA). RNA sequencing (RNAseq) data were evaluated for differential expression analysis and dispersion, and differential metabolite abundance was determined. Both RNA and MetRNA clustered together by principal component analysis, indicating that inter-individual differences were the largest source of variance. Over 85% of LCMV versus Veh differentially expressed genes were shared between extraction methods, with the remaining 15% evenly and randomly divided between groups. Differentially expressed genes unique to the extraction method were attributed to randomness around the 0.05 FDR cut-off and stochastic changes in variance and mean expression. In addition, analysis using the mean absolute difference showed no difference in the dispersion of transcripts between extraction methods. Altogether, our data show that prior metabolite extraction preserves RNAseq data quality, which enables us to confidently perform integrated pathway enrichment analysis on metabolomics and RNAseq data from a single sample. This analysis revealed pyrimidine metabolism as the most LCMV-impacted pathway. Combined analysis of genes and metabolites in the pathway exposed a pattern in the degradation of pyrimidine nucleotides leading to uracil generation. In support of this, uracil was among the most differentially abundant metabolites in serum upon LCMV infection. Our data suggest that hepatic uracil export is a novel phenotypic feature of acute infection and highlight the usefulness of our integrated single-sample multi-'omics approach.


Asunto(s)
Metabolómica , Virosis , Animales , Ratones , Análisis de Secuencia de ARN , Hígado , ARN
2.
JCI Insight ; 7(20)2022 10 24.
Artículo en Inglés | MEDLINE | ID: mdl-36066972

RESUMEN

Uterine fibroids (leiomyomas) affect Black women disproportionately compared with women of other races and ethnicities in terms of prevalence, incidence, and severity of symptoms. The causes of this racial disparity are essentially unknown. We hypothesized that myometria of Black women are more susceptible to developing fibroids, and we examined the transcriptomic and DNA methylation profiles of myometria and fibroids from Black and White women for comparison. Myometrial samples cluster by race in both their transcriptome and DNA methylation profiles, whereas fibroid samples only cluster by race in the latter. More differentially expressed genes (DEGs) were detected in the Black and White myometrial sample comparison than in the fibroid comparison. Leiomyoma gene set expression analysis identified 4 clusters of DEGs, including a cluster of 24 genes with higher expression in myometrial samples from Black women. One of the DEGs in this group, von Willibrands factor (VWF), was significantly hypomethylated in both myometrial samples from Black women and in all fibroids at 2 CpG probes that are near a putative enhancer site and that are correlated with VWF expression levels. These results suggest that the molecular basis for the disparity in fibroid disease between Black and White women could be found in the myometria before fibroid development and not in the fibroids themselves.


Asunto(s)
Leiomioma , Neoplasias Uterinas , Femenino , Humanos , Neoplasias Uterinas/genética , Neoplasias Uterinas/epidemiología , Neoplasias Uterinas/metabolismo , Transcriptoma , Epigenoma , Factor de von Willebrand/genética , Leiomioma/genética , Leiomioma/epidemiología , Leiomioma/metabolismo
3.
Horm Behav ; 121: 104719, 2020 05.
Artículo en Inglés | MEDLINE | ID: mdl-32081742

RESUMEN

Aromatase catalyzes conversion of testosterone to estradiol and is expressed in a variety of tissues, including the brain. Suppression of aromatase adversely affects metabolism and physical activity behavior, but mechanisms remain uncertain. The hypothesis tested herein was that whole body aromatase deletion would cause gene expression changes in the nucleus accumbens (NAc), a brain regulating motivated behaviors such as physical activity, which is suppressed with loss of estradiol. Metabolic and behavioral assessments were performed in male and female wild-type (WT) and aromatase knockout (ArKO) mice. NAc-specific differentially expressed genes (DEGs) were identified with RNAseq, and associations between the measured phenotypic traits were determined. Female ArKO mice had greater percent body fat, reduced spontaneous physical activity (SPA), consumed less energy, and had lower relative resting energy expenditure (REE) than WT females. Such differences were not observed in ArKO males. However, in both sexes, a top DEG was Pts, a gene encoding an enzyme necessary for catecholamine (e.g., dopamine) biosynthesis. In comparing male and female WT mice, top DEGs were related to sexual development/fertility, immune regulation, obesity, dopamine signaling, and circadian regulation. SPA correlated strongly with Per3, a gene regulating circadian function, thermoregulation, and metabolism (r = -0.64, P = .002), which also correlated with adiposity (r = 0.54, P = .01). In conclusion, aromatase ablation leads to gene expression changes in NAc, which may in turn result in reduced SPA and related metabolic abnormalities. These findings may have significance to post-menopausal women and those treated with an aromatase inhibitor.


Asunto(s)
Aromatasa/genética , Actividad Motora/genética , Núcleo Accumbens/metabolismo , Animales , Aromatasa/metabolismo , Metabolismo Energético/genética , Estradiol/metabolismo , Femenino , Expresión Génica , Regulación Enzimológica de la Expresión Génica , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Caracteres Sexuales , Testosterona/metabolismo
4.
BMC Res Notes ; 12(1): 522, 2019 Aug 19.
Artículo en Inglés | MEDLINE | ID: mdl-31426836

RESUMEN

OBJECTIVE: Soybean seed development is negatively impacted by elevated temperatures during seed fill, which can decrease seed quality and economic value. Prior germplasm screens identified an exotic landrace able to maintain ~ 95% seed germination under stress conditions that reduce germination dramatically (> 50%) for typical soybean seeds. Seed transcriptomic analysis was performed for two soybean lines (a heat-tolerant landrace and a typical high-yielding adapted line) for dry, mature seed, 6-h imbibed seed and germinated seed. Seeds were produced in two environments: a typical Midwestern field and a heat stressed field located in the Midsouth soybean production region. RESULTS: Transcriptomic analysis revealed 23-30K expressed genes in each seed tissue sample, and differentially expressed genes (DEGs) with ≥ twofold gene expression differences (at q-value < 0.05) comprised ~ 5-44% of expressed genes. Gene ontology (GO) enrichment analysis on DEGs revealed enrichment in heat-tolerant seeds for genes annotated for general and temperature-specific stress, as well as protein-refolding. DEGs were also clustered in modules using weighted co-expressed gene network analysis, which were examined for enrichment of GO biological process terms. Collectively, our results provide new and valuable insights into this unique form of genetic abiotic stress tolerance and to soybean seed physiological responses to elevated temperatures.


Asunto(s)
Adaptación Fisiológica/genética , Germinación/genética , Glycine max/genética , Calor , Semillas/genética , Transcriptoma/genética , Perfilación de la Expresión Génica/métodos , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Ontología de Genes , Redes Reguladoras de Genes , Genotipo , Peso Molecular , Semillas/crecimiento & desarrollo , Proteínas de Soja/química , Proteínas de Soja/genética , Glycine max/clasificación , Glycine max/crecimiento & desarrollo , Especificidad de la Especie
5.
Microbiol Resour Announc ; 8(14)2019 Apr 04.
Artículo en Inglés | MEDLINE | ID: mdl-30948465

RESUMEN

Here, we present the draft genome sequence of Bradyrhizobium sp. strain LVM 105, a soil bacterium that forms nitrogen-fixing nodules on the roots of partridge pea. The genome size was 8,386,213 bp, with a GC content of 63.8%. The draft assembly contained 97 scaffolds with an N 50 value of 415,026 bp.

6.
BMC Genomics ; 20(1): 87, 2019 Jan 25.
Artículo en Inglés | MEDLINE | ID: mdl-30683046

RESUMEN

BACKGROUND: Brain sexual differentiation is sculpted by precise coordination of steroid hormones during development. Programming of several brain regions in males depends upon aromatase conversion of testosterone to estrogen. However, it is not clear the direct contribution that Y chromosome associated genes, especially sex-determining region Y (Sry), might exert on brain sexual differentiation in therian mammals. Two species of spiny rats: Amami spiny rat (Tokudaia osimensis) and Tokunoshima spiny rat (T. tokunoshimensis) lack a Y chromosome/Sry, and these individuals possess an XO chromosome system in both sexes. Both Tokudaia species are highly endangered. To assess the neural transcriptome profile in male and female Amami spiny rats, RNA was isolated from brain samples of adult male and female spiny rats that had died accidentally and used for RNAseq analyses. RESULTS: RNAseq analyses confirmed that several genes and individual transcripts were differentially expressed between males and females. In males, seminal vesicle secretory protein 5 (Svs5) and cytochrome P450 1B1 (Cyp1b1) genes were significantly elevated compared to females, whereas serine (or cysteine) peptidase inhibitor, clade A, member 3 N (Serpina3n) was upregulated in females. Many individual transcripts elevated in males included those encoding for zinc finger proteins, e.g. zinc finger protein X-linked (Zfx). CONCLUSIONS: This method successfully identified several genes and transcripts that showed expression differences in the brain of adult male and female Amami spiny rat. The functional significance of these findings, especially differential expression of transcripts encoding zinc finger proteins, in this unusual rodent species remains to be determined.


Asunto(s)
Encéfalo/metabolismo , Murinae/genética , Caracteres Sexuales , Transcriptoma , Animales , Femenino , Perfilación de la Expresión Génica , Masculino , Murinae/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Análisis de Secuencia de ARN , Cromosoma Y
7.
Genome Biol ; 19(1): 122, 2018 08 23.
Artículo en Inglés | MEDLINE | ID: mdl-30134966

RESUMEN

The original version [1] of this article unfortunately contained a mistake. The additive effects of the eQTLs of lncRNAs were flipped, meaning that the base allele in the contrast to derive the additive effects should have been B73, rather than Mo17, due to the original coding of biallele SNPs as "0s" and "1s". Going through the entire analysis procedure, it was determined that the mistake was made while tabulating the eQTL results from QTL Cartographer.

8.
New Phytol ; 219(2): 697-713, 2018 07.
Artículo en Inglés | MEDLINE | ID: mdl-29726613

RESUMEN

Cyst nematodes deliver effector proteins into host cells to manipulate cellular processes and establish a metabolically hyperactive feeding site. The novel 30D08 effector protein is produced in the dorsal gland of parasitic juveniles, but its function has remained unknown. We demonstrate that expression of 30D08 contributes to nematode parasitism, the protein is packaged into secretory granules and it is targeted to the plant nucleus where it interacts with SMU2 (homolog of suppressor of mec-8 and unc-52 2), an auxiliary spliceosomal protein. We show that SMU2 is expressed in feeding sites and an smu2 mutant is less susceptible to nematode infection. In Arabidopsis expressing 30D08 under the SMU2 promoter, several genes were found to be alternatively spliced and the most abundant functional classes represented among differentially expressed genes were involved in RNA processing, transcription and binding, as well as in development, and hormone and secondary metabolism, representing key cellular processes known to be important for feeding site formation. In conclusion, we demonstrated that the 30D08 effector is secreted from the nematode and targeted to the plant nucleus where its interaction with a host auxiliary spliceosomal protein may alter the pre-mRNA splicing and expression of a subset of genes important for feeding site formation.


Asunto(s)
Arabidopsis/genética , Arabidopsis/parasitología , Núcleo Celular/metabolismo , Conducta Alimentaria , Regulación de la Expresión Génica de las Plantas , Proteínas del Helminto/metabolismo , Interacciones Huésped-Parásitos/genética , Tylenchoidea/metabolismo , Empalme Alternativo/genética , Secuencia de Aminoácidos , Animales , Genes de Plantas , Proteínas del Helminto/química , Estadios del Ciclo de Vida , Señales de Localización Nuclear , Parásitos/metabolismo , Células Vegetales/metabolismo , Hojas de la Planta/metabolismo , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo , Raíces de Plantas/parasitología , Regiones Promotoras Genéticas/genética , Unión Proteica , Interferencia de ARN , Plantones/metabolismo , Tylenchoidea/crecimiento & desarrollo , Regulación hacia Arriba
9.
Sci Rep ; 8(1): 4065, 2018 03 06.
Artículo en Inglés | MEDLINE | ID: mdl-29511208

RESUMEN

Countless studies have identified differences between the gut microbiota of humans affected with myriad conditions and healthy individuals, and animal models are commonly used to determine whether those differences are causative or correlative. Recently, concerns have arisen regarding the reproducibility of animal models between institutions and across time. To determine the influence of three common husbandry-associated factors that vary between institutions, groups of weanling mice were placed in either static or ventilated microisolator caging, with either aspen or paperchip bedding, and with one of three commonly used rodent chows, in a fully crossed study design. After thirteen weeks, samples were collected from multiple regions of the gastrointestinal tract and characterized using culture-independent sequencing methods. Results demonstrated that seemingly benign husbandry factors can interact to induce profound changes in the composition of the microbiota present in certain regions of the gut, most notably the cecum, and that those changes are muted during colonic transit. These findings indicate that differences in factors such as caging and bedding can interact to modulate the gut microbiota that in turn may affect reproducibility of some animal models, and that cecal samples might be optimal when screening environmental effects on the gut microbiota.


Asunto(s)
Crianza de Animales Domésticos/métodos , Dieta/métodos , Microbioma Gastrointestinal , Tracto Gastrointestinal/microbiología , Animales , Metagenómica , Ratones , Modelos Animales , Reproducibilidad de los Resultados
10.
Sci Rep ; 7(1): 2822, 2017 06 06.
Artículo en Inglés | MEDLINE | ID: mdl-28588204

RESUMEN

Due to their antimicrobial properties, silver nanoparticles (AgNPs) are being used in non-edible and edible consumer products. It is not clear though if exposure to these chemicals can exert toxic effects on the host and gut microbiome. Conflicting studies have been reported on whether AgNPs result in gut dysbiosis and other changes within the host. We sought to examine whether exposure of Sprague-Dawley male rats for two weeks to different shapes of AgNPs, cube (AgNC) and sphere (AgNS) affects gut microbiota, select behaviors, and induces histopathological changes in the gastrointestinal system and brain. In the elevated plus maze (EPM), AgNS-exposed rats showed greater number of entries into closed arms and center compared to controls and those exposed to AgNC. AgNS and AgNC treated groups had select reductions in gut microbiota relative to controls. Clostridium spp., Bacteroides uniformis, Christensenellaceae, and Coprococcus eutactus were decreased in AgNC exposed group, whereas, Oscillospira spp., Dehalobacterium spp., Peptococcaeceae, Corynebacterium spp., Aggregatibacter pneumotropica were reduced in AgNS exposed group. Bacterial reductions correlated with select behavioral changes measured in the EPM. No significant histopathological changes were evident in the gastrointestinal system or brain. Findings suggest short-term exposure to AgNS or AgNC can lead to behavioral and gut microbiome changes.


Asunto(s)
Disbiosis/microbiología , Microbioma Gastrointestinal/efectos de los fármacos , Nanopartículas del Metal/efectos adversos , Aggregatibacter/efectos de los fármacos , Animales , Bacteroides/efectos de los fármacos , Encéfalo/efectos de los fármacos , Encéfalo/fisiopatología , Clostridium/efectos de los fármacos , Corynebacterium/efectos de los fármacos , Disbiosis/inducido químicamente , Disbiosis/fisiopatología , Heces/microbiología , Microbioma Gastrointestinal/genética , Tracto Gastrointestinal/efectos de los fármacos , Tracto Gastrointestinal/fisiopatología , Humanos , Nanopartículas del Metal/administración & dosificación , Peptococcus/efectos de los fármacos , Ratas , Ratas Sprague-Dawley
11.
Physiol Genomics ; 49(4): 201-215, 2017 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-28159858

RESUMEN

Developmental exposure of turtles and other reptiles to endocrine-disrupting chemicals (EDCs), including bisphenol A (BPA) and ethinyl estradiol (EE), can stimulate partial to full gonadal sex-reversal in males. We have also recently shown that in ovo exposure to either EDC can induce similar sex-dependent behavioral changes typified by improved spatial learning and memory or possibly feminized brain responses. Observed behavioral changes are presumed to be due to BPA- and EE-induced brain transcriptomic alterations during development. To test this hypothesis, we treated painted turtles (Chrysemys picta) at developmental stage 17, incubated at 26°C (male-inducing temperature), with 1) BPA (1 ng/µl), 2) EE (4 ng/µl), or 3) vehicle ethanol (control group). Ten months after hatching and completion of the behavioral tests, juvenile turtles were euthanized, brains were collected and frozen in liquid nitrogen, and RNA was isolated for RNA-Seq analysis. Turtles exposed to BPA clustered separately from EE-exposed and control individuals. More transcripts and gene pathways were altered in BPA vs. EE individuals. The one transcript upregulated in both BPA- and EE-exposed individuals was the mitochondrial-associated gene, ND5, which is involved in oxidative phosphorylation. Early exposure of turtles to BPA increases transcripts linked with ribosomal and mitochondrial functions, especially bioenergetics, which has been previously linked with improved cognitive performance. In summary, even though both BPA and EE resulted in similar behavioral alterations, they diverge in the pattern of neural transcript alterations with early BPA significantly upregulating several genes involved in oxidative phosphorylation, mitochondrial activity, and ribosomal function, which could enhance cognitive performance.


Asunto(s)
Compuestos de Bencidrilo/toxicidad , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Etinilestradiol/toxicidad , Fenoles/toxicidad , Transcriptoma/efectos de los fármacos , Tortugas/genética , Animales , Femenino , Masculino
12.
Physiol Rep ; 5(3)2017 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-28196854

RESUMEN

Bisphenol A (BPA) is an endocrine-disrupting chemical (EDC) prevalent in many household items. Rodent models and human epidemiological studies have linked this chemical to neurobehavior impairments. In California mice, developmental exposure to BPA results in sociosexual disorders at adulthood, including communication and biparental care deficits, behaviors that are primarily regulated by the hypothalamus. Thus, we sought to examine the transcriptomic profile in this brain region of juvenile male and female California mice offspring exposed from periconception through lactation to BPA or ethinyl estradiol (EE, estrogen present in birth control pills and considered a positive estrogen control for BPA studies). Two weeks prior to breeding, P0 females were fed a control diet, or this diet supplemented with 50 mg BPA/kg feed weight or 0.1 ppb EE, and continued on the diets through lactation. At weaning, brains from male and female offspring were collected, hypothalamic RNA isolated, and RNA-seq analysis performed. Results indicate that BPA and EE groups clustered separately from controls with BPA and EE exposure leading to unique set of signature gene profiles. Kcnd3 was downregulated in the hypothalamus of BPA- and EE-exposed females, whereas Tbl2, Topors, Kif3a, and Phactr2 were upregulated in these groups. Comparison of transcripts differentially expressed in BPA and EE groups revealed significant enrichment of gene ontology terms associated with microtubule-based processes. Current results show that perinatal exposure to BPA or EE can result in several transcriptomic alterations, including those associated with microtubule functions, in the hypothalamus of California mice. It remains to be determined whether these genes mediate BPA-induced behavioral disruptions.


Asunto(s)
Compuestos de Bencidrilo/toxicidad , Disruptores Endocrinos/toxicidad , Etinilestradiol/toxicidad , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Fenoles/toxicidad , Efectos Tardíos de la Exposición Prenatal/metabolismo , Transcriptoma/efectos de los fármacos , Animales , Regulación hacia Abajo , Femenino , Masculino , Exposición Materna , Peromyscus , Embarazo , Efectos Tardíos de la Exposición Prenatal/inducido químicamente , Regulación hacia Arriba
13.
Sci Total Environ ; 579: 1804-1814, 2017 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-27932218

RESUMEN

Bisphenol A (BPA) is a widely present endocrine disruptor chemical found in many household items. Moreover, this chemical can bioaccumulate in various terrestrial and aquatic sources; thereby ensuring continual exposure of animals and humans. For most species, including humans, diet is considered the primary route of exposure. However, there has been little investigation whether commercial-brands of dog foods contain BPA and potential health ramifications of BPA-dietary exposure in dogs. We sought to determine BPA content within dog food, whether short-term consumption of these diets increases serum concentrations of BPA, and potential health consequences, as assessed by potential hematological, serum chemistry, cortisol, DNA methylation, and gut microbiome changes, in dogs associated with short-term dietary exposure to BPA. Fourteen healthy privately-owned dogs were used in this study. Blood and fecal samples were collected prior to dogs being placed for two-weeks on one of two diets (with one considered to be BPA-free), and blood and fecal samples were collected again. Serum/plasma samples were analyzed for chemistry and hematology profiles, cortisol concentrations, 5-methylcytosine in lymphocytes, and total BPA concentrations. Fecal samples were used for microbiome assessments. Both diets contained BPA, and after two-weeks of being on either diet, dogs had a significant increase in circulating BPA concentrations (pre-samples=0.7±0.15ng/mL, post-samples=2.2±0.15ng/mL, p<0.0001). Elevated BPA concentrations positively correlated with increased plasma bicarbonate concentrations and associated with fecal microbiome alterations. Short-term feeding of canned dog food increased circulating BPA concentrations in dogs comparable to amounts detected in humans, and greater BPA concentrations were associated with serum chemistry and microbiome changes. Dogs, who share our internal and external environments with us, are likely excellent indicators of potential human health concerns to BPA and other environmental chemicals. These findings may also have relevance to aquatic and terrestrial wildlife.


Asunto(s)
Compuestos de Bencidrilo/sangre , Exposición Dietética/análisis , Disruptores Endocrinos/sangre , Contaminación de Alimentos/análisis , Alimentos en Conserva/análisis , Fenoles/sangre , Animales , Compuestos de Bencidrilo/toxicidad , Perros/sangre , Disruptores Endocrinos/análisis , Disruptores Endocrinos/toxicidad , Contaminación de Alimentos/estadística & datos numéricos , Mascotas/sangre , Fenoles/toxicidad
14.
Reprod Fertil Dev ; 29(8): 1602-1612, 2017 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-27569192

RESUMEN

Our prior work showed that a novel microbiome resides in the seminal vesicles of wild-type and oestrogen receptor α (Esr1) knock-out mice and is impacted by the presence of functional Esr1 genes. The seminal fluid microbiome (SFM) may influence the health and reproductive status of the male, along with that of his partner and offspring. A high-fat diet (HFD) alters metabolites and other factors within seminal fluid and might affect the SFM. Adult (~15 weeks old) male mice were placed for 4 weeks on a control or high-fat diet and seminal fluid and fecal samples were collected, bacterial DNA isolated and subjected to 16s rRNA sequencing. Corynebacterium spp. were elevated in the seminal fluid of HFD males; however, Acinetobacter johnsonii, Streptophyta, Ammoniphilus spp., Bacillus spp. and Propionibacterium acnes were increased in control males. Rikenellaceae was more abundant in the fecal samples from HFD males. However, Bacteroides ovatus and another Bacteroides species, Bilophila, Sutterella spp., Parabacteroides, Bifidobacterium longum, Akkermansia muciniphila and Desulfovibrio spp. were greater in control males. Thus, short-term consumption of a HFD influences the seminal fluid and fecal microbiomes, which may have important health consequence for males and developmental origins of health and disease effects in resulting offspring.


Asunto(s)
Dieta Alta en Grasa , Microbioma Gastrointestinal/fisiología , Semen/fisiología , Animales , Heces/microbiología , Masculino , Ratones
15.
Gut Microbes ; 7(6): 471-485, 2016 11.
Artículo en Inglés | MEDLINE | ID: mdl-27624382

RESUMEN

Gut dysbiosis may result in various diseases, such as metabolic and neurobehavioral disorders. Exposure to endocrine disrupting chemicals (EDCs), including bisphenol A (BPA) and ethinyl estradiol (EE), especially during development, may also increase the risk for such disorders. An unexplored possibility is that EDC-exposure might alter the gut microbial composition. Gut flora and their products may thus be mediating factors for the disease-causing effects of these chemicals. To examine the effects of EDCs on the gut microbiome, female and male monogamous and biparental California mice (Peromyscus californicus) were exposed to BPA (50 mg/kg feed weight) or EE (0.1 ppb) or control diet from periconception through weaning. 16s rRNA sequencing was performed on bacterial DNA isolated from fecal samples, and analyses performed for P0 and F1 males and females. Both BPA and EE induced generational and sex-dependent gut microbiome changes. Many of the bacteria, e.g. Bacteroides, Mollicutes, Prevotellaceae, Erysipelotrichaceae, Akkermansia, Methanobrevibacter, Sutterella, whose proportions increase with exposure to BPA or EE in the P0 or F1 generation are associated with different disorders, such as inflammatory bowel disease (IBD), metabolic disorders, and colorectal cancer. However, the proportion of the beneficial bacterium, Bifidobacterium, was also elevated in fecal samples of BPA- and EE-exposed F1 females. Intestinal flora alterations were also linked to changes in various metabolic and other pathways. Thus, BPA and EE exposure may disrupt the normal gut flora, which may in turn result in systemic effects. Probiotic supplementation might be an effective means to mitigate disease-promoting effects of these chemicals.


Asunto(s)
Bacterias/efectos de los fármacos , Compuestos de Bencidrilo/toxicidad , Modelos Animales de Enfermedad , Etinilestradiol/toxicidad , Microbioma Gastrointestinal/efectos de los fármacos , Tracto Gastrointestinal/microbiología , Peromyscus/microbiología , Fenoles/toxicidad , Animales , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Femenino , Masculino
17.
J Appl Physiol (1985) ; 120(11): 1277-87, 2016 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-26968026

RESUMEN

5-HT neurons contribute to autoresuscitation and survival during intermittent severe hypoxia (IsH). In adults, catecholaminergic neurons in the ventrolateral medulla (VLM) contribute to the autonomic response to hypoxia. We hypothesized that 1) catecholaminergic neurons in the neonatal VLM are activated following IsH, 2) this activation is compromised following an acute loss of brain stem 5-HT, and 3) IsH induces cellular and/or transcriptomic plasticity within catecholaminergic and serotonergic neurons that are within or project to the VLM, respectively. To test these hypotheses, we treated rat pups with 6-fluorotryptophan, a tryptophan hydroxylase (TPH) inhibitor, and then exposed treated and vehicle controls to IsH or air. Along with immunohistochemistry to detect tyrosine hydroxylase (TH)- or Fos-positive neurons, we used RNA sequencing to resolve the effects of IsH and 5-HT deficiency on the expression of serotonergic and catecholaminergic system genes in the VLM. 5-HT deficiency compromised autoresuscitation and survival. IsH significantly increased the number of identifiable TH-positive VLM neurons, an effect enhanced by 5-HT deficiency (P = 0.003). Contrary to our hypothesis, 5-HT-deficient pups had significantly more Fos-positive neurons following IsH (P = 0.008) and more activated TH-positive neurons following IsH or air (P = 0.04). In both groups the expression of the 5-HT transporter and TPH2 was increased following IsH. In 5-HT-deficient pups, the expression of the inhibitory 5-HT1A receptor was decreased following IsH, while the expression of DOPA decarboxylase was increased. These data show that the serotonergic and catecholaminergic systems in the VLM of the neonatal rat are dynamically upregulated by IsH, potentially adapting cardiorespiratory responses to severe hypoxia.


Asunto(s)
Catecolaminas/metabolismo , Hipoxia/fisiopatología , Bulbo Raquídeo/fisiopatología , Plasticidad Neuronal/fisiología , Neuronas/fisiología , Receptor de Serotonina 5-HT1A/metabolismo , Animales , Hipoxia/metabolismo , Bulbo Raquídeo/efectos de los fármacos , Bulbo Raquídeo/metabolismo , Plasticidad Neuronal/efectos de los fármacos , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Proteínas Proto-Oncogénicas c-fos/metabolismo , Ratas , Ratas Sprague-Dawley , Transcriptoma/efectos de los fármacos , Transcriptoma/fisiología , Triptófano/análogos & derivados , Triptófano/farmacología , Tirosina 3-Monooxigenasa/metabolismo
18.
Sci Rep ; 6: 23027, 2016 Mar 14.
Artículo en Inglés | MEDLINE | ID: mdl-26971397

RESUMEN

Bacteria harbored in the male reproductive system may influence reproductive function and health of the male and result in developmental origins of adult health and disease (DOHaD) effects in his offspring. Such effects could be due to the seminal fluid, which is slightly basic and enriched with carbohydrates; thereby, creating an ideal habitat for microbes or a potential seminal fluid microbiome (SFM). Using wild-type (WT) and estrogen receptor-alpha (ESR1) knockout (KO) male mice, we describe a unique SFM whose inhabitants differ from gut microbes. The bacterial composition of the SFM is influenced according to whether mice have functional Esr1 genes. Propionibacterium acnes, causative agent of chronic prostatitis possibly culminating in prostate cancer, is reduced in SFM of ESR1 KO compared to WT mice (P ≤ 0.0007). In certain genetic backgrounds, WT mice show a greater incidence of prostate cancer than ESR1 KO, which may be due to increased abundance of P. acnes. Additionally, select gut microbiome residents in ESR1 KO males, such as Lachnospiraceae and Christensenellaceae, might contribute to previously identified phenotypes, especially obesity, in these mutant mice. Understanding how genetics and environmental factors influence the SFM may provide the next frontier in male reproductive disorders and possibly paternal-based DOHaD diseases.


Asunto(s)
Bacterias/metabolismo , Receptor alfa de Estrógeno/deficiencia , Microbiota/fisiología , Semen/microbiología , Animales , Bacterias/clasificación , Bacterias/genética , ADN Bacteriano/química , ADN Bacteriano/genética , Receptor alfa de Estrógeno/genética , Heces/microbiología , Firmicutes/fisiología , Genotipo , Interacciones Huésped-Patógeno , Masculino , Redes y Vías Metabólicas/genética , Ratones Noqueados , Microbiota/genética , Obesidad/genética , Obesidad/microbiología , Propionibacterium acnes/fisiología , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/microbiología , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN
19.
J Bacteriol ; 198(7): 1149-59, 2016 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-26833409

RESUMEN

UNLABELLED: The dimorphic alphaproteobacterium Prosthecomicrobium hirschii has both short-stalked and long-stalked morphotypes. Notably, these morphologies do not arise from transitions in a cell cycle. Instead, the maternal cell morphology is typically reproduced in daughter cells, which results in microcolonies of a single cell type. In this work, we further characterized the short-stalked cells and found that these cells have a Caulobacter-like life cycle in which cell division leads to the generation of two morphologically distinct daughter cells. Using a microfluidic device and total internal reflection fluorescence (TIRF) microscopy, we observed that motile short-stalked cells attach to a surface by means of a polar adhesin. Cells attached at their poles elongate and ultimately release motile daughter cells. Robust biofilm growth occurs in the microfluidic device, enabling the collection of synchronous motile cells and downstream analysis of cell growth and attachment. Analysis of a draft P. hirschii genome sequence indicates the presence of CtrA-dependent cell cycle regulation. This characterization of P. hirschii will enable future studies on the mechanisms underlying complex morphologies and polymorphic cell cycles. IMPORTANCE: Bacterial cell shape plays a critical role in regulating important behaviors, such as attachment to surfaces, motility, predation, and cellular differentiation; however, most studies on these behaviors focus on bacteria with relatively simple morphologies, such as rods and spheres. Notably, complex morphologies abound throughout the bacteria, with striking examples, such as P. hirschii, found within the stalked Alphaproteobacteria. P. hirschii is an outstanding candidate for studies of complex morphology generation and polymorphic cell cycles. Here, the cell cycle and genome of P. hirschii are characterized. This work sets the stage for future studies of the impact of complex cell shapes on bacterial behaviors.


Asunto(s)
Alphaproteobacteria/citología , Alphaproteobacteria/fisiología , Ciclo Celular/fisiología , Técnicas Bacteriológicas , Biopelículas/crecimiento & desarrollo
20.
Genome Announc ; 3(6)2015 Nov 19.
Artículo en Inglés | MEDLINE | ID: mdl-26586892

RESUMEN

We report the draft genome sequence of Prosthecomicrobium hirschii ATCC 27832(T), an alphaproteobacterium with remarkable cellular morphologies. The chromosome comprises 6,484,983 bp in six scaffolds with a G+C content of 69%, and 6,066 potential coding sequences.

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