scholarly journals Excess Folic Acid Supplementation before and during Pregnancy and Lactation Alters Behaviors and Brain Gene Expression in Female Mouse Offspring

Nutrients ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 66
Author(s):  
Xingyue Yang ◽  
Wenyan Sun ◽  
Qian Wu ◽  
Hongyan Lin ◽  
Zhixing Lu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Folic Acid ◽  
Motor Coordination ◽  
Female Offspring ◽  
Mrna Levels ◽  
Western Blots ◽  
Brain Gene Expression ◽  
Qrt Pcr ◽  
Morris Water Maze Task ◽  
Pregnancy And Lactation

Use of folic acid (FA) during early pregnancy protects against birth defects. However, excess FA has shown gender-specific neurodevelopmental toxicity. Previously, we fed the mice with 2.5 times the recommended amount of FA one week prior to mating and during the pregnancy and lactation periods, and detected the activated expression of Fos and related genes in the brains of weaning male offspring, as well as behavioral abnormalities in the adults. Here, we studied whether female offspring were affected by the same dosage of FA. An open field test, three-chamber social approach and social novelty test, an elevated plus-maze, rotarod test and the Morris water maze task were used to evaluate their behaviors. RNA sequencing was performed to identify differentially expressed genes in the brains. Quantitative real time-PCR (qRT-PCR) and Western blots were applied to verify the changes in gene expression. We found increased anxiety and impaired exploratory behavior, motor coordination and spatial memory in FA-exposed females. The brain transcriptome revealed 36 up-regulated and 79 down-regulated genes in their brains at weaning. The increase of Tlr1; Sult1a1; Tph2; Acacb; Etnppl; Angptl4 and Apold1, as well as a decrease of Ppara mRNA were confirmed by qRT-PCR. Among these genes; the mRNA levels of Etnppl; Angptl4andApold1 were increased in the both FA-exposed female and male brains. The elevation of Sult1a1 protein was confirmed by Western blots. Our data suggest that excess FA alteres brain gene expression and behaviors in female offspring, of which certain genes show apparent gender specificity.

scholarly journals Liver choline metabolism and gene expression in choline-deficient mice offspring differ with gender

2021 ◽  
Vol 85 (2) ◽  
pp. 447-451
Author(s):  
Yukino Miyachi ◽  
Kei Akiyama ◽  
Yoshiko Tsukuda ◽  
Thanutchaporn Kumrungsee ◽  
Noriyuki Yanaka
Keyword(s):  
Gene Expression ◽  
Female Offspring ◽  
Male Offspring ◽  
Mrna Levels ◽  
Choline Deficiency ◽  
Deficient Diet ◽  
Choline Metabolism ◽  
Pregnancy And Lactation ◽  
Gender Specific ◽  
Deficient Mice

ABSTRACT Choline is an important nutrient during pregnancy and lactation. Maternal choline deficiency in CD-1 mice lowers liver betaine levels in male offspring. By contrast, it increases elovl3 and vanin-1 mRNA levels in female offspring. Taken together, these observations suggest gender-specific responses to a choline-deficient diet.

scholarly journals Folic acid and its metabolites modulate IGF-I receptor gene expression in colon cancer cells in a p53-dependent manner

2006 ◽  
Vol 13 (2) ◽  
pp. 571-581 ◽  
Author(s):  
Z Attias ◽  
H Werner ◽  
N Vaisman
Keyword(s):  
Gene Expression ◽  
Colon Cancer ◽  
Folic Acid ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Western Blots ◽  
Primary Tumors ◽  
Chemopreventive Agent ◽  
Igf I ◽  
Apoptotic Activity

The insulin-like growth factor-I receptor (IGF-IR) has an important role in colorectal cancer development and progression. IGF-IR displays a potent anti-apoptotic activity and is overexpressed in primary tumors and colon cancer-derived cell lines. Folic acid, a member of the vitamin B family, is a chemopreventive agent whose deficiency has been linked to an enhanced colon cancer risk. The present study was aimed at testing the hypothesis that part of the modulatory effect of folic acid on malignant transformation may be attributed to its ability to regulate IGF-IR gene expression. Regulation of IGF-IR gene expression by folic acid was assessed using western blots, RT-PCR, transient transfections and chromatin immunoprecipitation assays. Activation of the IGF-IR signaling pathway was evaluated by measuring phosphorylation of ERK, and apoptosis was assayed using poly (ADP-ribose) polymerase cleavage and annexin V-FITC staining. Results obtained showed that folic acid induced a dose-dependent decrease in IGF-IR protein and mRNA levels in the HCT116 +/+ colon cancer cell line. This effect was associated with a significant reduction in IGF-IR promoter activity. Similar effects were elicited by the folic acid metabolites dihydrofolic acid and tetrahydrofolic acid. In addition, folic acid abrogated the IGF-I-stimulated phosphorylation of the downstream signaling molecule ERK1/2 and exhibited a pro-apoptotic activity. Moreover, folic acid induced a significant decrease in Sp1 binding to the IGF-IR promoter region. Finally, folic acid had no effect in wild-type p53-depleted HCT116 −/− and Caco-2 cells. In conclusion, the mechanism of action of folic acid involves regulation of IGF-IR gene expression. The ability of folic acid to downregulate the IGF-I signal transduction pathway may allow the micronutrient to function as a chemopreventive agent. Folic acid deficiency, on the other hand, may lead to increased IGF-IR gene expression, with ensuing pathological activation by endocrine and/or autocrine/paracrine IGF-I.

Developmental expression of pIgR gene in sheep mammary gland and hormonal regulation

2002 ◽  
Vol 69 (1) ◽  
pp. 13-26 ◽  
Author(s):  
AURORE RINCHEV-ALARNOLD ◽  
LUCETTE BELAIR ◽  
JEAN DJIANE
Keyword(s):  
Gene Expression ◽  
Mammary Gland ◽  
Hormonal Regulation ◽  
Mrna Level ◽  
Immunohistochemical Analysis ◽  
Developmental Expression ◽  
Secretory Iga ◽  
Mrna Levels ◽  
Immunoglobulin Receptor ◽  
Pregnancy And Lactation

Secretory IgA found in external secretions are constituted by polymeric IgA (pIgA) bound to the extra-cellular part of the polymeric immunoglobulin receptor (pIgR). The receptor mediates transcytosis of pIgA across epithelial cells. The aim of the present study was to analyse the evolution of pIgR expression in the sheep mammary gland during the development of the mammary gland and to analyse its hormonal regulation. Gene expression of the pIgR was analysed in sheep mammary gland during pregnancy and lactation. By Northern Blot analysis, we observed that low levels of pIgR mRNA are expressed until day 70 of pregnancy. Accumulation of pIgR mRNA started during the third part of pregnancy and intensified 3 d after parturition to reach highest levels during established lactation (day 70). In situ hybridization analysis was used to confirm the increase in pIgR gene expression per mammary epithelial cell. In order to examine the hormonal regulation of the pIgR expression, virgin ewes were hormonally treated. Treatment with oestradiol and progesterone increased pIgR mRNA levels slightly. Subsequent addition of glucocorticoids induced a significant accumulation of pIgR mRNA in the mammary gland of the treated animals. Immunohistochemical analysis was performed to verify that the increase of pIgR mRNA level was associated with enhancement of the pIgR protein in mammary cells. No increase of pIgR mRNA levels were observed if PRL secretion was blocked by bromocryptine injections throughout the hormonal procedure. In conclusion, the present experiments suggest that the enhancement of pIgR levels during lactation result from combined effects of both prolactin and glucocorticoids.

scholarly journals Gene Expression of Indoleamine 2,3 Dioxygenase 1, Insulin-Growth Factor 1 and Red/IK Cytokine in Alopecia Areata

2014 ◽  
Vol 6 (3) ◽  
pp. 263-266
Author(s):  
Simona Corina ȘENILĂ ◽  
Ovidiu BĂLĂCESCU ◽  
Loredana BĂLĂCESCU ◽  
Elisabeta CANDREA ◽  
Loredana UNGUREANU ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hair Follicle ◽  
Mhc Class I ◽  
Alopecia Areata ◽  
Fas Ligand ◽  
Immune Privilege ◽  
Mrna Levels ◽  
Insulin Growth Factor ◽  
Qrt Pcr ◽  
Anagen Hair

Alopecia areata (AA) is a chronic, T-cell mediated autoimmune disease directed against the hair follicle, which partially evolves due to a loss of the immune privilege of the anagen hair follicle. The immune privilege is maintained by several factors, including a downregulation of MHC class I and II, local immunosupressants and expression of Fas ligand. The purpose of the study was to evaluate several factors involved in the collapse and restoration of the immune privilege. We investigated IDO1, IGF1 and red/IK gene expression in lesional and perilesionalscalp biopsies from alopecia areata patients. Seven paired punch-biopsies were taken from the active edge of alopecic plaque and from the perilesional scalp. Expression of IDO1, IGF1 and red/IK genes was performed by qRT-PCR. In lesional tissue, IGF1, IDO1 and red/IK genes showed an increase in the mRNA levels as compared with the perilesional scalp. By comparing the pairs of data for the investigated genes, IDO1was statistically upregulated in the lesional area. No significant differences were observed between the gene expression in mild or severe AA, from the lesional or perilesional areas. IDO1 mRNA expression was higher in patients with a relapse duration of less than 6 months as compared to patients with a relapse duration of more than 6 months; levels of IGF1 and red/IK mRNA are increased in lesionals compared to perilesional scalp area.

scholarly journals Validation of Reference Genes for Quantitative Real-time PCR in Diaphanosoma Celebensis: For Chemical Exposure Conditions and Different Ages

2021 ◽  
Author(s):  
Young-Mi Lee ◽  
Soyeon In ◽  
Se-Joo Kim ◽  
Eun-Ji Won ◽  
Hayoung Cho ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Reference Gene ◽  
Reference Genes ◽  
Expression Profiles ◽  
Chemical Exposure ◽  
Mrna Levels ◽  
Qrt Pcr ◽  
Different Ages ◽  
Diaphanosoma Celebensis

Abstract Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), a primary approach for evaluating gene expression, requires an appropriate normalization strategy to rule out variations in gene expression among samples. The best option is to use a reference gene whose expression level is stable across various experimental conditions to compare the mRNA levels of a target gene. However, there is limited information on how the reference gene is differentially expressed at different ages (growth) in small invertebrates with notable changes such as molting. In this study, expression profiles of nine candidate reference genes from the brackish water flea, Diaphanosoma celebensis, were evaluated under diverse exposure to toxicants and according to growth. As a result, four different algorithms showed similar stabilities of genes for chemical exposures in the case of limited conditions using the same developmental stage (e.g., adult), while the results according to age showed a significantly different pattern in suite of candidate reference genes. This affected the results of genes EcRA and GST, which are involved in development and detoxification mechanisms, respectively. Our finding is the first step towards establishing a standardized real-time qRT-PCR analysis of this environmentally important invertebrate that has potential for aquatic ecotoxicology, particularly in estuarine environments.

A blood-based gene expression signature of early-stage non-small cell lung cancer.

2012 ◽  
Vol 30 (30_suppl) ◽  
pp. 69-69
Author(s):  
Melissa Rotunno ◽  
Nan Hu ◽  
Hua Su ◽  
Chaoyu Wang ◽  
Pier Alberto Bertazzi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Lung Cancer ◽  
Predictive Accuracy ◽  
Early Stage ◽  
Single Gene ◽  
Gene Expression Signature ◽  
Mrna Levels ◽  
Healthy Controls ◽  
Expression Signature ◽  
Qrt Pcr

69 Background: Accurate blood-based biomarker for early cancer detection could be an easier and more convenient screening option than monitoring the target organ via tissue or imaging. We recently identified and validated eight genetic biomarkers of early-stage lung adenocarcinoma detectable in both peripheral whole blood (PWB) and lung tissue of smokers. Since biomarkers distinguishing benign disease versus lung malignancy across all cell types are needed in the diagnostic clinical setting, it is important to test the identified biomarkers in other lung cancer histologies, particularly in squamous cell carcinoma (SQCC), the second most common lung cancer histology after adenocarcinoma (AD). Methods: Using Real-Time Quantitative PCR (qRT-PCR), we measured mRNA levels for the eight candidate genes in PWB of 48 randomly sampled stage I SQCC cases, in addition to previously analyzed 82 AD cases and 130 age, sex, and smoking frequency matched healthy controls from the Environment And Genetics in Lung cancer Etiology (EAGLE) case-control study. The qRT-PCR data were analyzed using the 2-ΔΔCtmethod to compare SQCC cases with controls. The area under the receiver operating characteristic curve (AUC) was computed to assess the predictive accuracy of the candidate biomarkers in SQCC separately, and in SQCC and AD together. Results: Expression of TGFBR3, RUNX3, TRGC2, TRGV9, TARP, and TSTA3 genes, significantly differentiated SQCC cases versus controls, while ACP1 and VCAN gene expression did not. The eight genes combined discriminated patients with lung cancer from healthy controls with similarly high accuracy in SQCC and overall (AUC = 0.80 ± 0.1). RUNX3 showed the highest single gene accuracy for SQCC (AUC = 0.78). Conclusions: We showed that the previously identified gene expression signature of early-stage lung AD also differentiated early stage SQCC from healthy controls and demonstrated its sensitivity and specificity as a potential diagnostic lung cancer biomarker. Since lung cancer is the most common cause of cancer mortality worldwide and current smokers are at very high risk, our smoking-specific findings, if confirmed and translated into screening approaches, have the potential to impact public health.

Identification of Genes Which Play a Crucial Role in C/EBPβ Downstream Signalling in ALK+ ALCL Cell Lines.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1943-1943
Author(s):  
Irina Bonzheim ◽  
Martin Irmler ◽  
Natasa Anastasov ◽  
Margit Klier-Richter ◽  
Sabine Schaefer ◽  
...  
Keyword(s):  
Gene Expression ◽  
Western Blot ◽  
Gene Expression Profiling ◽  
Cell Lines ◽  
Pathway Analysis ◽  
Expression Profiling ◽  
Cellular Proliferation ◽  
Compensatory Mechanism ◽  
Mrna Levels ◽  
Qrt Pcr

Abstract Abstract 1943 Poster Board I-966 Introduction: ALK+ anaplastic large cell lymphomas (ALCL) overexpress C/EBPβ, as a consequence of NPM-ALK kinase activity. We recently reported C/EBPβ as a transcription regulator of NPM-ALK induced cellular proliferation. To identify the downstream targets of C/EBPβ that might be responsible for cell proliferation and survival, we performed gene expression profiling and pathway analyses after C/EBPβ gene silencing Materials and Methods: C/EBPβ knockdown was done by lentiviral shRNA-transduction into two ALK+ ALCL cell lines with strong C/EBPβ expression – SUDHL1 and KiJK. At day three after infection, RNA was extracted and used for Gene Chip expression analysis (U133 Plus 2.0 arrays/ Affymetrix). Genes regulated in both cell lines were applied to Genomatix Bibliosphere Pathway analysis. Candidate genes were either strongly influenced by C/EBPβ knockdown, or had promoter binding sites for C/EBPβ, or showed remarkable pathway connections. The influence of C/EBPβ on these genes was validated by qRT-PCR and in part by Western blot. Results: Gene expression profiling analysis resulted in 167 genes being regulated in both cell lines, of which 26 genes were chosen for further analysis. Validation by qRT-PCR confirmed 23/26 genes. Pathway analysis revealed c-Jun, which is a member of the dimeric transcription factor AP-1, as a regulator of C/EBPβ expression. Silencing C/EBPβ led to a clear up-regulation of c-Jun mRNA. Western blot analysis demonstrated that C/EBPβ influenced not only the expression of c-Jun but also its phosphorylation on Ser63 and Ser73. In contrast to what has been reported, we found very low levels of c-Jun expression in ALK+ALCL cells lines and its expression correlated inversely with C/EBPβ mRNA levels. Although it has been shown that c-Jun regulates C/EBPβ expression directly, in ALK+ALCL the expression of C/EBPβ is clearly independent of c-Jun. Our data suggest that c-Jun up-regulation after C/EBPβ knockdown is a compensatory mechanism to maintain C/EBPβ expression. Additionally, of the 26 selected genes, Bibliosphere Analysis identified 12 genes, which might be transcriptionally regulated by C/EBPβ and are primary targets in C/EBPβ downstream signalling. Two of these genes are of particular interest. The anti-apoptotic protein BCL2A1 contains a promoter-binding site for C/EBPβ and has been shown previously to be both strongly regulated in ALK+ALCL and absolutely necessary for its transformation. The second is a DEAD box nucleolar RNA helicase protein involved in ribosomal RNA production and proliferation which we found to be strongly expressed in ALK+ALCL cell lines and primary cases. Conclusions: C/EBPβ silencing in ALK+ALCL cell lines showed 1) an inverse correlation between c-Jun and C/EBPβ mRNA expression levels, 2) the expression of C/EBPβ in ALK+ALCL is independent of c-Jun, 3) genes transcriptionally regulated by C/EBPβ seem to be essential for proliferation and survival in ALK+ALCL. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Localized Gene Expression in Pseudomonas aeruginosa Biofilms

2008 ◽  
Vol 74 (14) ◽  
pp. 4463-4471 ◽  
Author(s):  
Ailyn P. Lenz ◽  
Kerry S. Williamson ◽  
Betsey Pitts ◽  
Philip S. Stewart ◽  
Michael J. Franklin
Keyword(s):  
Gene Expression ◽  
Pseudomonas Aeruginosa ◽  
Green Fluorescent Protein ◽  
16S Rrna ◽  
Fluorescent Protein ◽  
Mrna Levels ◽  
Protein Fluorescence ◽  
Bacterial Gene ◽  
Qrt Pcr ◽  
Green Fluorescent

ABSTRACT Gene expression in biofilms is dependent on bacterial responses to the local environmental conditions. Most techniques for studying bacterial gene expression in biofilms characterize average values across the entire population. Here, we describe the use of laser capture microdissection microscopy (LCMM) combined with multiplex quantitative real-time reverse transcriptase PCR (qRT-PCR) to isolate and quantify RNA transcripts from small groups of cells at spatially resolved sites within biofilms. The approach was first tested and analytical parameters were determined for Pseudomonas aeruginosa containing an isopropyl-β-d-thiogalactopyranoside-inducible gene for the green fluorescent protein (gfp). The results show that the amounts of gfp mRNA were greatest in the top zones of the biofilms, and that gfp mRNA levels correlated with the zone of active green fluorescent protein fluorescence. The method then was used to quantify transcripts from wild-type P. aeruginosa biofilms for a housekeeping gene, acpP; the 16S rRNA; and two genes regulated by quorum sensing, phzA1 and aprA. The results demonstrated that the amount of acpP mRNA was greatest in the top 30 μm of the biofilm, with little or no mRNA for this gene at the base of the biofilms. In contrast, 16S rRNA amounts were relatively uniform throughout biofilm strata. Using this strategy, the RNA amounts of individual genes were determined, and therefore the results are dependent on both gene expression and the half-life of the transcripts. Therefore, the uniform amount of rRNA throughout the biofilms likely is due to the stability of the rRNA within ribosomes. The levels of aprA mRNA showed stratification, with the largest amounts in the upper 30-μm zone of these biofilms. The results demonstrate that mRNA levels for individual genes are not uniformly distributed throughout biofilms but may vary by orders of magnitude over small distances. The LCMM/qRT-PCR technique can be used to resolve and quantify this RNA variability at high spatial resolution.

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