mRNA expression in different developmental stages of the chicken bursa of Fabricius

AbstractGiven the strong trend to implement zebrafish (Danio rerio) embryos as translational model not only in ecotoxicological, but also toxicological testing strategies, there is an increasing need for a better understanding of their capacity for xenobiotic biotransformation. With respect to the extrapolation of toxicological data from zebrafish embryos to other life stages or even other organisms, qualitative and quantitative differences in biotransformation pathways, above all in cytochrome P450-dependent (CYP) phase I biotransformation, may lead to over- or underestimation of the hazard and risk certain xenobiotic compounds may pose to later developmental stages or other species. This review provides a comprehensive state-of-the-art overview of the scientific knowledge on the development of the CYP1-4 families and corresponding phase I biotransformation and bioactivation capacities in zebrafish. A total of 68 publications dealing with spatiotemporal CYP mRNA expression patterns, activities towards mammalian CYP-probe substrates, bioactivation and detoxification activities, as well as metabolite profiling were analyzed and included in this review. The main results allow for the following conclusions: (1) Extensive work has been done to document mRNA expression of CYP isoforms from earliest embryonic stages of zebrafish, but juvenile and adult zebrafish have been largely neglected so far. (2) There is insufficient understanding of how sex- and developmental stage-related differences in expression levels of certain CYP isoforms may impact biotransformation and bioactivation capacities in the respective sexes and in different developmental stages of zebrafish. (3) Albeit qualitatively often identical, many studies revealed quantitative differences in metabolic activities of zebrafish embryos and later developmental stages. However, the actual relevance of age-related differences on the outcome of toxicological studies still needs to be clarified. (4) With respect to current remaining gaps, there is still an urgent need for further studies systematically assessing metabolic profiles and capacities of CYP isoforms in zebrafish. Given the increasing importance of Adverse Outcome Pathway (AOP) concepts, an improved understanding of CYP capacities appears essential for the interpretation and outcome of (eco)toxicological studies.

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The Low Expression of DUSP1 and HSPA5 Inhibits Chicken Immune Response and Causes Decreased Immunity under Heat Stress

10.21203/rs.2.12530/v1 ◽

2019 ◽

Author(s):

Wenwu Zhang ◽

Jiguo Xu ◽

Hongmei Li ◽

Lianghui Zhou ◽

Qinghua Nie ◽

...

Keyword(s):

Immune Response ◽

Heat Stress ◽

Mrna Expression ◽

Production Performance ◽

Bursa Of Fabricius ◽

Mhc I ◽

Expression Levels ◽

Mhc Ii ◽

Stress Related Genes ◽

Immune Related Genes

Abstract Background As a major stressor, high temperatures negatively affect the poultry industry, through impairments to chicken immunity and production performance. The purpose of this study is to clarify how chicken immune systems responded to heat stress with and without immunization. In the present study, spleen and bursa of Fabricius of experimental chickens were subjected to RNA-seq. Key genes influencing immune response in heat-stressed chickens were identified and their functions validated. Results Immunized and heat-stressed chickens experienced a significant reduction in immune function. The expression of immune-related genes and heat stress-related genes in the spleen increased after immunization and decreased after heat stress, but in the bursa of Fabricius, few of these genes were differentially expressed after immunization and heat stress, indicating insensitivity to high temperature and the lack of vaccine processing. In the non-heat-stressed groups, spleen expression of DUSP1 and HSPA5 decreased significantly, suggesting their relationship to immunity. Upon DUSP1 or HSPA5 overexpression, the mRNA expression of MHC-I, MHC-II, CD80, CD86, CD1C, IL1B, IL6, and TLR4 was earlier than that under LPS stimulation only, indicating that DUSP1 or HSPA5 overexpression enhances HD11 recognition LPS. Inhibiting DUSP1 or HSPA5 expression, the mRNA expression levels of MHC-I, MHC-II, CD80, CD86, IL6 and TLR4 did not change significantly from LPS-stimulation-only levels but CD1C significantly decreased, suggesting that HD11 recognition of LPS is affected by DUSP1 or HSPA5 expression levels. Conclusions The inhibition of immune response due to lowly expressed DUSP1 and HSPA5 may be the cause of decreased immunity in chickens.

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Comprehensive analysis of lncRNA and mRNA expression changes in Tibetan chicken lung tissue between three developmental stages

Animal Genetics ◽

10.1111/age.12990 ◽

2020 ◽

Vol 51 (5) ◽

pp. 731-740

Author(s):

Ruixiang Tang ◽

Jiao Wang ◽

Min Zhou ◽

Yue Lan ◽

Lan Jiang ◽

...

Keyword(s):

Mrna Expression ◽

Lung Tissue ◽

Developmental Stages ◽

Comprehensive Analysis ◽

Tibetan Chicken

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Bis-(3-allyl-4-hydroxyphenyl) sulfone decreases embryonic viability and alters hepatic mRNA expression at two distinct developmental stages in chicken embryos exposed via egg injection

Environmental Toxicology and Chemistry ◽

10.1002/etc.3990 ◽

2017 ◽

Vol 37 (2) ◽

pp. 530-537 ◽

Cited By ~ 4

Author(s):

Doug Crump ◽

Suzanne Chiu ◽

Kim L. Williams

Keyword(s):

Mrna Expression ◽

Developmental Stages ◽

Chicken Embryos ◽

Egg Injection ◽

Embryonic Viability

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A quantitative RT-PCR study of the mRNA expression profile of the IGF axis during mammary gland development

Journal of Molecular Endocrinology ◽

10.1677/jme.0.0330195 ◽

2004 ◽

Vol 33 (1) ◽

pp. 195-207 ◽

Cited By ~ 44

Author(s):

M Boutinaud ◽

JH Shand ◽

MA Park ◽

K Phillips ◽

J Beattie ◽

...

Keyword(s):

Mammary Gland ◽

Mrna Expression ◽

Expression Profile ◽

Mammary Gland Development ◽

Developmental Stages ◽

Normal Mammary Gland ◽

Rt Pcr ◽

Mrna Expression Profile ◽

Igf I ◽

Gland Development

We have used quantitative RT-PCR to analyse the mRNA expression profile of the major components of the IGF axis in different stages of murine mammary gland development, including late pregnancy, lactation and involution. We have shown that all the genes studied, IGF-I, IGF-II, IGF receptor (IGFR) and IGF-binding protein (IGFBP)-1 to -6, were expressed in every stage, albeit at greatly differing levels and displaying unique expression profiles between developmental stages. IGF-I was always expressed at significantly higher levels than either IGF-II or IGFR. This suggests that IGF-I may be the more important IGF during mammary morphogenesis. Overall, IGFBP-3 demonstrated the highest level of expression of any of the IGFBP genes throughout all the developmental stages studied. However, within developmental stages, by far the highest level of expression of any of the IGFBPs was that of IGFBP-5 at day 2 of involution; this was almost an order of magnitude higher than any of the other IGFBP levels recorded. This corroborated our previous findings that the levels of IGFBP-5 protein are highly elevated in the involuting mammary gland, and demonstrated that this up-regulation of IGFBP-5 operates at the level of transcriptional control or message stability. Comparison of the expression profile for these different genes would strongly suggest that they are likely to have differential functions throughout mammary gland development, and also highlights potential interactions and co-regulation between different members of this axis. In addition, our results have identified some similarities and differences in the expression of IGFBPs between the mouse mammary epithelial cell line, HC11, and the normal mammary gland which are worthy of study, most notably the differential regulation of IGFBP-2 and the site of expression of IGFBP-4 and -6. Overall, this study has demonstrated the importance and complexity of the IGF axis during mammary gland development and provides a valuable resource for future research in this area.

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Regulation of growth hormone expression by thyrotropin-releasing hormone through the pituitary-specific transcription factor Pit-1 in chicken pituitary

Acta Veterinaria Hungarica ◽

10.1556/avet.52.2004.4.2 ◽

2004 ◽

Vol 52 (4) ◽

pp. 389-402 ◽

Cited By ~ 4

Author(s):

P. Van As ◽

C. Careghi ◽

V. Bruggeman ◽

O. M. Onagbesan ◽

S. Van der Geyten ◽

...

Keyword(s):

Growth Hormone ◽

Mrna Expression ◽

Developmental Stages ◽

Pituitary Hormones ◽

Thyroid Stimulating Hormone ◽

Thyrotropin Releasing Hormone ◽

Mrna Levels ◽

Rt Pcr ◽

Releasing Hormone ◽

Pou Domain

Pit-1 is a pituitary-specific POU-domain DNA binding factor, which binds to and trans-activates promoters of growth hormone- (GH), prolactin- (PRL) and thyroid stimulating hormone beta- (TSHβ) encoding genes. Pit-1 has been identified in several mammalian and avian species. Thyrotropin-releasing hormone (TRH) is located in the hypothalamus and it stimulates TSH, GH and PRL release from the pituitary gland. In the present study, we successfully developed a competitive RT-PCR for the detection of Pit-1 expression in the chicken pituitary, that was sensitive enough to detect picogram levels of Pit-1 mRNA. Applying this method, the effect of TRH injections on Pit-1 mRNA expression was determined in the pituitary of chick embryos and growing chicks. In both 18-day-old embryos and 10-day-old male chicks the Pit-1 mRNA expression was significantly increased following TRH injection, thereby indicating that the stimulatory effects of TRH on several pituitary hormones is mediated via its effect on Pit-1 expression. Therefore, a semi-quantitative RT-PCR method was used to detect possible changes in GH levels. TRH affected the GH mRNA levels at both developmental stages. These results, combined with the data on Pit-1 mRNA expression, indicate that Pit-1 has a role in mediating the stimulatory effects of TRH on pituitary hormones like GH.

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Transcriptome-Wide m6A Analysis Provides Novel Insights Into Testicular Development and Spermatogenesis in Xia-Nan Cattle

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.791221 ◽

2021 ◽

Vol 9 ◽

Author(s):

Shen-he Liu ◽

Xiao-ya Ma ◽

Ting-ting Yue ◽

Zi-chen Wang ◽

Kun-long Qi ◽

...

Keyword(s):

Mrna Expression ◽

Protein Interactions ◽

Molecular Mechanisms ◽

Reproductive Tract ◽

Developmental Stages ◽

Gene Expression Regulation ◽

Expression Profiles ◽

Testicular Development ◽

Production Traits ◽

Coding Region

Testis is the primary organ of the male reproductive tract in mammals that plays a substantial role in spermatogenesis. Improvement of our knowledge regarding the molecular mechanisms in testicular development and spermatogenesis will be reflected in producing spermatozoa of superior fertility. Evidence showed that N6-Methyladenosine (m6A) plays a dynamic role in post-transcription gene expression regulation and is strongly associated with production traits. However, the role of m6A in bovine testis has not been investigated yet. In this study, we conducted MeRIP-Seq analysis to explore the expression profiles of the m6A and its potential mechanism underlying spermatogenesis in nine bovine testes at three developmental stages (prepuberty, puberty and postpuberty). The experimental animals with triplicate in each stage were chosen based on their semen volume and sperm motility except for the prepuberty bulls and used for testes collection. By applying MeRIP-Seq analysis, a total of 8,774 m6A peaks and 6,206 m6A genes among the studied groups were identified. All the detected peaks were found to be mainly enriched in the coding region and 3′- untranslated regions. The cross-analysis of m6A and mRNA expression exhibited 502 genes with concomitant changes in the mRNA expression and m6A modification. Notably, 30 candidate genes were located in the largest network of protein-protein interactions. Interestingly, four key node genes (PLK4, PTEN, EGR1, and PSME4) were associated with the regulation of mammal testis development and spermatogenesis. This study is the first to present a map of RNA m6A modification in bovine testes at distinct ages, and provides new insights into m6A topology and related molecular mechanisms underlying bovine spermatogenesis, and establishes a basis for further studies on spermatogenesis in mammals.

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CD44 and CD27 Expression Delineates Stages of B Precursor Development Reflecting Recombination Status and Propensity to Leukemic Transformation.

Blood ◽

10.1182/blood.v108.11.1671.1671 ◽

2006 ◽

Vol 108 (11) ◽

pp. 1671-1671

Author(s):

Martina Vaskova ◽

Eva Fronkova ◽

Julia Starkova ◽

Tomas Kalina ◽

Ester Mejstrikova ◽

...

Keyword(s):

Mrna Expression ◽

Heavy Chain ◽

Light Chain ◽

Developmental Stages ◽

Early Stage ◽

Lymphoblastic Leukemia ◽

Late Phase ◽

Chain Gene ◽

Rag 1 ◽

Ig Heavy Chain

Abstract The expression of CD27, in the absence of CD44 is found in TEL/AML1+ acute lymphoblastic leukemia (ALL) but not in other B precursor subtypes of ALL (Vaskova et al., Leukemia 19(5), 2005). Since CD27 had not been shown in human B precursors, we searched for such cells among B precursors in nonmalignant bone marrow. In all 14 specimens of children without any evidence of malignant disease, we found CD27+CD44−CD10+CD19+ B precursors (6.3±3.9%). The subpopulation of CD27−CD44+ cells that corresponds to most other ALL subtypes was more frequent (26±14.4%). Two other subpopulations which rarely develop into B precursor leukemia were also present: double-positive (DP) (5.5±1.8%) and double-negative (DN) (61.8±14.9%). We asked, whether the 4 subpopulations represent consequent differentiation stages, therefore the expression of CD27 and CD44 combined with CD10 and CD19 was studied by polychromatic flow cytometry together with the differentiation markers CD34, terminal deoxyribonucleotidyl transferase (TdT), CD20, cytoplasmic IgM and cytoplasmic VpreB (CD179a). The percentage of CD34+ cells is the highest in CD27+CD44− and decreases gradually in DP, CD27−CD44+ and DN subpopulations (73.1±20.7%; 43±16.6%; 13.1±8.2%; 4.4±2.9%). A similar trend is found in the percentage of CD10bright cells, which become virtually missing in CD27− B precursor stages (41.3±15.4%; 7.9±8.1%; 1.4±1.1%; 2.7±1.8%). This sequence of developmental stages was further supported by a gradual loss of intracellular TdT and VpreB and by the increase of cytoplasmic IgM+ cells. We sorted these subpopulations to compare their recombination potential by measuring TdT and RAG-1 mRNA expression by RQ-RT-PCR. Similarly with the protein level, TdT mRNA expression decreases in concordance with the suggested developmental stages. Interestingly, the DP cells cease to transcript RAG-1, suggesting that these cells are in the stage of suppressed RAG-1 expression after completed immunoglobulin (Ig) heavy chain rearrangement. RAG-1 is re-expressed during Ig light chain rearrangement, seen as the reappearance in the CD27−CD44+ subpopulation. Since the cells with a downregulated RAG-1 are known to be frequent among the large proliferating cells we analyzed the percentage of large cells. The DP subpopulation contains the highest percentage of these cells (63.5±9.9%). In all four subpopulations, heavy chain gene (both segments VH1-3-JH and VH4-7-JH) rearrangements were detected, suggesting that heavy chain genes start to rearrange before or at the CD27+CD44− stage. We investigated the light chain rearrangements using the system detecting the intron RSS-Kde rearrangements, which appear in the late phase of Ig light chain rearrangement. Rearranged light chain genes appear at the CD27−CD44+ stage, whereas they are virtually missing at earlier stages. These results show that CD27 molecule, which is so far regarded a marker of memory B cells is expressed also in the early stage of B cell development during Ig heavy chain rearrangement completion. The expression of CD27 and CD44 define differentiation stages that very tightly correlate with Ig recombination maturity as well as with specific subtypes of B precursor leukemia.

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Goat PDGFRB : unique mRNA expression profile in gonad and significant association between genetic variation and litter size

Royal Society Open Science ◽

10.1098/rsos.180805 ◽

2019 ◽

Vol 6 (1) ◽

pp. 180805 ◽

Cited By ~ 6

Author(s):

Wenjing Yang ◽

Hailong Yan ◽

Ke Wang ◽

Yang Cui ◽

Tong Zhou ◽

...

Keyword(s):

Mrna Expression ◽

Litter Size ◽

Marker Assisted Selection ◽

Developmental Stages ◽

Growth Factor Receptor ◽

Platelet Derived Growth Factor ◽

Mrna Expression Profile ◽

Genetic Breeding ◽

Potential Association ◽

Cashmere Goats

β-Type platelet-derived growth factor receptor ( PDGFRB ) is a typical tyrosine kinase, as a candidate gene associated with reproduction. Its main roles include regulation of gonocytes (migration and proliferation) and of the cell cycle. The objectives of this study were to identify mRNA expression of the goat PDGFRB gene, as well as insertion/deletion (indel) variants and their association with litter size in 1122 healthy Shaanbei white cashmere goats. The results revealed that PDGFRB was widely expressed in all tested tissues, and the expression levels in testes at different developmental stages indicated a potential association with the mitosis-to-meiosis transition. Furthermore, the expression of PDGFRB was relatively higher in the ovary tissue of mothers of two lambs compared with mothers of single lamb. These results implied that PDGFRB was related to goat fertility. Meanwhile, two intronic indels, 5 bp ( n = 501) and 10 bp ( n = 1122), were identified. Statistical analysis revealed that only the 10 bp indel was associated with first-born litter size ( n = 1122, p = 6.030 × 10 −5 ), and that individuals of the genotype insertion/deletion had larger litter sizes than those of genotype insertion/insertion. Overall, these results indicated that the 10 bp indel of PDGFRB could be used in marker-assisted selection during goat genetic breeding.

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Identification of Suitable Internal Control Genes for Quantitative Real-Time PCR Expression Analyses in Peanut (Arachis hypogaea)

Peanut Science ◽

10.3146/ps09-014.1 ◽

2010 ◽

Vol 37 (1) ◽

pp. 12-19 ◽

Cited By ~ 18

Author(s):

Yael Brand ◽

Ran Hovav

Keyword(s):

Gene Expression ◽

Mrna Expression ◽

Real Time ◽

Internal Control ◽

Real Time Pcr ◽

Reference Genes ◽

Developmental Stages ◽

Peanut Kernel ◽

Tissue Samples ◽

The Stability

Abstract Real-time qPCR is currently the most sensitive technique available for the detection of low-level mRNA expression. For more reliable and precise gene expression analyses, real-time PCR data for a sequence of interest must be normalized against that of a control gene, which is uniformly expressed in various tissues and during different phases of development. So far, suitable internal controls for gene expression studies in peanut have not been identified. We assessed the expression of 10 frequently used housekeeping genes, specifically ubq10, gapdh, hel1, yls8, 14-3-3, 60s, ubc, ef-1α, act7, and adh3. Using the algorithms available through the GeNorm and NormFinder programs, the stability of their expression was estimated in a set of five diverse peanut tissue samples derived from a Virginia-type peanut cultivar (Shulamit). Collectively, the gene with the most stable expression across all of the examined tissues and both programs was adh3, followed by 60s and yls8, which had minimal estimated intra- and inter-tissue variation. The stability of two stable reference genes (adh3 and yls8) compared with two less stable (14-3-3 and ubq10) reference genes was validated in unpooled tissue samples from five peanut kernel developmental stages. Finally, the effect of the use of one or more reference genes on the observed relative expression levels of an important seed oil metabolism gene, diacylglycerol acyltransferase 1 (Dgat1), during kernel development was demonstrated. Based on findings, the suggestion is that adh3, or a combination of this gene with 60s and yls8 should be considered for use in quantitative mRNA expression analyses in Arachis, particularly in studies involving seed development; whereas ubq10 and gapdh should be avoided.

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