Histopathological evaluation and expression profile of PRM-1, TNP-2, 17B-HSD3, LHR, MHC-I, MIC-B, NC1 and NC3 genes in bovine testis

Histopathological and spermatogenesis classification by Johnsen is widely used in the germinal epithelium maturation analysis, besides identifying pathological alterations able to cause subfertility and even infertility. The aim of this study was to analyze cell-differentiation histopathological data and to correlate them with expression of PRM-1, TNP-2, 17β-HSD3, LHR, generic MHC-I, MIC-B, NC1 and NC3 genes, involved in bovine spermatogenesis using qRT-PCR from testicular parenchyma. Based on Johnsen’s criteria, the results showed normal spermatogenic activity in these animals, classified at 6, 7 and 8 scores. The qRT-PCR analysis expression showed that MHC-I (generic) gene was less expressed than all the other genes in evaluated scores (p < 0.05) and, PRM-1 and TNP-2 were the most expressed genes (p < 0.05). The PRM-1 gene expression was significantly higher than TNP-2 (p < 0.05). Comparing scores, 17β-HSD3 gene expression was lower (p < 0.05) in score 6 when compared to scores 7 and 8 animals. It was also observed that PRM-1 expression was lower in score 6 when compared to 7, as well as TNP-2 gene was less expressed in the score 6 (p < 0.05) when compared to 7 and 8 scores. Our results demonstrated that MHC I (generic), MIC-B, NC1, NC3, and LHR genes are poorly expressed in bovine testis, suggesting their marginal action on spermatogenesis. Instead, PRM-1, TNP-2, and 17β-HSD3 expression were higher, supporting the notion that these genes can act directly on the germ cells differential development during bovine spermatogenesis.

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Human Gene Expression Changes Following Diaphragm Unloading Via Controlled Mechanical Ventilation: QRT-PCR Analysis

10.1164/ajrccm-conference.2010.181.1_meetingabstracts.a5038 ◽

2010 ◽

Author(s):

Harsha Deoghare ◽

Tseng-Tien Huang ◽

D Martin

Keyword(s):

Gene Expression ◽

Mechanical Ventilation ◽

Human Gene ◽

Pcr Analysis ◽

Human Gene Expression ◽

Qrt Pcr ◽

Controlled Mechanical Ventilation

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Tuft Cells: A New Player in Hirschsprung's Disease

European Journal of Pediatric Surgery ◽

10.1055/s-0039-1700549 ◽

2019 ◽

Vol 30 (01) ◽

pp. 059-063 ◽

Cited By ~ 2

Author(s):

Anne Marie O'Donnell ◽

Hiroki Nakamura ◽

Prem Puri

Keyword(s):

Gene Expression ◽

Confocal Microscopy ◽

Hirschsprung's Disease ◽

Hirschsprung’S Disease ◽

Distal Colon ◽

Pcr Analysis ◽

Specific Marker ◽

Qrt Pcr ◽

Tuft Cell ◽

Tuft Cells

Abstract Introduction “Tuft” cells, also known as brush or caveolated cells, are characteristically fusiform shaped, with a distinct apical “tuft” of microvilli extending into the lumen. Double cortin-like kinase 1 (DCLK1) is a microtubule kinase and is a specific marker of intestinal tuft cells. DCLK1-positive tuft cells have been shown to play a key role in gastrointestinal chemosensation, inflammation, and neurotransmission. DCLK1 and Choline acetyltransferase (ChAT), the enzymes responsible for acetylcholine production, are reported to be coexpressed within the gastrointestinal tract. We designed this study to investigate the hypothesis that DCLK1 gene expression is altered in Hirschsprung's disease (HSCR). Materials and Methods HSCR tissue specimens (n = 6) were collected at the time of pull-through surgery, while control samples were obtained at the time of colostomy closure in patients with imperforate anus (n = 6). Quantitative real-time polymerase chain reaction (qRT-PCR) analysis was undertaken to quantify DCLK1 gene expression, and immunolabeling of DCLK1-positive tuft cells was visualized using confocal microscopy. Results qRT-PCR analysis revealed significant downregulation of the DCLK1 gene in both aganglionic and ganglionic HSCR specimens compared with controls (p < 0.05). Confocal microscopy revealed DCLK1-positive tuft cell expression within the colonic mucosa, with a reduction in expression in both aganglionic and ganglionic HSCR colon compared with controls. Conclusion DCLK1 is significantly downregulated in HSCR colon, suggesting a role for tuft cells in cholinergic neurotransmission of the distal colon. The marked decrease in DCLK1 expression within ganglionic specimens highlights the physiologically abnormal nature of this segment in HSCR patients.

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Comparison of Reliable Reference Genes Following Different Hormone Treatments by Various Algorithms for qRT-PCR Analysis of Metasequoia

International Journal of Molecular Sciences ◽

10.3390/ijms20010034 ◽

2018 ◽

Vol 20 (1) ◽

pp. 34 ◽

Cited By ~ 3

Author(s):

Jing-Jing Wang ◽

Shuo Han ◽

Weilun Yin ◽

Xinli Xia ◽

Chao Liu

Keyword(s):

Gene Expression ◽

Reference Genes ◽

Elongation Factor ◽

Pcr Analysis ◽

Qrt Pcr ◽

Gene Normalization ◽

Accurate Normalization ◽

Suitable Reference ◽

Different Tissues ◽

Gene Expression Levels

Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is the most sensitive technique for evaluating gene expression levels. Choosing appropriate reference genes for normalizing target gene expression is important for verifying expression changes. Metasequoia is a high-quality and economically important wood species. However, few systematic studies have examined reference genes in Metasequoia. Here, the expression stability of 14 candidate reference genes in different tissues and following different hormone treatments were analyzed using six algorithms. Candidate reference genes were used to normalize the expression pattern of FLOWERING LOCUS T and pyrabactin resistance-like 8. Analysis using the GrayNorm algorithm showed that ACT2 (Actin 2), HIS (histone superfamily protein H3) and TATA (TATA binding protein) were stably expressed in different tissues. ACT2, EF1α (elongation factor-1 alpha) and HIS were optimal for leaves treated with the flowering induction hormone solution, while Cpn60β (60-kDa chaperonin β-subunit), GAPDH (glyceraldehyde-3-phosphate dehydrogenase) and HIS were the best reference genes for treated buds. EF1α, HIS and TATA were useful reference genes for accurate normalization in abscisic acid-response signaling. Our results emphasize the importance of validating reference genes for qRT-PCR analysis in Metasequoia. To avoid errors, suitable reference genes should be used for different tissues and hormone treatments to increase normalization accuracy. Our study provides a foundation for reference gene normalization when analyzing gene expression in Metasequoia.

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Reference gene selection for qRT-PCR analysis of season- and tissue-specific gene expression profiles in the honey bee Apis mellifera

Scientific Reports ◽

10.1038/s41598-020-70965-4 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Ji Hyang Jeon ◽

KyungHwan Moon ◽

YeongHo Kim ◽

Young Ho Kim

Keyword(s):

Gene Expression ◽

Apis Mellifera ◽

Honey Bee ◽

Gene Selection ◽

Expression Profiles ◽

Gene Expression Profiles ◽

Pcr Analysis ◽

Specific Gene ◽

Qrt Pcr ◽

Selection For

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Comparison of Stemness and Gene Expression between Gingiva and Dental Follicles in Children

Stem Cells International ◽

10.1155/2016/8596520 ◽

2016 ◽

Vol 2016 ◽

pp. 1-11 ◽

Cited By ~ 1

Author(s):

Chung-Min Kang ◽

Seong-Oh Kim ◽

Mijeong Jeon ◽

Hyung-Jun Choi ◽

Han-Sung Jung ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Expression Profiles ◽

Gene Expression Profiles ◽

Pcr Analysis ◽

Expression Levels ◽

Qrt Pcr ◽

Stem Cell Source ◽

Regenerative Dentistry ◽

Induced Pluripotent

The aim of this study was to compare the differential gene expression and stemness in the human gingiva and dental follicles (DFs) according to their biological characteristics. Gingiva (n=9) and DFs (n=9) were collected from 18 children. Comparative gene expression profiles were collected using cDNA microarray. The expression of development, chemotaxis, mesenchymal stem cells (MSCs), and induced pluripotent stem cells (iPSs) related genes was assessed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Histological analysis was performed using hematoxylin-eosin and immunohistochemical staining. Gingiva had greater expression of genes related to keratinization, ectodermal development, and chemotaxis whereas DFs exhibited higher expression levels of genes related to tooth and embryo development. qRT-PCR analysis showed that the expression levels of iPSc factors includingSOX2,KLF4, andC-MYCwere58.5±26.3,12.4±3.5, and12.2±1.9times higher in gingiva andVCAM1(CD146) andALCAM(CD166) were33.5±6.9and4.3±0.8times higher in DFs. Genes related to MSCs markers includingCD13,CD34,CD73,CD90, andCD105were expressed at higher levels in DFs. The results of qRT-PCR and IHC staining supported the microarray analysis results. Interestingly, this study demonstrated transcription factors of iPS cells were expressed at higher levels in the gingiva. Given the minimal surgical discomfort and simple accessibility, gingiva is a good candidate stem cell source in regenerative dentistry.

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The Human Plasma Cell Gene Expression Program.

Blood ◽

10.1182/blood.v104.11.3242.3242 ◽

2004 ◽

Vol 104 (11) ◽

pp. 3242-3242

Author(s):

John De Vos ◽

Dirk Hose ◽

Thierry Reme ◽

Hartmut Goldschmidt ◽

Jean-Francois Rossi ◽

...

Keyword(s):

Gene Expression ◽

Transcription Factors ◽

Cell Differentiation ◽

Plasma Cell ◽

Expression Profile ◽

Plasma Cells ◽

Plasma Cell Differentiation ◽

Unfolded Protein ◽

The Mean ◽

Protein Response

Abstract Seven purified peripheral blood memory B-cells (BM), seven in-vitro-generated polyclonal plasmablastic cells (PPC) and seven purified bone marrow mature plasma cells (BMPC) were studied by oligonucleotide microarrays. All samples were obtained from healthy volunteers. The gene expression profiling of these samples was determined with Affymetrix pan genomic U133A + B arrays (44 928 oligonucleotide probesets). We determined that 2313 genes were differentially expressed between these three B cell categories (P 〈 0.01 by a Kruskal-Wallis test and a ratio between two categories 〉 3). These 2313 genes were classified into six categories, according to the expression profile: early plasma cell genes (EPC), late plasma cell genes (LPC), genes lost early during plasma cell differentiation (LEPC), genes lost late during plasma cell differentiation (LLPC), genes upregulated only in plasmablasts (PBO) and genes lost only in plasmablasts (LPBO). As expected, Ig transcripts where essentially classified as EPC. As a corollary, genes involved in protein synthesis or degradation, transmembrane transporters and metabolism genes were overrepresented in EPC genes. Interestingly, genes involved in intercellular communication and extracellular matrix were enriched in LPC, highlighting the fact that mature plasma cells develop tight interactions with the bone marrow environment. Of note, genes involved in cell cycle are upregulated mainly in plasmablasts, whereas antiapoptotic genes are lost in plasmablasts only. Mains genes known to be involved in plasma cell differentiation display an expression profile in agreement with published data, as illustrated for transcription factors in Figure 1, validating this DNA microarray dataset. However most of these 2313 genes have either never been described yet or have no yet been linked to plasma cell differentiation. The description of those genes among our genome whose expression vary most during plasma cell differentiation will be an essential step in understanding the biology of a cell type essential to immune defenses and involved in deadly diseases. Figure 1: Transcription factors involved in plasma cell differentiation. Color indicates the expression profile category. For each gene is given the ratio of the mean expression value in plasma cell samples (PPC and BMPC) to the mean expression value in BM. UPR: Unfolded Protein Response. Figure 1:. Transcription factors involved in plasma cell differentiation. Color indicates the expression profile category. For each gene is given the ratio of the mean expression value in plasma cell samples (PPC and BMPC) to the mean expression value in BM. UPR: Unfolded Protein Response.

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Gene Expression Profiling of Polycomb and Polycomb-Regulated Genes in Patients with Acute Myeloid Leukaemia - Significance and Relationship to Other Molecular Aberrations.

Blood ◽

10.1182/blood.v110.11.3165.3165 ◽

2007 ◽

Vol 110 (11) ◽

pp. 3165-3165

Author(s):

Lykke C. Grubach ◽

Caroline Juhl-Christensen ◽

Anita Rethmeier ◽

Lene H. Olesen ◽

Peter Hokland ◽

...

Keyword(s):

Gene Expression ◽

Multidrug Resistance ◽

Acute Myeloid Leukaemia ◽

Myeloid Leukaemia ◽

Expression Profile ◽

Expression Profiles ◽

The Other ◽

Expression Levels ◽

Healthy Donors ◽

Acute Myeloid

Abstract The Polycomb group (PcG) of genes is important for differentiation and X-chromosome silencing. Recently much attention has been afforded to the role of its aberrant expression in cancer, especially in relation to the inactivation of tumor suppressor genes. We hypothesized that a deregulation in the expression profile may contribute to the development of acute myeloid leukaemia (AML). To address this, we determined the RNA levels by RQ-PCR in diagnostic bone marrow samples from 126 patients and 20 healthy donors to delineate their expression profile of the PcG genes BMI-1, MEL18, SCML2, YY1 and EZH2. To address the interplay with downstream targets of PcG proteins, we also determined the expression of HOXA4, HOXA9 and MEIS1. These data were compared not only to the demographic and clinical data of the patients, but also to a large number of molecular assays already performed in these patients (Olesen LH et al. Br.J. Haematol. 2005 131(4):457–467; Rethmeier et al. Br.J. Haematol. 2006 133(3):276–283.). At first we noticed a striking heterogeneity in the expression profiles of the AML patients (Fig. 1). We also observed that HOXA9, MEIS1, SCML2, YY1, BMI-1 and EZH2 were significantly (p≤0.003) higher expressed in the patients compared to the healthy donors. Moreover, when patients were analyzed according to the three cytogenetic prognostic groups (normal, core-binding factor positive and complex), the expression profile of patients with the t(8,21) aberration was characterized by a significantly decreased expression of HOXA9 and MEIS1 and a higher one of SCML2, YY1 and BMI-1 than AML patients in general (p<0.003). When evaluating the impact of cytogenetic subgrouping, the expression levels of MEL18 and EZH2 significantly (p< 0.025) reflected highest expression in patients with adverse prognosis and lowest expression with patients exhibiting the most favourable prognosis. While the expression levels of the genes in focus did not correlate to course of disease, we observed that a direct relationship between transcript levels of PcG and PcG-related on the one hand and the DNA methyl transferases (DNMT’s), apoptosis and multidrug-resistance genes (p<0.001) on the other. In conclusion, in this study, which is the first to systematically analyze a series of PcG genes and genes regulated by PcG, we failed to demonstrate a correlation to the clinical outcome of patients with AML. On the other hand, our data strongly suggest that these genes might be involved in the leukaemogenic process by virtue of their relations to DNA methylation (DNMT1, DNMT3B), apoptosis (BAX, CASPASE 3) and multidrug resistance (MDR1, MRP1). Figure 1. Expression profiles of PcG or PcG-regulated genes in AML patients and healthy controls. A. Gene expression profile of all 126 AML patients included (black lines) compared to 20 healthy donors. B. Patients with CBF aberrations, t(8,21), n =7, or inv(16), n =12. The expression is calculated as 2−ΔCt *100), where ΔCt = CtTG−CtCG, CtTG is the Ct value of the target gene, and CtCG is the mean Ct value of the two control genes (B2M and ABL). Figure Figure

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