Specification and determination of limb identity: evidence for inhibitory regulation of Tbx gene expression

Development ◽  
2002 ◽  
Vol 129 (1) ◽  
pp. 211-220 ◽  
Author(s):  
Daisuke Saito ◽  
Sayuri Yonei-Tamura ◽  
Kohko Kano ◽  
Hiroyuki Ide ◽  
Koji Tamura
Keyword(s):  
Gene Expression ◽  
Culture Medium ◽  
Inhibitory Effects ◽  
Rt Pcr ◽  
Lateral Plate ◽  
Specific Expression ◽  
Type Identity ◽  
Transplantation Experiments ◽  
Inhibitory Regulation

Limb-type-specific expression of Tbx5/Tbx4 plays a key role in drawing distinction between a forelimb and a hindlimb. Here, we show insights into specification and determination during commitment of limb-type identity, in particular that median tissues regulate Tbx expressions. By using the RT-PCR technique on chick embryos, the onset of specific Tbx5/Tbx4 expression in the wing/leg region was estimated to be stage 13. Specification of the limb-type identity is thought to occur before stage 9, since all explants from stage 9 through 14 expressed the intrinsic Tbx gene autonomously in a simple culture medium. The results of transplantation experiments revealed that axial structures medial to the lateral plate mesoderm at the level of the wing region are capable of transforming leg identity to wing identity, suggesting that a factor(s) from the median tissues is involved in the limb-type determination. Nevertheless, the transplanted wing region was not converted to leg identity. The results of the transplantation experiments also suggested that wing-type identity is determined much earlier than is leg-type identity. Finally, we also found that inhibitory effects of median tissues mediate the specific expression of Tbx5/Tbx4 in the presumptive wing/leg region. We propose a model for limb-type identification in which inhibitory regulation is involved in restricting one Tbx gene expression by masking the other Tbx expression there.

scholarly journals Development of cDNA normalization system and preliminary transcription analysis of KCS genes in apple tissues

2011 ◽  
Vol 59 (3) ◽  
pp. 9-12 ◽  
Author(s):  
Zsolt Albert ◽  
Cs. Deák ◽  
A. Miskó ◽  
M. Tóth ◽  
I. Papp
Keyword(s):  
Gene Expression ◽  
Expression System ◽  
Expression Patterns ◽  
Housekeeping Genes ◽  
Apple Fruit ◽  
Rt Pcr ◽  
Specific Primers ◽  
Specific Expression ◽  
Transcription Analysis ◽  
Tissue Specific

Wax production is an important aspect of apple (Malus domestica Borkh.) fruit development from both theoretical and practical point of views. The complex molecular mechanism that controls wax biosynthesis is still widely unknown but many studies focused on this topic. We aimed to develop further the experimental framework of these efforts with a description of an improved reference genes expression system. Results in the literature show that similarities exist among the expression of some housekeeping genes of different plant species. Based on these considerations and on gene expression data from Arabidopsis thaliana, some genes in apple were assigned for analysis. EST sequences of apple were used to design specific primers for RT-PCR experiments. Isolation of intact RNA from different apple tissues and performing RT-PCR reaction were also key point in obtaining expression patterns. To monitor DNA contamination of the RNA samples, specific primers were used that amplify intron-containing sequences from the cDNA. We found that actin primers can be used for the detection of intron containing genomic DNA, and tubulin primers are good internal controls in RT-PCR experiments. We were able to make a difference between tissue-specific and tissue-independent gene-expression, furthermore we found tissue specific differences between the expression patterns of candidate genes, that are potentially involved in wax-biosynthesis. Our results show that KCS1 and KCS4 are overexpressed in the skin tissue, this could mean that these genes have skin-specific expression in apple fruit.

Lineage-specific defect in gene expression in human platelet phospholipase C-β2 deficiency

Blood ◽  
2002 ◽  
Vol 99 (3) ◽  
pp. 905-911 ◽  
Author(s):  
Guang Fen Mao ◽  
Vijender R. Vaidyula ◽  
Satya P. Kunapuli ◽  
A. Koneti Rao
Keyword(s):  
Gene Expression ◽  
Phospholipase C ◽  
Internal Control ◽  
Healthy Subjects ◽  
Leukotriene B4 ◽  
Calcium Mobilization ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Specific Expression ◽  
Platelet Factor

Abstract Phospholipase C (PLC)–β2 plays a major role in platelet activation. Previous studies have described a unique patient with impaired receptor-mediated platelet aggregation, secretion, calcium mobilization, and phospholipase C (PLC) activation associated with a selective decrease in platelet PLC-β2 isozyme. To identify the mechanisms leading to the defect, platelet RNA from the patient and healthy subjects was subjected to reverse transcription–polymerase chain reaction (RT-PCR) and the products sequenced. The PLC-β2 cDNA sequence in the patient showed no abnormalities. Platelet PLC-β2 and β-actin (internal control) mRNA levels were assessed by RT-PCR; the ratio of PLC-β2 to β-actin mRNA levels was 0.80 to 0.95 in 4 healthy subjects and 0.28 in the patient. PLC-β2 mRNA levels were similarly reduced compared with GPIIb and Gαq mRNA levels. PLC-γ2 and platelet factor 4 mRNA levels were normal. Calcium mobilization was studied in neutrophils upon activation with formyl-Met-Leu-Phe (fMLP), adenosine diphosphate (ADP), platelet-activating factor (PAF), interleukin-8 (IL-8), C5a, and leukotriene B4 (LTB4), and it was normal. Neutrophil elastase secretion upon activation with fMLP, ADP, PAF, IL-8, C5a, and LTB4 was normal, as were neutrophil PLC-β2 mRNA and PLC-β2 on immunoblotting. Thus, responses to activation, PLC-β2 protein, and PLC-β2 mRNA are decreased in patient platelets but not in neutrophils, providing evidence for a hitherto undescribed lineage (platelet)–specific defect in PLC-β2 gene expression. These studies provide a physiologically relevant model to delineate regulation of PLC-β2 gene and its tissue-specific expression.

scholarly journals Tumor quantification of several fluoropyrimidines resistance gene expression with a unique quantitative RT-PCR method. Implications for pretherapeutic determination of tumor resistance phenotype

2006 ◽  
Vol 242 (2) ◽  
pp. 168-179 ◽  
Author(s):  
Maud Barbado ◽  
Laurence Preisser ◽  
Michele Boisdron-Celle ◽  
Veronique Verriele ◽  
Gerard Lorimier ◽  
...  
Keyword(s):  
Gene Expression ◽  
Resistance Gene ◽  
Tumor Resistance ◽  
Rt Pcr ◽  
Resistance Phenotype ◽  
Pcr Method

scholarly journals Effects of acanthoic acid on TNF-α gene expression and haptoglobin synthesis

1998 ◽  
Vol 7 (4) ◽  
pp. 257-259 ◽  
Author(s):  
H-S. Kang ◽  
H. K. Song ◽  
J-J. Lee ◽  
K-H. Pyun ◽  
I. Choi
Keyword(s):  
Gene Expression ◽  
Biological Function ◽  
Acute Phase Protein ◽  
Inhibitory Effects ◽  
Rt Pcr ◽  
Tnf Α ◽  
Phase Protein ◽  
Factor Α ◽  
Necrosis Factor ◽  
Acanthoic Acid

Tumour necrosis factor-α (TNF-α) is a major proinflammatory cytokine inducing the synthesis and release of many inflammatory mediators. It is involved in immune regulation, autoim mune diseases, and inflammation. Our previous study demonstrated that acanthoic acid, (-)-pimara-9(11), 15-dien19-oic acid, a pimaradiene diterpene isolated fromAcanth opanax koreanum, inhibited TNF-α production. To extend our understanding of inhibitory effects of acanthoic acid on TNF-α production, its effects on TNF-α gene expression was tested. Based on the results from RT-PCR and promoter analysis of TNF-α, it was found that acanthoic acid suppressed TNF-α gene expression. But the same concentration of acanthoic acid had no effect on IL-6 gene expression. Haptoglobin is an acute phase protein which is induced by TNF-α. When liver cells were treated with acanthoic acid, haptoglobin synthesis was blocked by acanthoic acid. These data confirmed that acanthoic acid inhibited gene expression and biological function of TNF-α.

A real-time RT-PCR assay for the quantitiative determination of adenoviral gene expression in tumor cells

2006 ◽  
Vol 133 (1) ◽  
pp. 53-61 ◽  
Author(s):  
Mohamed A.I. Abou El Hassan ◽  
Stefan R. Braam ◽  
Frank A.E. Kruyt
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Tumor Cells ◽  
Rt Pcr ◽  
Pcr Assay

Quantitative Determination of p16 Gene Expression by RT-PCR

2004 ◽  
pp. 091-104
Author(s):  
Sylke Schneider ◽  
Kazumi Uchida ◽  
Dennis Salonga ◽  
Ji Min Yochim ◽  
Kathleen D. Danenberg ◽  
...  
Keyword(s):  
Gene Expression ◽  
Quantitative Determination ◽  
Rt Pcr ◽  
P16 Gene

710 Determination of Gene Expression in Endometrial Cancer Using Real-time RT-PCR

2012 ◽  
Vol 48 ◽  
pp. S168
Author(s):  
E. Pluciennik ◽  
K. Kosla ◽  
M. Pospiech ◽  
M. Nowakowska ◽  
K. Wojcik-Krowiranda ◽  
...  
Keyword(s):  
Gene Expression ◽  
Endometrial Cancer ◽  
Real Time ◽  
Rt Pcr

scholarly journals DD3PCA3 gene expression in cancer and prostatic hyperplasia

2009 ◽  
Vol 32 (6) ◽  
pp. 258 ◽  
Author(s):  
E Floriano-Sánchez ◽  
N Cárdenas-Rodríguez ◽  
M Castro-Marín ◽  
P Álvarez -Grave ◽  
E Lara-Padilla
Keyword(s):  
Gene Expression ◽  
Benign Prostatic Hyperplasia ◽  
Pearson Correlation ◽  
Housekeeping Gene ◽  
Prostatic Hyperplasia ◽  
Rt Pcr ◽  
Gapdh Gene ◽  
Polymerase Chain ◽  
Valuable Role

Purpose: DD3PCA3 is a novel gene with characteristics that indicate its potentially valuable role in early identification and diagnosis of malignancy and highly upregulated in transformed cells in PCa. The aim of this work was to validate and analyze, by real-time Reverse Transcription-Polymerase Chain Reaction (RT-PCR), the expression of the DD3PCA3 gene in a mexican population, both in intratumoral tissue with PCa and benign prostatic hyperplasia (BPH). Methods: Human samples from patients with PCa (40 cases) and benign prostatic hyperplasia (40 cases) were analyzed for the mRNA expression of DD3PCA3 by RT-PCR Results: The GAPDH gene showed better stability with a Pearson correlation of 0.953 (P < 0.007) for the determination of housekeeping gene. DD3PCA gene expression was 29.74 times higher in PCa tissue (P < 0.0001) than in BPH. The gene expression for the PCa and BPH was 1731±280 and 58.23±9.9 fold, respectively. Conclusions: Determination of DD3PCA3 gene expression by RT-PCR could be a potentially tool for the early detection of PCa in clinical specimens.

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