The murine cripto gene: expression during mesoderm induction and early heart morphogenesis

Development ◽  
1993 ◽  
Vol 118 (4) ◽  
pp. 1157-1168 ◽  
Author(s):  
R. Dono ◽  
L. Scalera ◽  
F. Pacifico ◽  
D. Acampora ◽  
M.G. Persico ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Pattern ◽  
Adult Animal ◽  
Mesoderm Induction ◽  
Coding Region ◽  
Developing Heart ◽  
Human Counterpart ◽  
Epidermal Growth

The murine cripto gene encodes a 171-aminoacid epidermal growth factor-related protein, with 93% similarity to its human counterpart in the ‘EGF-like’ domain. The murine cripto mRNA contains two B1 repeats in its 3′ non-coding region and a 163-nucleotide homology to the human mRNA. The mouse cripto gene is expressed at low level in specific organs of the adult animal such as spleen, heart, lung and brain. In situ hybridization analysis during murine embryogenesis (day 6.2 to day 10.5) reveals a very restricted expression pattern. cripto transcripts are first detected in a few epiblastic cells at day 6.5. During gastrulation, the transcripts are expressed in the forming mesoderm and later during development cripto gene expression is restricted to the truncus arteriosus of the developing heart. This expression pattern suggests a role for cripto gene in the determination of the epiblastic cells that subsequently give rise to the mesoderm.

scholarly journals High resolution spatial transcriptome analysis by photo-isolation chemistry

2020 ◽  
Author(s):  
Mizuki Honda ◽  
Shinya Oki ◽  
Akihito Harada ◽  
Kazumitsu Maehara ◽  
Kaori Tanaka ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcriptome Analysis ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Transcriptome Profiling ◽  
Tissue Sections ◽  
Complex Functions ◽  
Multicellular Organisms

ABSTRACTIn multicellular organisms, individual cells are characterized by their gene expression profiles and the spatial interactions among cells enable the elaboration of complex functions. Expression profiling in spatially defined regions is crucial to elucidate cell interactions and functions. Here, we established a transcriptome profiling method coupled with photo-isolation chemistry (PIC) that allows the determination of expression profiles specifically from photo-irradiated regions of whole tissues. PIC uses photo-caged oligodeoxynucleotides for in situ reverse transcription. After photo-irradiation of limited areas, gene expression was detected from at least 10 cells in the tissue sections. PIC transcriptome analysis detected genes specifically expressed in small distinct areas of the mouse embryo. Thus, PIC enables transcriptome profiles to be determined from limited regions at a spatial resolution up to the diffraction limit.

In situ localization of mRNA for epidermal growth factor in the submandibular gland of the mouse.

1985 ◽  
Vol 33 (12) ◽  
pp. 1235-1240 ◽  
Author(s):  
E W Gresik ◽  
R M Gubits ◽  
T Barka
Keyword(s):  
In Situ Hybridization ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Submandibular Gland ◽  
Cdna Probe ◽  
Coding Region ◽  
Specific Hybridization ◽  
Epidermal Growth ◽  
Convoluted Tubules

Epidermal growth factor (EGF) is a polypeptide originally isolated from the mouse submandibular gland, where it is localized immunocytochemically in cells of the granular convoluted tubules (GCT). cDNAs encoding the precursor of mouse submandibular EGF have been cloned (Scott et al. Science 221:236, 1983; Gray et al. Nature 303:722, 1983). A fragment of one of these clones, pmegf10, containing the EGF coding region, was tritium-labeled by nick-translation and used as a probe for in situ hybridization to EGF mRNA. A specific hybridization signal for EGF mRNA was seen only in mature or developing GCT cells. The intensity of the signal was stronger in glands of intact males than in females or in castrated males. In glands of castrates treated with testosterone, or of intact females treated with triiodothyronine (T3), the signal was comparable to that in intact males. In glands of males treated with T3 the intensity of the signal was stronger than in untreated males. A weak to moderate signal was seen in developing GCT cells of 20-day-old males but not females. Hybridization for 3 days gave a stronger signal than that for 1 day. No signal was seen in either sex at 10 days of age, or in control preparations exposed to labeled DNA of pBR322. The presence of EGF mRNA exclusively in GCT cells provides strong evidence that these cells are the only site of synthesis of EGF in the submandibular gland. In situ hybridization with this cDNA probe will provide a sensitive method to determine possible cellular sites of EGF production outside of the submandibular gland.

A GATA-dependent nkx-2.5 regulatory element activates early cardiac gene expression in transgenic mice

Development ◽  
1998 ◽  
Vol 125 (22) ◽  
pp. 4461-4470 ◽  
Author(s):  
R.D. Searcy ◽  
E.B. Vincent ◽  
C.M. Liberatore ◽  
K.E. Yutzey
Keyword(s):  
Gene Expression ◽  
Transgenic Mice ◽  
Expression Pattern ◽  
Binding Sites ◽  
Early Stage ◽  
Regulatory Element ◽  
Gene Activation ◽  
Flanking Sequence ◽  
Regulatory Factors ◽  
Developing Heart

nkx-2.5 is one of the first genes expressed in the developing heart of early stage vertebrate embryos. Cardiac expression of nkx-2.5 is maintained throughout development and nkx-2.5 also is expressed in the developing pharyngeal arches, spleen, thyroid and tongue. Genomic sequences flanking the mouse nkx-2.5 gene were analyzed for early developmental regulatory activity in transgenic mice. Approximately 3 kb of 5′ flanking sequence is sufficient to activate gene expression in the cardiac crescent as early as E7.25 and in limited regions of the developing heart at later stages. Expression also was detected in the developing spleen anlage at least 24 hours before the earliest reported spleen marker and in the pharyngeal pouches and their derivatives including the thyroid. The observed expression pattern from the −3 kb construct represents a subset of the endogenous nkx-2.5 expression pattern which is evidence for compartment-specific nkx-2.5 regulatory modules. A 505 bp regulatory element was identified that contains multiple GATA, NKE, bHLH, HMG and HOX consensus binding sites. This element is sufficient for gene activation in the cardiac crescent and in the heart outflow tract, pharynx and spleen when linked directly to lacZ or when positioned adjacent to the hsp68 promoter. Mutation of paired GATA sites within this element eliminates gene activation in the heart, pharynx and spleen primordia of transgenic embryos. The dependence of this nkx-2. 5 regulatory element on GATA sites for gene activity is evidence for a GATA-dependent regulatory mechanism controlling nkx-2.5 gene expression. The presence of consensus binding sites for other developmentally important regulatory factors within the 505 bp distal element suggests that combinatorial interactions between multiple regulatory factors are responsible for the initial activation of nkx-2.5 in the cardiac, thyroid and spleen primordia.

scholarly journals Human epidermal growth factor receptor 2 testing in breast cancer

2010 ◽  
Vol 63 (1-2) ◽  
pp. 69-74 ◽  
Author(s):  
Tatjana Ivkovic-Kapicl ◽  
Slavica Knezevic-Usaj
Keyword(s):  
Breast Cancer ◽  
Epidermal Growth Factor Receptor ◽  
In Situ Hybridization ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Gene Amplification ◽  
Growth Factor Receptor ◽  
Epidermal Growth

Introduction. Testing for human epidermal growth factor receptor-2 in breast cancer at the time of primary diagnosis is now the standard of care. Positivity for epidermal growth factor receptor-2 in breast cancer is a prognostic factor regarding tumor aggressiveness and a predictive factor for response to Herceptin. Accurate assessment is essential to ensure that all patients who may benefit from Herceptin are correctly identified. Assay method. The principal testing methods used for determination of epider?mal growth factor receptor-2 status are immunohistochemistry for protein over expression and in situ hybridization using either fluorescence or a chromogen. Immunohistochemical testing method allows identification of epidermal growth factor receptor-2 positive patients (3+) who may benefit from Herceptin therapy, whereas negative patients (0/1+) can be excluded. A proportion of specimens defined as equivocal by immunohistochemistry (2+) must be retested by in situ hybridization to determine their status. Chromogen in situ hybridization is a method for determination of gene amplification using a peroxidase-based chromogenic reaction, which can be viewed using a conventional bright field microscope and it determines the actual degree of gene amplification. Various factors can affect the results achieved with these assays, including the assay antibody/probe, the methodology and the experience of personnel. Many countries implemented national testing guidelines in an attempt to standardize testing procedures and make results more accurate. Conclusion. The key point underlined by this review is that whatever method is used to test HER2 status, the technology must be validated first, and there must be regular internal and external quality assurance procedures.

Determination of gene expression patterns using in situ hybridization to Drosophila testes

2009 ◽  
Vol 4 (12) ◽  
pp. 1807-1819 ◽  
Author(s):  
Ceri A Morris ◽  
Elizabeth Benson ◽  
Helen White-Cooper
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Expression Patterns ◽  
Gene Expression Patterns

scholarly journals Left-Right Asymmetry and Kinesin Superfamily Protein KIF3A: New Insights in Determination of Laterality and Mesoderm Induction by kif3A−/− Mice Analysis

1999 ◽  
Vol 145 (4) ◽  
pp. 825-836 ◽  
Author(s):  
Sen Takeda ◽  
Yoshiaki Yonekawa ◽  
Yosuke Tanaka ◽  
Yasushi Okada ◽  
Shigenori Nonaka ◽  
...  
Keyword(s):  
Sonic Hedgehog ◽  
Mesoderm Induction ◽  
Developmental Abnormalities ◽  
Neural Tissues ◽  
Left Right Asymmetry ◽  
Kinesin Superfamily ◽  
Anterior Posterior

KIF3A is a classical member of the kinesin superfamily proteins (KIFs), ubiquitously expressed although predominantly in neural tissues, and which forms a heterotrimeric KIF3 complex with KIF3B or KIF3C and an associated protein, KAP3. To elucidate the function of the kif3A gene in vivo, we made kif3A knockout mice. kif3A−/− embryos displayed severe developmental abnormalities characterized by neural tube degeneration and mesodermal and caudal dysgenesis and died during the midgestational period at ∼10.5 dpc (days post coitum), possibly resulting from cardiovascular insufficiency. Whole mount in situ hybridization of Pax6 revealed a normal pattern while staining by sonic hedgehog (shh) and Brachyury (T) exhibited abnormal patterns in the anterior-posterior (A-P) direction at both mesencephalic and thoracic levels. These results suggest that KIF3A might be involved in mesodermal patterning and in turn neurogenesis.

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