Methods on In Situ Hybridization, Immunohistochemistry and �-Galactosidase Reporter Gene Detection

Franco, Diego; Boer, P. A. de; Vries, Corrie de Gier-de; Lamers, Wouter H.; Moorman, A. F. M.

doi:10.1076/ejom.39.3.169.4670

Cited by 36 publications

(29 citation statements)

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“…We have previously documented that Kcnq1 and Kcnh2 are homogeneously distributed in the developing heart from early stages of development (Franco et al, 2001b), in line with the results obtained in this study. The role of these potassium channels in the cardiac repolarization phase of the action potential is widely acknowledged within the developing (Davies et al, 1996) and adult myocardium (Charpentier et al, 2004).…”

Section: K Channel Pore-forming Subunits In Developing Cardiovascularsupporting

confidence: 92%

“…We previously described the transcript expression profiles of Kcnq1, Kcnh2, Kcne1, Kcne2, and Kcne3 in the developing heart (Franco et al, 2001b). Our observations suggested a critical role of ␤-subunits in the chamber-specific cardiac repolarization based on their differential gene expression profile in the developing heart.…”

mentioning

confidence: 59%

“…Kcnq1 and Kcnh2 have been reported to be expressed in distinct adult organs by RNAse protection assays and RT-PCR techniques (London et al, 1997;Demolombe et al, 2001;Polvani et al, 2003). However, to date, a single study has described the protein expression profile of these potassium channel subunits at the tissue level (Franco et al, 2001b), whereas very little information is available on their onset of protein expression and tissue distribution during embryogenesis. In this study, we provide a comprehensive overview of the protein expression profile of Kcnq1 and Kcnh2 during embryogenesis.…”

Section: Discussionmentioning

confidence: 99%

“…However, Kcnq1 and Kcnh2 are much wider expressed than exclusively in the heart (Demolombe et al, 2001), making such a paradigm exclusively applicable to the heart. Second and more important, the complex Kcnq1/Kcne3 provides a constitutive open channel conformation when coexpressed in Xenopus oocytes and Kcne3 expression in the heart as well as in other organs is broadly documented (Franco et al, 2001b;Lan et al, 2005). Lundquist et al (2005) have recently provided some evidences that a fine balance of Kcne members can modulate the functional properties of Kcnq1 and Kcnh2.…”

Section: Role Of ␤-Subunits In K Channel Distribution and Functionmentioning

confidence: 99%

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Protein distribution of Kcnq1, Kcnh2, and Kcne3 potassium channel subunits during mouse embryonic development

2006

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Voltage-dependent potassium channels consist of a pore-forming ␣-subunit, which is modulated by additional ␤-ancillary or regulatory subunits. Kcnq1 and Kcnh2 ␣-channel subunits play pivotal roles in the developing and adult heart. However, Kcnq1 and Kcnh2 have a much wider expression profile than strictly confined to the myocardium, similar to their putative regulatory Kcne1-5 ␤-subunits. At present, the distribution of distinct potassium channel subunits has been partially mapped in adult tissues, whereas almost no information is available during embryonic development. In this study, we report a detailed analysis of Kcnq1, Kcnh2, and Kcne3 protein expression during mouse embryogenesis. Our results demonstrate that Kcnq1 and Kcnh2 are widely distributed. Coexpression of both ␣-subunits is observed in a wide variety of organs, such as heart and the skeletal muscle, whereas others display unique Kcnq1 or Knch2 expression. Interestingly, Kcne3 expression is also widely observed in distinct tissue layers during embryogenesis, supporting the notion that an exquisite balance of ␣-and ␤-subunit expression is required for modulating potassium conductance in distinct organs and tissue layers.

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Section: K Channel Pore-forming Subunits In Developing Cardiovascularsupporting

confidence: 92%

mentioning

confidence: 59%

Section: Discussionmentioning

confidence: 99%

Section: Role Of ␤-Subunits In K Channel Distribution and Functionmentioning

confidence: 99%

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Protein distribution of Kcnq1, Kcnh2, and Kcne3 potassium channel subunits during mouse embryonic development

2006

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“…Quantitative PCR was performed using SYBR Green I master mix on a LightCycler 480 (Roche Applied Science), with RPL7 as the housekeeping gene control. In situ hybridization was performed as described (20). The PRL2 probe was amplified from mouse PRL2 cDNA using primers listed in supplemental Table 1.…”

Section: Methodsmentioning

confidence: 99%

Phosphatase of Regenerating Liver 2 (PRL2) Is Essential for Placental Development by Down-regulating PTEN (Phosphatase and Tensin Homologue Deleted on Chromosome 10) and Activating Akt Protein

Dong

Zhang

et al. 2012

Journal of Biological Chemistry

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Background:The physiological functions of the PRL phosphatases are poorly understood. Results: PRL2 deficiency causes placental insufficiency, decreased spongiotrophoblast proliferation, and growth retardation. Conclusion: PRL2 plays an important role in placental development by down-regulating PTEN and activating Akt. Significance: This study provides the first evidence of an essential function for PRL2 and offers a biochemical basis for PRLs as oncoproteins to repress PTEN expression.

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Species‐specific differences of myosin content in the developing cardiac chambers of fish, birds, and mammals

et al. 2002

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Key morphogenetic events during heart ontogenesis are similar in different vertebrate species. We report that in primitive vertebrates, i.e., cartilaginous fishes, both the embryonic and the adult heart show a segmental subdivision similar to that of the embryonic mammalian heart. Early morphogenetic events during cardiac development in the dogfish are long-lasting, providing a suitable model to study changes in pattern of gene expression during these stages. We performed a comparative study among dogfish, chicken, rat, and mouse to assess whether species-specific qualitative and/or quantitative differences in myosin heavy chain (MyHC) distribution arise during development, indicative of functional differences between species. MyHC RNA content was investigated by means of in situ hybridisation using an MyHC probe specific for a highly conserved domain, and MyHC protein content was assessed by immunohistochemistry. MyHC transcripts were found to be homogeneously distributed in the myocardium of the tubular and embryonic heart of dogfish and rodents. A difference between atrial and ventricular MyHC content (mRNA and protein) was observed in the adult stage. Interestingly, differences in the MyHC content were observed at the tubular heart stage in chicken. These differences in MyHC content illustrate the distinct developmental profiles of avian and mammalian species, which might be ascribed to distinct functional requirements of the myocardial segments during ontogenesis. The atrial myocardium showed the highest MyHC content in the adult heart of all species analysed (dogfish (S. canicula), mouse (M. musculus), rat (R. norvegicus), and chicken (G. gallus)). These observations indicate that in the adult heart of vertebrates the atrial myocardium contains more myosin than the ventricular myocardium. Anat Rec 268: 27-37, 2002.

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Methods on In Situ Hybridization, Immunohistochemistry and �-Galactosidase Reporter Gene Detection

Cited by 36 publications

References 0 publications

Protein distribution of Kcnq1, Kcnh2, and Kcne3 potassium channel subunits during mouse embryonic development

Protein distribution of Kcnq1, Kcnh2, and Kcne3 potassium channel subunits during mouse embryonic development

Phosphatase of Regenerating Liver 2 (PRL2) Is Essential for Placental Development by Down-regulating PTEN (Phosphatase and Tensin Homologue Deleted on Chromosome 10) and Activating Akt Protein

Species‐specific differences of myosin content in the developing cardiac chambers of fish, birds, and mammals

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