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  1. Article ; Online: Murine-Derived Glioma Organoids and Cell Line Culture Systems.

    Ayala-Sarmiento, Alberto E / Pacheco, David Rincon Fernandez / Breunig, Joshua J

    Current protocols

    2023  Volume 3, Issue 2, Page(s) e665

    Abstract: Research models in cancer have greatly evolved in the last decade, with the advent of several new methods both in vitro and in vivo. While in vivo models remain the gold standard for preclinical studies, these methods present a series of disadvantages ... ...

    Abstract Research models in cancer have greatly evolved in the last decade, with the advent of several new methods both in vitro and in vivo. While in vivo models remain the gold standard for preclinical studies, these methods present a series of disadvantages such as a high cost and long periods of time to produce results compared with in vitro models. We have previously developed a method named Mosaic Analysis by Dual Recombinase-mediated cassette exchange (MADR) that generates autochthonous gliomas in immunocompetent mice through the transgenesis of personalized driver mutations, which highly mimic the spatial and temporal tumor development of their human counterparts. Due to the control of single-copy expression of transgenes, it allows for comparing the visualization of tumor cells and non-tumor cells. Here we describe a method to generate murine-derived glioma organoids (MGOs) and cell line cultures from these murine models by physical and enzymatic methods for in vitro downstream applications. Tumor cells can be readily distinguished from non-tumor cell populations, in both organoids and monolayer cell cultures, and isolated due to the use of personalized fluorescent reporter transgenes. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Generation of 3D murine-derived glioma organoids Basic Protocol 2: Generation of 2D glioma monolayer cell lines.
    MeSH term(s) Mice ; Humans ; Animals ; Glioma/genetics ; Glioma/pathology ; Cell Line ; Cell Culture Techniques/methods ; Transgenes ; Organoids/pathology
    Language English
    Publishing date 2023-02-03
    Publishing country United States
    Document type Journal Article
    ISSN 2691-1299
    ISSN (online) 2691-1299
    DOI 10.1002/cpz1.665
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: De Novo

    Ayala-Sarmiento, Alberto E / Kobritz, Naomi / Breunig, Joshua J

    STAR protocols

    2020  Volume 1, Issue 3, Page(s) 100184

    Abstract: Mosaic analysis by dual recombinase-mediated cassette exchange (MADR) is a technology that allows stable and locus-specific integration of transgenic elements into recipient cells carrying loxP and FRT sites. Nevertheless, most cell lines lack these ... ...

    Abstract Mosaic analysis by dual recombinase-mediated cassette exchange (MADR) is a technology that allows stable and locus-specific integration of transgenic elements into recipient cells carrying loxP and FRT sites. Nevertheless, most cell lines lack these recombination-specific sites. This protocol describes a method to introduce the minimum requirements into cells, leading to the generation of
    MeSH term(s) Animals ; Cell Culture Techniques/methods ; Cell Line ; Genetic Loci ; HEK293 Cells ; Humans ; Mice ; Recombinases/metabolism ; Reproducibility of Results ; Transgenes
    Chemical Substances Recombinases
    Language English
    Publishing date 2020-11-25
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ISSN 2666-1667
    ISSN (online) 2666-1667
    DOI 10.1016/j.xpro.2020.100184
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  3. Article ; Online: The Internalization of Neurotensin by the Low-Affinity Neurotensin Receptors (NTSR2 and vNTSR2) Activates ERK 1/2 in Glioma Cells and Allows Neurotensin-Polyplex Transfection of tGAS1.

    Ayala-Sarmiento, Alberto E / Martinez-Fong, Daniel / Segovia, José

    Cellular and molecular neurobiology

    2015  Volume 35, Issue 6, Page(s) 785–795

    Abstract: Glioblastoma is the most malignant primary brain tumor and is very resistant to treatment; hence, it has a poor prognosis. Neurotensin receptor type 1 (NTSR1) plays a key role in cancer malignancy and has potential therapeutic applications. However, the ... ...

    Abstract Glioblastoma is the most malignant primary brain tumor and is very resistant to treatment; hence, it has a poor prognosis. Neurotensin receptor type 1 (NTSR1) plays a key role in cancer malignancy and has potential therapeutic applications. However, the presence and function of neurotensin (NTS) receptors in glioblastoma is not clearly established. RT-PCR assays showed that healthy (non-tumor) astroglial cells and C6 glioma cells express NTSR2 and its isoform (vNTSR2) rather than NTSR1. In glioma cells, NTS promotes the phosphorylation of extracellular signal-regulated kinases 1/2 (ERK 1/2), an effect that was completely abolished by blocking the internalization of the NTS/NTSR complex. We demonstrated pharmacologically that the internalization is dependent on the activation of NTSR2 receptors and it was prevented by levocabastine, a NTSR2 receptor antagonist. The internalization of NTSR2 and vNTSR2 was further demonstrated by its ability to mediate gene transfer (transfection) via the NTS-polyplex system. Expression of reporter transgenes and of the pro-apoptotic soluble form of growth arrest specific 1 (tGAS1) was observed in glioma cells. A significant reduction on the viability of C6 cells was determined when tGAS1 was transfected into glioma cells. Conversely, astroglial cells could neither internalize NTS nor activate ERK 1/2 and could not be transfected by the NTS-polyplex. These results demonstrate that the internalization process of NTSR2 receptors is a key regulator necessary to trigger the activation of the ERK 1/2. Our data support a new internalization pathway in glioma C6 cells that involve NTSR2/vNTSR2, which can be used to selectively transfer therapeutic genes using the NTS-polyplex system.
    MeSH term(s) Animals ; Animals, Newborn ; CHO Cells ; Cell Cycle Proteins/genetics ; Cells, Cultured ; Cricetinae ; Cricetulus ; Endocytosis/genetics ; GPI-Linked Proteins/genetics ; Glioma/metabolism ; Glioma/pathology ; MAP Kinase Signaling System/physiology ; Mice ; Nanoparticles/metabolism ; Neurotensin/metabolism ; Polymers ; Protein Binding ; Protein Isoforms/metabolism ; Rats ; Receptors, Neurotensin/metabolism ; Transfection/methods
    Chemical Substances Cell Cycle Proteins ; GPI-Linked Proteins ; Gas1 protein, rat ; Ntsr2 protein, rat ; Polymers ; Protein Isoforms ; Receptors, Neurotensin ; Neurotensin (39379-15-2)
    Language English
    Publishing date 2015-08
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 283404-2
    ISSN 1573-6830 ; 0272-4340
    ISSN (online) 1573-6830
    ISSN 0272-4340
    DOI 10.1007/s10571-015-0172-z
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1727: Show issues Location:
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    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
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  4. Article ; Online: GAS1 is present in the cerebrospinal fluid and is expressed in the choroid plexus of the adult rat.

    Ayala-Sarmiento, Alberto E / Estudillo, Enrique / Pérez-Sánchez, Gilberto / Sierra-Sánchez, Arturo / González-Mariscal, Lorenza / Martínez-Fong, Daniel / Segovia, José

    Histochemistry and cell biology

    2016  Volume 146, Issue 3, Page(s) 325–336

    Abstract: Growth arrest specific 1 (GAS1) is a GPI-anchored protein that inhibits proliferation when overexpressed in tumors but during development it promotes proliferation and survival of different organs and tissues. This dual ability is caused by its capacity ... ...

    Abstract Growth arrest specific 1 (GAS1) is a GPI-anchored protein that inhibits proliferation when overexpressed in tumors but during development it promotes proliferation and survival of different organs and tissues. This dual ability is caused by its capacity to interact both by inhibiting the signaling induced by the glial cell line-derived neurotrophic factor and by facilitating the activity of the sonic hedgehog pathway. GAS1 is expressed as membrane bound in different organs and as a secreted form by glomerular mesangial cells. In the developing central nervous system, GAS1 is found in neural progenitors; however, it continues to be expressed in the adult brain. Here, we demonstrate that soluble GAS1 is present in the cerebrospinal fluid (CSF) and it is expressed in the choroid plexus (CP) of the adult rat, the main producer of CSF. Additionally, we confirm the presence of GAS1 in blood plasma and liver of the adult rat, the principal source of blood plasma proteins. The pattern of expression of GAS1 is perivascular in both the CP and the liver. In vitro studies show that the fibroblast cell line NIH/3T3 expresses one form of GAS1 and releases two soluble forms into the supernatant. Briefly, in the present work, we show the presence of GAS1 in adult rat body fluids focusing in the CSF and the CP, and suggest that secreted GAS1 exists as two different isoforms.
    MeSH term(s) Animals ; Cell Cycle Proteins/cerebrospinal fluid ; Cell Cycle Proteins/metabolism ; Cells, Cultured ; Choroid Plexus/metabolism ; GPI-Linked Proteins/cerebrospinal fluid ; GPI-Linked Proteins/metabolism ; Mice ; NIH 3T3 Cells ; Rats ; Rats, Wistar
    Chemical Substances Cell Cycle Proteins ; GPI-Linked Proteins ; Gas1 protein, rat
    Language English
    Publishing date 2016-05-25
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 1222930-1
    ISSN 1432-119X ; 0301-5564 ; 0948-6143
    ISSN (online) 1432-119X
    ISSN 0301-5564 ; 0948-6143
    DOI 10.1007/s00418-016-1449-0
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    Ub I Zs.94/a: Show issues Location:
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  5. Article ; Online: Annexin A1, Annexin A2, and Dyrk 1B are upregulated during GAS1-induced cell cycle arrest.

    Pérez-Sánchez, Gilberto / Jiménez, Adriana / Quezada-Ramírez, Marco A / Estudillo, Enrique / Ayala-Sarmiento, Alberto E / Mendoza-Hernández, Guillermo / Hernández-Soto, Justino / Hernández-Hernández, Fidel C / Cázares-Raga, Febe E / Segovia, Jose

    Journal of cellular physiology

    2017  Volume 233, Issue 5, Page(s) 4166–4182

    Abstract: GAS1 is a pleiotropic protein that has been investigated because of its ability to induce cell proliferation, cell arrest, and apoptosis, depending on the cellular or the physiological context in which it is expressed. At this point, we have information ... ...

    Abstract GAS1 is a pleiotropic protein that has been investigated because of its ability to induce cell proliferation, cell arrest, and apoptosis, depending on the cellular or the physiological context in which it is expressed. At this point, we have information about the molecular mechanisms by which GAS1 induces proliferation and apoptosis; but very few studies have been focused on elucidating the mechanisms by which GAS1 induces cell arrest. With the aim of expanding our knowledge on this subject, we first focused our research on finding proteins that were preferentially expressed in cells arrested by serum deprivation. By using a proteomics approach and mass spectrometry analysis, we identified 17 proteins in the 2-DE protein profile of serum deprived NIH3T3 cells. Among them, Annexin A1 (Anxa1), Annexin A2 (Anxa2), dual specificity tyrosine-phosphorylation-regulated kinase 1B (Dyrk1B), and Eukaryotic translation initiation factor 3, F (eIf3f) were upregulated at transcriptional the level in proliferative NIH3T3 cells. Moreover, we demonstrated that Anxa1, Anxa2, and Dyrk1b are upregulated at both the transcriptional and translational levels by the overexpression of GAS1. Thus, our results suggest that the upregulation of Anxa1, Anxa2, and Dyrk1b could be related to the ability of GAS1 to induce cell arrest and maintain cell viability. Finally, we provided further evidence showing that GAS1 through Dyrk 1B leads not only to the arrest of NIH3T3 cells but also maintains cell viability.
    MeSH term(s) Animals ; Annexin A1/genetics ; Annexin A2/genetics ; Apoptosis/genetics ; Cell Cycle Checkpoints/genetics ; Cell Cycle Proteins/genetics ; Cell Proliferation/genetics ; Eukaryotic Initiation Factor-3/genetics ; GPI-Linked Proteins/genetics ; Gene Expression Regulation/genetics ; Humans ; Mice ; NIH 3T3 Cells ; Protein Serine-Threonine Kinases/genetics ; Protein-Tyrosine Kinases/genetics ; Transcriptional Activation ; Dyrk Kinases
    Chemical Substances Annexin A1 ; Annexin A2 ; Cell Cycle Proteins ; Eukaryotic Initiation Factor-3 ; GPI-Linked Proteins ; Gas1 protein, mouse ; annexin A1, mouse ; Protein-Tyrosine Kinases (EC 2.7.10.1) ; Protein Serine-Threonine Kinases (EC 2.7.11.1)
    Language English
    Publishing date 2017-12-04
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 3116-1
    ISSN 1097-4652 ; 0021-9541
    ISSN (online) 1097-4652
    ISSN 0021-9541
    DOI 10.1002/jcp.26226
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    Uc I Zs.212: Show issues Location:
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