کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5506895 | 1400305 | 2016 | 30 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Real-time imaging of mast cell degranulation in vitro and in vivo
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کلمات کلیدی
RFPVAMPIgEGFPSNARETNPOVA2,4,6-trinitrophenol - 2،4،6-ترنیتروفنلOvalbumin - اوبلبومینExocytosis - اگزوسیتوز IgE, immunoglobulin E - ایمونوگلوبولین EImaging - تصویربرداریpH sensor - سنسور pHDegranulation - قرمزی چشمMast cell - ماستسلVesicle-associated membrane protein - پروتئین غشاء مرتبط با Vesiclegreen fluorescent protein - پروتئین فلورسنت سبزred fluorescent protein - پروتئین فلورسنت قرمزsoluble N-ethylmaleimide-sensitive factor attachment protein receptor - گیرنده پروتئین دلبستگی حساس به پروتئین محلول N-ethylmaleimide
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Mast cells undergo degranulation in response to various stimuli and rapidly release pre-formed mediators present in secretory granules, leading to immediate-type allergic reactions. Mast cell degranulation is commonly detected and quantified in vitro by measuring histamine or β-hexosaminidase released to culture medium. However, this type of assay cannot monitor degranulation of individual cells in real time, and it is not suitable for in vivo detection of degranulation. At the aim of real time imaging of mast cell degranulation at single cell level, we here developed a fluorescent protein-based indicator of degranulation, designated immuno-pHluorin (impH). When expressed in mast cells, impH is located in the membrane of secretory granules and non-fluorescent under homeostatic conditions while it turns fluorescent following degranulation, due to the pH change inside of granules during exocytosis. impH enabled us to detect polarized degranulation within one single cell when mast cells were stimulated via the small area of cell surface. Transplantation of impH-expressing mast cells into mast cell-deficient mice demonstrated that impH could function as a real-time indicator of degranulation in vivo. Thus, impH is a useful tool for imaging of mast cell activation and degranulation in vitro and in vivo, and may be applied for screening of reagents regulating mast cell degranulation.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical and Biophysical Research Communications - Volume 479, Issue 3, 21 October 2016, Pages 517-522
Journal: Biochemical and Biophysical Research Communications - Volume 479, Issue 3, 21 October 2016, Pages 517-522
نویسندگان
Kayo Horiguchi, Soichiro Yoshikawa, Asuka Saito, Salma Haddad, Takuya Ohta, Kensuke Miyake, Yoshinori Yamanishi, Hajime Karasuyama,