کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1924367 1048958 2007 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Improved cryosections and specific immunohistochemical methods for detecting hypoxia in mouse and rat cochleae
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Improved cryosections and specific immunohistochemical methods for detecting hypoxia in mouse and rat cochleae
چکیده انگلیسی

SummaryThe present study was undertaken to develop an improved cryoembedding method for analysis of mice and rat cochleae, which permits high-quality cryosections and preserves overall structure and cellular resolution as shown by hematoxylin/eosin staining. The preservation of morphology and antigenicity is mandatory to achieve optimal results. A total of 20 male cd/1 mice and 14 male Sprague-Dawley rats were used in experiments for optimization of preservation, fixative, decalcification, embedding and cryosectioning of cochleae from adult and aged rodents. In addition, a novel immunohistochemical procedure (using Hydroxyprobe™-1 kit) was developed for detecting regions of hypoxia in mice and rat cochlea. This method employs a primary fluorescent-conjugated monoclonal antibody directed against pimonidazole protein adducts that are created in hypoxic tissues. Subsequent studies of hypoxia inducible factor-1α (HIF-1α) by immunofluorescence in the cochlea of these animals were performed in order to confirm that immunochemical detection of pimonidazole protein is representative of a hypoxic environment. We conclude that the present method results in high-quality cryosections of cochlear tissues presenting good anatomical and histological preservation. Furthermore, our optimized procedures provide novel tools for the investigation of neuro-sensory-epithelium in physio-pathological situations associated with hypoxia and/or ischemia, such as inner ear development, plasticity, regeneration and senescence.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Acta Histochemica - Volume 109, Issue 3, 18 June 2007, Pages 177–184
نویسندگان
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