کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1993784 1064706 2012 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Purification of cardiac cells from Drosophila embryos
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Purification of cardiac cells from Drosophila embryos
چکیده انگلیسی

Recent genome-level innovations have enabled the identification of the entire cadre of genes that are expressed in specific tissues at particular developmental times. However, to be informative as to how individual cell types develop, this process relies upon the successful and efficient purification of cells for a particular tissue. Here, we describe a method to isolate cardiac cells from Drosophila embryos. We generated transgenic embryos in which a cardiac-specific enhancer of the Sulphonylurea receptor (Sur) gene drove expression of the green fluorescent protein (GFP) gene. Homogenized embryos were subjected to fluorescence activated cell sorting (FACS), resulting in approximately 50,000 cardiac cells purified. The prevalence of cardiac cells in the purified population was high, based upon a significant enrichment for cardiac-specific marker genes, including Sur and Toll. This enrichment also enabled the identification of cardiac-specific alternatively spliced isoforms of the Zasp66 gene. In the future, this approach can be used to describe the cardiac transcriptome of Drosophila at distinct stages of embryonic development.


► A cardiac-specific GFP-expressing line was generated.
► Homogenized embryos were subjected to cell sorting to enrich for cardiac cells.
► Up to 50,000 cardiac cells could be isolated from whole-embryo homogenates.
► The purified cells showed strong enrichment for cardiac markers.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Methods - Volume 56, Issue 1, January 2012, Pages 44–49
نویسندگان
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