Calcium phosphate transfection of primary hippocampal neurons.
نویسندگان
چکیده
Calcium phosphate precipitation is a convenient and economical method for transfection of cultured cells. With optimization, it is possible to use this method on hard-to-transfect cells like primary neurons. Here we describe our detailed protocol for calcium phosphate transfection of hippocampal neurons cocultured with astroglial cells.
منابع مشابه
Chemically controlled formation of a DNA/calcium phosphate coprecipitate: application for transfection of mature hippocampal neurons.
Numerous methods exist for transfecting postmitotic neurons, for example, DNA/calcium phosphate coprecipitation, cationic lipids, viruses, and physical methods such as microinjection, electroporation, and biolistics. Most methods, however, are either toxic to the cell, yield only poor transfection efficiencies, or cells have to be electroporated before plating. In this article, we present a sta...
متن کاملTime course of dysregulation of calcium homeostasis in acutely isolated CA1 hippocampal pyramidal neurons after pilocarpine-induced Status Epilepticus
Glutamate induces excitotoxic damage to hippocampal pyramidal neurons in Status Epilepticus (SE) and epilepsy. In this study, we investigated time course of dysregulation of calcium homeostasis at various intervals after an episode of SE in acutely isolated CA1hippocampal pyramidal. For this purpose, male Sprague-Dawley rats (200 g) were subjected to pilocarpine-induced SE. The SE was blocked a...
متن کاملTime course of dysregulation of calcium homeostasis in acutely isolated CA1 hippocampal pyramidal neurons after pilocarpine-induced Status Epilepticus
Glutamate induces excitotoxic damage to hippocampal pyramidal neurons in Status Epilepticus (SE) and epilepsy. In this study, we investigated time course of dysregulation of calcium homeostasis at various intervals after an episode of SE in acutely isolated CA1hippocampal pyramidal. For this purpose, male Sprague-Dawley rats (200 g) were subjected to pilocarpine-induced SE. The SE was blocked a...
متن کاملAntibiotic Supplements Affect Electrophysiological Properties and Excitability of Rat Hippocampal Pyramidal Neurons in Primary Culture
Introduction: Antibiotic supplements are regularly used in neuronal culture media to control contamination however, they can interfere with the neuronal excitability and affect electrophysiological properties. Therefore, in this study, the effect of penicillin/streptomycin supplements on the spontaneous electrophysiological activity of hippocampal pyramidal neurons was examined. Methods: Electr...
متن کاملInvolvement of the proximal C terminus of the AMPA receptor subunit GluR1 in dendritic sorting.
Studies on dendritic sorting of transmembrane proteins in hippocampal neurons in culture have shown that these cells use similar mechanisms as epithelial cells to sort transmembrane proteins to the basolateral membrane domain. However, information is still scarce with regard to which amino acidic sequences are required for dendritic sorting in neurons. The glutamate receptor 1 (GluR1) subunit o...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Journal of visualized experiments : JoVE
دوره 81 شماره
صفحات -
تاریخ انتشار 2013