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10.3389/fendo.2017.00068 http://scihub22266oqcxt.onion/10.3389/fendo.2017.00068 C5383706!5383706!28439257     free     free     free Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Front+Endocrinol+(Lausanne) 2017 ; 8 (ä): ä Nephropedia Template TP {{tp|p=28439257|t=2017. High-Resolution Recording of the Circadian Oscillator in Primary Mouse ?- and ?-Cell Culture |pdf=|usr=}}{{28439257}} gab.com Text High-Resolution Recording of the Circadian Oscillator in Primary Mouse - and -Cell Culture JO: Front Endocrinol (Lausanne) http://www.kidney.de/pget.php?&tw=1&pmid=28439257 #FOAvip Circadian clocks have been developed in evolution as an anticipatory mechanism allowing for adaptation to the constantly changing light environment due to rotation of the Earth. This mechanism is functional in all light-sensitive organisms. There is a considerable body of evidence on the tight connection between the circadian clock and most aspects of physiology and metabolism. Clocks, operative in the pancreatic islets, have caught particular attention in the last years due to recent reports on their critical roles in regulation of insulin secretion and etiology of type 2 diabetes. While ?-cell clocks have been extensively studied during the last years, ?-cell clocks and their role in islet function and orchestration of glucose metabolism stayed unexplored, largely due to the difficulty to isolate ?-cells, which represents a considerable technical challenge. Here, we provide a detailed description of an experimental approach for the isolation of separate mouse ?- and ?-cell population, culture of isolated primary ?- and ?-cells, and their subsequent long-term high-resolution circadian bioluminescence recording. For this purpose, a triple reporter ProGlucagon-Venus/RIP-Cherry/Per2:Luciferase mouse line was established, carrying specific fluorescent reporters for ?- and ?-cells, and luciferase reporter for monitoring the molecular clockwork. Flow cytometry fluorescence-activated cell sorting allowed separating pure ?- and ?-cell populations from isolated islets. Experimental conditions, developed by us for the culture of functional primary mouse ?- and ?-cells for at least 10?days, will be highlighted. Importantly, temporal analysis of freshly isolated ?- and ?-cells around-the-clock revealed preserved rhythmicity of core clock genes expression. Finally, we describe the setting to assess circadian rhythm in cultured ?- and ?-cells synchronized in vitro. The here-described methodology allows to analyze the functional properties of primary ?- and ?-cells under physiological or pathophysiological conditions and to assess the islet cellular clock properties. Twit Text FOAVip High-Resolution Recording of the Circadian Oscillator in Primary Mouse - and -Cell Culture ????Front Endocrinol (Lausanne) http://www.kidney.de/pget.php?&tw=1&pmid=28439257 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=28439257 #FOAvip English Wikipedia <ref>{{Cite pmid|28439257}}</ref> High-Resolution Recording of the Circadian Oscillator in Primary Mouse ?- and ?-Cell Culture #MMPMID28439257 Petrenko V; Gosmain Y; Dibner C Front Endocrinol (Lausanne) 2017[]; 8 (ä): ä PMID28439257show ga Circadian clocks have been developed in evolution as an anticipatory mechanism allowing for adaptation to the constantly changing light environment due to rotation of the Earth. This mechanism is functional in all light-sensitive organisms. There is a considerable body of evidence on the tight connection between the circadian clock and most aspects of physiology and metabolism. Clocks, operative in the pancreatic islets, have caught particular attention in the last years due to recent reports on their critical roles in regulation of insulin secretion and etiology of type 2 diabetes. While ?-cell clocks have been extensively studied during the last years, ?-cell clocks and their role in islet function and orchestration of glucose metabolism stayed unexplored, largely due to the difficulty to isolate ?-cells, which represents a considerable technical challenge. Here, we provide a detailed description of an experimental approach for the isolation of separate mouse ?- and ?-cell population, culture of isolated primary ?- and ?-cells, and their subsequent long-term high-resolution circadian bioluminescence recording. For this purpose, a triple reporter ProGlucagon-Venus/RIP-Cherry/Per2:Luciferase mouse line was established, carrying specific fluorescent reporters for ?- and ?-cells, and luciferase reporter for monitoring the molecular clockwork. Flow cytometry fluorescence-activated cell sorting allowed separating pure ?- and ?-cell populations from isolated islets. Experimental conditions, developed by us for the culture of functional primary mouse ?- and ?-cells for at least 10?days, will be highlighted. Importantly, temporal analysis of freshly isolated ?- and ?-cells around-the-clock revealed preserved rhythmicity of core clock genes expression. Finally, we describe the setting to assess circadian rhythm in cultured ?- and ?-cells synchronized in vitro. The here-described methodology allows to analyze the functional properties of primary ?- and ?-cells under physiological or pathophysiological conditions and to assess the islet cellular clock properties. äHigh-Resolution Recording of the Circadian Oscillator in Primary Mouse(epmc).pdf DeepDyve Pubget Overpricing