کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2169632 1092954 2006 14 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Trehalose uptake through P2X7 purinergic channels provides dehydration protection
موضوعات مرتبط
علوم زیستی و بیوفناوری علوم کشاورزی و بیولوژیک علوم کشاورزی و بیولوژیک (عمومی)
پیش نمایش صفحه اول مقاله
Trehalose uptake through P2X7 purinergic channels provides dehydration protection
چکیده انگلیسی

The tetra-anionic form of ATP (ATP4−) is known to induce monovalent and divalent ion fluxes in cells that express purinergic P2X7 receptors and with sustained application of ATP it has been shown that dyes as large as 831 Da can permeate the cell membrane. The current study explores the kinetics of loading α,α-trehalose (342 Da) into ATP stimulated J774.A1 cells, which are known to express the purinergic P2X7 receptor. Cells that were incubated at 37 °C in a 50 mM phosphate buffer (pH 7.0) containing 225 mM trehalose and 5 mM ATP, were shown to load trehalose linearly over time. Concentrations of ∼50 mM were reached within 90 min of incubation. Cells incubated in the same solution at 4 °C loaded minimally, consistent with the inactivity of the receptor at low temperatures. However, extended incubation at 37 °C (>60 min) resulted in zero next-day survival, with adverse effects appearing even with incubation periods as short as 30 min. By using a two-step protocol with a short time period at 37 °C to allow pore formation, followed by an extended loading period on ice, cells could be loaded with up to 50 mM trehalose while maintaining good next day recovery (49 ± 12% by Trypan blue exclusion, 56 ± 20% by alamarBlue assay). Cells porated by this method and allowed an overnight recovery period exhibited improved dehydration tolerance suggesting a role for ATP poration in the anhydrous preservation of cells.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Cryobiology - Volume 52, Issue 1, February 2006, Pages 114–127
نویسندگان
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