CALL FOR PAPERS Neurohypophyseal Hormones: From Genomics and Physioogy to Disease Visually guided whole cell patch clamp of mouse supraoptic nucleus neurons in cultured and acute conditions

نویسندگان

  • Tevye J. E. Stachniak
  • Charles W. Bourque
چکیده

Stachniak, Tevye J. E., and Charles W. Bourque. Visually guided whole cell patch clamp of mouse supraoptic nucleus neurons in cultured and acute conditions. Am J Physiol Regul Integr Comp Physiol 291: R68–R76, 2006. First published February 9, 2006; doi:10.1152/ajpregu.00830.2005.—Recent advances in neuronal culturing techniques have supplied a new set of tools for studying neural tissue, providing effective means to study molecular aspects of regulatory elements in the supraoptic nucleus of the hypothalamus (SON). To combine molecular biology techniques with electrophysiological recording, we modified an organotypic culture protocol to permit transfection and whole cell patch-clamp recordings from SON cells. Neonatal mouse brain coronal sections containing the SON were dissected out, placed on a filter insert in culture medium, and incubated for at least 4 days to allow attachment to the insert. The SON was identifiable using gross anatomical landmarks, which remained intact throughout the culturing period. Immunohistochemical staining identified both vasopressinergic and oxytocinergic cells present in the cultures, typically appearing in well-defined clusters. Whole cell recordings from these cultures demonstrated that certain properties of the neonatal mouse SON were comparable to adult mouse magnocellular neurons. SON neurons in both neonatal cultures and acute adult slices showed similar sustained outward rectification above 60 mV and action potential broadening during evoked activity. Membrane potential, input resistance, and rapidly inactivating potassium current density (IA) were reduced in the cultures, whereas whole cell capacitance and spontaneous synaptic excitation were increased, perhaps reflecting developmental changes in cell physiology that warrant further study. The use of the outlined organotypic culturing procedures will allow the study of such electrophysiological properties of mouse SON using whole cell patch-clamp, in addition to various molecular, techniques that require longer incubation times.

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Visually guided whole cell patch clamp of mouse supraoptic nucleus neurons in cultured and acute conditions.

Recent advances in neuronal culturing techniques have supplied a new set of tools for studying neural tissue, providing effective means to study molecular aspects of regulatory elements in the supraoptic nucleus of the hypothalamus (SON). To combine molecular biology techniques with electrophysiological recording, we modified an organotypic culture protocol to permit transfection and whole cell...

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The activity of the magnocellular neurons (MCNs) of supraoptic nucleus (SON) is regulated by a variety of excitatory and inhibitory inputs. Opioids are one of the important compounds that affect these inputs at SON synapses. In this study, whole-cell patch clamp recording of SON neurons was used to investigate the effect of acute and repeated morphine administration on spontaneous inhibitory an...

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Potassium channels are trans-membrane proteins, which selectively transport K+ ions across cell membranes and play a key role in regulating the physiology of excitability cells and signal transduction pathways. Bothutous Schach (BS) scorpion venom consists of several polypeptides that could modulate ion channels. In this study, the effects of BS crude venom on passive and active electrophysiolo...

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تاریخ انتشار 2006