Secretory responses of intact glomus cells in thin slices of rat carotid body to hypoxia and tetraethylammonium
We have developed a thin-slice preparation of whole rat carotid body that allows us to perform patch-clamp recording of membrane ionic currents and to monitor catecholamine secretion by amperometry in single glomus cells under direct visual control. In normoxic conditions (PO2 ' 140 mmHg; 1 mmH...
| Autores: | , , , |
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| Tipo de recurso: | artículo |
| Fecha de publicación: | 2000 |
| País: | España |
| Institución: | Universidad de Sevilla (US) |
| Repositorio: | idUS. Depósito de Investigación de la Universidad de Sevilla |
| OAI Identifier: | oai:idus.us.es:11441/17785 |
| Acceso en línea: | http://hdl.handle.net/11441/17785 https://doi.org/10.1073/pnas.030522297 |
| Access Level: | acceso abierto |
| Palabra clave: | Carotid-body slice O2 sensing Glomus cell secretion |
| Sumario: | We have developed a thin-slice preparation of whole rat carotid body that allows us to perform patch-clamp recording of membrane ionic currents and to monitor catecholamine secretion by amperometry in single glomus cells under direct visual control. In normoxic conditions (PO2 ' 140 mmHg; 1 mmHg 5 133 Pa), most glomus cells did not have measurable secretory activity, but exposure to hypoxia (PO2 ' 20 mmHg) elicited the appearance of a large number of spike-like exocytotic events. This neurosecretory response to hypoxia was fully reversible and required extracellular Ca21 influx. The average charge of single quantal events was 46 6 25 fC (n 5 218), which yields an estimate of '140,000 catecholamine molecules per vesicle. Addition of tetraethylammonium (TEA; 2–5 mM) to the extracellular solution induced in most (>95%) cells tested (n 5 32) a secretory response similar to that elicited by low PO2. Cells nonresponsive to hypoxia but activated by exposure to high external K1 were also stimulated by TEA. A secretory response similar to the responses to hypoxia and TEA was also observed after treatment of the cells with iberiotoxin to block selectively Ca21- and voltage-activated maxi-K1 channels. Our data further show that membrane ion channels are critically involved in sensory transduction in the carotid body. We also show that in intact glomus cells inhibition of voltage-dependent K1 channels can contribute to initiation of the secretory response to low PO2. |
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