Multiple components of voltage-dependent potassium current in normal rat anterior pituitary cells

1. Voltage-dependent K+ currents were studied in normal rat anterior pituitary cells using the patch-clamp technique. To obtain cultures enriched for lactotrophs, density gradient centrifugation was performed on pituitary cells isolated from lactating rats. 2. Depolarizations to about -30 mV from a holding potential of -80 mV activate a rapidly inactivating [time constant (tau) approximately 15–20 ms at -20 mV]K+ current. This transient current activated at low voltages (termed IA) is abolished by 5 mM external 4-aminopyridine (4-AP) but is largely resistant to external tetraethylammonium (TEA) (< or = 30 mM). 3. Recovery from inactivation of IA is fast, with a tau of 100–200 ms at -80 mV. Deactivation is also fast (tau approximately 2.2 ms at -50 mV). The voltage of half-activation of IA is approximately -20 mV. The current is completely inactivated at a holding potential of -40 mV. 4. Voltage-dependent K+ current activated by depolarizations from a holding potential of -40 mV was first detectable at about -20 mV (high voltage-activated) and had a time course that varied among cells. 5. Deactivation of high voltage-activated K+ current was best described by the sum of two exponentials, with tau of about 3.7 and 30 ms at -50 mV. Both components reversed close to the equilibrium potential for K+. 6. The amplitudes of the two tail currents were independent of each other when variable-duration commands were used to activate current. The amplitude of the fast component was largest with 10- to 20-ms commands to +40 mV and was reduced (< or = 50%) with 136-ms commands. The slow component amplitude reached a peak by 40 ms and remained constant for commands < or = 136 ms at +40 mV. 7. The contribution of each component to the total high voltage-activated tail current was variable among cells, with the amount of fast component correlating with the amount of inactivation produced by commands to +40 mV. 8. The two components of tail current activated by depolarizations from the -40 mV holding potential were abolished by external TEA (10 mM). 4-AP (5 mM externally) selectively abolished the fast component of high voltage-activated tail current while only partially reducing the slow component. 9. These results suggest that normal rat anterior pituitary cells possess at least three distinct types of voltage-dependent K+ current: a low voltage-activated, transient current (IA) and two high voltage-activated currents.(ABSTRACT TRUNCATED AT 400 WORDS)

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Control of secretion in anterior pituitary cells--linking ion channels, messengers and exocytosis

Journal of Experimental Biology ◽

10.1242/jeb.139.1.287 ◽

1988 ◽

Vol 139 (1) ◽

pp. 287-316

Author(s):

W. T. Mason ◽

S. R. Rawlings ◽

P. Cobbett ◽

S. K. Sikdar ◽

R. Zorec ◽

...

Keyword(s):

Ion Channels ◽

Intracellular Calcium ◽

Anterior Pituitary ◽

Hormone Secretion ◽

Cell Types ◽

Pituitary Cell ◽

Pituitary Cells ◽

Calcium Activity ◽

Anterior Pituitary Cells ◽

Voltage Dependent

Normal anterior pituitary cells, in their diversity and heterogeneity, provide a rich source of models for secretory function. However, until recently they have largely been neglected in favour of neoplastic, clonal tumour cell lines of pituitary origin, which have enabled a number of studies on supposedly homogeneous cell types. Because many of these lines appear to lack key peptide and neurotransmitter receptors, as well as being degranulated with accompanying abnormal levels of secretion, we have developed a range of normal primary anterior pituitary cell cultures using dispersion and enrichment techniques. By studying lactotrophs, somatotrophs and gonadotrophs we have revealed a number of possible transduction mechanisms by which receptors for hypothalamic peptides and neurotransmitters may control secretion. In particular, the transduction events controlling secretion from pituitary cells may differ fundamentally from those found in other cell types. Patch-clamp recordings in these various pituitary cell preparations have revealed substantial populations of voltage-dependent Na+, Ca2+ and K+ channels which may support action potentials in these cells. Although activation of these channels may gate Ca2+ entry to the cells under some conditions, our evidence taken with that of other laboratories suggests that peptide-receptor interactions leading to hormone secretion occur independently of significant membrane depolarization. Rather, secretion of hormone and rises in intracellular calcium measured with new probes for intracellular calcium activity, can occur in response to hypothalamic peptide activation in the absence of substantial changes in membrane potential. These changes in intracellular calcium activity almost certainly depend on both intracellular and extracellular calcium sources. In addition, strong evidence of a role for multiple intracellular receptors and modulators in the secretory event suggests we should consider the plasma membrane channels important for regulation of hormone secretion to be predominantly agonist-activated, rather than of the more conventional voltage-dependent type. Likewise, evidence from new methods for recording single ion channels suggests the existence of intracellular sites for channel modulation, implying they too may play an important role in secretory regulation. We shall consider new data and new technology which we hope will provide key answers to the many intriguing questions surrounding the control of pituitary hormone secretion. We shall highlight our work with recordings of single ion channels activated by peptides, and recent experiments using imaging of intracellular ionized free calcium.(ABSTRACT TRUNCATED AT 250 WORDS)

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Blockade by lithium ions of potassium channels in rat anterior pituitary cells

AJP Cell Physiology ◽

10.1152/ajpcell.1991.261.2.c218 ◽

1991 ◽

Vol 261 (2) ◽

pp. C218-C223 ◽

Cited By ~ 17

Author(s):

M. Kato ◽

P. M. Lledo ◽

J. D. Vincent

Keyword(s):

Growth Hormone ◽

Anterior Pituitary ◽

Hormone Secretion ◽

Male Rats ◽

Delayed Rectifier ◽

Pituitary Cells ◽

Patch Clamp Technique ◽

Anterior Pituitary Cells ◽

K Current ◽

K Currents

Extracellular Li+ has been known to facilitate the basal secretion of growth hormone from anterior pituitary cells and of catecholamine from chromaffin cells. In both cases, the intracellular accumulation of Li+ seems to be the prerequisite, and the presence of extracellular Ca2+ is indispensable. In this series of experiments, we examined whether Li+ blocked K+ currents by using primary cultured anterior pituitary cells from male rats. K+ currents were measured in the whole cell configuration of the patch-clamp technique. Extracellular Li+ (140 mM) suppressed both the delayed rectifier K+ current (IK) and the transient outward K+ current to 71 and 69% of control, respectively, in a reversible manner. IK elicited by a voltage step to +70 mV from holding potential of -70 mV was suppressed by 32.5 mM internal Li+ to 28% of control. Half-maximal suppression of K+ conductance by internal Li+ was 16 mM. Furthermore, Ca(2+)-channel blocker methoxyverapamil potently suppressed Li(+)-induced growth hormone secretion. From these results we propose that the blockade by Li+ of K+ channels could depolarize the cells and activate Ca2+ channels, thereby promoting the influx of Ca2+ and hormone secretion as a mechanism of Li(+)-induced hormone secretion.

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