tetrodotoxin and Adenoma

The effects of pituitary adenylate cyclase activating polypeptide (PACAP) on ion channels were examined in GH3 cells human pituitary adenoma cells. In GH3 cells, PACAP-38 (10-9 M) reversibly activated tetrodotoxin-sensitive NA+ channels but had little effect on nicardipine-sensitive Ca2+ channels. PACAP-induced increase in Na+ currents was inhibited by PACAP (6-38), a specific PACAP receptor antagonist, and Rp-cAMPs, an inhibitor for protein kinase A, and mimicked by 8-bromo-cAMP. In human pituitary adenoma cells, PACAP also activated tetrodotoxin-sensitive Na+ channels and growth hormone secretion. These results suggest the possibility that PACAP can activate voltage-gated Na+ channels via adenylate cyclase-protein kinase A pathway in the pituitary.

Topics: 8-Bromo Cyclic Adenosine Monophosphate; Adenoma; Cell Line; Human Growth Hormone; Humans; Middle Aged; Neuropeptides; Pituitary Adenylate Cyclase-Activating Polypeptide; Pituitary Gland; Sodium Channels; Tetrodotoxin; Tumor Cells, Cultured

Electrophysiology of human growth hormone secreting tumour cells and its modification by hGRF has been studied using on-cell and Nystatin-perforated whole-cell recording configurations. Local application of hGRF (10 nM) produced an increase in the frequency of action potentials. Ca2+ currents were isolated by a ramp depolarizing pulse from -120 mV to +60 mV in the presence of tetrodotoxin (1 microM). Human GRF increased the Ca2+ currents which could be blocked by Ni+ (300 microM). We conclude that an increase in Ca2+ current is integral to the action of hGRF on these cells.

Topics: Action Potentials; Adenoma; Calcium Channels; Growth Hormone; Growth Hormone-Releasing Hormone; Humans; Membrane Potentials; Pituitary Neoplasms; Sodium Channels; Tetrodotoxin; Tumor Cells, Cultured

The mechanism of the inhibitory effect of local anesthetics on hormone secretion was studied in the GH4C1 line of rat pituitary tumor-derived cells. Lidocaine between 0.1 and 5 mM exerted significant dose-dependent inhibition on the increment in cytosol Ca2+ concentration ([Ca2+]i) and prolactin (PRL) secretion induced by 30 mM K+. For both effects the IC50 was 0.25 mM and maximal inhibition occurred at 5 mM. A normal response returned within 20 min after removal of lidocaine from the incubation medium. 1 microM tetrodotoxin had no effect on the 30 mM K+ induced [Ca2+]i transient or PRL secretion, indicating that Na+ channels are not involved in the inhibitory effect of lidocaine. Lidocaine similarly inhibited the [Ca2+]i increment and PRL secretion induced by 30% medium hyposmolarity and 1 microM Bay K 8644. Lidocaine was much less effective in inhibiting secretion induced by 1 microM phorbol 12-myristate 13-acetate (TPA) or 5 microM forskolin. 5 mM procaine produced effects similar to those of lidocaine. Our data suggest that in GH4C1 cells local anesthetics depress secretagogue-induced PRL secretion primarily by blocking Ca2+ influx, probably through L-type Ca2+ channels.

Topics: 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; Adenoma; Animals; Calcium; Colforsin; Lidocaine; Pituitary Neoplasms; Potassium; Procaine; Prolactin; Rats; Tetradecanoylphorbol Acetate; Tetrodotoxin; Tumor Cells, Cultured

Spontaneous and CRF-stimulated changes in the cytosolic free calcium concentration ([Ca2+]i) were studied in two types of corticotrophs: 1) cultured human ACTH-secreting pituitary adenoma cells (hACTH cells), and 2) identified small ovoid corticotrophs cultured from normal rat pituitaries. [Ca2+]i was monitored in individual corticotrophs by dual emission microspectrofluorimetry using indo-1 as the intracellular fluorescent Ca2+ probe. In hACTH cells, [Ca2+]i measurements were carried out in combination with electrophysiological recordings obtained using whole cell patch-clamp techniques. It was shown that a single spontaneous Ca(2+)-dependent action potential led to a marked transient increase in [Ca2+]i in human tumoral corticotrophs. Spontaneous fluctuations in [Ca2+]i were also observed in unpatched corticotrophs whether derived from human pituitary tumors or normal rat tissue. Based on their striking kinetic features and their sensitivity to external Ca2+, we suggest that these spontaneous [Ca2+]i transients were the consequence of action potential firing. Under separate voltage-clamp (patch-clamp) conditions, tumor corticotrophs showed two Ca2+ current components: a low threshold, rapidly inactivating (T-type) current, and a higher threshold, slowly inactivating (L-type) current. The dihydropyridine Ca2+ channel blocker PN 200-110 (100 nM) abolished the L-type current without affecting the T-type current, while the inorganic Ca2+ channel blocker Cd2+ (200 microM) suppressed both Ca2+ currents. The Na+ channel blocker tetrodotoxin (5 microM) did not affect inward currents in tumor corticotrophs. Both L- and T-type voltage-gated Ca2+ channels were involved in controlling [Ca2+]i transients in both tumor and normal corticotrophs, inasmuch as Cd2+ (200 microM) abolished [Ca2+]i) transients, while PN 200-110 (100 nM) greatly diminished, but did not completely abolish, [Ca2+]i transients. The latter did not appear to depend on a voltage-dependent Na+ influx, since they were unaffected by tetrodotoxin (5 microM). Corticotrophs generate [Ca2+]i transients in response to the hypothalamic secretagogue CRF by acting on their membrane excitability. Indeed, we demonstrated in combined fluorescent and electrophysiological experiments that CRF (100 nM) had a coordinate action on human tumoral corticotrophs comprised of a modest depolarization and an increase in the frequency of both action potentials and subsequent [Ca2+]i transients. A coincident increase in the

Topics: Action Potentials; Adenoma; Adrenocorticotropic Hormone; Animals; Calcium; Calcium Channel Blockers; Calcium Channels; Corticotropin-Releasing Hormone; Cytosol; Egtazic Acid; Female; Humans; Isradipine; Membrane Potentials; Oxadiazoles; Pituitary Gland; Pituitary Neoplasms; Rats; Rats, Inbred Strains; Spectrometry, Fluorescence; Tetrodotoxin; Tumor Cells, Cultured

Human anterior pituitary cells derived from a somatotropin-secreting adenoma were capable of generating action potentials with Ca2+ and tetrodotoxin-sensitive Na+ components. A major fraction of the action current was carried by Na ions.

Topics: Action Potentials; Adenoma; Calcium; Cells, Cultured; Female; Humans; Middle Aged; Pituitary Gland, Anterior; Sodium; Tetrodotoxin