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Biomedical subjects

O Sand

Publications and source records attributed to O Sand.

At least 55 records · Page 3Linked to original sources
PubMed1988

The effect of fructose 1,6 diphosphate on pyruvate kinase from the liver of the flounder (Platichthys flesus L.).

1. Pyruvate kinase purified from flounder liver in two forms, i.e. PKI and PKII, is activated by fructose 1,6 diphosphate. 2. Two or more binding sites for FDP are demonstrated for PKII, the binding to which is influenced by the levels of substrates. 3. FDP reduces or abolishes the cooperative effect of PEP. 4. FDP increases the maximal activity. 5. The inhibition observed at higher levels of ADP is not abolished by FDP.

Adenosine Diphosphate↗
PubMed1988

Somatostatin inhibits prolactin secretion by multiple mechanisms involving a site of action distal to increased cyclic adenosine 3',5'-monophosphate and elevated cytosolic Ca2+ in rat lactotrophs.

The release of prolactin (PRL) from a clonal cell-line of anterior pituitary cells (GH4C1) was inhibited by somatostatin (SRIH) in a dose-dependent manner (ED50 nM). The inhibition (20% of control levels) was detectable within 50 s and maximal within 90 s. Thyroliberin (TRH) enhancement of PRL secretion was biphasic. SRIH inhibited both phases equally. Ionomycin in combination with the phorbol ester, TPA, mimics the TRH-elicited PRL release, and SRIH partly inhibited this effect. SRIH had no effect on TRH-stimulated formation of inositol trisphosphate, and only small effects on TRH-activated adenylate cyclase. Vasoactive intestinal peptide (VIP) and forskolin stimulated cAMP formation and PRL release potently. SRIH inhibited both effects of VIP and forskolin, and there was a close correlation between the inhibition of PRL secretion and cAMP accumulation. 8-Bromo-cAMP enhanced PRL release, an effect that was also partly reduced by SRIH. The Ca2+ channel activator, BAY-K-8644 and high extracellular K+ increased PRL release, and SRIH caused a partial reduction in the release response to both secretagogues. SRIH lowered [Ca2+]i, and markedly reduced the rise in [Ca2+]i elicited by TRH, VIP and K+. SRIH did not influence the Ca2+ spikes recorded in Na+-free solution, and had no effect on the TRH-induced membrane potential changes. Our results demonstrate that SRIH may inhibit PRL release from GH4C1 cells by (1) inhibiting hormone-sensitive adenylate cyclase, (2) blocking the effect of cAMP and (3) lowering [Ca2+]i. None of these effects is, however, sufficient to explain all the effects of SRIH, suggesting that SRIH also exerts a major action at a step subsequent to cAMP accumulation and [Ca2+]i elevation. Since the GH4C1 cells possess one single class of binding sites, this implies that the same SRIH receptor is coupled to several cellular signalling systems.

Animals↗
PubMed1987

Bombesin stimulates prolactin secretion from cultured rat pituitary tumour cells (GH4C1) via activation of phospholipase C.

Bombesin (BBS) stimulated prolactin (PRL) secretion from monolayer cultures of rat pituitary tumour cells (GH4C1) in a dose-dependent manner with half maximal and maximal effect at 2 nM and 100 nM, respectively. No additional stimulatory effect on PRL secretion was seen when BBS was combined with thyroliberin (TRH) used in concentrations known to give maximal effects, while the effects of BBS and vasoactive intestinal peptide (VIP) were additive. Using a parafusion system, BBS (1 microM) was found to increase PRL secretion within 4 s and the secretion profiles elicited by BBS and TRH (1 microM) were similar. Both BBS and TRH increased inositoltrisphosphate (IP3) as well as inositolbisphosphate (IP2) formation within 2 s. BBS also induced the same biphasic changes in the electrical membrane properties of GH4C1 cells as TRH, and both peptides caused a rapid and sustained increase in intracellular [Ca2+]. These results suggest that BBS stimulates PRL secretion from the GH4C1 cells via a mechanism involving the immediate formation of IP3 thus resembling the action of TRH.

Animals↗
PubMed1987

Vasoactive intestinal peptide and peptide with N-terminal histidine and C-terminal isoleucine increase prolactin secretion in cultured rat pituitary cells (GH4C1) via a cAMP-dependent mechanism which involves transient elevation of intracellular Ca2+.

Vasoactive intestinal peptide (VIP) and peptide (P) with N-terminal histidine and C-terminal isoleucine (PHI) stimulated prolactin (PRL) secretion from GH4C1 cells equipotent with ED50 values of 30-50 nM. In a parafusion system optimized to give high time resolution both VIP and PHI increased PRL secretion with a delay of about 60 s and subsequent to the activation of the adenylate cyclase. Thyroliberin (TRH) increased PRL secretion within 4 s. The dose-response curves for VIP- and PHI-stimulated cAMP accumulation were superimposable on those for PRL secretion. At submaximal concentrations the effects of VIP and PHI on both cAMP accumulation and PRL secretion were additive, whereas the effects were not additive at concentrations giving maximal effects. VIP and PHI increased [Ca2+]i measured by quin-2 in a different way than TRH, without inducing changes in the electrophysiological membrane properties of the GH4C1 cells. We conclude that both VIP and PHI stimulate PRL secretion via a cAMP-dependent process involving an increase in [Ca2+]i.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1987

Effects of calmodulin antagonists on hormone release and cyclic AMP levels in GH3 pituitary cells.

In GH3 cells the calmodulin antagonists trifluoperazine and N-(6-aminohexyl)-5-chloro-1-napthalene sulphonamide hydrochloride (W-7) showed a dose-dependent, biphasic effect on the release of growth hormone (GH) and prolactin (PRL). Hormone release was inhibited with 15-30 microM trifluoperazine and with 30-80 microM W-7, while stimulation was observed with 50-100 microM trifluoperazine and with 150 microM W-7. Trifluoperazine (greater than or equal to 30 microM) and W-7 (greater than or equal to 80 microM) increased the concentration of cellular cyclic AMP. Sulphoxides of trifluoperazine and chlorpromazine (less than or equal to 150 microM were without effect on hormone release and cellular cyclic AMP. Hydrolysis of cyclic AMP by GH3 cytosol was reduced after incubation of intact GH3 cells with trifluoperazine (15-60 microM). When trifluoperazine was incubated with cytosol, both the high and low affinity forms of cyclic AMP phosphodiesterase were inhibited competitively with calculated Ki of 4.5 and 56 microM, respectively. Stimulation of cyclic AMP phosphodiesterase caused by endogenous calmodulin was blocked by trifluoperazine. Particulate bound adenylyl cyclase activity was inhibited by trifluoperazine, and this effect was counteracted by endogenous calmodulin.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1986

Requirement of a transmembrane pH gradient for the entry of diphtheria toxin into cells at low pH.

The effects of acidification of the cytosol and of electrical depolarization on the entry of diphtheria toxin were studied. Entry of the toxin from the cell surface was induced by low pH, and the presence of the toxin in the cytosol was monitored as toxin-induced inhibition of protein synthesis. To reduce the membrane potential the cells were incubated in a buffer containing a high concentration of potassium. The cytosol was acidified either by incubating the cells with acetic acid, by incubating them with ammonium chloride which was subsequently removed in the presence of amiloride to prevent pH regulation by the Na+/H+ exchanger, or by incubating the cells in isotonic KCl in the presence of nigericin and valinomycin. The results showed that when the cytosol was acidified by either method toxin entry was inhibited, while a reduction in the membrane potential did not strongly interfere with the entry. A pH gradient across the membrane of at least 1 pH unit was required for entry. Possibly this gradient acts as a driving force for diphtheria toxin entry.

Animals↗
PubMed1986

Kinetic properties of liver pyruvate kinase from the flounder (Platichthys flesus L.).

Pyruvate kinase, purified from flounder liver, in two forms, i.e. PK I and PK II, is characterized by sigmoid kinetics with phosphoenolpyruvate as substrate at pH 6.3, 6.7 and 7.7. K0.5 for PEP increases with increasing pH. PK I and PK II show hyperbolic kinetics with ADP, but are inhibited by ADP concentrations above 1-2 mM. K0.5 for ADP decreases with increasing pH. PK I and PK II differ in their K0.5 values for PEP with a factor of at least 2, showing the highest figures for the latter. K0.5 for ADP is about the same for the two enzyme forms. Other nucleotide diphosphates can replace ADP as the substrate. When the nucleoside diphosphates are arranged in a rank order showing decreasing effectiveness as substrate, different rank orders are obtained for PK I and PK II.

Adenosine Diphosphate↗
PubMed1986

Electrophysiological properties of calcitonin-secreting cells derived from human medullary thyroid carcinoma.

Primary cultures of human medullary thyroid carcinoma tissue were prepared from lymph node metastases in two patients. The parenchymal, cultured cells displayed positive immunocytochemical staining for CT, and the cells also released the hormone into the culture medium. The membrane potential and resistance of the CT-producing cells were 50.1 +/- 8.9 mV and 634 +/- 154 M omega (mean +/- SD, n = 46). TTX sensitive action potentials with maximum rate of rise up to 51 V s-1 were evoked by current injection in Na+-containing solution, whereas TTX insensitive action potentials with maximum rate of rise up to 9 V s-1 were generated in Na+-free solution. These action potentials were reversibly blocked by D-600. We conclude that the action potentials of the human MTC cells have both a Na+ and a Ca2+ component. Ejection of CA2+-free solution close to the cells caused membrane hyperpolarization associated with decreased membrane resistance. The reversal potential of this response was -66.2 +/- 10.9 mV (n = 10), indicating that a permeability increase to Cl- and/or K+ may be involved. We suggest that elevated plasma Ca2+ concentration in vivo may cause increased excitability due to membrane depolarization and resistance increase, thus leading to enhanced Ca2+ influx and hormone secretion.

Action Potentials↗
PubMed1986

The phorbol ester TPA induces hormone release and electrical activity in clonal rat pituitary cells.

The phorbol ester TPA activates the protein kinase C in a similar way as 1,2-diacylglycerol. The effect of TPA on prolactin (PRL) secretion and electrical properties of rat pituitary cells in culture (GH4C1 cells) were compared with the effects of thyroliberin (TRH) on the corresponding parameters. The rate of hormone release was measured using a parafusion system optimized to give high time resolution. Samples for PRL measurements were taken every 4 s. The TRH evoked a biphasic PRL release, with a transient peak after about 30 s followed by a lower but sustained enhancement of the secretion. The TPA mimicked the late phase of the secretory response to TRH. The TPA analogue, 4 alpha-PDD, had no effect on the PRL release. The TRH also evoked biphasic membrane potential changes in the GH4C1 cells; the late phase consisting of membrane depolarisation associated with increased input resistance and enhanced firing of Ca2+ dependent action potentials. The TPA mimicked to a great extent these late phase effects of TRH, whereas the inactive analogue 4 alpha-PDD was ineffective. Continuous exposure to TPA masked the late phase of the electrophysiological response to TRH, suggesting that TPA and TRH share common mechanisms in their action on GH4C1 cells. We suggest that TRH enhances the electrical activity in these cells due to protein phosphorylation induced by diacylglycerol activation of protein kinase C, which in turn suppresses the membrane permeability to K+.

Animals↗
PubMed1986

Detection of infrasound by the Atlantic cod.

Below about 50 kHz the level of ambient noise in the sea increases continuously towards lower frequencies. In the infrasound range the spectral slope is particularly steep. This low-frequency noise may propagate long distances with little attenuation, causing a directional pattern of infrasound in the sea. Using a standing-wave acoustic tube, we have studied the sensitivity of cod to infrasound down to 0.1 Hz by means of the cardiac conditioning technique. The threshold values, measured as particle acceleration, showed a steady decline towards lower frequencies below 10 Hz, reaching a value close to 10(-5)ms-2 at 0.1 Hz. The spectrum level at 0.1 Hz in the sea ranges between 120 and 180 dB (re 1 microPa), with corresponding particle accelerations from less than 10(-6) to more than 10(-4)ms-2. The sensitivity of cod is thus sufficient to detect the highest levels of ambient infrasound, and we put forward the hypothesis that fish may utilize information about the infrasound pattern in the sea for orientation during migration, probably in addition to an array of other sensory inputs.

Acoustic Stimulation↗
PubMed1986

Effects of bromocriptine on prolactin release, electrical membrane properties and transmembrane Ca2+ fluxes in cultured rat pituitary adenoma cells.

The effects of the dopamine (DA) agonist bromocriptine on prolactin (Prl) release, electrical membrane properties and transmembrane Ca2+ fluxes have been studied in a clonal strain of rat pituitary adenoma cells (GH3). These cells generate Ca2+ dependent action potentials, and produce and secrete spontaneously both Prl and growth hormone. Prl release stimulated by thyroliberin (TRH) and elevated extracellular K+ concentration was completely blocked by bromocriptine, whereas the basal release was only moderately affected. The TRH and K+ evoked Prl release were half maximally inhibited by bromocriptine at 5-10 and 10-50 microM, respectively. The normal biphasic membrane response to TRH and the depolarizing effect of elevated K+ concentration were not altered by bromocriptine, whereas the Ca2+- spikes in Na+-free solution were suppressed by the drug. We therefore suggest that bromocriptine blocks the voltage sensitive Ca2+-channels of GH3 cell. In agreement with this notion, bromocriptine also suppressed the basal and TRH induced 45Ca2+ efflux from preloaded cells. We conclude that the inhibitory effect of bromocriptine on the voltage dependent Ca2+- channels is an important mechanism responsible for suppression of Prl release.

Action Potentials↗
PubMed1985

Characterization of human pregnancy zone protein. Comparison with human alpha 2-macroglobulin.

Native human pregnancy zone protein (PZP), a close homolog of alpha 2-macroglobulin (alpha 2M), can be obtained in approximately 20% yield from pooled late pregnancy plasma or serum by a combination of polyethylene glycol precipitation, euglobulin precipitation, DEAE-Sephacel chromatography, zinc-chelate affinity chromatography, and negative affinity chromatography on insolubilized antibodies against human serum proteins. Both proteins are similarly organized as disulfide-bridged dimers of 360 kDa containing 180-kDa subunits. These dimers constitute the proteinase-binding units of PZP, and in contrast to alpha 2M, they appear to be only loosely associated, indicating a subtle difference in the quaternary structure of these alpha-macroglobulins. The preparations contain functionally intact beta-cysteinyl-gamma-glutamyl thiol esters, located in the same nonapeptide sequence as found in alpha 2M, and form complexes with a variety of proteinases in which a large fraction of the proteinase is bound covalently. Proteinases bound to PZP are still active and poorly accessible to reaction with large inhibitors like alpha 1-proteinase inhibitor. The structural and functional features of PZP indicate that PZP and alpha 2M, although extremely similar, may have different yet overlapping sets of proteinases as targets. It is possible that PZP mainly controls the activity of cellular proteinases released under conditions of increased cellular turnover and that PZP could be the human equivalent to the acute phase alpha-macroglobulins known in other species.

Chemical Precipitation↗
PubMed1984

Vasoactive intestinal peptide causes a calcium-dependent stimulation of prolactin secretion in GH4C1 cells.

We have studied the effects of vasoactive intestinal peptide (VIP) on PRL secretion in a Bio-Gel column parafusion system containing rat pituitary tumour cells (GH4C1). A dose-dependent increase in PRL release was observed with half-maximal and maximal effect (2.1-fold) at 8 X 10(-8) and 5 X 10(-6) M, respectively. The PRL-stimulatory effect of VIP was instantaneous and maintained during the parafusion experiments (up to 60 min). On a molar basis VIP was always less effective than thyroliberin (THR), and the maximum stimulation of PRL release obtained with TRH was 1.2-3.0-fold higher (n = 12) than the maximum effect seen after VIP administration. The PRL-releasing effects of VIP, THR and high extracellular K+ were almost completely abolished in the presence of two inhibitors of voltage-sensitive Ca2+ channels, CoCl2 (10(-3) M) and verapamil (10(-4) M). In Ca2+-free buffer VIP, TRH and high extracellular K+ had only negligible effects, but the responses were fully restored in the presence of normal concentrations of extracellular Ca2+. In contrast to TRH, VIP had no demonstrable effect on the Ca2+-dependent action potentials of the GH4 cells.

Animals↗
PubMed1984

Purification and properties of pyruvate kinase from liver of the flounder (Platichthys flesus L.).

Two forms of pyruvate kinase, PK I and PK II, have been demonstrated in flounder liver. PK I, purified 991-fold to a specific activity of 105 units per mg of protein, has an unusually high molecular weight of about 2 X 10(6). PK II, purified 172-fold to a specific activity of 16.5 units per mg of protein, has a molecular weight of 210,000 when determined on a sucrose gradient but of 300,000 when derived from gel chromatography. PK I and PK II differ in sensitivity to the inhibitor L-phenylalanine, a fact which is used to evaluate the amount of each of them in a mixture. pH optimum for both forms is 6-6.6. PK I and PK II behave different in an Arrhenius plot--PK II showing a transition at 21 degrees C.

Animals↗
PubMed1983

A time course study of the effect of repetitive doses of estradiol-17 beta on serum glucose and lipids, liver glycogen and some carbohydrate metabolizing enzymes in liver of male flounder (Platichtys flesus L.).

1. Repeated treatment of male flounder with 5 and 100 microgram doses of estradiol-17 beta increases the level of phospholipid, triglyceride, free fatty acids and total lipid in serum as a function of time and dose, during a period of 17 days. 2. The glucose level in serum is increased and decreased respectively by doses of 5 and 100 micrograms. 3. Five and 100 microgram doses decrease the level of glycogen in liver. 4. Five microgram doses do not affect the activity of the measured enzymes considerably, with the exception of phosphorylase a. 5. One hundred microgram doses increase the activity of glucose-6-phosphate dehydrogenase after 11 days. 6. One hundred microgram doses increase the activity of pyruvate kinase continuously during the experimental period and decrease phosphorylase a and glucose-6-phosphatase activity.

Animals↗
PubMed1982

Trifluoperazine blocks calcium-dependent action potentials and inhibits hormone release from rat pituitary tumour cells.

The effects of trifluoperazine (TFP) on basal and stimulated release of prolactin (PRL) and growth hormone (GH) and on the electrical properties of the membrane were studied in clonal rat pituitary tumour cells in culture (GH3 cells). The basal GH release was inhibited maximally 50% by TFP (13-30 microM) and the K+- and thyroliberin (TRH)-induced stimulation of both PRL and GH release was blocked significantly. The sustained depolarization caused by elevated extracellular K+ concentration and the biphasic membrane potential response to TRH (normally leading to spontaneous action potentials) were not affected by TFP. However, TFP inhibited the Ca2+-dependent action potentials, probably by blocking the voltage sensitive Ca2+ channels in the membrane. We therefore suggest that TFP inhibits hormone release by blocking the uptake of extracellular Ca2+. This action of TFP is probably due to direct membrane effects which are independent of calmodulin.

Action Potentials↗
PubMed1981

Sodium and calcium action potentials in cells derived from a rat medullary thyroid carcinoma.

Primary monolayer cultures were made from a rat medullary thyroid carcinoma. All the cultures produced calcitonin throughout the 4 week experimental period. Two different parenchymal cell types could be distinguished, based on a marked size difference. Only the group of smaller cells was electrically excitable. The membrane potential and resistance of these cells were 55.3 +/- 15.4 mV and 623 +/- 275 M omega (mean +2- S.D., n = 23), and current injections produced regenerative responses in all the cells. All-or-none action potentials were recorded in both normal and Na+-free solution. In Na+-free solution the maximum rate of rise of the action potential was reduced by about 70%, and the action potential was reversibly blocked by D-600. It is concluded that the action potentials have both a Na+ and a Ca2+ component, and circumstantial evidence suggests that our recordings were obtained from the calcitonin-producing cells.

Action Potentials↗