Showing papers in "Experimental Physiology in 1990"

Stimulation of ovine placental transport of calcium and magnesium by mid-molecule fragments of human parathyroid hormone-related protein.

Abstract: Perfusion in situ of the placenta of intact or previously parathyroidectomized fetal lambs has been used to assess the ability of three mid-molecule fragments of the human parathyroid hormone-related protein (PTHrP) molecule to stimulate the placental transport of calcium and magnesium. PTHrP(67-86 amide) was most effective but some activity was also shown by PTHrP(75-86 amide) and by PTHrP (75-84) in decreasing order. This placental action of PTHrP(67-86 amide) was rapid and could be observed using the placenta from an intact fetus, whereas it was necessary to use the placenta from a previously parathyroidectomized fetus to demonstrate stimulation of placental calcium transport by PTHrP(1-84). PTHrP(67-86 amide) may resemble the molecule that activates the placental calcium pump.

Prenatal maturation, the timing of birth and how it may be regulated in domestic animals

Abstract: In this review, the maturational changes wich occur in the fetus and mother towards the end of gestation are examined in relation to the timing of birth. Methodology. Prepartum maturational events in the fetus and the mother. Prepartum signals and the timing of birth in the ruminant, horse and pig ; role of the fetus, maternal influences and environmental factors. Artificial induction of labour.

The identification of neutral amino acid transport systems

Abstract: Dans cet article de synthese on donne les caracteristiques des systemes de transport des acides amines neutres dans les cellules non epitheliales et dans les cellules epitheliales de l'intestin grele, du rein, du placenta, des glandes salivaires et de la barriere hemato-encephalique. Comparaison et identification de la specificite du substrat, des substrats preferentiels, des proprietes d'echange, de la stereospecificite, de la dependance ionique et des regulations.

Basic mechanisms of focal epilepsies.

Abstract: CONTENTS PAGE Introduction 127 Neuronal discharge patterns in epileptic foci 130 Intrinsic burst mechanisms and the paroxysmal depolarization shift (PDS) 130 Synaptic transmission and the PDS 132 Excitatory amino acids as transmitters; implications for novel anticonvulsants 133 Synchronization and epileptic discharges 134 Synchronization by excitatory synapses 135 Recurrent excitatory collaterals between hippocampal CA3 pyramidal cells 135 Role of recurrent excitation in epileptic discharges in disinhibited hippocampal slices 137 Neocortical seizures 140 Chronic, subacute and clinical epilepsies 140 Synchronization by excitatory synapses in the presence of intact inhibition 143 Synchronization by inhibitory synapses 145 Synchronization by non-synaptic mechanisms 146 Electrical interactions between neurones 146 Fluctuations in extracellular ions 148 Durations of epileptic discharges: seizures and interictal spikes 149 What causes clinical epilepsies? 151 Conclusions 152 References 153

Amylin-amide: a new bone-conserving peptide from the pancreas.

Abstract: Amylin-amide is a new member of the family of peptides encoded by the calcitonin multigene complex. In the present study, we have compared directly, the hypocalcaemic potency and duration of action of human amylin-amide and human calcitonin in an in vivo rat bioassay and an in vitro osteoclast bone resorption assay. Amylin-amide was found to have a potency approximately 40-fold lower than human calcitonin, whilst both peptides followed the same time course. This suggests that amylin-amide is the most potent non-calcitonin hypocalcaemic peptide so far reported. An important physiological implication follows. It would seem that amylin-amide can play a central role in the maintenance of the skeleton by virtue of its inhibitory influence on osteoclastic function.

Inhibition of nitric oxide synthesis potentiates hypoxic vasoconstriction in isolated rat lungs.

Abstract: We have investigated the influence of endogenous nitric oxide (NO) on the vascular resistance of isolated rat lungs by inhibiting its synthesis with the false substrate N-monomethyl-L-arginine (L-NMMA). When perfused with blood at constant flow the addition of L-NMMA (10(-3) M) did not affect pulmonary arterial pressure in hyperoxia but did increase the response to hypoxia (PO2 25-35 mmHg) by 2.5 +/- 0.2 fold (mean +/- S.E.M.). The effect of L-NMMA was reversed by 3 x 10(-3) M-L-arginine, the true substrate for NO synthesis. Thus NO is an important pulmonary vasodilator but hypoxic vasoconstriction does not result from a reduction of its background release.

Intracellular free magnesium and its regulation, studied in isolated ferret ventricular muscle with ion-selective microelectrodes.

Abstract: Intracellular free magnesium ([Mg2+]i) was measured in isolated ferret papillary muscles using ion-selective microelectrodes filled with the new magnesium sensor ETH 5214. This new sensor, unlike its predecessor ETH 1117, does not react to marked changes in K+, Na+ or pH. Reducing Ca2+ from 20 microM to around 10 nM also did not affect the response so these electrodes are ideally suited to study intracellular Mg2+ and its regulation. The mean value for the [Mg2+]i from thirty-two experiments (forty-two impalements) was 0.85 mM, confirming previous estimates from this laboratory. Intracellular Mg2+ is not passively distributed and the possibility that Mg2+ is transported out of the cell by a Na(+)-Mg2+ exchanger was investigated. An increase in [Mg2+]o caused an increase in [Mg2+]i, as did stepwise reduction in the [Na+]o. However, this increase in [Mg2+]i on Na+ reduction also occurred in Mg2(+)-free solution suggesting that the increase in [Mg2+]i was due to the increase in intracellular Ca2+ on Na+ reduction. Moreover, increasing [Na+]i by strophanthidin did not change the [Mg2+]i and on increasing [Mg2+]o there was no reduction in the [Na+]i. Blocking ATP production lead to small increases in the [Mg2+]i. These results are not consistent with a Na(+)-Mg2+ exchanger as being the main outward transport mechanism for Mg2+ in this tissue.

Effects of exercise intensity on absorption of ingested fluids in man.

Abstract: Plasma 2H accumulation was measured in six male volunteers after ingestion of drinks containing trace amounts of 2H2O. Subjects fasted overnight and remained seated at rest or exercised on a cycle ergometer at 42, 61 or 80% of their maximum oxygen uptake (VO2, max). The rate of plasma 2H accumulation was faster at rest than during exercise at 61 or 80% of VO2, max (P less than 0.05), and was faster at 42 and 61% than at 80% of VO2, max (P less than 0.05). The time to peak plasma 2H concentration was longer during exercise than at rest. This suggests that strenuous exercise may reduce the availability of fluid ingested during exercise.

Analysis and presentation of intracellular measurements obtained with ion-selective microelectrodes

Abstract: We have considered the manner in which data obtained with ion-selective electrodes should be evaluated. The potential difference recorded by such electrodes, with respect to a stable reference, is converted to a concentration by a non-linear transformation--the Nernst or Nikolsky equation. The mean and standard deviation of such estimations of concentration are then usually presented, which assumes that the latter variable is normally distributed. If, however, the recorded potential difference (PD) is the normally distributed variable and the mean value calculated, then a different value of mean concentration will be obtained. We show here that the recorded PD is indeed the normally distributed variable using data from a variety of ion-selective electrode measurements and conclude that the mean values of quoted ion concentrations have been overestimated by 6-43%.

The effects of changes in laryngeal airway CO2 concentration on genioglossus muscle activity in the anaesthetized cat.

Abstract: In the anaesthetized cat the larynx was isolated in situ, artificially ventilated and used to assess reflex effects exerted by respiration-related laryngeal stimuli on genioglossus electromyographic activity (Gg EMG) and respiratory frequency (RF). Phasic Gg EMG was not observed when the larynx was unventilated but was evoked, with a concurrent decrease in RF, when negative pressures or oscillatory pressures similar to those of normal ventilation were applied to the larynx. Increases in laryngeal airway CO2 concentration also enhanced Gg EMG and reduced RF. All reflex effects were abolished by bilateral section of the superior laryngeal nerves. We propose that negative intralaryngeal pressure and CO2 may act together to restore pharyngeal patency during obstructive apnoea.

Neurones in the dorsal motor vagal nucleus of the cat with non-myelinated axons projecting to the heart and lungs.

Abstract: Extracellular recordings were made from ninety-four single motoneurones in the dorsal motor vagal nucleus of chloralose-anaesthetized or decerebrate cats. Fifty-five neurones had axons in cardiac vagal branches and thirty-nine had axons in pulmonary vagal branches; the conduction velocities of the axons were in the C fibre range, i.e. the axons were non-myelinated. The neurons exhibited little or no spontaneous activity. Excitatory and inhibitory synaptic inputs were demonstrated by electrical stimulation of the vagus nerve or its branches. Twenty-four neurones were tested by carotid sinus distension but only one was excited. Iontophoretic excitation of neurones projecting to cardiac vagal branches had no effect on heart rate. The properties of these neurones and their possible functions are discussed and contrasted with those of cardiac and pulmonary vagal motoneurones in the nucleus ambiguus.

The secretion of organic acids and bases by the ovine fetal kidney.

Abstract: Excretion of organic acids and bases was studied in twelve fetal sheep aged 120-140 days. There was no significant plasma protein binding of the organic anion, p-aminohippurate (PAH), nor of the organic cation, [14C]tetraethylammonium (TEA). There was a significant amount of acetyl-PAH (20 +/- 3%) in fetal urine but none could be detected in fetal plasma. The fractional excretion of unconjugated PAH was less than one, i.e. there was net reabsorption of 31.7 +/- 3.9% of the filtered load of unconjugated PAH. Since there was no acetyl-PAH in fetal plasma it is concluded that all acetyl-PAH in fetal urine occurred as a result of metabolism of PAH and secretion of the metabolite into the tubular lumen. The rate of excretion of acetyl-PAH in fetal urine varied from 0 to 14.0 micrograms min-1. Thus unconjugated PAH is filtered and there is net reabsorption; in addition, PAH is metabolized and enters the urine via tubular mechanisms. The fractional excretion of PAH was unaffected by I.V. administration of penicillin either acutely or chronically. The clearance of [14C]TEA was significantly greater than the glomerular filtration rate (GFR). The mean fractional excretion of [14C]TEA was 5.4 +/- 0.17. Thus 80.7 +/- 0.63% of the excreted TEA was secreted. The clearance of TEA was related to body weight (P less than 0.001) but the fractional excretion of TEA declined with gestation age, probably because GFR increased at a greater rate than the rate at which the secretory pathways increase their activity. It is concluded that those pathways that excrete organic anions like PAH into the urine mature much later (probably after birth) than those pathways responsible for the tubular secretion of organic bases.

A cultured human renal epithelioid cell line responsive to vasoactive intestinal peptide.

Abstract: A permanently transformed cell line derived from human embryo renal cortical cells (HEK293) has been investigated for the retention of renal-specific properties. The cell line is epithelioid in growth on plastic, and transmission electron microscopy demonstrates the formation of apical zonae occludentes. There is no prominent brush-border. The response of HEK293 cell adenylate cyclase is noteworthy for the response to vasoactive intestinal peptide (half-maximal activation at 0.9 nM). The HEK adenylate cyclase response to VIP is specific, with related peptides such as glucagon and secretin being ineffective. The response to VIP is competitively antagonized by the VIP receptor antagonist (4Cl-D-Phe6,Leu17)-VIP.

Adaptation of the mechanisms controlling gastric motility following chronic vagotomy in the ferret

Abstract: Changes in gastric motility were studied in the urethane-anaesthetized ferret following acute or chronic (3 weeks) vagotomy. The stomach was divided into the corpus and antrum and the effects of vagotomy on tone, frequency and contraction amplitude were investigated separately in the two gastric regions. In the corpus tone is kept at low levels by vagal activation of nonadrenergic, non-cholinergic (NANC) inhibitory neurones and also tonic sympathetic inhibition of intramural cholinergic activity. Frequency of contractions is also low due to tonic inhibition of cholinergic neurones by the vagus but not the sympathetic nervous system. There appears to be little vagal involvement in contraction amplitude but there is sympathetic inhibition of this parameter again via inhibition of cholinergic neurones. In the antrum there is no vagally driven inhibition of tone but a sympathetic inhibition of cholinergic neurones tends to reduce tone in the intact animal. Frequency of contractions does not appear to be extrinsically modulated. The vagus is tonically excitatory with regard to contraction amplitude in the antrum whereas the sympathetic nervous system is inhibitory, again via inhibition of cholinergic neurones. After chronic vagotomy some adaptation appears to take place within the surviving control systems in both the corpus and the antrum. Changes in cholinergic function have been suggested previously and are corroborated in this study. In addition novel alterations in intrinsic NANC systems and the remaining sympathetic innervation have been demonstrated in both regions of the stomach which tended to reduce the effects of vagotomy and return values for the parameters measured toward those observed in intact animals. The contribution of the cholinergic, adrenergic and NANC neurotransmitter systems to the post-vagotomy motility patterns differed in the corpus and antrum.

The Wellcome prize lecture. Intracellular sodium in cardiac muscle: effects on contraction.

Abstract: Article de synthese sur la regulation de la concentration en sodium intracellulaire (Na + i) et sur ses effets sur la contraction du muscle cardiaque chez les vertebres. Synthese sur les relations entre la concentration en Na + i et le taux de calcium au repos, pendant la systole et les contractions. Synthese sur les mecanismes de liberation du calcium du reticulum sarcoplasmique

Mechanism of action of magnesium on acetylcholine‐evoked secretory responses in isolated rat pancreas

Abstract: This study investigates the effects of magnesium (Mg2+) on acetylcholine (ACh)-evoked secretory responses and calcium (Ca2+) mobilization in the isolated rat pancreas. ACh induced marked dose-dependent increases in total protein output and amylase release from superfused pancreatic segments in zero, normal (1 x 1 mM) and elevated (10 mM) extracellular Mg2+. Elevated Mg2+ attenuated the ACh-evoked secretory responses compared to zero and normal Mg2+. In the absence of extracellular Ca2+, but presence of 1 mM-EGTA (ethylene glycol bis(beta-aminoethylether)-N,N,N',N''-tetraacetic acid), ACh elicited a small transient release of protein from pancreatic segments compared to a larger and more sustained secretion in the absence of both Ca2+ and Mg2+. Incubation of pancreatic segments with 45Ca2+ resulted in time-dependent uptake with maximum influx of 45Ca2+ occurring after 20 min of incubation period. ACh stimulated markedly the 45Ca2+ uptake compared to control tissues. In elevated extracellular Mg2+ the ACh-induced 45Ca2+ influx was significantly (P less than 0.001) reduced compared to zero and normal Mg2+. ACh also evoked dose-dependent increases in cytosolic free Ca2+ concentrations ([Ca2+]i) in pancreatic acinar cells loaded with the fluorescent dye Fura-2 AM. In elevated Mg2+ the ACh-induced cytosolic [Ca2+]i was significantly (P less than 0.001) reduced compared to zero and normal Mg2+. These results indicate that Mg2+ can influence ACh-evoked secretory responses possibly by controlling both Ca2+ influx and release in pancreatic acinar cells.

Catecholamine release from bovine chromaffin cells: the role of sodium-calcium exchange in ouabain-evoked release

Abstract: Spontaneous catecholamine (CA) release from bovine chromaffin cells maintained in primary tissue culture has been measured after pre-loading the cells with [3H]noradrenaline. Ouabain inhibited 86Rb+ uptake and increased 3H release in a concentration-dependent manner during a 60 min incubation period. Low external Na+ (5 mM: Li+ substitution) also increased 3H release. Whereas the 3H-releasing action of ouabain was maintained, the Li(+)-evoked release decreased with time. The effects of both ouabain and low Na+ solution on 3H release were completely inhibited by removal of Ca2+ from the external medium even though in Ca2(+)-free solution ouabain further inhibited 86Rb+ uptake into the cells. Readmission of Ca2+ to Na(+)-loaded cells (10-4 M-ouabain in Ca2(+)-free-1 mM-EGTA solution for 60 min) markedly increased the release of 3H. In the additional presence of diphenylhydantoin (DPH, 10-4 M) 3H release was significantly less on Ca2+ readmission. The 3H release from Na(+)-loaded cells was proportional to the concentration of Ca2+ readmitted. The 3H release was further increased from Na(+)-loaded cells in response to Ca2+ readmission when [Na+]o was lowered from 149 to 5 mM (Li+, choline+, Tris+ or sucrose substitution) though Li+ was less effective than the other Na+ substitutes. Potassium removal from the external medium significantly inhibited the 3H release evoked by Ca2+ readmission to Na(+)-loaded cells, even when [Ca2+]o was greater than normal (7.5 mM) or if Ca2+ was readmitted in low [Na+]o solution. Rb+, Cs+ or Li+ could substitute for K+ with the order of potency: Rb+ greater than or equal to K+ greater than Cs+ greater than Li+. A slight increase of external K+ (10.8 mM) potentiated the 3H release from Na(+)-loaded cells on Ca2+ readmission, but a higher concentration of K+ (149.4 mM) had the opposite action. The data is consistent with the hypothesis that ouabain-evoked CA release from bovine chromaffin cells is, in part, a consequence of an internal Na(+)-dependent Ca2+ influx. The evidence also suggests that there is Na(+)-Ca2+ competition at the external arm of the exchanger together with a monovalent cation activation site.

Phenylbiguanide not phenyldiguanide is used to evoke the pulmonary chemoreflex in anaesthetized rabbits.

Abstract: The pulmonary chemoreflexes evoked by phenylbiguanide (PBG), phenyldiguanide (PDG), phenylguanidine (PG) and 5-hydroxytryptamine (5-HT) have been investigated in the pentobarbitone-anaesthetized rabbit. The rank order of potency was 5-HT greater than PBG greater than PG greater than PDG. The responses evoked by all agonists were antagonized by pre-treatment with the selective 5-HT3 receptor antagonist MDL 72222, suggesting that their reflex effects are mediated by 5-HT3 receptors located on pulmonary vagal afferents. Furthermore, considering the low potency and long injection response time of PDG compared to PBG, we conclude that previous workers have used PBG and not PDG to identify non-myelinated pulmonary vagal afferents and to evoke the pulmonary chemoreflex.

Fetal magnesium homeostasis in the sheep

Abstract: The fetal plasma magnesium concentration exceeds that of the mother but the difference is small compared to that of ionized calcium concentration. Although not fully independent of changes in maternal magnesaemia, fetal magnesaemia showed a high degree of autonomy during both hypermagnesaemic and hypomagnesaemic changes induced in the ewe. As with calcium, the placental gradient is reversed after fetal thyroparathyroidectomy (TXPTX) with thyroxine replacement. During perfusion in situ of the placenta from such TXPTX fetuses isolated from the fetus itself, a stable positive placental gradient of magnesium concentration could be re-established between the perfusing blood and the maternal circulation. As with calcium, this gradient could be increased by fetal calf parathyroid extract, parathyroid hormone-related protein (PTHrP 1-141), PTHrP (1-84) but not by PTHrP (1-34). It was concluded that a mid-molecule portion of PTHrP can stimulate a putative placental pump which is responsible for the gradients of both calcium ions and magnesium across the ovine placenta.

A sodium‐activated potassium current in intact ventricular myocytes isolated from the guinea‐pig heart

Abstract: A current generated by the Na-activated K channel has been identified in whole cell currents recorded from isolated guinea-pig ventricular myocytes. A partial activation of this current can be achieved near to the physiological range of intracellular sodium concentration [( Na+]i) when it contributes significantly to the global outward current. The decline of the Na-activated K current, the lengthening of the action potential duration and the recovery of [Na+]i occur with a similar time course during recovery from Na loading.

Sympathetic stimulation causes increased output of lymphocytes from the popliteal node in anaesthetized sheep.

Abstract: Stimulation of the lumbar sympathetic chain at a frequency of 4 Hz caused a threefold increase in the lymphocyte output in efferent lymph from the popliteal node in anaesthetized sheep. The increase was accounted for by a rise in both lymph flow and lymphocyte count. Intra-arterial infusion of phentolamine (10 micrograms kg-1 min-1) abolished the nerve-mediated increases in flow and cell count.

The intracellular concentration of free magnesium in extensor digitorum longus muscles of the rat.

Abstract: Magnesium-sensitive microelectrodes were used to measure the intracellular concentration of free Mg2+, [Mg2+]f, in rat extensor digitorum longus muscles in vitro at 30 degrees C. The intracellular activities of Na+ and K+ were also determined so that allowance could be made for the interference from these ions with the Mg2+ electrode response. The mean value for [Mg2+]f based on twenty-six measurements in twelve muscles was 0.47 mM.

Abnormal expression of dipeptidylpeptidase IV activity in enterocyte brush-border membranes of children suffering from coeliac disease.

Abstract: Dipeptidylpeptidase IV (DPP IV) activity has been shown cytochemically to decrease significantly in enterocytes of children suffering from coeliac disease. This decrease is due to a halving of the time available for enterocytes to express DPP IV in their brush-border membranes during development. This effect is compared with previous results showing coeliac disease to inhibit disaccharidase activities selectively.

The responses of superior laryngeal nerve afferent fibres to laryngeal airway CO2 concentration in the anaesthetized cat

Abstract: In anaesthetized cats, the isolated, in situ, larynx was subjected to a simulated respiratory cycle and the responses of fifty-six superior laryngeal nerve (SLN) afferent fibres to respiration-related stimuli were examined during changes in the fractional CO2 concentration of the laryngeal airway (Faw, CO2). Sensory SLN fibres which displayed low rates of discharge when the larynx was unventilated (quiescent fibres) and which responded to negative laryngeal airway pressure were excited by elevations in Faw, CO2 whereas quiescent fibres responsive to positive laryngeal pressure were inhibited by the same procedure. We propose that changes in airway CO2 levels may play a role in maintaining upper airway patency, especially during sleep.

Water and sodium movements across the ruminal epithelium in fed and food-deprived sheep.

Abstract: Fluid and electrolyte movements across the ruminal epithelium of sheep were studied using the temporarily isolated rumen technique. The sheep were all subjected to the following treatments: (1) fed sheep (fed twice daily), after rumen emptying, received rumen contents from a fed sheep; (2) food-deprived sheep (two meals were omitted), after rumen emptying, received rumen contents from a food-deprived sheep; (3) fed sheep, after rumen emptying, received rumen contents from a food-deprived sheep; and (4) food-deprived sheep, after rumen emptying, received rumen contents from a fed sheep. Food deprivation led to an increased Na concentration of the rumen fluid while K and Cl concentrations, as well as osmolality, decreased. Plasma Na and osmolality decreased. During the 40 min after the rumen contents were exchanged no net movements of water occurred. Then the sheep were given an intraruminal load of saline which gave rise to a significant net absorption of fluid from the rumens of those sheep which had received rumen contents from fed sheep. The change in composition of the rumen contents after food deprivation impaired the absorption of Na and water from the rumen. Furthermore food deprivation reduced the Na absorptive function of the ruminal epithelium, but not the water permeability.

Hepatic branch vagotomy attenuates the feeding response to 2-deoxy-D-glucose in rats.

Abstract: The effect of hepatic branch vagotomy on the feeding response induced in rats by intraperitoneally injected 2-deoxy-D-glucose (2-DG) was tested. Injections were given 1 h after onset of the dark phase, and immediately [corrected] after the rats had ingested a meal. 2-DG produced a smaller feeding response in hepatic branch-vagotomized rats compared with that in sham-vagotomized rats. This finding suggests that hepatic glucoreceptors with vagal afferent fibres are involved in the feeding response to glucose deprivation.

Interactions between K+ and beta 2-adrenoreceptors in determining muscle vasodilatation induced in the rat by systemic hypoxia.

Abstract: In spontaneously breathing anaesthetized rats, both moderate and severe hypoxia caused increases in [K+] in venous efflux from hindlimb muscle, from 4.3 to 4.6 and from 3.8 to 4.4 mM respectively; the increases were accentuated to 5.2 and 5.7 mM after beta 2-receptor blockade with I.V. sotalol. Sotalol also potentiated the vasodilatation evoked in hindlimb muscle by moderate hypoxia, but reduced that evoked by severe hypoxia. We propose that K+ released from muscle during hypoxia contributed to the local vasodilatation. Further, we suggest that this effect was enhanced in moderate hypoxia by blockade of the beta 2-mediated uptake mechanism for K+ in skeletal muscle, but outweighed in severe hypoxia by blockade of the beta 2-mediated dilator action of circulating catecholamines on vascular muscle.

Autonomic and somatic responses evoked by stimulation of the cerebellar uvula in the conscious rabbit.

Abstract: The present study has investigated the effects of electrical stimulation of the cerebellar posterior cortex in the conscious rabbit. Stimulation of lobule IX (the uvula) elicited an increase in mean arterial blood pressure and heart rate, accompanied by EEG desynchronization, pupillary dilatation and a specific motor reaction, consisting of pricking of the ears, neck stiffening and running movements. Stimulation of lobule VIII was ineffective in evoking equivalent responses. The administration of barbiturates reversed the cardiovascular response to uvula stimulation, producing a fall in mean arterial pressure. The characteristics of the pattern of responses obtained, together with the anatomical and electrophysical observations provided in the preceding paper (Bradley, Ghelarducci, La Noce, Paton, Spyer & Withington-Wray, 1990), suggest a possible involvement of uvula cortex in the expression of the alerting reaction in the rabbit.

Actions of steroids and bemegride on the GABAA receptor of mouse spinal neurones in culture.

Abstract: The effects of a synthetic and an endogenous steroid were studied on the GABAA receptors of isolated mouse spinal neurones, maintained in culture. Low doses of alphaxalone reversibly increased GABA-evoked whole-cell currents. Alphaxalone at higher doses (10-50 microM), when pressure ejected onto spinal neurones, also directly evoked a membrane chloride current. Such currents were reversibly suppressed by bicuculline (a GABAA antagonist) and enhanced by phenobarbitone. 5 beta-Pregnan-3 alpha-ol-20-one, a progesterone metabolite, dose-dependently potentiated the amplitude of GABA-evoked whole-cell currents. The mechanism of potentiation was examined at the single-channel level using outside-out patches from spinal neurones. The main action of the steroid on the GABAA receptor appears to be similar to that found for barbiturates, in that they prolonged GABA-activated bursts of channel openings. Bemegride had an antagonistic action on the GABAA receptor, suppressing both GABA- and pentobarbitone-evoked whole-cell currents to similar extents.

Secretory agonists stimulate a rise in intracellular cyclic AMP but not Ca2+ and inositol phosphates in cultured rat epididymal epithelium.

Abstract: Primary monolayer cultures of rat epididymal cells have been shown to secrete chloride and bicarbonate when stimulated with beta-adrenergic agents, humoral agents and vasoactive peptides The intracellular messengers mediating the secretory response are unknown In this study intracellular AMP, Ca2+ and inositol phosphates were measured in epididymal monolayers at rest and upon stimulation with various secretory agonists Adrenaline, forskolin, lysylbradykinin, prostaglandin, endothelin, angiotensin II, antidiuretic hormone and vasoactive intestinal peptide at concentrations that stimulate anion secretion caused a rise in intracellular cyclic AMP The increase in cyclic AMP by adrenaline was blocked by propranolol but not by phentolamine Studies of the concentration-effect relationships showed that for adrenaline and endothelin the EC50 for stimulation of cyclic AMP was higher than that for stimulation of anion secretion None of these agonists affects intracellular Ca2+ concentration and inositol phosphate contents in epididymal monolayers Ca2+ ionophores A21387, ionomycin and erythrosin B (with irradiation with white light), at concentrations that stimulate anion secretion, also stimulated a rise in intracellular cyclic AMP and concomitantly increased intracellular Ca2+ The increase in cyclic AMP was dependent on extracellular Ca2+ It is not known whether the secretory response to Ca2+ ionophores was mediated by an increase in cell Ca2+ per se, or cyclic AMP However, it can be concluded that cyclic AMP is the second messenger which mediates the secretory responses to physiological stimuli