Ethanol influence on insulin secretion from isolated rat islets

This study was done to delineate the role of alpha- and beta-adrenergic receptors and cyclic AMP in the mechanism of ethanol effects on insulin release from isolated islets. Rats were given an alpha-adrenergic blocker, phentolamine, or a beta-adrenergic blocker, propranolol. In addition, ethanol 1 g/kg was given intragastrically 1 h prior to sacrifice. Glucose mediated insulin release from isolated islets was enhanced by phentolamine and decreased by propranolol. Ethanol treatment inhibited glucose-induced insulin release from isolated islets of control rats as well as those given phentolamine and/or propranolol. Insulin release from isolated islets in response to dibutyryl-cyclic AMP was attenuated by ethanol. Theophylline enhanced glucose mediated insulin release from control islets but ethanol treatment produced a significant inhibition of insulin response. The data suggest that the site of action of the deleterious effects of ethanol on insulin release from isolated islets in rat does not involve adrenergic system and cyclic AMP.     

Alternative methods of animal sacrifice: The effect on intestinal function in vitro

Summary The total fluid uptake of everted sacs of rat jejunum was compared in animals killed by stunning and decapitation, or anaesthetised with ether. Fluid transport was significantly higher in the tissue prepared from stunned and decapitated animals. It is suggested that etherization may have adverse effects on the physiological viability of subsequently isolated tissues.     

Alternative methods of animal sacrifice: the effect on intestinal function in vitro.

The total fluid uptake of everted sacs of rat jejunum was compared in animals killed by stunning and decapitation, or anaesthetised with ether. Fluid transport was significantly higher in the tissue prepared from stunned and decapitated animals. It is suggested that etherization may have adverse effects on the physiological viability of subsequently isolated tissues.     

Alpha-receptor subsensitivity of isolated atria from rats following repeated injections of phenylephrine or isoprenaline

Summary Daily injections of phenylephrine or isoprenaline for 4 or 7 days lowered the chronotropic sensitivity of -adrenoreceptors in isolated rat atria.This study was supported by grants from the National Science Council of Finland.     

An adrenergic participation subserving a positive inotropism and chonotropism of prostacyclin on isolated rat atria.

The effects of prostacyclin (PGI2) on the contractile frequency and on the isometric developed tension of spontaneously beating or paced isolated rat atria were explored. PGI2 enhanced frequency and contractile tension, both effects being blocked by the presence of propranolol, or following a pretreatment with 6-OHDopamine.     

Mechanical responses of the isolated cervix of the day-22 pregnant rat to field stimulation.

Field stimulation of isolated, spirally-cut cervix from day-22 pregnant rats produced contractions which could be inhibited by tetrodotoxin or hyoscine and potentiated by propranolol. The rat cervix would appear to receive both cholinergic and noradrenergic innervations whose transmitters activate muscarinic cholinoceptors and beta-adrenoceptors respectively.     

Chemical sympathectomy reveals pre-and postsynaptic effects of neuropeptide Y (NPY) in the cardiovascular system

Summary Intravenous injection of neuropeptide Y (NPY) caused short-lasting dose-dependent pressor responses in anesthetized rats. NPY was equipotent with noradrenaline in producing proportional pressor effects. Chemical sympathectomy, following the administration of 100 mg/kg 6-hydroxydopamine (6-OHDA), significantly potentiated the systemic pressor effects elicited by NPY or noradrenaline. Pretreatment with 2 nmol NPY enhanced the noradrenaline-induced pressor response in control rats. NPY did not change the basal tension of isolated rat aortic strips but significantly potentiated the contractile activity induced by 16 nM noradrenaline. This effect of NPY was not observed in aortic strips from rats pretreated with 6-OHDA. The presence of pre-and postsynaptic sites of action for NPY in the cardiovascular system of the rat is discussed.     

Chemical sympathectomy reveals pre- and postsynaptic effects of neuropeptide Y (NPY) in the cardiovascular system

Intravenous injection of neuropeptide Y (NPY) caused short-lasting dose-dependent pressor responses in anesthetized rats. NPY was equipotent with noradrenaline in producing proportional pressor effects. Chemical sympathectomy, following the administration of 100 mg/kg 6-hydroxydopamine (6-OHDA), significantly potentiated the systemic pressor effects elicited by NPY or noradrenaline. Pretreatment with 2 nmol NPY enhanced the noradrenaline-induced pressor response in control rats. NPY did not change the basal tension of isolated rat aortic strips but significantly potentiated the contractile activity induced by 16 nM noradrenaline. This effect of NPY was not observed in aortic strips from rats pretreated with 6-OHDA. The presence of pre- and postsynaptic sites of action for NPY in the cardiovascular system of the rat is discussed.     

Inhibition of the ureteral contractions induced by rat urine with kallikrein antibodies

Summary Purified urinary kallikrein induces contractions of the rat ureter in vitro. Antibodies against kallikrein block the contractile response of the isolated ureter to rat urine.This work was supported by the Deutsche Forschungsgemeinschaft (SFB 90). The technical assistance of J. Kopatsch, R. Marpoder and H. Seeger is gratefully acknowledged.     

Stimulating effect of thromboxane B2 on isolated rat atria

Summary Thromboxane B₂ enhanced the beating frequency and the contractile tension of isolated rat atria. Both effects were inhibited by propranolol, by pretreatment with 6-hydroxydopamine, or after inhibition of prostaglandins synthesis with indomethacin and acetylsalicylic acid.This work has been supported by grant 6638 from CONICET (Argentina).     

Chemical studies on the silver staining of nucleoli

The uptake of Ag-ions by isolated nucleoli of rat liver cells was studied. Nucleolar proteins separated by electrophoresis were examined for selective silver-staining. A mechanism for preferential Ag-staining of the nucleoli is discussed.     

Specificity of action of phospholipases of rat intestinal mucosa]

The total homogenate of rat intestine is devoid of C or D phospholipase activity. At pH 6,5 in the presence of phosphatidylcholines, it exhibits a B phospholipase activity and in the presence of exogen lysophosphatidylcholines an A1 lysophospholipase activity. At pH 8,5 the intestinal mucous membrane of a rat shows an isolated A2 phospholipase activity.     

Alpha-adrenergic subsensitivity of isolated femoral arteria following short-term cold acclimatisation in rats

Summary Cold acclimatisation lasting 2 and 4 days lowered the response of isolated femoral arteria to noradrenaline and phenylephrine in rat.This study was supported by the Academy of Finland.     

Effect of lipophilic cations on thiamine transport system in isolated rat hepatocytes

The effect of lipophilic cations such as triphenylmethylphosphonium, tetraphenylphosphonium and tetraphenylarsonium in addition to dibenzyldimethylammonium on thiamine transport in isolated rat hepatocytes was studied. Lipophilic cations at the concentration 10 microM almost completely inhibited thiamine uptake. Kinetic studies showed that these compounds were competitive inhibitors with a very high affinity. These results suggest that lipophilic cations in addition to quaternary ammonium compounds also share a common binding site for thiamine in isolated rat hepatocytes.     

Selective killing of smooth muscle cells in culture by the ricin A-chain conjugated with monoclonal antibodies to a cell surface antigen via a dextran bridge

Monoclonal antibodies to a surface antigen of the modulated smooth muscle cells originally isolated from the rat aorta media were conjugated with ricin A-chain via an oxidized dextran bridge. The interaction of cultured cells with the conjugates obtained and with control substances was monitored following incorporation of 14C-leucine radioactivity. It was found that 14C-leucine incorporation was suppressed by 80-90% at a conjugate concentration of 10(-6)-10(-7) M. Antigen-negative cells (line IAR; rat hepatocytes) were insensitive to the conjugate at any concentration used. Control use of purified ricin A-chain, native or oxidized dextran, specific and nonspecific IgG did not affect normal 14C-leucine incorporation. The data obtained may be useful for designing targeted drug transport systems and for selective screening of modulated smooth cells in vascular pathology models in vivo.     

High shortening velocity of isolated single arterial muscle cells

Summary Surprisingly high shortening velocities (<200 msec contraction-relaxation cycles) were found in isolated vascular muscle cells cultured from rat or chick aorta. All of the small fraction of cells with such quick contractions had membrane excitation by short duration spikes, rather than the slower graded depolarization of the other cells which produced 20-fold slower contractions.This research was supported by grant HL16328 from the National Institutes of Health. I wish to thank Aurita Puga and Paige DeCino for their excellent contributions to the cell culture aspects of these experiments.     

High shortening velocity of isolated single arterial muscle cells

Surprisingly high shortening velocities (less than 200 msec contraction-relaxation cycles) were found in isolated vascular muscle cells cultured from rat or chick aorta. All of the small fraction cells with such quick contractions had membrane excitation by short duration spikes, rather than the slower graded depolarization of the other cells which produced 20-fold slower contractions.     

GABA-A-mediated gastrin release induced by baclofen in the isolated vascularly perfused rat stomach

Summary In order to investigate the role of peripheral GABA-B receptors, the effects of the putative GABA-B agonist baclofen on immunoreactive gastrin release from an isolated vascularly perfused rat stomach preparation were examined. The vascular infusion of baclofen at graded concentrations induced a dose-dependent increase in gastrin release; this was unaffected by the GABA-B antagonist delta-aminovaleric acid, but was fully prevented by the selective GABA-A antagonist bicuculline as well as by atropine or tetrodotoxin. These results suggest that the stimulant effects of baclofen are mediated by nervous cholinergic structures, associated with GABA-A receptors, and indicate that this GABA-B agonist must be regarded as a partial agonist of peripheral GABA-A receptors.     

GABA-A-mediated gastrin release induced by baclofen in the isolated vascularly perfused rat stomach

In order to investigate the role of peripheral GABA-B receptors, the effects of the putative GABA-B agonist baclofen on immunoreactive gastrin release from an isolated vascularly perfused rat stomach preparation were examined. The vascular infusion of baclofen at graded concentrations induced a dose-dependent increase in gastrin release; this was unaffected by the GABA-B antagonist delta-aminovaleric acid, but was fully prevented by the selective GABA-A antagonist bicuculline as well as by atropine or tetrodotoxin. These results suggest that the stimulant effects of baclofen are mediated by nervous cholinergic structures associated with GABA-A receptors, and indicate that this GABA-B agonist must be regarded as a partial agonist of peripheral GABA-A receptors.     

Is tubulin involved in the electrically-induced mechanical activity of the isolated rat sciatic nerve?

Summary The electrically-induced mechanical activity of isolated segments of rat sciatic nerves remains unaffected following incubation with 10^–4 M colchicine, vinblastine or melatonin. Vinblastine depressed tubulin levels in incubated nerves. These results suggest that microtubules are not involved in nerve mechanical activity in vitro.This work was supported by grant No. 6638 from the CONICET (Argentina). It is dedicated to the memory of C.E. Valenti, decreased November 1979.