B2-kininergic action of linear and cyclic tryptophan6- and tyrosine6-kallidin.

This study was undertaken to determine the importance of the central aromatic moiety in the kallidin and cyclokallidin molecules, using the relaxation of the isolated duodenum of the rat. Replacement in kallidin of the central phenylalanine by tryptophan increased the potency from an EC50 of 3 x 10(-10)M to 2 x 10(-12)M. Replacement by tyrosine decreased the potency to an EC50 of 8 x 10(-8)M. In cyclo-kallidin (EC50: 10(-8)M) the potencies were decreased: cyclo-Trp6-kallidin showed an EC50 of 10(-6)M and cyclo-Tyr6-kallidin of 3 x 10(-7)M. The relaxation of the rat duodenum by linear and cyclic kinins was potentiated by the bradykinin potentiating peptide BPP5a and antagonized by the B2 antagonist HOE-140. At a concentration of 10(-9)M, HOE-140 significantly decreased the potencies of bradykinin and cyclo-kallidin, but not of the B1 agonist desArg9-bradykinin.

Philanthotoxins block glutamatergic transmission in rat hippocampus--I. Reduction of high affinity glutamate uptake demonstrated by light microscopy surface autoradiography.

1. Effects of structural philanthotoxin (PTX) analogues on the high affinity uptake of [3H]-L-glutamate by slices of the rat hippocampus were studied using light microscopy surface autoradiography. 2. Considering the various effects of the PTX-analogues it can be concluded that the presence of an aromatic nucleus in the molecule is an important moiety, while the number and distribution of positive charges in the polyamine chain seems to be of a still not understood importancy.

Structure-activity relationship of philanthotoxins--I. Pre- and postsynaptic inhibition of the locust neuromuscular transmission.

1. Like the natural toxin, synthetic delta-philanthotoxin, now called PTX-4.3.3 acts as a reversible postsynaptic open ion-channel blocker of the glutamatergic neuromuscular system of the locust. 2. It also inhibits the high-affinity re-uptake of glutamate in the nerve endings and glial cells. 3. To study the structure-activity relationship, three parts of the PTX-4.3.3 molecule were changed. 4. One of these PTX-analogues, trifluoromethyl-PTX-4.3.3, proved to be a more potent postsynaptic blocker. 5. Moreover, compared with PTX-4.3.3 a delayed recovery period is seen with trifluoromethyl-PTX-4.3.3. 6. A number of PTX-analogues were equipotent to PTX-4.3.3 regarding the inhibition of iontophoretically evoked, postsynaptic glutamate potentials. 7. However, complete inactivation was achieved by reducing the length of the polyamine chain, moreover dideaza-PTX-12 was nearly completely inactive and a reduced activity was seen with dephenol-PTX-4.3.3. 8. A decrease of the decay time constant of glutamate potentials, normally seen by open ion-channel blockers in Con A pretreated preparations, was unaffected during application of the latter two analogues. 9. Possibly these two toxins act as weak receptor antagonists. 10. The presynaptic inhibition of the glutamate re-uptake, seemed to be a very specific property of PTX-4.3.3. Only one of the tested analogues (dehydroxy-PTX-4.3.3) exhibited this capacity.

Presynaptic effects of spider toxins: increase of high affinity uptake in the arthropod peripheral glutamatergic system.

In contrast to the reported effects of polyamines on the high affinity neurotransmitter uptake, two polyamine-like spider toxins significantly increase the high affinity uptake of glutamate as demonstrated with high resolution autoradiography. The effects of both spider toxins were compared to those of a polyamine toxin from the wasp Philanthus triangulum, which is known to inhibit the high affinity glutamate uptake.

Dexamethasone reduces the high-affinity binding of hemicholinium-3 to rat phrenic nerve endings in vitro: an autoradiographic study.

Receptor autoradiography has revealed high-affinity binding sites for hemicholinium-3 (HC-3) in the phrenic nerve endings of rat diaphragm. It has been demonstrated that 200 nM dexamethasone (Dex) in vitro reduces the binding of HC-3 to these high-affinity sites. It seems likely that the decrease in inhibitory effects of HC-3 on choline uptake and acetylcholine synthesis in rat diaphragm caused by Dex, should be the result of an interference of Dex with the binding of HC-3 to its high affinity sites.

Reduction of high affinity glutamate uptake in rat hippocampus by two polyamine-like toxins isolated from the venom of the predatory wasp Philanthus triangulum F.

Two components of the venom of the predatory wasp Philanthus triangulum F. significantly reduce--to a greater or less extent--the high affinity uptake of glutamate in rat hippocampus. A concentration of 10 microM delta-PTX caused a reduction of 74%, while the other component, beta-PTX, at the same concentration, caused a reduction of 18%. Hence the effect of delta-PTX on high affinity glutamate uptake in the hippocampus is comparable with its effect on high affinity glutamate uptake in insect neuromuscular junctions. Contrary to our previous findings that beta-PTX has no effect on high affinity glutamate uptake in insect glutamatergic terminal axons, however, beta-PTX significantly reduces high affinity glutamate uptake in the hippocampus, albeit less effectively than delta-PTX.

Selectivity of the uptake of glutamate and GABA in two morphologically distinct insect neuromuscular synapses.

The common inhibitor (CI) and slow excitor tibiae (SETi) innervated slow muscles 135cd of the locust Schistocerca gregaria were incubated under high-affinity uptake conditions either in [3H]GABA or in [3H]glutamate. [3H]GABA is accumulated in the glia of the nerve endings of the CI as well as the SETi; however, it is accumulated only in the terminal axons of the CI, not in the terminal axons of the SETi. The grain densities above the glia and above the CI terminal axons are approximately 2 grains/micron2. After incubation in [3H]glutamate the grain densities above the CI terminal axons and the SETi terminal axons are approximately 4 grains/micron2; the grain densities above the glia of both types of nerve endings are approximately 17 grains/micron2. The relatively high labeling (3 grains/micron2) of the muscles after incubation in the presence of glutamate is ascribed to the high metabolic requirements of slow muscles. The conclusion is drawn that a high-affinity uptake system for GABA is present in the CI terminal axons and in the glia of both the CI and SETi nerve endings. However, while the glutamate uptake in the CI and SETi nerve endings of the slow 135cd is comparable to the high-affinity uptake of glutamate in the fast excitor tibiae (FETi) nerve endings of the fast retractor unguis muscle, a high-affinity uptake of glutamate was only demonstrated in the glia of both types of nerve endings. A high-affinity uptake in the terminal axons of the CI and SETi may be masked by an extensively low-affinity uptake of glutamate by the muscles.

Dexamethasone affects radioactive choline uptake in rat diaphragm nerve endings and not in muscle fibres.

The initial rate of radioactive choline (Ch) uptake in the endplate-rich area (EPA) of both stimulated and unstimulated hemidiaphragms is significantly increased by 0.2 microM dexamethasone (Dex) in the presence of 10 microM Ch. In autoradiographs, the mean grain densities above the muscle fibres are not altered by Dex. The mean grain densities above the nerve endings are significantly increased in the presence of Dex in stimulated tissue, and slightly but not significantly increased in unstimulated tissue. There is a positive correlation between the initial rate of Ch uptake in the EPA and the amount of isotope in the nerve terminals, in the absence and presence of Dex. Without correcting for the large amount of diffusion which occurs, the ratio of the grain densities above the nerve terminals to that above the muscle fibres in the presence of Dex is 2.12 in stimulated tissue, and 1.40 in unstimulated tissue. The ratio in the stimulated tissue is significantly greater than the control ratio in the absence of Dex (1.66). Therefore, Dex affects radioactive Ch uptake in nerve endings and not in muscle fibres in the rat diaphragm. The stimulation-induced increase in the uptake of isotope into the nerve endings is abolished in a Na+-depleted medium, and in the absence of Ca2+. Dex has no effect on this abolition. We conclude that relatively low concentrations of Dex affect Ch transport in rat diaphragm nerve endings by a mechanism as yet to be defined.

Radioactive choline uptake in the isolated rat phrenic nerve-hemidiaphragm preparation. A biochemical and autoradiographic study.

When hemidiaphragms are stimulated via the phrenic nerve in the presence of 10 microM radioactive choline (Ch), the rate of radioactive Ch uptake in the endplate-rich area (EPA) is greater than that in the endplate-poor muscle (M). Ch uptake in the EPA is temperature-dependent, with a Q10 of 2.9 and an activation energy of 19.5 kcal/mol. It is inhibited in a Na+-depleted medium, in the absence of Ca2+, and by 10-20 microM hemicholinium-3 (HC-3) and it is not inhibited by alpha-bungarotoxin even when the muscle is completely paralyzed. In the absence of stimulation the rate of uptake in the EPA is slightly, but not significantly, greater than in M. Using autoradiography, we find an enhanced amount of isotope in the nerve terminals and their immediate vicinities compared with the muscle fibres, in both stimulated and unstimulated hemidiaphragms. There is no enhanced uptake of isotope into the nerve terminals in stimulated tissues in the presence of 26 microM HC-3. The uptake of isotope into the muscle is not altered by any of these treatments. There is a positive correlation between the initial rate of radioactive Ch uptake in the EPA and the amount of isotope in the nerve terminals (the mean corrected grain density above the nerve terminals). Without correcting for the large amount of diffusion that occurs, the ratio of the grain density above the synapses to that above the muscle fibres is 1.66 in tissue stimulated at 1 Hz, 1.04 in stimulated tissues in the presence of 26 microM HC-3, and 1.31 in unstimulated tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

Inhibition of the glutamate uptake in the excitatory neuromuscular synapse of the locust by delta-philanthotoxin; a component of the venom of the solitary wasp Philanthus triangulum F. A high resolution autoradiographic study.

Glutamate uptake in insect neuromuscular junctions is inhibited by delta-philanthotoxin. Two other components of the philanthus venom (beta- and gamma-philanthotoxin) do not affect this glutamate uptake. No synergism between the active delta-philanthotoxin and the two inactive components had to be assumed to explain the inhibition by the Philanthus venom.

Properties of a catecholaminergic system in some coelenterates. A histochemical and autoradiographic study.

Properties of a catecholaminergic nervous system in a coelenterate nervous system are described. The catecholamine present shows similarities to noradrenalin but it is not identical with noradrenalin. Contrary to known vertebrate catecholaminergic systems, neither a reuptake system nor a degrading system is present in the coelenterate catecholaminergic nervous system.

Estrogen receptors in human breast cancer. I. Specificity of the histochemical localization of estrogen receptors using an estrogen-albumin FITC complex.

An investigation of the specific localization of estrogen receptors in frozen sections of mammary tumors using an estrogen-BSA-FITC complex is presented. The results are discussed in relation to the existing literature. Although the fluorescence is certainly not due to interaction with type I receptors, the reaction is not as non-specific as is suggested sometimes in the literature. It is likely that type II receptors are involved, which are probably localized at the same site as type I receptors.

Deglycyrrhizinised liquorice (DGL) and the renewal of rat stomach epithelium.

Deglycyrrhizinised liquorice (DGL) stimulated proliferation in the forestomach of the rat but did not stimulate and possibly even inhibited proliferation in the glandular part. DGL increased the number of fundus glands in which labelled mucus secreting cells occur as well as the total number of labelled mucus cells per gland. The mechanism of action proposed for DGL is that DGL stimulates and/or accelerates the differentiation to glandular cells as well as mucus formation and secretion. The accelerated proliferation observed in the forestomach is ascribed to an improved environment in the stomach as a consequence of the enhanced mucus secretion under the influence of DGL.

Variations of the labelling index in vitro of rat mammary gland in pregnancy and early lactation.

The labelling index of rat mammary gland during oestrus, pregnancy and early lactation was studied in vitro. The implications concerning the existence of a critical cell division are discussed.