Evidence for sedative effects of low doses of morphine in mice involving receptors insensitive to naloxone

Low doses of morphine (0.30–2.5 mg/kg) decrease in a dose-dependent manner spontaneous climbing behaviour in mice. This effect is not modified by administration of naloxone at doses up to 1.25 mg/kg. These morphine doses do not modify the locomotor activity but, when they are associated with naloxone (0.5 mg/kg), an obvious inhibition occurs. In rats, a hyperactivity follows the akinesia produced by a morphine administration (10 mg/kg). This hyperactivity is changed into a significant hypokinesia when the animals are treated with naloxone (0.05 mg/kg). These results might reveal a dual effect of low doses of morphine, the excitatory effect of morphine being antagonized by naloxone whereas no action on the sedative effect is observed.     

Ultrastructural immunocytochemical localization of prolactin and growth hormone in the porcine pituitary

Summary The immunocytochemical peroxidase-antiperoxidase technique was used to identify prolactin- and growth hormone-producing cells in the porcine pituitary at the ultrastructural level. The growth hormone-producing cells contain round secretory granules (300 nm to 500 nm in diameter). The prolactin-producing cells can be identified by their distinct round and ovoid secretory granules which vary in size. Most of these cells contain large granules (450 nm to 750 nm in diameter), but some prolactin-producing cells display smaller secretory granules (250 nm to 500 nm). The two hormones were localized exclusively in the secretory granules. Staining for prolactin was observed in round and ovoid granules, as well as in small and polymorphic granules within the Golgi complex. This study confirmed (i) that the two hormones are located in different cells, and (ii) that under normal physiological conditions no one cell can synthesize and store both hormones simultaneously.     

The effects of membrane lipid order and cholesterol on the internal and external cationic sites of the Na+−K+ pump in erythrocytes

Cholesterol depletion alters the apparent affinity of the internal cationic sites and the maximal translocation rate but not the affinity of the external cationic sites of the $\text{Na}{}^{\mathrm{+}}\text{?K}{}^{\mathrm{+}}$ pump in human erythrocytes. To test whether these effects were mediated by a direct cholesterol-internal site interaction or by a change in membrane lipid order, the effects of five fluidizing amphiphiles (chlorpromazine, imipramine, benzyl alcohol, sodium oleate and sodium benzenesulphonate) on the kinetic parameters of the $\text{Na}{}^{\mathrm{+}}\text{?K}{}^{\mathrm{+}}$ pump were determined. The cholesterol removal and all the agents used induced dose-response decreases in membrane lipid order as measured by fluorescence polarization or ESR. Positive and neutral amphiphiles mimicked the effects of cholesterol removal on the affinity of the internal sites of the pump and to a lesser extent on the maximal translocation rate. Anionic amphiphiles had no effect on internal sites, probably because they distributed preferentially within the outer leaflet on the membrane. These results indicate that cholesterol controls the affinity of the internal sites of the $\text{Na}{}^{\mathrm{+}}\text{?K}{}^{\mathrm{+}}$ pump by altering the membrane lipid order. In contrast, neither cholesterol depletion nor the agents used altered the affinity of the external sites of the $\text{Na}{}^{\mathrm{+}}\text{?K}{}^{\mathrm{+}}$ pump. This difference in sensitivity to membrane lipid order suggests that internal and external cationic sites, although borne by the same protein, are in different lipid environments.     

Muscle and nerve in muscular dysgenesis in the mouse at birth: Sprouting and multiple innervation

Muscular dysgenesis (mdg) in the mouse is a recessive autosomal mutation affecting the striated musculature: during the whole gestation period, the muscles never show any sign of contractile activity. They are cytologically immature at birth, although the diaphragm is more mature than limb muscles, as confirmed by the levels of creatine phosphokinase. In both limb muscles and diaphragm the cytochemical localization of acetylcholinesterase demonstrates focal accumulations on the entire surface of $\text{mdg}\text{mdg}$ muscles, whereas such foci of acetylcholinesterase activity are restricted to a narrow end plate-rich region in $\text{+}\text{mdg}$? diaphragms. Teased single $\text{mdg}\text{mdg}$ myofiber preparations show that one myofiber can possess several foci of acetylcholinesterase, generally presenting aspects of very immature motor end plates. A study of the motor innervation, after silver nitrate impregnation, provides evidence for a spectacular overgrowth and a generalized sprouting of $\text{mdg}\text{mdg}$ nerves and axons. The $\text{mdg}\text{mdg}$ nerve terminals are generally very immature-looking, with an intense ultraterminal sprouting. Aspects suggesting a denser multiple innervation of $\text{mdg}\text{mdg}$ than $\text{+}\text{mdg}$? myofibers have been observed and choline acetyltransferase activity is increased in $\text{mdg}\text{mdg}$ tissues. Acetylcholinesterase specific activity and the number of α-bungarotoxin binding sites per milligram protein increased in $\text{mdg}\text{mdg}$ compared to $\text{+}\text{mdg}$? diaphragms. The very low amount of 16 S (and 12 S) acetylcholinesterase is probably related to $\text{mdg}\text{mdg}$ muscle inactivity. If the cytological and biochemical data are compared, it seems possible to propose that $\text{mdg}\text{mdg}$ myofibers and axons are in contact in several regions of the same myofiber, in variably mature appositions, and with a very dense multi-innervation.     

A fast and simple radiometric assay for adenosine deaminase using reversed-phase thin-layer chromatography

A new quantitative radiometric assay for adenosine deaminase is described. The reaction conditions are similar to those used in other radioassays and are shown to result in an activity which increases linearly with time and with enzyme concentration. An original feature of the technique resides in the use of reversed-phase thin-layer chromatography to separate adenosine from inosine. The separation is complete, fast, and reproducible. Both compounds can be recovered almost quantitatively from the plates. The assay is very simple and allows the determination of up to 36 samples in 3 h.     

Iron-porphyrin catalysis of the oxidative dealkylation of para-nitro-anisole and 7-ethoxycoumarin by cumylhydroperoxide: a possible model for the corresponding cytochrome P 450-dependent reactions

A biphasic system containing an iron porphyrin, Fe (TPP) (C1)¹ or [Fe(TPP)]₂O, efficiently catalyzes the cumyl-or tertiobutyl-hydroperoxide-supported dealkylation of p-nitroanisole and 7-ethoxycoumarin to the corresponding phenol and formaldehyde. Stoichiometric amounts of iron porphyrin and hydroperoxide give a quantitative reaction. Catalytic amounts of iron porphyrin give reaction rates and yields which are proportional to substrate concentration. With increasing hydroperoxide concentrations, the rates level offto limit values and the yield rapidly decreases. The maximum rates obtained approach those of the reactions mediated by cytochrome P 450-dependent monooxygenases.