Characteristics of the nuclear translocation of progesterone receptor in fetal guinea pig uterus "in vivo", "in vitro" and in organ culture

The translocation of progesterone receptor from the cytosol into the nucleus was studied under "in vivo" and "in vitro" conditions in the uteri of guinea pig fetuses exposed to progesterone or a synthetic progestin, R5020. Progesterone treatment of estrogen-primed fetuses leads to a rapid (before 1h) transfer of cytosol progesterone receptor into the nucleus which is, however, short-lived (less than 3h). A rapid decrease in the retention of the estrogen receptor in the nucleus also occurs. In the "in vitro" incubations of whole fetal uteri, translocation of progesterone receptor is temperature-dependent and specific for progesterone and R5020; estradiol and cortisol have no effect. Putative progesterone receptors can also be induced in explants of fetal guinea pig uteri in organ culture which translocate from the cytosol into the nucleus under the same "in vitro" conditions as in whole uteri. Fetal uterine progesterone receptor, either stimulated "in vivo" by estrogen-priming or induced in organ culture, translocates from the cytosol into the nucleus and this process seems to be accompanied by a decrease in retention of the estrogen receptor in the nucleus which appears to be the mechanism by which progesterone antagonises estrogen action in fetal guinea pig uterus.     

Interpretation of light scattering associated with prolonged neural activity and temperature changes

Changes in light scattering from lobster giant axon which accompany the action potential were observed during periods of prolonged stimulation and as a function of temperature. At an initial temperature of 10°C most (more than 90%) axons produced positive light scattering signals which increased in amplitude when the temperature was lowered. At 2 and 5°C approximately half of the axons produced positive scattering signals. The remaining half produced negative scattering signals which became positive when the temperature was raised to 10°C. The amplitude of the negative signals followed sigmoid transition to positive values as a function of time. The time and temperature dependence of the signal are interpreted in terms of differential changes between the indices of refraction of the membrane matrix and the open or closed early activation channel.     

Neonatal administration of beta-endorphin produces "chronic" insensitivity to thermal stimuli

Male and female rat pups were injected with β-endorphin, naloxone or a saline control solution during days 2–7 postnatally. At 90 days of age the rats were tested for analgesia with the tail flick test. Testing was conducted during the first 2 hours of the light and the dark cycle. In both sexes and during both phases of the light cycle rats treated with β-endorphin as infants evidenced a significant elevation in threshold for painful thermal stimuli. Early treatment with naloxone also resulted in elevated threshold for thermal stimuli. Administration of naloxone to these rats as adults did not reverse the analgesic effect. It was concluded that early exposure to β-endorphin results in permanent changes in behavior perhaps by altering the interaction of endogenous opiates with their binding sites during a ciritcal period of opiate receptor development.     

Aryl hydrocarbon hydroxylase of rat brain mitochondria: Properties of,and effect of inhibitors and inducers on,enzyme activity

Aryl hydrocarbon hydroxylase (AHH), a typical example of mixed-function oxidase system, was studied in rat brain mitochondria. The enzyme was found to require oxygen and NADH for optimal expression of the activity. Coaddition of NADPH in the incubation system containing NADH resulted in an additive effect on the enzyme activity. NADH- and NADPH-dependent mitochondrial AHH activity was linear with respect to protein concentration and incubation time. The enzyme exhibited a sharp optima at pH 7.6. Specific activity of NADH-dependent mitochondrial AHH in rat brain was 3–4 and 8–11 times higher than that of NADPH-dependent mitochondrial and microsomal enzyme activity, respectively. Of the species investigated, NADH-dependent mitochondrial AHH followed the order: mice ? guinea pig > rat, while NADPH-supported mitochondrial AHH was in the order: rat > guinea pig ? mice. Specific activity of NADH-dependent mitochondrial AHH in various rat brain regions was similar with the exception of olfactory lobes which exhibited 60% higher activity than other region. When total region activities were added approximately whole brain activity was recovered. The apparent K_m value of NADH-dependent mitochondrial AHH was 1.18 μm with benzo(a)pyrene as a substrate. This K_m value was five to six times lower than that of NADPH-dependent microsomal AHH in rat brain (6.66 μm). NADH-dependent mitochondrial AHH was inhibited by KCN in a concentration-dependent manner while NADPH-supported mitochondrial AHH did not reveal any sensitivity to cyanide. Brain microsomal NADH as well as NADPH-supported AHH was also inhibited by KCN in a concentration-dependent manner. Carbon monoxide inhibited NADH-dependent mitochondrial AHH activity (48%) and had no effect on NADPH-dependent mitochondrial enzyme. Mitochondrial NADH and NADPH-dependent AHH activities were induced by 3-methylcholanthrene (64–73%) and benzo(a)pyrene (91–92%) pretreatments while no induction occurred with phenobarbital administration. 1-Benzylimidazole, SKF 525 A, metyrapone, and α-naphthoflavone inhibited both basal and 3-methylcholanthreneinduced NADH-dependent mitochondrial AHH activity. α-Naphthoflavone was more effective in inhibiting 3-methylcholanthrene-stimulated rat brain NADH-dependent mitochondrial AHH. Mitochondrial NADH-dependent AHH activity increased gradually with the onset of development and attained a steady state after 49–56 days of age. An increase of eight- to ninefold in the specific enzyme activity was observed between 7- and 56-day-old rats. No significant increase in brain mitochondrial AHH activity was observed between 56- and 91-day-old rats.     

Configurational, conformational, and solvent effects on the reduction of a Schiff base by reduced pyridine nucleotide analogs

The rate of reduction of the Schiff base, delta 1-pyrroline-2-carboxylic acid, by beta-NADH has previously been shown to be 6.8 times smaller than that calculated from the substituent effects in 1-substituted 1,4-dihydronicotinamides. The factors governing this anomalous rate have been studied by measuring the reduction rate with a number of reduced pyridine nucleotides in water and water-methanol mixtures. The beta-configuration for the nicotinamide-ribosidic linkage was found to be responsible for 75% of the anomaly; the same reduced coenzyme with alpha-linkage, however, behaves normally. It was also shown that the low reactivities of the reduced coenzymes with beta-configuration are entirely the result of their abnormal thermodynamic stabilities. Conformational effects (the folding of beta-NADH) account for only 20% of the reduction rate anomaly. Addition of organic solvents causes only a modest decrease in the overall rate. These solvent effects are interpreted in terms of the opposing effects of solvents on the basicity of the Schiff base and on the reduction step itself. Specific solvation effects appear important in controlling the reduction rates.