Apolipoprotein E polymorphism in Pick's disease and in Huntington's disease.

The polymorphism of apolipoprotein E (apoE) has been recognized as a genetic risk factor in different neurodegenerative disorders, with or without tau protein- related neuropathology, but few published epidemiological data are available as concerns the association of different apoE alleles with two relatively rare forms of dementia, Pick's disease (PiD) and Huntington's disease (HD). In this study the frequency of the apoE4 allele was examined in 36 persons with histopathologically proven PiD and compared with that of the apoE genotype in 28 HD probands and 79 aged healthy controls. The E4 allele was overrepresented selectively in PiD (42%) as compared with the control population (7%). No such association was found for HD probands (9%). This finding lends further support to the hypothesis that the E4 genotype is not an Alzheimer's disease specific susceptibility factor, and that it could be present in diverse dementing disorders with tau protein related neuropathology, such as PiD.

Field-specific changes in hippocampal opioid mRNA, peptides, and receptors due to prenatal morphine exposure in adult male rats.

Alterations in the opioid system in the hippocampal formation and some of the possible functional consequences were investigated in adult male rats that were prenatally exposed to either saline or morphine (10 mg/kg twice daily on gestational days 11-18). In situ hybridization and Northern blots were used to measure proenkephalin and prodynorphin mRNA, and radioimmunoassays quantified proenkephalin- and prodynorphin-derived peptide levels in the dentate gyrus, CA3, and CA1 subfields of the hippocampal formation. Prenatal morphine exposure in male rats decreases proenkephalin and increases prodynorphin mRNA selectively in the granule cell layer of the dentate gyrus. Similarly, met-enkephalin peptide levels are decreased and dynorphin B peptide levels are increased in the dentate gyrus but not CA3 or CA1 of prenatally morphine-exposed males. In addition, there are decreases in dynorphin-derived peptides in the CA3 subfield. Receptor autoradiography revealed increases in the density of micro but not delta receptor labeling in discrete strata of specific hippocampal subfields in morphine-exposed males. Because alterations in the hippocampal opioid system suggest possible alterations in the excitability of the hippocampal formation, changes in opioid regulation of seizures were examined. Morphine exposure, however, does not alter the latency to onset or number of episodes of wet dog shakes or clonic seizures induced by infusion of 10 nmol [D-Ala2, MePhe4, Gly-ol5]enkephalin into the ventral hippocampal formation. Interestingly, a naloxone (5 mg/kg) injection 30 min before bicuculline administration reverses the increased latency to onset of clonic and tonic-clonic seizures in morphine-exposed males. Thus, the present study suggests that exposure of rats to morphine during early development alters the hippocampal opioid system, suggesting possible consequences for hippocampal-mediated functions.

The influence of external sodium and potassium on lithium uptake by primary brain cell cultures at "therapeutic" lithium concentration.

The ionic regulating of lithium homeostasis and steady-state intra:extracellular lithium distribution in the brain can be approached by experimental methods using intact nerve cells in vitro. Primary cultures prepared from chick embryonic brain were applied to study the effect of extracellular sodium and potassium on the lithium uptake of nerve cells at 'therapeutic' lithium concentration (1.5 mM). Lithium influx and the level of steady-state intracellular lithium were significantly reduced by increasing the external sodium concentration. At physiological extracellular sodium level, the steady-state content of lithium in the brain cells was about half of that observed in the presence of 10 mM sodium in the incubation media and the value of the intra:extracellular lithium distribution ratio was below 1. External potassium (0.5 - 3mM) strongly inhibited lithium uptake of the nerve cells. Ouabain (10(-4)M) had no effect on this potassium-sensitive lithium uptake in Tyrode media. Sodium influx studied by isotope tracer methodology was higher in cultures preloaded with lithium as compared to that of the controls. It can be concluded that sodium and potassium ions, at physiological concentrations, significantly influence lithium uptake as well as the intra:extracellular lithium distribution in brain cell cultures.

On a possible new intracellular signal-system in rat gastric mucosa.

It is known that cAMP and cGMP, as an "intracellular second messenger system" play a significant role as a signal system, in the mechanism of action of anti-ulcerogenic (cytoprotective) drugs. According to our present, preliminary investigations it seems that during different experimental circumstances the gastric mucosal 3'-5'-cyclic-cytidine-mono-phosphate (cCMP) 3'-5'-cyclic-uridine-monophosphate and (cUMP) levels were changed--similarly to CAMP and cGMP--and these changes might be a possible indicator of a further, most probably secondary, signal- system role.

Gastric mucosal endogenous prostacyclin levels are different in Brattleboro rats compared with Wistar strain.

Homozygous Brattleboro rats were investigated and compared to normal (physiological) Wistar strain rats regarding their gastric mucosal endogenous prostacyclin (PG-I(2)) level. It seems that the Brattleboro animals have a significantly lower level of this important protective material. Wistar rats having an artificial pituitary stalk lesion (which is the artificial equivalent of homozygous Brattleboro animals) showed no differences in endogenous mucosal prostacyclin level compared to normal Wistar rats. Therefore, we concluded that this hitherto unknown property of the homozygous Brattleboro rats is genetically determined.

Prenatal exposure to morphine alters brain mu opioid receptor characteristics in rats.

Prenatal morphine exposure alters neither the binding capacity nor the affinity of ligand binding to mu opioid receptors of adult male brains. However, males have significantly higher Bmax in the hypothalamus than ovariectomized females. In females, prenatal exposure to morphine reduces the Bmax of mu opioid receptors 25% in the hypothalamus and preoptic area. Estrogen treatment increases the Bmax of mu opioid receptors in the striatum of all ovariectomized females but in the hypothalamus only of morphine-exposed females, thereby eliminating the sex difference observed in control animals.

Prenatal morphine exposure alters estrogen regulation of kappa receptors in the cortex and POA of adult female rats but has no effects on these receptors in adult male rats.

The binding characteristics of kappa receptors were assessed in the frontal cortex (CX), striatum, hypothalamus, preoptic area (POA), cerebellum, and ventral tegmental area of adult male and female rats exposed prenatally to morphine or saline. Prenatal morphine exposure altered estrogen regulation of kappa receptors in the CX and POA of females, but had no effects on kappa receptors in any of the examined brain regions in male rats.

Modulation of catecholamine turnover rate in brain regions of rats exposed prenatally to morphine.

The concentration and turnover rate of brain catecholamines were measured in the hypothalamus, preoptic area (POA), frontal cortex, striatum and cerebellum of adult male and female rats exposed in utero to morphine (5-10 mg/kg/twice a day) during gestation days 11-18. Norepinephrine (NE) and dopamine (DA) turnover rates were estimated following alpha-methylparatyrosine (AMPT) administration. Prenatal morphine altered NE content and turnover in male and female rats in a regionally specific, sexually dimorphic manner. Basal NE content increased approximately 60% in the hypothalamus of male rats, but it decreased about 30% in the hypothalamus of female rats. NE turnover in the hypothalamus of morphine-exposed rats increased 50% in males and decreased 50% in females. Prenatal morphine had no effects on NE turnover in the male POA, but in female rats NE turnover decreased approximately 60%. Alterations in the frontal cortex of morphine-exposed male and female rats resembled the pattern in the hypothalamus; however, the differences did not reach statistical significance. In addition, prenatal morphine had no effect on striatal or cerebellar NE or on basal levels or turnover of DA in any brain regions. These results demonstrate that prenatal morphine alters the content and turnover of NE in a sexually dimorphic manner in specific brain regions of male and female rats, suggesting alterations in the density of terminals and/or utilization of NE. These sexually dimorphic alterations in hypothalamic NE induced by prenatal morphine may be related to the changes observed in adult male and female sexual behavior in our previous work.

Na-dependent Li-transport in primary nerve cell cultures.

Primary cultures from chick embryonic brain were used to study the steady state distribution of lithium. The intra/extracellular Li+ ratio decreased by enhancing the external Na+ concentration. Ouabain did not influence this unequality. A phloretin-sensitive component was revealed in the Li uptake at low Na+ concentration. The findings might suggest the existence of a Na+-dependent Li+ countertransport system in these brain cell cultures.

Prenatal exposure to morphine differentially alters gonadal hormone regulation of delta-opioid receptor binding in male and female rats.

The present study tested the hypothesis that exposure to morphine on gestation days 11-18 differentially alters delta-opioid receptors in the brain of adult male and female rats. In Experiment 1, the binding characteristics of delta-opioid receptors were examined in membrane homogenates from six brain regions, including the hypothalamus (HYP), preoptic area, frontal cortex (CX), ventral tegmental area, striatum (STR) and cerebellum of adult male and female rats. In Experiment 2, the density of delta-opioid receptors was assessed in the CX and STR using receptor autoradiography. Prenatal morphine exposure has no effects on delta-opioid receptors in the brain of gonadally intact, adult male rats regardless of methodology. However, when male rats were gonadectomized in Experiment 2, morphine-exposed males have fewer delta-opioid receptors than controls in the CX but not in the STR. These reductions in cortical delta-opioid receptors are restored by testosterone replacement, demonstrating that prenatal morphine exposure alters testosterone regulation in the CX of male rats. In ovariectomized (OVX) female rats, prenatal morphine exposure increases the density of delta-opioid receptors in the frontal CX. Interestingly, this up-regulation of delta-opioid receptors is not present when the CX is investigated by autoradiography. Moreover, progesterone given alone or in combination with estrogen reduces the density of delta-opioid receptors in the CX and STR of both saline- and morphine-exposed, OVX females. Thus, mid to late gestational morphine exposure differentially alters the influence of adult gonadal hormones on delta-opioid receptors in the CX, decreasing the sensitivity in females and increasing it in males. This is also the first report to demonstrate that gonadal hormones regulate delta receptor densities in brain regions other than the HYP of OVX females.

Sex dimorphic alterations in postnatal brain catecholamines after gestational morphine.

The concentration of brain catecholamines was measured in the hypothalamus, preoptic area (POA), frontal cortex, cerebellum, and striatum of rats exposed in utero to morphine (5-10 mg/kg/twice daily) during gestation days 11-18. Prenatal morphine induced regionally specific, sexually dimorphic alterations in male and female norepinephrine (NE), and dopamine (DA) content at different postnatal ages. Prenatal morphine significantly increased NE content in the hypothalamus of both sexes at postnatal day (PND) 23. In the POA, on the other hand, morphine increased NE content in exposed males at PND 23 and in females at PND 33. In the cerebellum, the NE content of both sexes was significantly elevated at PND 45. In the striatum, NE content was increased by the prenatal morphine only in females at PND 16. The concentration of DA was also affected in a sexually dimorphic manner. At PND 16, prenatal morphine increased the levels of hypothalamic DA only in males, and it reduced the content of DA in female but not male POA. At PND 45, prenatal morphine increased DA in the hypothalamus of females and decreased it in males. In the cerebellum of 16-day-old morphine-exposed animals, DA levels were increased only in males; at PND 45, the levels of DA were still increased in males but had not changed in females. In the striatum, the DA content was reduced only in males at PND 16. Thus, prenatal morphine alters the development of both NE and DA neurotransmitter systems in the hypothalamus, POA, striatum, and cerebellum in a sexually dimorphic manner.

Prenatal morphine exposure differentially alters TH-immunoreactivity in the stress-sensitive brain circuitry of adult male and female rats.

Previously, we demonstrated that exposure to morphine during gestation increases hypothalamic norepinephrine (NE) content and turnover rate in adult male rats and decreases these measures in adult females. To investigate the basis of these alterations, the present study examined the effects of prenatal exposure to morphine on tyrosine hydroxylase immunoreactivity (TH-IR) in the brains of adult male and female progeny. In male rats, prenatal morphine exposure significantly increased the density of TH-IR in cells and fibers in the caudal paraventricular nucleus of the hypothalamus (PVN) and locus coeruleus (LC), but had no effects in the lateral hypothalamus (LH). In female rats that were ovariectomized (OVX), prenatal morphine exposure significantly decreased the density of TH-IR in cells and fibers in the LC. Interestingly, an injection of estrogen in OVX control females reduced the mean optical density of TH-IR in the LC, but it was ineffective in drug-exposed females in the same brain region. Estrogen injections also reduced the mean optical density of TH-IR in the LH but not in the PVN of females, regardless of prenatal drug exposure. Thus, the present study suggests that prenatal morphine exposure induces long-term, sex-specific alterations in TH-IR in the PVN and LC of adult progeny.

Alteration of erythrocyte phosphate transport in primary depressive disorders.

Plasma and erythrocyte phosphate levels, concentrations of red cell organic phosphate ester fractions (acid labile and acid resistant phosphate pools) and intracellular ATP content were determined in 16 patients with bipolar depression, in 14 subjects with unipolar depression as a group of primary affective disorders, in 15 patients suffering from neurotic depression and in 45 healthy controls. Simultaneous in vitro measurements were carried out to detect the exchange rates of inorganic phosphate between the extracellular phosphate pool and the different intracellular phosphate fractions, by use of 32P and applying tracer kinetic analysis. Plasma and red cell inorganic phosphate levels, passive phosphate transport and transfer of inorganic phosphate into the membrane ATP pool were significantly lowered in the primary depressive group as compared to the biochemical values observed in the neurotic group and in the healthy controls. The results suggest an alteration in anion transport across the red cell membrane in primary depressive disorders.

The effect of osmolality changes on gastric mucosal endogenous prostacyclin levels in drug-induced experimental ulcer model of rat.

Osmolality changes evoked with intragastric administration of natural honey or mannitol, significantly decreased the gastric ulceration of rats induced by indomethacin. Together with this effect, a parallel increase was detectable in the mucosal level of endogenous prostacyclin. Although many processes may be involved in this phenomenon, the authors explain their data with a stimulating effect on gastric mucosal microcirculation due to osmolality changes.

Absorption of leucine, alanine and lysine from the rumen.

The absorption of three amino acids (leucine, alanine and lysine) from the washed, closed rumen was studied in a short-term (75 min) experiment in situ. The concentration of leucine and alanine did not change in the rumen during the experiment, while that of lysine continuously decreased, and 40% of the total lysine placed in the rumen was absorbed during the experimental period. The rate of absorption decreased in proportion to the fall of amino acid concentration.

Steady-state distribution of lithium during cultivation of dissociated brain cells.

The formation of a steady-state intracellular lithium level was studied in the course of cultivation of dissociated nerve cell cultures obtained from chick embryonic brains. When lithium was given at a concentration of 2 mM, in the nutrient medium, at day 5, a steady state intracellular lithium content was achieved after about 30 min of incubation and it did not change significantly during the time of cultivation up to the 13th day in vitro.

Uptake, metabolism, and release of [3H]-histamine by glial cells in primary cultures of chicken cerebral hemispheres.

Labelled histamine was taken up into cultured glial cells of chick embryonic brain by a system with high affinity for histamine and diffusion. The active uptake, occurring at low concentrations of the amine, was Na+ dependent and gave an apparent Km of 0.24 microM and a Vmax of 0.31 pmol x mg protein-1 x min-1. The uptake was completely blocked by desmethylimipramine (Ki = 2.5 microM) and partially by the histamine agonists and histamine-N-methyltransferase blockers 4-methylhistamine and 2-methylhistamine (I30 values obtained were 2 microM and 5 microM). Other psychoactive drugs were either ineffective (imipramine) or they showed moderate inhibitory effects (amitriptyline and cocaine). Ouabain (100 microM) inhibited uptake by approximately 50%. Diffusion occurred at high concentrations of the amine, was insensitive to extracellular Na+, and was proportional to histamine concentration up to 1 mM. [3H]-Histamine, taken up into the cells, was metabolized and/or released. The spontaneous efflux of the radioactivity measured after 10 min of exposure to [3H]-histamine (when most of it was still unmetabolized), was moderately Ca++ dependent, accelerated by both reduced concentrations of extracellular Na+ and enhanced concentrations of K+ and inhibited by desmethylimipramine. After prolonged (60 min) incubation, histamine metabolites detected in the cells presented 78% of the chromatogram radioactivity and consisted of N tau-methylhistamine and N tau-methylimidazole acetic acid. These results indicate that at low nM concentrations, histamine is taken up and metabolized by (and released from) glial cells by an Na(+)-dependent system, and the intracellular metabolism seems to serve an increased uptake of the amine.

Effects of lithium on morphological characteristics of dissociated brain cells in culture.

Lithium chloride was added in 5 and 10 mM concentrations for different periods of exposition time to dissociated cultures obtained from chicken embryonic brain. When supplementing lithium at day 1 in vitro for five days, a dose-dependent decrease in total protein was observed in the cultures as compared to the sodium-treated controls. Profound reduction was revealed in the length of neuronal processes and in the number of neuronal cell bodies by phase contrast microscopy and by morphometric means. After exposition of lithium in 10 mM concentration for 48 h, beside a slight decrease in number of perikaryons, a marked reduction in process length of neural elements was observed in the 6-day-old tissue cultures. Ultrastructurally, swollen and degenerating nerve processes have been found after lithium treatment suggesting a particular sensitivity of these structures to lithium ion.