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G Almazan

Publications and source records attributed to G Almazan.

54 records · Page 3Linked to original sources
PubMed1986

Regulation of vasopressin gene expression in rat hypothalamic neurons. Response to osmotic stimulation.

Although the steps involved in biosynthesis and secretion of the neuropeptide vasopressin (AVP) have been extensively studied, the factors which regulate AVP gene expression remain unknown. Therefore, we sought to determine the dynamics of AVP mRNA accumulation in response to a strong stimulus for AVP release, i.e. during salt imbibition and the ensuing period of rehydration. AVP mRNA levels were determined in terms of absolute amounts by a novel quantitative densitometric hybridization assay, using in vitro synthesized sense-strand RNA as a quantitative standard and complementary anti-sense RNA as a specific probe. The template used for RNA transcription consisted of a 196-base pair genomic DNA fragment corresponding to exon C of the rat AVP gene. Determination of basal hypothalamic AVP mRNA levels yielded 12.5 +/- 2.7 fmol/hypothalamus. Salt imbibition, which induced a 6% rise in blood osmolality and an 82% loss of pituitary AVP, resulted in a 3-fold increase of AVP mRNA to 35 +/- 5 fmol/hypothalamus. Following rehydration, plasma osmolality returned to control levels by day 2, pituitary AVP by day 6, and hypothalamic AVP by day 14. By contrast, AVP mRNA levels remained significantly elevated throughout the 30-day rehydration period. Furthermore, pituitary AVP reached a level of 177% of control by day 14 of rehydration. These data show that osmotic stimulation results in a long-lasting elevation of hypothalamic AVP mRNA; during rehydration, these elevated mRNA levels direct AVP biosynthesis at a rate which surpasses secretory demands; AVP mRNA accumulation does not appear to be directly regulated by either pituitary or hypothalamic AVP. Therefore, either an unusually long half-life of greater than or equal to 7 days must be assumed for AVP mRNA or, alternatively, a continued stimulation of AVP gene transcription must occur, even in the absence of a secretory stimulus and following complete repletion of cellular AVP stores.

Animals↗
PubMed1986

Myelin instability and oligodendrocyte metabolism in myelin-deficient mutant mice.

During the active phase of myelination in myelin-deficient mutant mice (mld), myelin basic protein (MBP) synthesis is defective and the myelin lamellae are uncompacted. In these mutants, we found a fast metabolism of the myelin-associated glycoprotein (MAG) and of sulfatides, and the presence of cholesterol esters and a degradation product of MAG, dMAG, indicating that mld myelin was unstable. The increased synthesis of MAG and Wolfgram protein, two proteins present in uncompacted myelin sheath and paranodal loops, was demonstrated by high levels of messengers. Simultaneously, we found an accumulation of inclusion bodies, vacuoles, and rough endoplasmic reticulum in mld oligodendrocytes. This material was heavily immunostained for MAG. Furthermore, the developmental change between the two molecular forms of MAG (p72MAG/p67MAG) was delayed in mld mice. In 85-d-old mld mice, the MBP content increased and myelin lamellae became better compacted. In these mutants, dMAG was absent and MAG mRNAs were found in normal amounts. Furthermore, the fine structure of mld oligodendrocytes was normal and the MAG immunostaining was similar to age-matched controls. These results support a functional role for MBP in maintaining the metabolic stability and the compact structure of myelin. Furthermore, in the absence of MBP and myelin compaction, the regulation of the synthesis of at least two membrane proteins related to myelin cannot proceed.

Age Factors↗
PubMed1986

Dexamethasone stimulates the biochemical differentiation of fetal forebrain cells in reaggregating cultures.

The influence of dexamethasone on the development of neurons and oligodendrocytes was studied in serum-free, aggregating rat brain cell cultures. Synaptogenesis and myelination occur in this culture system. The concentration of myelin basic protein and the activity of 2',3'-cyclic nucleotide 3'-phosphodiesterase were used as oligodendroglia and myelin markers. Choline acetyltransferase and acetylcholinesterase served as neuronal markers, glutamine synthetase reflected astrocyte differentiation, while ornithine decarboxylase served as a general marker for cell growth and maturation. This study showed that dexamethasone stimulated the differentiation of cholinergic neurons and astrocytes. The effect of dexamethasone on oligodendroglial differentiation and myelination depended on the stage of development: during the early phase of myelination dexamethasone had a stimulatory effect, whereas at a later stage it showed a significant inhibition.

Acetylcholinesterase↗
PubMed1985

Epidermal growth factor and bovine growth hormone stimulate differentiation and myelination of brain cell aggregates in culture.

Bovine growth hormone (bGH) and epidermal growth factor (EGF) increased the activity of ornithine decarboxylase (ODC) in brain cell aggregates cultured in a serum-free chemically defined medium. ODC is considered as a marker of cell growth and differentiation. The effect of bGH and EGF on myelination was investigated by measuring two myelin markers, 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP) and myelin basic protein (MBP). EGF treatment at days 2 and 5 caused a dose-dependent increase of both myelin markers at culture day 12. This increase could still be observed at culture day 19, indicating a prolonged action of EGF. The continual presence of bGH in the culture medium produced a large accumulation of MBP at day 19. This effect was dose-dependent and required the presence of triiodothyronine (T3). In contrast, the effect of bGH on CNP activity did not require the presence of T3. This is the first report showing a direct effect of bGH on CNS myelination in vitro and of EGF on both MBP accumulation and ODC activity.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗
PubMed1985

Triiodothyronine stimulation of oligodendroglial differentiation and myelination. A developmental study.

The biochemical development of rotation-mediated aggregating brain cell cultures was studied in a serum-free chemically defined medium in the presence (complete medium) or the absence of triiodothyronine (T3). The expression of 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP) and myelin basic protein (MBP), two myelin components, was temporally dissociated in brain cell aggregating cultures grown in a complete medium. CNP increased from day 8 and reached a plateau around day 25. MBP accumulated rapidly from the third until the fourth week in culture. The total protein content increased gradually until day 25. The activity of ornithine decarboxylase (ODC) used as an index of cell growth and differentiation, showed two well-defined peaks of activity. The first peak reached a maximum at day 6 and correlated with both the highest DNA content and the peak of [3H]-thymidine incorporation. The second peak of ODC activity (from day 19 to 35) coincided with the differentiation of oligodendrocytes. These results confirm that aggregating fetal rat brain cells cultured in a serum-free chemically defined medium undergo extensive differentiation. Addition of T3 to the culture medium doubled the CNP activity by day 16. In contrast, MBP was only slightly increased by day 16, reaching at 25 and 35 days 8 to 10-fold higher values than the untreated cultures. When T3 was removed between day 16 and 25, CNP decreased almost to control values and MBP failed to accumulate. Moreover, when T3 was reintroduced into the medium (between day 25 and 35), CNP activity was restored and MBP content was partially corrected. T3 treatment produced a concentration-dependent increase in ODC activity which was observed only around day 19. The first peak of ODC activity observed at culture day 6 was independent of the presence of T3. These results obtained in brain cell cultures emphasize the direct effect of T3 on myelination.

2',3'-Cyclic Nucleotide 3'-Phosphodiesterase↗
PubMed1984

Anticonvulsive treatment of myelin-deficient (mld) mice improves survival and confirms the delayed increase of myelin basic protein.

Myelin-deficient (mld) mutant mice were treated with phenobarbital between 60 and 90 d of age. The survival rate at 90 d increased from 1.4% in untreated mutants to 46% in those who received phenobarbital. This is evidence that apneic spells during tonic seizures are a major cause of death in mld mice. Myelin basic protein (MBP) content of brain homogenates from treated mld mice increased significantly between 30 and 90 d. MBP was present in myelin purified from the 90-d-old treated mld mice. These results demonstrate that the MBP increase, which occurs after the active phase of myelin formation is completed, is a general phenomenon and is not caused by the selection of a small and mildly affected subpopulation of mutants.

Animals↗
PubMed1984

Effects of collagenase on the release of [3H]-noradrenaline from bovine cultured adrenal chromaffin cells.

Bovine isolated adrenal chromaffin cells maintained in culture at 37 degrees C for 1-7 days become polygonal and bipolar, with typical varicosity-like extensions. Catecholamine levels and dopamine beta-hydroxylase activity decreased after 24-48 h of culture, but recovered to normal levels 3-7 days later. Incubation of 1-7 day-old cells in the presence of increasing concentrations of [3H]-noradrenaline (3.91 to 125 nM) resulted in the retention by the cells of amounts of radioactivity directly proportional to the amine present in the media. One day-old cells took up and retained only one third of the radioactivity found in 2-7 day-old cells. The addition of collagenase to cultured cells caused a decrease in the uptake of tritium. However, the enzyme treatment did not affect the amine taken up by the cell before collagenase treatment. Release of tritium from cultured cells evoked by nicotine, acetylcholine (ACh) or 59 mM K+ was very poor in 24 h-old cells; the secretory response to nicotine, ACh or K+ was dramatically increased after 2-7 days of culture. Bethanecol did not cause any secretory response. When treated with collagenase, cultured cells which had recovered fully their secretory response, lost again the ability to release tritium evoked by ACh or nicotine. However, the responses to high K+, veratridine or ionophore X537A were not affected. The nicotinic response was recovered two days after collagenase treatment. The data suggest that the use of collagenase to disperse the adrenomedullary tissue during the isolation procedure might be responsible for the lost secretory response of young cultured chromaffin cells. Since collagenase specifically impairs the nicotinic cholinoceptor-mediated catecholamine release, it seems likely that the enzyme is exerting its action on the ACh receptor complex. It is unlikely that either voltage-sensitive Na+ or Ca2+ channels are affected by collagenase as the responses induced by high K+ or veratridine were unaffected by this enzyme.

Absorption↗
PubMed1983

Neurotransmitter interaction in regulation of adrenocortical ornithine decarboxylase.

Administration of the dopamine receptor agonists apomorphine (APM) and piribedil increases adrenocortical ornithine decarboxylase (ODC) activity. In this paper alterations in the sympathoadrenal system, and in the central dopaminergic and serotonergic systems have been produced to study the possible mechanism of action of APM on adrenocortical ODC. Unilateral splanchnicotomy, unilateral rhizotomy, and bilateral demedullation each attenuated the response of adrenocortical ODC to APM. Intracerebroventricular 6-hydroxydopamine and 5,6-dihydroxytryptamine, intraperitoneal p-chlorophenylalanine and electrolytic lesion of the medial raphe nucleus reduced the APM-induced increase. None of these treatments produced any changes in the endogenous ODC activity of the adrenal cortex. It is postulated that dopaminergic brain structures participate directly in the stimulatory effect on the hypophyseo-adrenal system to increase adrenocortical ODC activity. An intact central serotonergic system seems to be necessary for APM to exert its effect on adrenocortical ODC activity, particularly the medial raphe nucleus.

Adrenal Cortex↗
PubMed1983

Neuroendocrine control of adrenocortical ornithine decarboxylase activity.

The increase in activity of adrenocortical ornithine decarboxylase (ODC) elicited by the administration of apomorphine (APM) was studied in rats, four days after transection of the spinal cord or 24 h after various types of brain surgery: transection of the mesencephalon or of the diencephalon, or hypothalamic deafferentation (creation of a "hypothalamic island"). Section of the cord elevated endogenous adrenocortical ODC activity and potentiated the induction of the enzyme by APM. Incomplete sections of the brain at the level of the mesencephalon or diencephalon produced no change in either endogenous or induced ODC activity. In contrast to this, interruption of the mesencephalic-diencephalic connections by complete diencephalic transection produced profound decreases of endogenous ODC and of the response to APM. Deafferentation of the hypothalamus raised endogenous ODC concentrations of the adrenal cortex and potentiated the response to APM. The results strongly suggest that APM acts at the level of the diencephalon (hypothalamus) to increase adrenocortical ODC activity. Diencephalic-mesencephalic connections must be intact for this to occur. Peripheral and extrahypothalamic influences play a modulatory role in this effect.

Adrenal Cortex↗
PubMed1982

Adrenomedullary ornithine decarboxylase activity: its use in biochemical mapping of the origins of the splanchnic nerve in the rat.

The activity of ornithine decarboxylase in the adrenal medulla of the rat can be induced transsynaptically by the repeated administration of apomorphine. Unilateral section of one to four ventral spinal cord roots from T4 to T12 partially prevents this effects. Interruption of the intercostal nerves (T7--T13), with preservation of the autonomic innervation of the adrenal medulla, does not produce any alteration in the response of the medullary ODC to APO in the operated side as compared to the intact side. Dorsal root section at T7--T10 leads to a small reduction, while section at T2--T4 has no effect at all. Thus, selective surgical interruption of spinal cord roots indicates that the bulk of splanchnic fibers mediating the transsynaptic induction of adrenomedullary ornithine decarboxylase course in the ventral roots between T7 and T10. Dorsal rhizotomy demonstrates a modulatory role in this induction of afferent information to sympathoadrenal preganglionic neurons involved in innervation of the chromaffin cells.

Adrenal Medulla↗
PubMed1982

Effect of apomorphine, piribedil and haloperidol on adrenal ornithine decarboxylase activity of the rat.

The administration of the dopaminergic drugs, apomorphine and piribedil to rats resulted in an increase in the activity of ornithine decarboxylase of the adrenal medulla and cortex. Pretreatment of the rats with the dopamine-receptor antagonist haloperidol caused a partial blockade of the apomorphine-induced effect at 4 hr in both medulla and cortex. At 6 hr, however, haloperidol did not block the effect of apomorphine and produced an increase in ornithine decarboxylase activity of both structures when administered alone. Hypophysectomy abolished the cortical ornithine decarboxylase response to apomorphine and haloperidol and the medullary response to haloperidol. The results suggest that the response of cortical ornithine decarboxylase activity to apomorphine and haloperidol is entirely mediated by the hypophysis and that the effect of apomorphine and the antagonistic action of haloperidol towards apomorphine in regard to the induction of adrenal medullary ornithine decarboxylase must be taking place at some central site independent of the hypothalamic-hypophyseal system.

Adrenal Glands↗
PubMed1981

Effect of haloperidol on adrenal ornithine decarboxylase activity of the rat.

The administration of the dopamine antagonist haloperidol (HLP) to rats produced a temporary increase in adrenomedullary and cortical ornithine decarboxylase (ODC) activity. The time-course of stimulation of ODC activity by HLP showed different patterns in both structures. Medullary ODC activity was highest at 2.5 h, decreasing at later times; cortical ODC activity was not affected by the drug at 2.5 h, but then increased up to at least 6.5 h. The medullary increase observed at 2.5 h was dose-related and could be prevented by splanchnicotomy. Hypophysectomized rats, on the contrary, showed an enhanced response to HLP. The results suggest that haloperidol-induced increase of adrenomedullary ODC activity is caused by a reflex increase in preganglionic nerve activity, and that the pituitary gland can modulate this response. Cortical ODC response to HLP, as previously demonstrated, is mediated entirely by the hypophysis.

Adrenal Cortex↗
PubMedDate not supplied

Intrinsic myelin proteins are normally synthesized in vitro in the myelin-deficient (mld) mutant mouse.

The synthesis of myelin basic protein is severely reduced in the central and peripheral nervous system of myelin-deficient (mld) mutant mice. Using an in vitro system and immunoprecipitation, we found a normal rate of synthesis in mld mice for two intrinsic myelin proteins: proteolipid protein in central nervous system myelin and P0 in peripheral nervous system myelin. These results indicate that protein synthesis on membrane-bound ribosomes is not affected by the mutation.

Animals↗