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Biomedical subjects

K E Moore

Publications and source records attributed to K E Moore.

At least 109 records · Page 6Linked to original sources

A method to detect rabbit neutrophil phagocytosis of Pasteurella multocida.

A method was developed to detect neutrophil phagocytosis of bacteria by determining whether neutrophil-associated bacteria were intra- or extracellular. Neutrophils were treated with 8-(N,N-diethylamino)-octyl-3,4,5-trimethoxybenzoate hydrochloride to inhibit degranulation and, consequently, killing of bacteria. Treated neutrophils and opsonized Pasteurella multocida were combined. Following phagocytosis, the suspensions were centrifuged and the pellets were washed to remove non-cell-associated bacteria. The pellets were resuspended and heparin was added to prevent further phagocytosis. Samples were removed, and the number of viable bacteria was determined by a dilution and plate count technique. Streptomycin, an antibiotic that is poorly taken up by neutrophils, was added to kill extracellular bacteria, and the suspensions were incubated for 20 minutes at 37 C, and samples were removed again and bacterial numbers were determined. Percentage killing of bacteria by streptomycin was calculated. Phagocytosed bacteria were protected from the bactericidal action of streptomycin.

Animals↗

Electrical stimulation of the arcuate nucleus increases the metabolism of dopamine in terminals of tuberoinfundibular neurons in the median eminence.

The purpose of the present study was to examine the effects of procedures that alter impulse flow in tuberoinfundibular dopamine (DA) neurons on the metabolism of DA in the median eminence and on the secretion of prolactin from the anterior pituitary. Twenty min following the administration of gamma-butyrolactone (GBL, 1000 mg/kg, i.p.) there was a marked decrease in 3,4-dihydroxyphenylacetic acid (DOPAC) concentrations in the median eminence and an increase in prolactin concentrations in the serum, indicating that a decrease in activity of tuberoinfundibular DA neurons is accompanied by a decrease in DA metabolism in the median eminence and a loss of tonic inhibition of pituitary prolactin secretion. Activation of tuberoinfundibular DA neurons by bilateral stimulation of the arcuate nucleus in GBL-treated rats produced a rapid increase in median eminence DOPAC concentrations and a time-dependent decrease in serum prolactin concentrations. Nomifensine (25 mg/kg, i.p., 30 min), a DA uptake inhibitor, had no effect on median eminence DOPAC concentrations in sham- or arcuate nucleus-stimulated rats, indicating that regardless of the level of activity of tuberoinfundibular neurons, very little DA is recaptured and metabolized in terminals of these neurons in the median eminence. Taken together these results reveal that alterations in impulse flow in tuberoinfundibular DA neurons are accompanied by corresponding changes in the metabolism of DA in the median eminence.

3,4-Dihydroxyphenylacetic Acid↗

Effect of electrical stimulation of the arcuate nucleus on neurochemical estimates of tuberoinfundibular and tuberohypophysial dopaminergic neuronal activities.

The activity of nigrostriatal dopaminergic neurons has been estimated biochemically by measuring the rates of dopamine (DA) synthesis (accumulation of dihydroxyphenylalanine (DOPA) after NSD 1015) and turnover (decline of DA concentrations after alpha-methyltyrosine) in the striatum. It has been assumed that the activities of tuberoinfundibular dopaminergic (TIDA) and tuberohypophysial dopaminergic (THDA) neurons can also be estimated by making the same measurements in the terminals of these neurons in the median eminence and the posterior pituitary, respectively. In the present study, this assumption was tested directly by measuring the rates of DA synthesis and turnover in the median eminence and posterior pituitary following electrical stimulation of TIDS and THDA cell bodies in the arcuate nucleus. Electrical stimulation of the arcuate nucleus increased the rate of DOPA accumulation and the alpha-methyltyrosine-induced decline of DA concentrations in the median eminence and in the neural and intermediate lobes of the posterior pituitary. gamma-Butyrolactone (GBL), an anesthetic that selectively inhibits DA impulse flow, reduced the rates of DA synthesis and turnover in the median eminence. GBL also increased prolactin secretion which is tonically inhibited by DA released from TIDA neurons. Serum prolactin levels were significantly decreased by arcuate nucleus stimulation in GBL-anesthetized rats. These results indicate that the rates of DA synthesis and turnover within the median eminence and posterior pituitary reflect the activities of TIDA and THDA neurons, respectively.

4-Butyrolactone↗

The metabolism of dopamine in the median eminence reflects the activity of tuberoinfundibular neurons.

The purpose of the present study was to characterize the metabolism of dopamine (DA) in tuberoinfundibular (TI) neurons terminating in the median eminence and to examine the effects of procedures that alter the synthesis and turnover of DA in these neurons on the concentrations of dihydroxyphenylacetic acid (DOPAC) in the median eminence. The DA uptake inhibitor nomifensine (25 mg/kg, i.p.; 30 min) failed to alter median eminence DOPAC concentrations indicating that very little released DA is recaptured and metabolized by TIDA neurons. Within 5 min following the administration of the monoamine oxidase inhibitor pargyline (50 mg/kg, i.v.) median eminence DOPAC concentrations declined to 15% of control demonstrating that this metabolite has a high turnover rate and is rapidly removed from the median eminence. Median eminence DOPAC concentrations in diestrous female rats, whose TIDA neuronal activity is higher than in the male, were two-fold greater than in male rats. Prolactin (10 micrograms/rat, i.c.v.; 12 h), which increases TIDA neuronal activity, produced a corresponding increase in median eminence DOPAC concentrations in male rats. Restraint stress (30 min), which decreases TIDA neuronal activity, produced a corresponding decrease in median eminence DOPAC concentrations in diestrous female rats. The results from the present study suggest that DOPAC concentrations in the median eminence can be used as an index of TIDA neuronal activity.

3,4-Dihydroxyphenylacetic Acid↗

Determination of the source of 5-hydroxytryptaminergic neuronal projections to the neural and intermediate lobes of the rat pituitary gland through the use of electrical stimulation and lesioning experiments.

5-Hydroxytryptamine (5-HT)-containing axons and terminals have been visualized in the neural and intermediate lobes of the rat pituitary gland, but the origin of these fibers remains in question. This study was designed to determine if 5-HT cell bodies in the brainstem or in the dorsomedial nucleus of the hypothalamus project to either of these pituitary lobes. Since lesions and electrical stimulation of 5-HT cell bodies decrease and increase, respectively, the rate of 5-HT synthesis in regions innervated by these cells, these techniques were employed. The in vivo rate of 5-HT synthesis was determined by quantifying the rate of accumulation of the 5-HT precursor, 5-hydroxytryptophan (5-HTP), in the neural and intermediate lobes of the pituitary gland 30 min after the administration of a decarboxylase inhibitor (NSD 1015, 100 mg/kg, i.p.). The application of 30 min of stimulating current (monophasic cathodal pulses of 1 ms duration and 0.3 mA current delivered at a frequency of 10 Hz) to electrodes implanted in the dorsal and median raphe nuclei increased the rate of 5-HT synthesis in both the neural and intermediate lobes of the pituitary gland. 5,7-Dihydroxytryptamine lesions of these nuclei altered neither 5-HTP accumulation nor 5-HT concentrations in the neural and intermediate lobes, but similar lesions of the nuclei raphe pontis and raphe magnus decreased both the concentration of 5-HT and the accumulation of 5-HTP in these pituitary regions.(ABSTRACT TRUNCATED AT 250 WORDS)

5,7-Dihydroxytryptamine↗

A small hydrophobic domain anchors leader peptidase to the cytoplasmic membrane of Escherichia coli.

Leader peptidase is an enzyme of the Escherichia coli cytoplasmic membrane which removes amino-terminal leader sequences from many secreted and membrane proteins. Three potential membrane-spanning segments exist in the first 98 amino acids of leader peptidase. We have characterized the topology of leader peptidase based on its sensitivity to protease digestion. Proteinase K and trypsin treatment of right-side-out inner membrane vesicles and spheroplasts yields protected fragments of approximately 80 and 105 amino acid residues, respectively. We have shown that both fragments are derived from the amino terminus of the protein and that the smaller protected peptide can be derived from the larger. Removal of the third potential membrane-spanning segment (residues 82-98) does not affect the size of the proteinase K-protected fragment but does reduce the size of the trypsin-protected peptide. Because the proteinase K-protected fragment is about 9000 daltons, is derived from the amino terminus of leader peptidase, and its size is not affected when amino acids 82-98 are removed from the protein, it must extend from the amino terminus to approximately residue 80. Likewise, the trypsin-protected fragment must extend from the amino terminus to about residue 105. These data suggest a model for the orientation of leader peptidase in which the second hydrophobic stretch (residues 62-76) spans the cytoplasmic membrane and the third hydrophobic stretch resides in the periplasmic space.

Amino Acid Sequence↗

5-Hydroxytryptamine is synthesized in neurons terminating in the neural and intermediate lobes of the rat pituitary gland.

Immunocytochemistry has revealed that nerve fibers within the neural and intermediate lobes of the rat pituitary gland contain 5-hydroxytryptamine (5-HT). Recent anatomical evidence suggests that the content of this amine in the intermediate but not the neural lobe of the pituitary gland may represent 5-HT that has been taken up from the blood rather than synthesized intraneuronally. The purpose of this study was to determine if 5-HT is synthesized in neurons of the neurointermediate lobe of the pituitary gland. 5-HT synthesis was estimated by measuring the accumulation of the 5-HT precursor, 5-hydroxytryptophan (5-HTP), in the neurointermediate lobe of male Long-Evans rats following the administration of NSD 1015, an inhibitor of aromatic L-amino acid decarboxylase. Thirty min following the injection of NSD 1015 (100 mg/kg, i.p.), 5-HTP accumulated in the neurointermediate lobe and the rate of this accumulation was increased by the administration of the 5-HTP precursor, tryptophan, and by electrical stimulation of the pituitary stalk. In addition, repeated injections of the 5-HT uptake inhibitor, fluoxetine (10 mg/kg, i.p., every 12 h for a total of 7 injections), induced a marked depletion of platelet 5-HT but did not alter the concentration of 5-HT in either the neural or intermediate lobes of the pituitary gland. Taken together these results indicate that much of the 5-HT in the neurointermediate lobe of the pituitary gland does not represent 5-HT taken up from the blood, but rather the amine is synthesized in neurons projecting to this region.(ABSTRACT TRUNCATED AT 250 WORDS)

5-Hydroxytryptophan↗

Differential action of bromocriptine on nigrostriatal versus mesolimbic dopaminergic neurons.

The present study was undertaken to compare the abilities of the dopaminergic agonists apomorphine, bromocriptine, and lergotrile to inhibit the synthesis of dopamine (DA) in terminals of nigrostriatal and mesolimbic DA neurons. The in vivo synthesis of DA was estimated by measuring the rate of accumulation of dihydroxyphenylalanine (DOPA) in terminals of nigrostriatal (striatum) and mesolimbic (nucleus accumbens, olfactory tubercle) neurons 30 min after the administration of NSD 1015, a decarboxylase inhibitor. The activation of DA autoreceptors in these regions was evaluated by measuring the abilities of the DA agonists to inhibit DA synthesis in brain regions of rats pretreated with gamma-butyrolactone (GBL). Apomorphine (0.03-1.0 mg/kg for 45 min) and bromocriptine (0.1-10 mg/kg for 90 min) produced dose-dependent decreases in the rate of DA synthesis in all three brain regions of both vehicle- and GBL-treated rats. A time course of the effects of the highest dose of bromocriptine (10 mg/kg), however, demonstrated dramatic regional differences in the ability of this drug to inhibit DA synthesis in saline-versus GBL-pretreated rats. Bromocriptine inhibited the GBL-induced increase in DA synthesis for 6 hours in all regions examined. In the striatum of saline-treated rats the decrease in DA synthesis was evident only at 1.5 hours after bromocriptine administration, while in the nucleus accumbens and olfactory tubercle DA synthesis remained inhibited for 6 hours. By contrast, lergotrile reduced DA synthesis to a similar extent in all three regions for at least 6 hours in both vehicle- and GBL-treated rats. These results suggest that there is no regional difference in the ability of bromocriptine to inhibit DA synthesis via DA autoreceptor mechanisms, but there appear to be differences in postsynaptic DA receptor-mediated mechanisms which regulate nigrostriatal versus mesolimbic DA neurons.

4-Butyrolactone↗

Stress, prolactin and hypothalamic dopaminergic neurons.

Most of what is known about dopamine (DA)-containing neurons in the brain has been learned from studies on the nigrostriatal system, although it is misleading to consider a neuron in this system as a "model" for other DA neurons in the brain. In this presentation the properties of nigrostriatal DA neurons are compared with those hypothalamic DA neurons that comprise the tuberoinfundibular system. These latter neurons, which have cell bodies in the arcuate nucleus and short axons which terminate in the median eminence, function to inhibit the release of prolactin from the anterior pituitary. The activities of the nigrostriatal and tuberoinfundibular DA neurons were estimated biochemically by measuring the rates of synthesis (accumulation of DOPA after the administration of a decarboxylase inhibitor, NSD 1015), turnover (decline of DA after the administration of a tyrosine hydroxylase inhibitor, alpha-methyltyrosine) and metabolism (concentrations of dihydroxyphenylacetic acid) of DA in regions of the brain that contain the terminals of these neurons (striatum and median eminence, respectively). Tuberoinfundibular DA neurons differ from nigrostriatal DA neurons in that the former neurons: (1) are not directly regulated by DA receptor-mediated mechanisms, (2) are stimulated by prolactin, (3) exhibit a sexual difference with activity being 2-3 times greater in females and (4) are inhibited by afferent neuronal circuits that are activated by suckling and restraint stress.

3,4-Dihydroxyphenylacetic Acid↗

Interactions between prolactin and dopaminergic neurons.

The secretion of prolactin from the adenohypophysis is tonically inhibited by dopamine that is released into the hypophysial portal blood from terminals of tuberoinfundibular neurons located in the external layer of the median eminence. These tuberoinfundibular neurons are unique among other dopaminergic neurons in the brain (including the well-characterized nigrostriatal neurons) in that they are not directly regulated by dopaminergic receptor-mediated mechanisms, but instead are selectively responsive to changes in prolactin concentrations in blood and cerebrospinal fluid. In the rat, the activity of the tuberoinfundibular dopaminergic neurons is higher in the female than in the male, exhibits a characteristic cyclical pattern during the first half of pregnancy and is constantly high as a result of stimulation by placental lactogen during the last 9 days of pregnancy, and is reduced in lactating animals and acutely inhibited during suckling.

Animals↗

The 20-Hz signals of finback whales (Balaenoptera physalus).

The 20-Hz signals of finback whales (Balaenoptera physalus) were analyzed from more than 25 years of recordings at a variety of geographic locations on near-surface hydrophones close to whales and on deep hydrophone systems. These signals were composed of 1-s pulses of sinusoidal waveform with downward sweeping frequency from approximately 23 to 18 Hz at variable source levels up to 186 dB (re: 1 microPa at 1 m), usually with slightly lower levels for the pulses at the beginning and end of sequences. These "20-Hz" pulses were produced in signal bouts (separated by more than 2 h) lasting as long as 32.5 h. Bouts were composed of regularly repeated pulses at intervals of 7-26 s (typically), either at one nominal pulse rate or at two alternating (doublet) pulse intervals. Signal bouts were interrupted by rests of 1-20 min at roughly 15-min intervals and by irregular gaps lasting between 20 and 120 min. The distribution of these signals throughout the year and their temporal sequence were analyzed from the continuous drum records of the Bermuda SOFAR Station. Signal bouts occurred during winter, sometimes beginning in September and ending in May. The sound sequences were never exactly replicated. Direct association of the bouts with the reproductive season for this species points to the 20-Hz signals as possible reproductive displays by finback whales.

Acoustics↗

Responsiveness of tuberoinfundibular dopamine neurons in the aged female rat to the stimulatory actions of prolactin.

The autoregulatory feedback control of prolactin, which is mediated by tuberoinfundibular dopamine (DA) neurons, is altered in the aged rat; this is evidenced by increased circulating concentrations of prolactin and decreased activity of these neurons. In the present study the action of prolactin on tuberoinfundibular DA neurons in young and aged female rats was estimated by measuring the rate of DA synthesis (dopa accumulation following the administration of a decarboxylase inhibitor) in the median eminence. The rate of dopa accumulation in the median eminence of the aged (26 months) rat was reduced to 50-60% of that in the young (3 months) rat. The acute systemic administration of haloperidol, a DA antagonist which increases serum concentrations of prolactin or intracerebroventricular infusions of prolactin increased the rate of dopa accumulation in the median eminence of both young and aged rats by the same relative amount. The administration of haloperidol and prolactin increased the rate of DA synthesis to a greater extent in young than in aged rats. The administration of bromocriptine, a DA agonist which reduces serum concentrations of prolactin, decreased the rate of dopa accumulation in the median eminence of both young and aged rats. In young animals the intracerebroventricular administration of prolactin reversed the bromocriptine-induced decrease in DA synthesis in the median eminence after 4 h and caused a further increase after 12 h. Qualitatively similar effects were seen in the aged rats; however, prolactin-treated young rats had much higher levels of DA synthesis than aged rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Restraint stress decreases the neurosecretory activity of tuberoinfundibular dopaminergic neurons in young but not in aged female rats.

The regulation of prolactin secretion by tuberoinfundibular dopamine (DA) neurons appears to be altered in the aged rat: the concentration of prolactin in the serum increases and the activity of the tuberoinfundibular DA neurons decreases. In the young female rat a brief period of stress reduces the tuberoinfundibular DA neurosecretory activity and increases the secretion of prolactin. The purpose of the present study was to determine if the responsiveness of tuberoinfundibular DA neurons to restraint stress is altered in the aged female rat. The activity of these neurons was estimated from the rate of DA synthesis in their terminals in the median eminence, as measured by the rate of accumulation of dihydroxyphenylalanine (DOPA) after the administration of a decarboxylase inhibitor. Thirty minutes of restraint stress increased serum prolactin concentrations in both young (3 months) and aged (26 months) constant estrous rats, but reduced the rate of DOPA accumulation in the median eminence of only the young rats. Restraint also decreased the rate of DOPA accumulation in the median eminence of intermediate-aged rats (14 months) independently of whether the rats were exhibiting normal ovarian cycles (measured on the day of estrus) or were in a constant estrus. This suggests that the loss of ovarian cyclicity per se is not associated with the age-related change in the response of tuberoinfundibular DA neurons to restraint stress.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Comparative effects of the neurotoxins N-chloroethyl-N-ethyl-2-bromobenzylamine hydrochloride (DSP4) and 6-hydroxydopamine on hypothalamic noradrenergic, dopaminergic and 5-hydroxytryptaminergic neurons in the male rat.

The intracerebroventricular (i.c.v.) administration of 6-hydroxydopamine (6-OHDA; 50 micrograms X 3) and the systemic administration of DSP4 (50 mg/kg X 2; i.p.), alone and in combination, were compared for their abilities to alter the concentrations of norepinephrine (NE), dopamine (DA), dihydroxyphenylacetic acid (DOPAC) and 5-hydroxytryptamine (5-HT) in selected hypothalamic and extra-hypothalamic (striatum, frontal cortex, hippocampus) regions of the male rat brain. DSP4 markedly lowered NE concentrations in extrahypothalamic regions, and within the hypothalamus produced a mild and variable reduction of NE without altering concentrations of DA, DOPAC or 5-HT. 6-OHDA markedly lowered NE concentrations in all brain regions, but was without effect on DA, DOPAC and 5-HT concentrations in any region analyzed. Combined treatment with DSP4 and 6-OHDA did not produce additional effects on levels of NE, DA and DOPAC over either drug alone, but did cause a mild reduction of 5-HT in several brain regions. These results indicate that systemic treatments with DSP4 per se are not as effective as i.c.v. 6-OHDA in depleting NE in the hypothalamus, and that when the two neurotoxins are administered there appears to be some destruction of 5-HT neurons.

3,4-Dihydroxyphenylacetic Acid↗

Effects of acute and chronic bupropion on locomotor activity and dopaminergic neurons.

Acute administration of bupropion (10 or 30 mg/kg) to rats increased locomotor activity in a dose-related manner. The highest dose increased the dopamine (DA) concentration while both doses reduced the concentration of dihydroxyphenylacetic acid (DOPAC) in the striatum. The enhancement of locomotor activity and the decrease of striatal DOPAC concentrations were increased with chronic administration (up to 40 days) of bupropion. The rate of DA synthesis in the striatum was increased by the acute administration of d-amphetamine but was not altered by acute or chronic administration of bupropion.

3,4-Dihydroxyphenylacetic Acid↗

A comparison of biochemical indices of 5-hydroxytryptaminergic neuronal activity following electrical stimulation of the dorsal raphe nucleus.

The activity of 5-hydroxytryptaminergic neurons has been estimated from measurements of: concentrations of 5-hydroxyindoleacetic acid; the ratio of the concentrations of 5-hydroxyindoleacetic acid to 5-hydroxytryptamine; the rate of accumulation of 5-hydroxytryptophan following the administration of an aromatic L-amino acid decarboxylase inhibitor (e.g., NSD 1015); the rate of accumulation of 5-hydroxytryptamine, and the rate of decline of 5-hydroxyindoleacetic acid following the administration of a monoamine oxidase inhibitor (e.g., pargyline). The purpose of the present study was to compare these different methods under conditions of changing neuronal impulse traffic produced by electrical stimulation of 5-hydroxytryptaminergic neurons. Male rats anesthetized with chloral hydrate were killed following 0, 15, or 30 min of electrical stimulation of the dorsal raphe nucleus at a frequency of 0, 5, or 10 Hz. The concentrations of 5-hydroxytryptamine, 5-hydroxyindoleacetic acid, and 5-hydroxytryptophan in nucleus accumbens, amygdala, suprachiasmatic nucleus, and dorsomedial nucleus were measured using HPLC coupled to an electrochemical detector. In each brain region, stimulation elicited an increase in the concentration of 5-hydroxyindoleacetic acid and the 5-hydroxyindoleacetic acid/5-hydroxytryptamine concentration ratio in saline-treated animals and an increase in 5-hydroxytryptophan accumulation in NSD 1015-treated animals, but did not alter the concentration of 5-hydroxytryptamine or 5-hydroxyindoleacetic acid in pargyline-treated rats. The results o f this study indicate that although the first three methods serve as valid indices of 5-hydroxytryptaminergic neuronal activity, the pargyline-dependent techniques are not responsive to changes in the rate of 5-hydroxytryptamine nerve firing.

5-Hydroxytryptophan↗

Exogenous tryptophan increases synthesis, storage, and intraneuronal metabolism of 5-hydroxytryptamine in the rat hypothalamus.

The effects of tryptophan administration on neurochemical estimates of synthesis [5-hydroxytryptophan (5-HTP) accumulation following administration of a decarboxylase inhibitor], storage [5-hydroxytryptamine (5-HT) concentrations], and metabolism [5-hydroxyindoleacetic acid (5-HIAA) concentrations] of 5-HT in selected regions of the hypothalamus were determined using HPLC coupled to an electrochemical detector. Tryptophan methyl ester HCl (30-300 mg/kg i.p.) produced a dose-dependent increase in the rate of 5-HTP accumulation throughout the hypothalamus but had no effect on the rate of accumulation of 3,4-dihydroxyphenylalanine. Peak 5-HTP levels were attained by 30 min following administration of tryptophan (100 mg/kg i.p.) and were maintained for an additional 60 min. Tryptophan also produced concomitant dose-dependent increases in 5-HT and 5-HIAA concentrations in these same regions without changes in the 5-HIAA/5-HT ratio. These results indicate that exogenous tryptophan administration selectively increases the synthesis, storage, and metabolism of 5-HT in the hypothalamus without altering the synthesis of catecholamines. Inhibition of 5-HT uptake with chlorimipramine or fluoxetine produced modest (10-40%) reductions in 5-HIAA concentrations throughout the hypothalamus, revealing that only a minor portion of 5-HIAA is derived from released and recaptured 5-HT, whereas the major portion of this metabolite reflects intraneuronal metabolism of unreleased 5-HT. In both chlorimipramine- and fluoxetine-treated rats, 5-HIAA concentrations were significantly increased by tryptophan administration, indicating that the increase in synthesis of 5-HT following precursor loading is accompanied by an increase in the intraneuronal metabolism of 5-HT.

Animals↗