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N R Zahniser

Publications and source records attributed to N R Zahniser.

89 records · Page 5Linked to original sources
PubMed1985

Binding characteristics of the dopamine uptake inhibitor [3H]nomifensine to striatal membranes.

Binding of the radiolabeled antidepressant [3H]nomifensine to rat and rabbit striatal membranes has been characterized. The specific binding of [3H]nomifensine to striatal membranes was stable, reversible and saturable. Saturation experiments indicated that [3H]nomifensine labeled a single site with an affinity (Kd) of 80 nM and a total number of binding sites (Bmax) of 6.5 pmoles/mg protein both in rat and rabbit striatal membranes. The affinity constants obtained from kinetic analyses and competition experiments were in fairly good agreement with those obtained in saturation experiments. Compounds known to inhibit [3H]dopamine uptake in vitro, such as nomifensine, 4-hydroxy-nomifensine, mazindol, amfonelic acid and benztropine, were the most potent competitors of nomifensine binding. Additionally, the absolute potencies of various drugs in competing for [3H]nomifensine binding to rat and rabbit striatal membranes correlated closely with their potencies in inhibiting [3H]dopamine uptake into striatal synaptosomes. Specific [3H]nomifensine binding was dependent on the presence of NaCl which is also consistent with its association with the dopamine uptake pump. The number, but not the affinity, of striatal [3H]nomifensine binding sites was reduced significantly following in vivo lesions with 6-hydroxydopamine. The number of [3H]nomifensine binding sites was found to be highest in areas rich in dopamine nerve terminals such as the striatum and olfactory tubercle. These results suggest that [3H]nomifensine binds to a site on dopaminergic nerve terminals associated with the dopamine uptake pump.

Animals↗
PubMed1985

Quantitative autoradiography of 3H-nomifensine binding sites in rat brain.

The distribution of 3H-nomifensine binding sites in the rat brain has been studied by quantitative autoradiography. The binding of 3H-nomifensine to caudate putamen sections was saturable, specific, of a high affinity (Kd = 56 nM) and sodium-dependent. The dopamine uptake inhibitors benztropine, nomifensine, cocaine, bupropion and amfonelic acid were the most potent competitors of 3H-nomifensine binding to striatal sections. The highest levels of (benztropine-displaceable) 3H-nomifensine binding sites were found in the caudate-putamen, the olfactory tubercle and the nucleus accumbens. 6-Hydroxydopamine-induced lesion of the ascending dopaminergic bundle resulted in a marked decrease in the 3H-ligand binding in these areas. Moderately high concentrations of the 3H-ligand were observed in the bed nucleus of the stria terminalis, the anteroventral thalamic nucleus, the cingulate cortex, the lateral septum, the hippocampus, the amygdala, the zona incerta and some hypothalamic nuclei. There were low levels of the binding sites in the habenula, the dorsolateral geniculate body, the substantia nigra, the ventral tegmental area and the periaqueductal gray matter. These autoradiographic data are consistent with the hypothesis that 3H-nomifensine binds primarily to the presynaptic uptake site for dopamine but also labels the norepinephrine uptake site.

Animals↗
PubMed1984

Electrophysiological effects of cocaine in the rat hippocampus in vitro.

The effects of cocaine on electrophysiological activity were examined in the CA1 region of the in vitro hippocampal slice. Low concentrations of cocaine (0.25-2.5 microM) had no direct effect on evoked potentials, but potentiated responses to threshold concentrations of norepinephrine (NE; 0.5 microM); this effect was also seen with other inhibitors of catecholamine uptake such as desipramine. The ability of low doses of cocaine to augment the changes in population spike amplitude elicited by NE appeared to be related to the ability of cocaine to block the uptake of NE into the noradrenergic nerve terminals in the rat hippocampus. Higher concentrations of cocaine (5-200 microM) elicited responses consistent with local anesthetic actions on neuronal membranes.

Animals↗
PubMed1984

Characterization and regulation of insulin receptors in rat brain.

An in vitro receptor binding assay, using filtration to separate bound from free [125I]insulin, was developed and used to characterize insulin receptors on membranes isolated from specific areas of rat brain. The kinetic and equilibrium binding properties of central receptors were similar to those of hepatic receptors. The binding profiles in all tissues were complex and were consistent with binding in multiple steps or to multiple sites. Similar binding properties were found among receptors in olfactory tubercle/bulb, cerebral cortex, hippocampus, striatum, hypothalamus, and cerebellum. High affinity [125I]insulin binding sites (KD = 3-11 nM) were distributed evenly between membranes isolated from P1 and P2 fractions of these brain areas, with the exception of the olfactory tubercle in which binding to P2 membranes was four-fold greater (Bmax = 150 fmol/mg protein). One difference between insulin receptors in brain and peripheral target tissues, however, was observed. Following exposure to 0.17 microM insulin for 3 h at 37 degrees C, the number of specific [125I]insulin binding sites on adipocytes decreased by 40%, while the number of binding sites on minces of cerebral cortex/olfactory tubercle remained constant. The results suggest that although the binding characteristics of central and peripheral insulin receptors are similar, these receptors do not appear to be regulated in the same manner.

Animals↗
PubMed1983

Structural differences between insulin receptors in the brain and peripheral target tissues.

Insulin receptors in various brain regions (olfactory tubercle, hippocampus, and hypothalamus) were photoaffinity labeled using the photoreactive analogue of insulin B2(2-nitro,4-azidophenylacetyl)-des-PheB1-insulin (NAPA-DP-insulin). A protein with an apparent Mr of 400,000 was specifically labeled with 125I-NAPA-DP-insulin in all three brain regions. When radiolabeled proteins were reduced with dithiothreitol prior to electrophoresis, specific labeling occurred predominantly in a protein with an apparent Mr of 115,000 and to a much lesser extent in a protein with an apparent Mr of 83,000. The size of these receptor proteins, based on their electrophoretic mobilities, was consistently smaller than insulin receptor proteins in adipocytes. The covalent labeling of insulin receptors in brain by 125I-NAPA-DP-insulin was not blocked by anti-insulin receptor antiserum. Additionally, in contrast to effects observed in peripheral target tissues, this antisera did not inhibit the binding of 125I-insulin to brain membranes. Neuraminidase treatment resulted in an increase in the electrophoretic mobilities of insulin receptor subunits in adipocytes, but, had no effect on receptor subunits in brain. Solubilized insulin receptors from adipocytes were retained by wheat germ agglutinin columns and specifically eluted with N-acetylglucosamine. In contrast, solubilized insulin receptors from brain did not bind to these columns. The results from this study indicate that structural differences, including molecular weight, antigenicity, and carbohydrate composition exist between insulin receptors in brain and peripheral target tissues.

Adipose Tissue↗
PubMed1983

Electrophysiological and biochemical sequelae of the destruction of hippocampal noradrenergic afferents by DSP4.

The effects of DSP4 lesions were examined 20-53 days postlesion in the rat hippocampus. A single treatment with DSP4 produced decreases of 42-94% in the norepinephrine (NE) content of this brain region. There was, however, no effect of DSP4 treatment on either the number or affinity of beta-adrenergic receptor sites as determined by radioligand binding studies with (-)-[125I]pindolol; furthermore, there was no relationship between the concentrations of NE and the number of receptor sites in individual hippocampi. The DSP4-induced depletion of functionally releasable NE was confirmed by the loss of electrophysiological responsiveness to amphetamine in the in vitro hippocampus following such lesions. In contrast, electrophysiological responses to direct acting beta-adrenergic or alpha-adrenergic agonists were unchanged following DSP4 treatment. This finding again suggests the lack of any change in postsynaptic sensitivity. The results of this study demonstrate that while the potent noradrenergic neurotoxin DSP4 is able to reduce NE concentrations significantly in noradrenergic target regions in brain, these lesions are not necessarily associated with postsynaptic changes in adrenergic systems.

Afferent Pathways↗
PubMed1983

Comparison of dopamine receptor sites labeled by [3H]-S-sulpiride and [3H]-spiperone in striatum.

Binding of the radiolabeled active isomer of the neuroleptic sulpiride, [3H]-S-sulpiride, to rat and rabbit striatal membranes was characterized. Regardless of whether the specific binding of [3H]-S-sulpiride was defined with spiperone or the active isomers of butaclamol or flupenthixol, a single homogeneous++ population of binding sites (rat: Kd = 5.6 nM, maximum binding = 590 fmol/mg of protein; rabbit: Kd = 8.3 nM, maximum binding = 540 fmol/mg of protein) was detected. The pharmacological profile of these sites was characteristic of that described for the dopaminergic D-2 receptor subtype. To determine whether [3H]-S-sulpiride and [3H]spiperone label common sites in the striatum, the binding of these two radioligands was compared under similar assay conditions. When specific binding of [3H]spiperone was defined with S-sulpiride, [3H]spiperone labeled the same number of binding sites as [3H]-S-sulpiride despite the fact that the affinity of the sites for [3H]spiperone was 80- to 90-fold higher than for [3H]-S-sulpiride. When specific binding of [3H]spiperone was defined with either (+)-butaclamol or (alpha)-flupenthixol, however, approximately 30% more sites were labeled. The predominant site labeled by [3H]spiperone also possessed the characteristics of the D-2 receptor. It is concluded that [3H]-S-sulpiride under the conditions used is a selective radioligand with which dopamine receptors of the D-2 subtype can be directly measured and localized. [3H]Spiperone can be used to detect the same sites only if specific binding is defined with S-sulpiride.

Animals↗
PubMed1983

Thermodynamic differences between agonist and antagonist interactions with binding sites for [3H]spiroperidol in rat striatum.

The characteristics of the binding of the dopamine receptor antagonist [3H]spiroperidol to rat striatal membranes were examined at six different incubation temperatures ranging from 1 degree to 37 degrees. Although the number of receptors labeled at each temperature was identical, the affinity of the receptor for [3H]spiroperidol decreased 10-fold as the incubation temperature was lowered from 37 degrees to 1 degree. The binding of [3H]spiroperidol was entropy-driven (delta S degree = +80 cal/mole-deg), endothermic (delta H degree = +10 kcal/mole), and exergonic (delta G degree = -13 kcal/mole). Qualitatively similar results were found for (+)-butaclamol, another dopamine receptor antagonist. The binding of the agonists dopamine and (+/-)-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene to sites labeled by [3H]spiroperidol in the striatum also appeared to be entropy-driven (delta S degree = +35 cal/mole-deg). In contrast to the results obtained in studies with antagonists, however, the affinity of the receptor for agonists was independent of the incubation temperature between 8 degrees and 37 degrees. Competition curves for the inhibition of [3H]spiroperidol binding by agonists became increasingly complex as the incubation temperature was lowered. The addition of GTP reduced the affinity of the receptor for agonists at all temperatures but did not simplify interpretation of these complex curves. At 1 degree there was a decrease in the affinity of the receptor for dopamine, and the effect of GTP was abolished.

Animals↗
PubMed1980

Effects of ethanol administration and withdrawal on neurotransmitter receptor systems in C57 mice.

C57BL/6 mice were treated with 7% (v/v) ethanol in a Bio-Serve liquid diet for 7 days. Some animals were then allowed to withdraw from ethanol for a period of 24 hr. The severity of the ethanol withdrawal was assessed by monitoring behavioral changes and by quantitating the decrease in body temperature that occurred during the first 16 hr of withdrawal. Animals withdrawn from ethanol for 24 hr showed a decreased hypothermic response to apomorphine suggesting that changes in dopaminergic systems had occurred. This possibility was further examined in homogenates of striatum by measuring dopamine-stimulated adenylate cyclase activity and the binding of [3H]spiroperidol. However, there were no changes observed in either basal- or dopamine-stimulated adenylate cyclase activity or in the density or affinity or receptors for [3H]spiroperidol. The affinity of apomorphine for the dopamine receptor was also unchanged. In other experiments, alpha and beta adrenergic receptor-mediated increases in cyclic AMP accumulation were assessed in slices of cerebral cortex. There was no change in cyclic AMP accumulation due to either alpha or beta adrenergic receptors. There was, however, a significant decrease in the density of beta adrenergic receptors in both the ethanol-treated mice and in the withdrawn mice. This decrease was restricted to the beta-2 receptor subtype with no change being observed in the density of beta-1 adrenergic receptors. Ethanol administration was also associated with a significant increase in the density of muscarinic cholinergic receptors in the hippocampus and cerebral cortex. The effect was not observed in animals allowed to withdraw for 24 hr.

Alcoholism↗
PubMed1977

Is 2-dimethylaminoethanol (deanol) indeed a precursor of brain acetylcholine? A gas chromatographic evaluation.

Acute administration of deanol-p-acetamidobenzoate (Deaner; deanol) has been reported to elevate brain choline (CH) and acetylcholine (ACh) levels. We have developed a specific and sensitive gas chromatographic assay to measure deanol levels in tissue and have applied this assay to our studies of the effect of acute deanol administration on deanol, ACh and Ch levels in rodent brains. Details of the method are described in this text. This procedure is quantitative and yields reproducible results over a wide range of deanol concentrations (0.30-200 nmol). Seven endogenous and pharmacological parameters have been studied using this procedure. In control rodent brain, liver, heart, lung and plasma, we detected no free endogenous deanol (less than 1 nmol/g). After deanol administration, we were able to detect deanol in tissue and have attempted to determine a relationship between these levels and values of ACh in the same tissue. Regardless of deanol pretreatment time (1-30 minutes) or doses (33.3-3000 mg/kg i.p.) used, we detected no increase in mouse whole brain ACh levels. Likewise, there was no detectable elevation in ACh levels in rat whole brain, cortex, striatum or hippocampus after a 15-minute pretreatment with 550 mg/kg of deanol (i.p.). The only elevation in ACh levels which we detected occurred selectively in the striatum of mice pretreated with a massive dose (900 mg/kg i.p.) of deanol for 30 minutes. This selective increase in striatal ACh levels oculd not, however, be related to levels of deanol in the striatum because there was no greater accumulation of deanol in the striatum than in other brain areas tested or in whole brain. These data do not confirm the results of other investigators who reported elevations in whole brain or striatal ACh levels after acute administration of lower doses of deanol. The data emphasize the need for further investigation into the mode of action of deanol and question its suggested role as an immediate precursor of ACh synthesis in the central nervous system.

Acetylcholine↗
PubMedDate not supplied

Quantitative autoradiographic analysis of 125I-pindolol binding in Fischer 344 rat brain: changes in beta-adrenergic receptor density with aging.

Age-related changes in beta-adrenergic receptor density in Fischer 344 rat brain were examined using in vitro 125I-pindolol (IPIN) binding and quantitative autoradiographic analysis. Localized protein concentrations were determined using a new quantitative histological technique, and these were used to normalize the densities of receptors. Saturation binding studies in brain sections revealed 40-50% decreases in beta-adrenergic receptor density in the thalamus of 23-25-month-old and the cerebellum and brainstem of both 18-19-month-old and 23-25-month-old compared to 4-6-month-old rats. The loss of cerebellar beta-adrenergic receptors may be correlated with reports of deficits in sensitivity to beta-adrenergic-mediated transmission in the cerebellum of aged rats. No changes in specific IPIN binding with age were observed in rat cortex or hippocampus. In all areas examined no age-related differences were observed in receptor affinity. No changes in protein concentration were found in any of the areas examined in the different aged animals. These results demonstrate a region-specific loss of beta-adrenergic receptors with age in the brain of Fischer 344 rats.

Aging↗
PubMedDate not supplied

Long-term treatment of male F344 rats with deprenyl: assessment of effects on longevity, behavior, and brain function.

L-Deprenyl (selegiline) was chronically administered to male Fischer 344 rats via their drinking water beginning at 54 weeks of age (estimated daily dose: 0.5 mg/kg/day). Beginning at 84 weeks of age, the rats were behaviorally evaluated using a sensorimotor battery, a motor-learning task, and the Morris water maze. At 118 weeks of age, cerebellar noradrenergic function was evaluated in the surviving rats using in vivo electrochemistry. The rats were then sacrificed to measure brain monoamine oxidase activity and perform quantitative autoradiography to evaluate the effect of chronic deprenyl treatment on beta-adrenergic receptors in the cerebellum, alpha 2-adrenergic receptors several brain regions, and D1 and D2 dopamine receptors in the striatum. Deprenyl treatment reduced brain monoamine oxidase B activity by 85%, but had no effect on brain monoamine oxidase A. A clear effect of chronic deprenyl treatment upon longevity was not observed. Several measures of CNS function were altered in the deprenyl-treated animals: 1) spatial learning in the Morris water maze was improved; 2) electrochemical signals recorded following local application of NE were reduced, and the responsiveness to the reuptake blocker nomifensine was enhanced, in the cerebellum; 3) beta-adrenergic receptor binding affinity was increased in the cerebellum; 4) alpha 2-adrenergic receptor density was increased in the inferior colliculus; and 5) striatal D1 dopamine receptor density was reduced but binding affinity was enhanced. In contrast, chronic deprenyl treatment did not cause changes in: 1) sensorimotor function, as evaluated by balance beam, inclined screen, or wire hang tasks; 2) motor learning; 3) alpha 2-adrenergic receptor density in any region examined except for the inferior colliculus, or binding affinity in any region examined; or 4) striatal D2 dopamine receptor number or affinity. Thus, long-term oral administration of deprenyl extended the functional life span of rats with respect to cognitive, but not motor, performance.

Animals↗