GABAB binding sites in early adult and aging rat brain.

The effect of aging on GABAB binding was investigated in rat brain. Receptor autoradiography was used to investigate both GABAB and GABAA binding at 2 months, 3 months, 13 months, and 23 months. GABAB binding decreases significantly between 2 months and 23 months of age, as does GABAA binding, with was investigated in rat brain. Receptor autoradiography was used to investigate both GABAB and GABAA binding at 2 months, 3 months, 13 months, and 23 months. GABAB binding decreases significantly between 2 months and 23 months of age, as does GABAA binding, with the greatest decrease between 2 and 3 months. The decrease in GABAB binding appears to be due to a decrease in binding site affinity rather than a decrease in receptor density. The noncompetitive GABAB antagonist zinc, the competitive GABAB antagonist CGP 35348, and the guanyl nucleotide analogue GTP-gamma-S all inhibit GABAB binding identically in 2 month and 23 month brain. These data indicate subtle age-related changes in the GABAB binding in early adult life but little change with senescence.

Postnatal ontogeny of GABAB binding in rat brain.

The postnatal development of GABAB binding sites in rat brain was studied by quantitative receptor autoradiography using [3H]GABA under selective conditions. Binding levels peak at regionally specific times during the first three weeks of life and then decline to adult levels. GABAB binding peaked in the globus pallidus, vestibular and spinal trigeminal nuclei, and the CA3 region of the hippocampus at postnatal day 3; in the striatum, nucleus accumbens, inferior olive, septum, dentate gyrus and CA1 region of the hippocampus at postnatal day 7; in the neocortex and thalamus at postnatal day 14; and in the medial geniculate at postnatal day 21. Following these regionally specific peaks, binding decreased to postnatal day 28 levels. Further significant decreases in binding were observed in all regions examined between postnatal day 28 and adulthood. Comparisons of binding site pharmacology reveal equipotent displacement of GABAB binding by several competitive agonists and antagonists in postnatal day 7 and adult rat brain, indicating that immature and adult binding sites have similar pharmacological properties with regard to these compounds. The GABAB receptor antagonist CGP 54626A, however, inhibited binding more potently in the postnatal day 7 thalamus and neocortex than in these areas in the adult brain. The guanyl nucleotide analogue guanosine 5'-O-(3-thiotriphasphate) inhibited GABAB binding extensively in both postnatal day 7 and adult brain. The non-competitive antagonist zinc also inhibited GABAB binding at both ages and was more potent in postnatal day 7 brain than in adult brain. Saturation analyses reveal two binding sites with similar affinities in both immature and adult rat brain, indicating that postnatal modulation of GABAB binding reflects changes in binding site density rather than modulation of binding site affinity. While immature GABAB binding sites share most pharmacological characteristics with adult binding sites and appear to be coupled to G-proteins at an early age, their interactions with zinc and CGP 54626A suggest that GABAB binding sites in immature brain may have a distinct pharmacological profile. Our data suggest significant regional and pharmacological changes in GABAB binding during development. The implications of these findings are discussed with regards to a possible role of GABAB receptors in the development of the central nervous system.

The delayed effects of phencyclidine enhance amphetamine-induced behavior and striatal C-Fos expression in the rat.

The ability for the delayed effects of phencyclidine to model schizophrenia-like symptomatology was investigated by assessing the effects of phencyclidine pretreatment on amphetamine-induced behavior. Corresponding changes in striatal, nucleus accumbens and anterior cingulate cortex c-Fos induction were also assessed in order to test the hypothesis that alterations in the neurochemistry of these regions accompany phencyclidine-induced changes in amphetamine-induced behaviors. Rats were treated with 15.0 mg/kg phencyclidine or vehicle 24 h prior to behavioral testing following vehicle, 0.5, 2.5 or 5.0 mg/kg amphetamine. Phencyclidine pretreatment significantly increased amphetamine-induced locomotion and rearing in response to 0.5 mg/kg amphetamine. Likewise, phencyclidine pretreatment produced an increase in the number of striatal cells expressing c-Fos following treatment with 0.5 mg/kg amphetamine. Phencyclidine pretreatment did not alter c-Fos induction in the nucleus accumbens, but did decrease the basal number of c-Fos-containing cells in the anterior cingulate cortex. While stereotypy rating revealed that phencyclidine pretreatment enhanced the behavioral response to 5.0 mg/kg amphetamine over time, no other alterations in behavior or c-Fos expression in response to the higher doses of amphetamine were induced by phencyclidine pretreatment. These data demonstrate that the delayed effects of a single dose of phencyclidine alter anterior cingulate cortex neurochemistry, and enhance the behavioral and striatal c-Fos response to a low dose of amphetamine. These findings suggest that the delayed effects of a single dose of phencyclidine may produce a reasonable animal model for schizophrenia.

Pharmacology, distribution, cellular localization, and development of GABAB binding in rodent cerebellum.

Quantitative receptor autoradiography using [3H]GABA under selective conditions was used to characterize the pharmacology, distribution, cellular localization, and development of GABAB binding sites in rodent cerebellum. Pharmacologic analysis of [3H]GABA binding showed that drugs active at GABAB receptors displaced [3H]GABA with the following order of potency: 3-aminopropylphosphonous acid > CGP 35348 = 2-hydroxysaclofen > phaclofen. GTP-gamma-S and GDP-beta-S also diminished potently [3H]GABA binding in a dose-dependent manner. The pattern of [3H]GABA binding to GABAB binding sites was systematically mapped throughout the rat cerebellum. GABAB binding was greatest in the molecular layer and a pattern of parasagittal zonation was observed in the molecular layer of lobules VII-X in adult rats. The cellular localization of GABAB binding was investigated using lesion techniques. Neither methyl azoxymethanol lesions of cerebellar granule cells nor 3-acetylpyridine lesions of climbing fibers resulted in a decrease in [3H]GABA binding. Homozygote stumbler mutant mice, deficient in Purkinje cell dendrites, had a significant decrease in [3H]GABA binding in the molecular layer. These results suggest that the majority of cerebellar molecular layer GABAB binding sites detected by [3H]GABA autoradiography are located on Purkinje cell dendrites. Examination of [3H]GABA binding to GABAB binding sites during development revealed that binding in the molecular layer peaks between postnatal day 14 and postnatal day 28 and then decreases to adult levels. Transient expression of high levels of GABAB binding was observed in the deep cerebellar nuclei, peaking at postnatal day 3 and decreasing to adult levels by postnatal day 21. Our investigation of GABAB pharmacology yielded data in agreement with previously reported results. We have described a parasagittal pattern of GABAB binding in the cerebellar molecular layer and assigned the majority of cerebellar GABAB binding sites to Purkinje cell dendrites. Finally, development studies reveal transient peaks in GABAB binding in the cerebellar molecular layer and deep cerebellar nuclei.

Apomorphine priming alters the response of striatal outflow pathways to D2 agonist stimulation in 6-hydroxydopamine-lesioned rats.

Chronic treatment with dopaminergic agonists is associated with response fluctuations to L-dihydroxyphenylalanine in Parkinson's disease and enhanced motor activity to D1 and D2 dopamine agonists in rats with 6-hydroxydopamine lesions of the nigrostriatal pathway. In dopamine-depleted rodents this phenomenon has been referred to as "priming" or reverse tolerance. The neurochemical changes that underlie "priming" of dopaminergic agonist responses are poorly understood. Some aspects of priming of D1 agonist-mediated rotation in the 6-hydroxydopamine-lesioned rat have been characterized, but priming of D2-agonist-dependent motor responses has been less thoroughly studied. In this study, examination of rotational behaviour and induction of Fos-like immunoreactivity were used to investigate changes in the striatal outflow systems in response to treatment with the D2 agonist quinpirole in 6-hydroxydopamine-lesioned rats that had been primed with apomorphine. Administration of apomorphine (0.5 mg/kg; three injections at three to six day intervals) permitted an otherwise inactive dose of quinpirole (0.25 mg/kg) to produce robust contralateral rotation and to induce the expression of Fos in striatal neurons belonging to the striato-nigro-entopeduncular ("direct") pathway. The increase in contralateral rotation and ipsilateral striatal Fos expression following administration of quinpirole to apomorphine-primed rats was mediated by a D2-like receptor and did not appear to be due to a change in sensitivity of D2 receptors. Apomorphine priming also enhanced the ability of quinpirole to induce Fos expression in the globus pallidus, a target of the striatopallidal ("indirect") pathway. Western blot analysis confirmed that treatment with quinpirole induced the expression of c-Fos protein with no change in the expression of 35-37,000 mol. wt Fos-related antigens in apomorphine-primed rats treated with water or quinpirole. Induction of Fos expression in the striatum generally results from blockade of D2 receptors and the striato-nigro-entopeduncular pathway preferentially expresses D1 receptors. Thus, the quinpirole-dependent induction of striatal Fos in apomorphine-primed 6-hydroxydopamine-lesioned rats represents a qualitative alteration in striatal outflow. These studies demonstrate that pretreatment of 6-hydroxydopamine-lesioned rats with apomorphine increases the activity of the "direct" and "indirect" striatal outflow pathways in response to D2 receptor stimulation. These changes have the net result of enhancing thalamocortical activity and likely underlie the enhanced contralateral rotation produced by quinpirole in apomorphine-primed rats. Changes in striatal outflow, particularly in the striato-nigro-entopeduncular pathway, may contribute to alterations in D2-dependent motor responses observed after chronic dopaminergic stimulation in the dopamine-depleted striatum.

GABAA receptor binding in the aging rat inferior colliculus.

The inhibitory neurotransmitter GABA has been shown to be critically involved in shaping neuronal responses to simple and complex acoustic stimuli in the inferior colliculus. Studies in the rat and human inferior colliculus have suggested significant changes in functions related to GABA neurotransmission occur in the aged. These changes include significant decreases in GABA content, GABA release, GABA neurons, glutamate decarboxylase enzymatic activity, and GABAB receptor binding. Such changes within the inferior colliculus may affect the ability of elderly listeners to process complex acoustic signals, particularly in the presence of background noise. The present study was designed to examine the regional distribution and effects of aging on GABAA receptor binding sites in the Fischer 344 rat inferior colliculus using in vitro quantitative receptor autoradiography. [3H]GABA binding to GABAA receptors was significantly reduced in the inferior colliculus of young adult (3 months) and aged (18-26 months) rats when compared to 2-month animals. However, no significant changes were observed after 3 months of age. Single concentrations of tritiated GABAA receptor ligands (muscimol, t-butylbicycloorthobenzoate, and flunitrazepam) revealed no significant age-related changes in receptor binding in the inferior colliculus between 3 and 26 months of age. To characterize further the pharmacology of the GABAA receptor in the inferior colliculus, GABA modulation of the picrotoxin binding site was examined using [3H]t-butylbicycloorthobenzoate. When increasing concentrations of GABA were added to the incubation buffer, a significant decrease in binding was observed in the inferior colliculus of rats in each age group. In aged rats, the dose-response curve was shifted to the left, indicating an increase in the potency of GABA to inhibit [3H]t-butylbicycloorthobenzoate binding. Although no changes in GABAA receptor binding were detected in the inferior colliculus after 3 months of age, a significant alteration in interaction between the GABA and picrotoxin binding sites was observed in the inferior colliculus of aged rats when compared to 3-month-old young adults. This difference appears to reflect an increased sensitivity of the receptor to GABA modulation in aged rats and, thus, may serve as a compensatory mechanism to enhance GABAA receptor function in response to a presynaptic loss of inhibition.

Zinc modulates GABAB binding in rat brain.

The effects of ZnCl2 on [3H]GABA binding to GABAA and GABAA binding sites were investigated using receptor autoradiography. At concentrations exceeding 100 microM, zinc non-competitively inhibited GABAB binding in a dose dependent fashion. GABAA binding was not inhibited significantly by zinc eliminating the possibility of a non-specific effect of zinc. Increased calcium concentrations up to 10 mM enhanced total GABAB binding but did not prevent zinc induced inhibition of GABAB binding, indicating a separate site of action for these cations at the GABAB binding site. In some regions, zinc modulates GABAB binding in a biphasic manner as concentrations of 10-100 microM zinc significantly enhanced GABAB binding in the hippocampus and the molecular layer of the cerebellum but not in the thalamus. These results provide further evidence for a neuromodulatory role for zinc in the central nervous system.

The effects of phencyclidine pretreatment on amphetamine-induced behavior and c-Fos expression in the rat.

Previous data demonstrate that a single injection of phencyclidine enhances amphetamine-induced behaviors 24 h later, suggesting that the delayed effects of a single dose of phencyclidine may produce a schizophrenia-like state in animals. These behavioral changes were accompanied by altered patterns of c-Fos induction, suggesting possible neurochemical correlates to the observed behaviors. Because investigations into PCP's ability to model schizophrenia have found that the effects of repeated, or subchronic, PCP administration differ according to the dose and administration paradigm, this study sought to determine whether single and subchronic PCP exposure produce different effects on amphetamine-induced behaviors and c-Fos induction. No differences were observed between these administration paradigms; both single and subchronic PCP exposure enhanced amphetamine-induced c-Fos in the striatum, decreased c-Fos in the prefrontal cortex, and decreased the number of cage-crossings. However, the observation that PCP pretreatment affected c-Fos induction in the same manner observed previously while having an opposite effect on amphetamine-induced behavior suggests that these behavioral and neurochemical effects are dissociated.

Electrolytic lesions of the medial septum enhance latent inhibition in a conditioned taste aversion paradigm.

The effects of medial septal lesions on latent inhibition (LI) were assessed in a conditioned taste aversion paradigm. Animals were tested in a LI paradigm 2 weeks after receiving medial septal or sham lesions. The LI paradigm involved a pre-exposure phase in which water-deprived rats were allowed access to either water (non-pre-exposed; NPE) or 5% sucrose (pre-exposed; PE), followed by a conditioning phase in which animals were allowed access to sucrose and subsequently injected with lithium chloride, and a test phase in which animals were allowed access to both sucrose and water. LI was assessed by comparing the %-sucrose consumed in PE and NPE groups on the test day. There was a significantly greater LI effect in the lesion group than in the sham group, suggesting that electrolytic lesions to the medial septum can enhance LI in a CTA paradigm.

Effects of selective adenosine A1 and A2a agonists on amphetamine-induced locomotion and c-Fos in striatum and nucleus accumbens.

Low to moderate doses of amphetamine produce locomotion which is dependent on release of dopamine in the anteromedial striatum and nucleus accumbens. The effects of selective adenosine A1 and A2a receptor agonists on locomotion and c-Fos induction following a moderate dose of amphetamine was assessed in rats. Pretreatment with the adenosine A1 receptor agonist N6-cyclohexyladenosine (CHA) or the adenosine A2a receptor agonist 2-[(2-aminoethylamino)carbonylethylphenylethylamino]-5'-N- ethylcarboxamidoadenosine (APEC) inhibited locomotion following an injection of amphetamine (1.5 mg/kg). This dose of amphetamine induced Fos-like immunoreactivity in an antero-dorsomedial distribution in the caudate-putamen and uniformly in the core and shell of the nucleus accumbens. Pretreatment with the adenosine A2a receptor agonist APEC, but not the adenosine A1 receptor agonist CHA, attenuated c-Fos induction in caudate-putamen and nucleus accumbens by amphetamine. These findings indicate that amphetamine-induced behavior is subject to modulation by adenosine receptors through mechanisms which are both related to and independent of c-Fos induction.

Enhanced CREB phosphorylation and changes in c-Fos and FRA expression in striatum accompany amphetamine sensitization.

Expression in striatum of c-Fos, a 35 kDa Fos-related antigen (FRA) and the phosphorylated form of cyclic AMP response element binding protein (phosphoCREB) was assessed using Western blots in rats that developed behavioral sensitization following repeated amphetamine administration. Treatment with d-amphetamine (5 mg/kg) for 5 consecutive days produced behavioral sensitization. Similar to previous observations using chronic cocaine administration, amphetamine sensitized animals had decreased c-Fos and increased FRA proteins in striatum. Supershift analysis with antisera to c-Fos and FRA proteins demonstrated that 4-Fos and the 35 kDa FRA are components of the striatal AP-1 binding complex from sensitized rats. Thus, amphetamine sensitization is accompanied by alterations in the composition of the AP-1 DNA binding complex. An increased amount of phosphoCREB protein was also present in the striatum of amphetamine sensitized rats. These results suggest that alterations in Fos, FRA and CREB transcription factors are common neuronal responses to chronic psychostimulant administration and may contribute to regulation of genes important to the neuroplastic changes underlying psychostimulant sensitization.

Behavioral sensitization to amphetamine is not accompanied by a decrease in the number of c-Fos containing cells in the striatum.

The expression of c-Fos-like immunoreactivity (FLI) and chronic Fos-related antigen-like immunoreactivity (FRALI) accompanying behavioral sensitization to amphetamine was assessed in male rat striatum. Animals were treated for four days with amphetamine (A; 5 mg/kg) or vehicle (V) and challenged with A or V on the fifth day. The number of FLI-positive cells in the striatum was enhanced in V-A and A-A groups as compared to control (V-V), while the number of FRALI-positive cells in the striatum was enhanced in the A-V and A-A groups as compared to control. These results suggest that the absence of a decrease in the number of striatal FLI-positive cells accompanying chronic amphetamine treatment is not due to antibody cross-reactivity with chronic FRAs, and that behavioral sensitization to amphetamine is not accompanied by a change in the number of striatal cells expressing c-Fos.

Phencyclidine (PCP) reduces the intensity of caffeine-induced convulsions in rats.

The effects of phencyclidine (PCP) on the threshold and intensity of caffeine-induced convulsions in rats were examined. There was a dose-dependent effect of PCP on convulsion intensity with significant reduction in intensity at 4.0 and 8.0 mg/kg PCP. At 16.0 mg/kg PCP, convulsant intensity was reduced in 50% of subjects but potentiated to the point of death in the remaining 50%. PCP had no significant effect on threshold for caffeine-induced convulsions. These results suggest that PCP antagonizes caffeine-induced convulsions and further suggests that the mechanisms involved in onset of caffeine-induced convulsions and the decrease of convulsion intensity are pharmacologically dissociable.

The delayed effects of phencyclidine (PCP) disrupt latent inhibition in a conditioned taste aversion paradigm.

The acute effects of a low dose of phencyclidine (PCP) and the delayed effects of a high dose of PCP on latent inhibition (LI) were assessed in a series of experiments using conditioned taste aversion paradigms. Each paradigm involved a preexposure phase in which water-deprived male rats were allowed access to either water (nonpreexposed; NPE) or 5% sucrose (preexposed; PE), followed by a conditioning phase in which animals were allowed access to sucrose and subsequently injected with the negative reinforcer lithium chloride, and a test phase in which animals were allowed access to both sucrose and water. LI was assessed by comparing the %-sucrose consumed in PE and NPE groups on the test day. The effects of low-dose PCP (2.5 mg/kg) were assessed by comparing LI in animals treated with vehicle or PCP 15 min prior to the onset of the preexposure and conditioning phases. A 4-day paradigm involved 2 days of preexposure followed by a day of conditioning and a test day. This paradigm produced comparable levels of LI in vehicle and PCP-treated animals. A 5-day extinction paradigm involved 2 days of preexposure followed by 2 days of conditioning and a test day. This paradigm abolished LI in vehicle and PCP-treated animals. A 3-day paradigm involved 1 day of preexposure followed by a day of conditioning and a test day. One day of preexposure induced a modified LI effect in both in vehicle and PCP-treated animals. The delayed effects of high dose PCP (8.6 mg/kg) were assessed by comparing LI in animals treated with vehicle or PCP 20 h prior to the onset of the preexposure and conditioning phases in the 4-day paradigm. PCP disrupted latent inhibition in this paradigm. The results are discussed in the context of their relevance to the ability for PCP to model schizophrenic symptomatology.

The delayed effects of DTG and MK-801 on latent inhibition in a conditioned taste-aversion paradigm.

The delayed effects of phencyclidine (PCP) have been shown to disrupt latent inhibition (LI) in a conditioned taste-aversion paradigm. In an attempt to understand the mechanism of this disruption, the delayed effects of the selective sigma receptor agonist 1,3-Di(2-tolyl)guanidine (DTG) and the selective NMDA receptor antagonist MK-801 on latent inhibition were assessed in the same paradigm. Water-deprived male rats were allowed access to either water (nonpreexposed; NPE) or 5% sucrose (preexposed; PE) for 30 min on 2 consecutive days. On the third day, animals were allowed access to sucrose and subsequently injected with lithium chloride. On the forth day, animals were allowed access to both sucrose and water. LI was assessed by comparing the percent sucrose consumed in PE and NPE groups on the fourth day. DTG (1.0, 5.0, or 10.0 mg/kg), MK-801 (0.5, 1.0, or 2.0 mg/kg), or vehicle was administered IP 20 h before preexposure (days 1 and 2) and conditioning (day 3). In vehicle-treated groups, PE animals consumed a significantly higher percent sucrose on the test day than NPE animals, indicating the presence of LI. DTG (10.0 mg/kg) and MK-801 (2.0 mg/kg) decreased the percent sucrose consumed by animals in the PE group to the level observed in the NPE group, indicating disrupted LI. However, this dose of MK-801 was found to produce a decrease in percent sucrose consumed in PE animals not treated with lithium chloride, indicating that the decrease observed in the LI paradigm could be due to MK-801-induced decrease in taste preference for sucrose rather than a disruption of LI. Lower doses of MK-801 that did not produce a decrease in taste preference for sucrose did not significantly disrupt LI. None of the doses of DTG tested altered taste preference for sucrose. These data suggest a role for sigma receptors in the previously observed PCP-induced disruption of LI. Published by Elsevier Science Inc., 2000