Jantiena B. Sebens

Cyclic AMP in the rat cerebral cortex after stimulation of the locus coeruleus: decrease by antidepressant drugs.

The study concerned the effect of repeated treatment with antidepressant drugs on the elevation of cyclic AMP levels in the rat cerebral cortex following electrical stimulation of the locus coeruleus. Some of the tricyclic and tetracyclic antidepressant drugs inhibited the cyclic AMP response. Desmethylimipramine was the most potent (effective when given 5 mg/kg/day for 2 weeks). Imipramine and nomifensine (daily dose 10 mg/kg for 2 weeks) produced slight decreases, while iprindol and clomipramine were ineffective. After 6 weeks of treatment (daily 10 mg/kg) iprindol, clomipramine and mianserin were without effect. The cyclic AMP response was suppressed by higher doses of the latter two drugs (2 weeks, 20 mg/kg/day). These results indicate that tricyclic and tetracyclic antidepressant drugs are able to decrease cerebral noradrenergic neurotransmission of locus coeruleus neurons, as far as this is mediated by cyclic AMP. It is not clear, however, whether such modification is related to the therapeutic action of antidepressant drugs.

DIFFERENTIAL FOS-PROTEIN INDUCTION IN RAT FOREBRAIN REGIONS AFTER ACUTE AND LONG-TERM HALOPERIDOL AND CLOZAPINE TREATMENT

Both acute and long-term effects of haloperidol and clozapine on Fos-like immunoreactive nuclei in several rat forebrain areas were quantified. Rats were treated with saline (1 ml/kg.day, control), haloperidol (1 mg/kg.day) and clozapine (20 mg/kg.day) i.p. for 21 days. Two hours before perfusion fixation a single (acute treatment) or last (long-term treatment) dose of the drug was given. Drug-induced catalepsy and gain in body weight were also measured. A single dose of haloperidol produced large increases in Fos-like immunoreactive nuclei in the striatum, the nucleus accumbens and central amygdala. Following long-term treatment these increases were reduced in all nuclei studied, except the lateral septum. Acute clozapine treatment had slight (if any) effects on the number of Fos-like immunoreactivity-expressing nuclei in the striatum, but the increases in the nucleus accumbens, the lateral septum, the paraventricular and supraoptic nuclei of the hypothalamus and the central amygdala were substantial. Long-term clozapine treatment reduced the acute response significantly in all the areas except the nucleus accumbens. Both haloperidol and clozapine treatment reduced the weight gain of the rats. Haloperidol, but not clozapine, induced catalepsy that remained maximal during the long-term haloperidol treatment. These results indicate that in most brain areas high Fos-protein levels are not necessary to maintain antipsychotic activity or side-effects. The persisting effect of clozapine in the nucleus accumbens may be of significance to the efficacy of this drug in treatment-refractory schizophrenia.

In vivo binding of N-n-propylnorapomorphine in the rat brain: Regional localization, quantification in striatum and lack of correlation with dopamine metabolism

The accumulation and retention of radioactivity in rat brain were studied after intravenous injection of the dopamine (DA) agonist [3H]N-n-propylnorapomorphine ( [3H]NPA). Dose-dependent saturable accumulation of label was found in the striatum, nucleus accumbens and olfactory tubercle. DA agonists (apomorphine, N,N-dipropyl-5,6-ADTN) and antagonists (haloperidol, cis-flupenthixol) prevented this accumulation. Enhanced accumulation of radioactivity in the striatum was found after 6-OHDA lesions and short- and long-term treatment with reserpine. These results are an indication of specific NPA binding to presumably postsynaptically situated DA receptors. One hour after administration of the drug, the effect of NPA on striatal DA metabolism was not correlated with receptor saturation. Maximal numbers of in vivo NPA binding sites (about 30 and 22 pmol . g-1) in striatal tissue were calculated from independent measurements at 15 and 60 min after NPA injection. Regional distribution of radioactivity after a tracer dose of [3H]NPA was assessed in 35 brain areas and parts of the spinal cord. In addition to the already mentioned DA-rich areas receptor-specific NPA binding was also found in several other brain parts.

Cyclic AMP in cerebrospinal fluid: Accumulation following probenecid and biogenic amines

Abstract Levels of cyclic AMP in rabbit cisternal cerebrospinal fluid (CSF) was determined under a number of conditions. Cyclic AMP in CSF was increased by probenecid (intraperitoneally); this increase was not influenced by tricyclic antidepressant drugs, haloperidol, isoprenaline or L-DOPA. Cyclic AMP, administered intravenously, did not penetrate into CSF. Intracisternally introduced noradrenaline, isoprenaline, dopamine, histamine, and intravenously injected isoprenaline all increased the CSF levels of cyclic AMP. The results indicate that cyclic AMP in the CSF is of central origin and is eliminated by a probenecid sensitive transport mechanism. Because of the effects of biogenic amines, there seems to be a close relationship between cyclic AMP of brain tissue and of CSF. Central beta-adrenergic receptors may be measured in the intact animal by measuring the release of the nucleotide into CSF after intravenously isoprenaline. This observation may lead to a clinical applicable test for measuring properties of central adrenergic receptors.

A MOSAIC-LIKE DISTRIBUTION OF DOPAMINE-RECEPTORS IN RAT NEOSTRIATUM AND ITS RELATIONSHIP TO STRIOSOMES

The distribution of dopamine receptors in rat neostriatum was determined by means of light microscopic autoradiography of in vivo labeled binding sites for [3H]N-n-propylnorapomorphine and compared with the distribution of acetylcholinesterase histochemical staining. The distribution of dopamine receptors was non-homogeneous and patches of low receptor density were in register with acetylcholinesterase-poor striosomes. This suggests that the distribution pattern of dopamine receptors is specifically related to various input systems.

Prolonged c-Jun expression in the basolateral amygdala following bulbectomy: possible implications for antidepressant activity and time of onset.

Olfactory bulbectomy is a well established animal model of depression. Neurochemical and behavioral alterations observed following olfactory bulbectomy, are due, in part, to the neurodegeneration of specific brain structures. Amygdaloid dysfunction in particular, is known to play a substantial role in the syndrome of the olfactory bulbectomized rat. The present study examined both short- and long-term alterations in immediate early gene expression, tyrosine hydroxylase and serotonin immunoreactivity, and classical silver staining, following olfactory bulbectomy in the basolateral amygdala. The results indicated no consistent change in Fos expression observed over the experimental period. Following bulbectomy, long term (up to 64 days post-lesion) Jun expression, not coincident with silver staining, was observed in the basolateral nucleus. The basolateral nucleus was also intensely immunoreactive for serotonin at this timepoint post-bulbectomy. Thus, following bulbectomy long term alterations in Jun expression occurs in the serotonin rich basolateral amygdala. As a site of action for antidepressant compounds, alterations at the immediate early gene level in this region may have implications both for the model, and antidepressant drug action therein.

Reduction of adenosine A1-receptors in the perforant pathway terminal zone in Alzheimer hippocampus

The cells of origin of the perforant pathway are destroyed in Alzheimers disease (AD). In rat the adenosine A1-receptors are specifically localized on the perforant path terminals in the molecular layer of the dentate gyrus. In the present study the density of A1-receptors in the hippocampus of Alzheimers disease (AD) patients (n = 9) and non-dement controls (n = 3) has been investigated autoradiographically with [3H]8-cyclopentyl-1,3-dipropylxanthine ([3H]CPDPX) as the ligand probe. In AD hippocampi binding of [3H]CPDPX was greatly reduced in the outer two thirds of the dentate gyrus molecular layer, likely due to the degeneration of the perforant path. Binding of [3H]CPDPX was not significantly altered in other parts of the AD hippocampus, e.g. the CA1 and the CA3, in spite of a pronounced cellular pathology and reduced N-methyl-D-aspartate (NMDA) receptor densities, assessed as strychnine insensitive [3H]glycine autoradiography. This contrasts with the presumed localization on dendrites of pyramidal neurons of A1 receptors within the CA1 and the CA3.

Loss of dopamine receptors in the olfactory bulb of patients with Alzheimer's disease

The presence and localization of dopamine D2 receptors was studied by means of in vitro autoradiography with [3H]N-n-propylnorapomorphine in olfactory bulbs obtained postmortem from Alzheimer patients and age-matched controls. It appeared that, in 5 age-matched controls, the greatest density of dopamine D2 receptors was found in the glomerular layer of the bulb. In 6 of 7 Alzheimer patients, the labeling of the glomerular layer was decreased so that glomerular and granular layer did not differ in labeling density. Tangles, as revealed by Thioflavin S staining, were present in all different layers of the bulbus in Alzheimer patients. Observation of Nissl-stained preparations revealed that mitral cell bodies in bulbs of these patients were not present. Since mitral cells are projection neurons with targets in the entorhinal and piriform cortex, the observation of loss of these cells supports the hypothesis of early involvement of the olfactory system in Alzheimers disease and the spread from the olfactory mucosa and bulb to the cerebral cortex and hippocampus via degeneration of interconnecting neurons. Moreover, in vivo detection of bulbar dopamine receptors might in the future provide a diagnostic tool for the early detection of senile dementia of the Alzheimer type.

Localization of NMDA and AMPA receptors in rat barrel field

The aim of this study was to assess the distribution of N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-S-methyl-4-isoxazole propionic acid (AMPA) receptors in the barrel field of rat primary somatosensory (SI) cortex using light-microscopic in vitro autoradiography. NMDA receptors were labeled with the competitive antagonist [3H]CGP39653 or with [3H]glycine in the presence of strychnine, and AMPA receptors with [3H]AMPA. In the SI cortex high densities of the NMDA receptor occurred in the supragranular layers and in layer Va. In layer IV high NMDA receptor densities were specifically confined to the barrel hollows. The AMPA binding sites showed less intralaminar variation and no apparent density differences between the barrel hollows and sides in layer IV. It can be concluded that the distribution of NMDA (but not AMPA) receptors in the rat barrel field shows a strong coincidence with the zone of termination of the specific sensory afferents from the ventral posterior nucleus of the dorsal thalamus.

Quantification of in vivo spiperone binding in the rat striatum after lesions produced by kainate or decortication

The potential of in vivo spiperone binding as a tool for the detection and quantitative analysis of striatal dopamine (DA) receptor alterations was studied in rat brain lesioned in several ways. Two weeks after kainate (KA) injection a significantly higher radioactivity accumulation was observed in the lesioned striatum than in the contralateral structure after a tracer dose of [3H]spiperone. The difference was maximal 2 days after surgery and it was present for at least 4 weeks while it was reversed 11 weeks after KA injection. The radioactivity uptake (tracer dose of [3H]spiperone) measured 2 weeks after surgery could be specifically prevented in both KA-lesioned and contralateral striatum by haloperidol and N-n-propylnorapomorphine while non-dopaminergic drugs were almost without effect. More than 80% of the radioactivity accumulation was saturable in both contralateral (unlesioned) and KA-lesioned striatum, leaving a slightly higher non-saturable radioactivity level in the latter. One week after unilateral ablation of the cerebral cortex overlying the striatum only minor bilateral differences in striatal radioactivity content were found after a tracer dose of [3H]spiperone. No differences were present after 6-OHDA lesion of the nigrostriatal pathway. Striatal DA receptor densities (Bmax) were determined from the dose-dependency of total striatal spiperone accumulation. This relationship was assessed using cerebellar spiperone accumulation instead of dose. Thus a Bmax of about 75 fmol X mg-1 tissue was found in the striatum of control (unoperated) rats and contralateral to the striatal KA lesion while 2 weeks after surgery it was approximately 33 fmol X mg-1 in the KA-lesioned striatum. One week after unilateral decortication Bmax values of about 50 and 65 fmol X mg-1 were found ipsi- and contralaterally to the lesion respectively.