Marta Kowalska

Preliminary comparison of behavioral and biochemical effects of chronic transcranial magnetic stimulation and electroconvulsive shock in the rat

To confirm the assumption that repetitive rapid-rate transcranial magnetic stimulation (TMS) induces the functional and structural changes analogous to those which are evoked during electroconvulsive shock (ECS), we compared now the effects of treatments with TMS and ECS on the behavioral responses in rats. We also tested the reactivity of the cyclic adenosine monophosphate (AMP) generating system in cerebral cortical slices. TMS similarly to ECS shortened the immobility time in the forced swimming test and produced a depression of responsiveness of the noradrenaline-stimulated cyclic AMP generating system, although the significance of the latter effect was borderline. In contrast to ECT, TMS produced no such immediate behavioral effects as analgesia and depression of the early phase of locomotor activity. The data suggest that TMS produces in rats some responses that are regarded as predictive for antidepressant activity, similar to those produced by ECS, but less adverse effects.

Formalin hindpaw injection induces changes in the [3H]prazosin binding to α1-adrenoceptors in specific regions of the mouse brain and spinal cord

Summary.Involvement of the α1-adrenoceptor subtypes in early and late phases of formalin pain was investigated by quantitative in vitro autoradiography in the spinal cord and brain structures of CD-1 mice. Total α1-adrenoceptors binding (including all α1-adrenoceptor subtypes) was assessed with [3H]prazosin; α1B-adrenoceptor was assessed with [3H]prazosin in the presence of 10 nM WB4101 to mask remaining α1-adrenoceptor subtypes. Early after formalin injection the α1-adrenoceptors (mainly α1B receptor) binding was reduced in the contralateral hind limb area of the somatosensory cortex and in the secondary motor cortex. A reduction occurred also in the ipsilateral laminae I–III of the spinal cord (both α1B- and non-α1B-adrenoceptors). Lately an increase of α1-adrenoceptors binding (mostly subtypes other than α1B) appeared in discrete amygdaloid and thalamic nuclei. These results provide the first description of changes at the level of central α1-adrenoceptors’ binding during the formalin-induced pain in mice. Their distribution suggests that they may have a functional meaning.

Effects of the noradrenergic neurotoxin DSP-4 on the expression of α1-adrenoceptor subtypes after antidepressant treatment

We have previously reported that chronic imipramine and electroconvulsive treatments increase the α(1A)-adrenoceptor (but not the α(1B) subtype) mRNA level and the receptor density in the rat cerebral cortex. Furthermore, we have also shown that chronic treatment with citalopram does not affect the expression of either the α(1A)- or the α(1B)-adrenoceptor, indicating that the previously observed up-regulation of α(1A)-adrenoceptor may depend on the noradrenergic component of the pharmacological mechanism of action of these antidepressants. Here, we report that previous noradrenergic depletion with DSP-4 (50 mg/kg) (a neurotoxin selective for the noradrenergic nerve terminals) significantly attenuated the increase of α(1A)-adrenoceptor mRNA induced by a 14-day treatment with imipramine (IMI, 20 mg/kg, ip) and abolished the effect of electroconvulsive shock (ECS, 150 mA, 0.5 s) in the prefrontal cortex of the rat brain. The changes in the receptor protein expression (as reflected by its density) that were induced by IMI and ECS treatments were differently modulated by DSP-4 lesioning, and only the ECS-induced increase in α(1A)-adrenoceptor level was abolished. This study provides further evidence corroborating our initial hypothesis that the noradrenergic component of the action of antidepressant agents plays an essential role in the modulation of α(1A)-adrenoceptor in the rat cerebral cortex.

Does Ca2+ channel blockade modulate the antidepressant-induced changes in mechanisms of adrenergic transduction?

SummaryWe investigated how the L-type calcium channel blockade (CCB) with nifedipine affects the cyclic AMP responses to noradrenaline or isoproterenol in cerebral cortical slices from rats receiving antidepressant treatments that induce (electroconvulsive shock, imipramine) or do not induce (amitriptyline) β-downregulation. To assess the role of protein kinase C (PKC) in receptor crosstalk under CCB conditions, the cyclic AMP responses were tested also in the presence of a PKC activator, TPA. CCB alone induced no changes, but modulated the action of those antidepressants that down regulate the β-adrenergic system. Chronic ECS and imipramine treatments were differently affected. ECS, under conditions of CCB, down regulated the response to isoproterenol in the presence of TPA, while imipramine ceased to block the TPA-potentiation of cyclic AMP responses. Thus, CCB affects the processes related to the antidepressant-induced changes on the crosstalk between α1- and β-adrenergic receptors, depending on the specific properties of the antidepressant.

Disruption of glucocorticoid receptors in the noradrenergic system leads to BDNF up-regulation and altered serotonergic transmission associated with a depressive-like phenotype in female GRDBHCre mice

Recently, we have demonstrated that conditional inactivation of glucocorticoid receptors (GRs) in the noradrenergic system, may evoke depressive-like behavior in female but not male mutant mice (GR(DBHCre) mice). The aim of the current study was to dissect how selective ablation of glucocorticoid signaling in the noradrenergic system influences the previously reported depressive-like phenotype and whether it might be linked to neurotrophic alterations or secondary changes in the serotonergic system. We demonstrated that selective depletion of GRs enhances brain derived neurotrophic factor (BDNF) expression in female but not male GR(DBHCre) mice on both the mRNA and protein levels. The possible impact of the mutation on brain noradrenergic and serotonergic systems was addressed by investigating the tissue neurotransmitter levels under basal conditions and after acute restraint stress. The findings indicated a stress-provoked differential response in tissue noradrenaline content in the GR(DBHCre) female but not male mutant mice. An analogous gender-specific effect was identified in the diminished content of 5-hydroxyindoleacetic acid, the main metabolite of serotonin, in the prefrontal cortex, which suggests down-regulation of this monoamine system in female GR(DBHCre) mice. The lack of GR also resulted in an up-regulation of alpha2-adrenergic receptor (α2-AR) density in the female but not male mutants in the locus coeruleus. We have also confirmed the utility of the investigated model in pharmacological studies, which demonstrates that the depressive-like phenotype of GR(DBHCre) female mice can be reversed by antidepressant treatment with desipramine or fluoxetine, with the latter drug evoking more pronounced effects. Overall, our study validates the use of female GR(DBHCre) mice as an interesting and novel genetic tool for the investigation of the cross-connected mechanisms of depression that is not only based on behavioral phenotypes.

Depressive-like immobility behavior and genotype × stress interactions in male mice of selected strains

Abstract In this study, we investigated whether basal immobility time of C57BL/6J mice, which are commonly used in transgenesis, interferes with detection of depressive-like behavior in the tail suspension test (TST) after chronic restraint stress (CRS). We included in the study mice of the C57BL/6N strain, not previously compared with C57BL/6J for behavior in the TST, and contrasted both strains with NMRI mice which exhibit low basal immobility. NMRI, C57BL/6J, and C57BL/6N male mice (n = 20 per strain) were tested under basal conditions and after CRS (2 h daily for 14 d). NMRI and C57BL/6J mice were differentiated in the TST by low and high basal immobility times, respectively, while the C57BL/6N and NMRI mice showed similar levels of basal immobility. CRSextended the immobility time of NMRI mice in the TST, whereas both C57BL/6J and C57BL/6N mice were unaffected regardless of their initial phenotype. We explored whether detailed analysis of activity microstructure revealed effects of CRS in the TST, which are not apparent in the overall comparison of total immobility time. Interestingly, unlike C57BL/6J and/6N strains which showed no sensitivity to CRS, stressed NRMI mice displayed distinct activity microstructure. In contrast to behavioral differences, all stressed mice showed significant retardation in body weight gain, decreased thymus weight and increased adrenal cortex size. However, after CRS, enlargement of the adrenal medulla was observed in both C57BL/6J and C57BL/6N mice, suggesting similar sympatho-medullary activation and stress coping mechanism in these substrains.

Paroxetine pretreatment does not change the effects induced in the rat cortical β-adrenergic receptor system by repetitive transcranial magnetic stimulation and electroconvulsive shock

Repetitive transcranial magnetic stimulation (rTMS) has been proposed as a clinically effective antidepressant treatment, but meta-analysis suggests that its efficacy is marginal. We investigated whether the administration of rTMS together with paroxetine would enhance its effects on the beta-adrenergic system of the rat. We compared our results with the effects of electroconvulsive shock therapy (ECS). The experiment was performed for 12 d on male Wistar rats that received a physical treatment of either rTMS (B=1.4 T, f=10 Hz, 300 s) or ECS (I=130 mA, f=50 Hz, t=500 ms), preceded by sterile water or paroxetine (10 mg/kg i.p. 30 min earlier). All rats were decapitated 24 h after the final treatment. Cyclic AMP (cAMP) was measured in cortical slices prelabelled with [3H]adenine and stimulated with noradrenaline. beta-adrenoceptor parameters (Bmax and KD) were assessed in the P2 fraction of cortical homogenates using [3H]CGP 12177 as a ligand. ECS resulted in down-regulation of both the cAMP response and beta-adrenoceptor density, while rTMS depressed only the responsiveness of the cAMP-generating system. Paroxetine, which was only effective in dampening the cAMP response, did not change the effects of either physical treatment. The data suggest that any possible interaction between paroxetine and rTMS or ECS does not involve the beta-adrenergic mechanisms.

A lack of α1A-adrenergic receptor-mediated antidepressant-like effects of S-(+)-niguldipine and B8805-033 in the forced swim test.

The &agr;1-adrenergic receptors (&agr;1-ARs), which belong to a G protein-coupled receptor family, consist of three highly homologous subtypes known as &agr;1A-ARs, &agr;1B-ARs, and &agr;1D-ARs. Our previous findings suggested that &agr;1A-ARs are an important target for imipramine and electroconvulsive therapy. The current study sought to evaluate whether S-(+)-niguldipine and B8805-033, two selective antagonists of &agr;1A-ARs, can evoke antidepressant-like effects in the forced swim test in rats. Both compounds were administered at three time points (24, 5, and 1 h before testing), and the effects of three doses (2, 5, and 10 mg/kg) of each compound were investigated. S-(+)-Niguldipine produced no antidepressant-like effects other than a 14% reduction in immobility time at the highest dose. Although B8805-033 at a dose of 2 mg/kg did not influence the rats’ behavior, higher B8805-033 doses (5 and 10 mg/kg) produced significant reductions in immobility time (approximately 42 and 44% vs. controls, respectively; P<0.01). However, this effect was abolished by the concomitant administration of WAY100135, a serotonin receptor antagonist, suggesting that the observed antidepressant-like effects of B8805-033 are unrelated to &agr;1A-ARs. Nevertheless, given the current dearth of selective &agr;1A-AR agonists, the question of whether this particular subtype could be involved in antidepressant therapy mechanisms remains unresolved.

P.6.d.009 Effects of spontaneous and naloxone-precipitated morphine withdrawal on G(q/11) and G(12) protein level in rat brain

Supported by the statutory funds from the Institute of Pharmacology, Polish Academy of Sciences, Krakow, Poland Acknowledgements Morphine, an agonist of opioid receptors, is a potent analgesic drug with known addictive properties. Guanine nucleotide-binding proteins (G proteins) composed of Gα and G subunits, play critical role in brain signal transduction. G proteins, attached to the cell plasma membrane, that connect receptors to effectors and thus to intracellular signaling pathways. On the basis of sequence similarity, the Gα subunits have been divided into four families: G(s), G(i/o), G(q/11), G(12/13) [Tab. 1]. Opioid receptors are coupled with the G(i/o) family. Recently, we have found that chronic treatment with morphine affects the mRNA level of Gα(q/11) and Gα(12) subunits, which are known to be linked to other than opioid receptors. Morphine induced changes were especially visible after the period of drug withdrawal (Ref.1). The identification of morphine-induced changes in the expression of Gα subunits is of critical importance for understanding of addictive behavior Introduction