Haleh Razani

Galanin Is a Potent In Vivo Modulator of Mesencephalic Serotonergic Neurotransmission

Neurochemical, molecular, immunohistochemical and behavioral methods were used to examine the in vivo effects of the neuropeptide galanin on central 5-HT neurotransmission and on 5-HT1A receptor-mediated responses. Intraventricularly infused galanin caused a long-lasting and dose-dependent reduction of basal extracellular 5-HT levels in the ventral hippocampus of awake rats as measured by microdialysis. Infusion of galanin into the dorsal raphe nucleus (DRN), but not intrahippocampally, reduced 5-HT release. The effect of i.c.v. galanin on 5-HT release was blocked by the galanin receptor antagonist M35, acting most likely via galanin receptors at the level of the DRN. Galanin also reduced the levels of tryptophanhydroxylase mRNA in the DRN. Therefore, the effects of galanin on 5-HT1A receptor-mediated responses were further investigated. Surprisingly, galanin significantly attenuated the reduction of hippocampal 5-HT release induced by systemic injection of the 5-HT1A receptor agonist 8-OH-DPAT. Galanin also attenuated 8-OH-DPAT-induced hypothermia and locomotor activity in rats. These results indicate that galanin has important inhibitory actions on central 5-HT neurotransmission and on 5-HT1A receptor-mediated events.

Galanin Modulates 5‐Hydroxytryptamine Functions: Focus on Galanin and Galanin Fragment/5‐Hydroxytryptamine1A Receptor Interactions in the Braina

Abstract: The reciprocal interactions between galanin and 5‐HT1A receptors in the rat brain are presented. Galanin and its NH2‐terminal fragments antagonize 5‐HT1A receptor‐mediated transmission at the postjunctional level, whereas galanin receptor activation mimics the inhibitory action of 5‐HT1A receptor activation at the soma‐dendritic level, leading to reductions of 5‐HT metabolism and release. These interactions have been shown in both receptor binding studies and functional studies. In view of the present findings, galanin antagonists may represent a new type of antidepressant drug, based on the 5‐HT hypothesis of depression, by enhancing 5‐HT release and postjunctional 5‐HT1A‐mediated transmission. Moreover, following intracerebroventricular injection galanin was found to be internalized in a population of hippocampal nerve cells mainly representing GABA, somatostatin, and/or NPY‐immunoreactive nerve cells. The relevance of these findings is discussed in relation to the concept of volume transmission.

Intraventricular galanin modulates a 5-HT1A receptor-mediated behavioural response in the rat

The present studies have examined whether the neuropeptide galanin can modulate brain serotoninergic (5‐HT) neurotransmission in vivo and, particularly, 5‐HT1A receptor‐mediated transmission. For that purpose, we studied the ability of galanin (given bilaterally into the lateral ventricle, i.c.v.) to modify the impairment of passive avoidance retention induced by the selective 5‐HT1A agonist 8‐hydroxy‐2‐(di‐n‐propyloamino)tetralin (8‐OH‐DPAT) when injected prior to training. This impairment appears to be mainly related to activation of 5‐HT1A receptors in the CNS. Galanin dose‐dependently (significant at 3.0 nmol/rat) attenuated the passive avoidance impairment (examined 24 h after training) induced by the 0.2 mg/kg dose of 8‐OH‐DPAT. This 8‐OH‐DPAT dose produced signs of the 5‐HT syndrome indicating a postsynaptic 5‐HT1A receptor activation. Furthermore, both the impairment of passive avoidance and the 5‐HT syndrome were completely blocked by the 5‐HT1A receptor antagonist WAY 100635 (0.1 mg/kg). Galanin (0.3 or 3.0 nmol) or WAY 100635 (0.1 mg/kg) failed by themselves to affect passive avoidance retention. 8‐OH‐DPAT given at a low dose 0.03 mg/kg, which presumably stimulates somatodendritic 5‐HT1A autoreceptors in vivo, did not alter passive avoidance retention or induce any visually detectable signs of the 5‐HT syndrome. Galanin (0.3 or 3.0 nmol) given i.c.v. in combination with the 0.03 mg/kg dose of 8‐OH‐DPAT, did not modify passive avoidance. The immunohistochemical study of the distribution of i.c.v. administered galanin (10 min after infusion) showed a strong diffuse labelling in the periventricular zone (100–200 μm) of the lateral ventricle. Furthermore, in the dorsal and ventral hippocampus galanin‐immunoreactive nerve cells appeared both in the dentate gyrus and the CA1, CA2 and CA3 layers of the hippocampus. In the septum only endogenous fibres could be seen while in the caudal amygdala also galanin‐immunoreactive nerve cells were visualized far away from the labelled periventricular zone. At the level of the dorsal raphe nucleus a thin periventricular zone of galanin immunoreactivity was seen but no labelling of cells. These results suggest that galanin can modulate postsynaptic 5‐HT1A receptor transmission in vivo in discrete cell populations in forebrain regions such as the dorsal and ventral hippocampus and parts of the amygdala. The indication that galanin administered intracerebroventrically may be taken up in certain populations of nerve terminals in the periventricular zone for retrograde transport suggests that this peptide may also affect intracellular events.

Galanin and learning

A number of studies indicate that galanin (GAL) is a potent modulator of basal acetylcholine release in the rat forebrain e.g. in the cholinergic neurons of the septo-hippocampal projections. Thus, GAL perfused through the microdialysis probe decreased basal acetylcholine release in the ventral hippocampus, while it enhanced acetylcholine release in the dorsal hippocampus. This finding indicates that GAL may act via different mechanisms within the subsystems of the hippocampus. This hypothesis has received support from studies using the Morris swim maze, a learning task dependent on hippocampal mechanisms. GAL (3 nmol/rat) infused into the ventral hippocampus impaired spatial learning acquisition, while it tended to facilitate when injected into the dorsal hippocampus. However, the effects of GAL on acetylcholine release and on spatial learning, which are due to activation of GAL-receptors, appear to be indirectly mediated possibly via noradrenaline transmission. GAL is also a potent inhibitor of mesencephalic 5-HT neurotransmission in vivo. These findings are discussed in relation to the role of acetylcholine and serotonin in cognition.

Intraventricular galanin produces a time-dependent modulation of 5-HT1A receptors in the dorsal raphe of the rat

Galanin given i.c.v. increased the KD values of 5-HT1A receptors in the DR by 100% 10 min after administration. This effect had disappeared 2 h and 5 h after galanin administration. In contrast, i.c.v. galanin caused a 40% increase in the Bmax values at 2 h without affecting the Bmax values 10 min and 5 h after i.c.v. galanin. The mRNA levels of 5-HT1A receptors and of galanin in the DR were significantly decreased by 33% and 56% respectively 2 h but not 5 h after i.c.v. galanin. These results suggest that in vivo galanin can modulate somatodendritic 5-HT1A receptors in the DR in a time-dependent manner via galanin-5-HT1A receptor interactions. The findings implicate a potential role of galanin in affective disorders.

Prolonged effects of intraventricular galanin on a 5-hydroxytryptamine1A receptor mediated function in the rat

Galanin (3 nmol/rat), 2 h after its intracerebroventricular (i.c.v.) administration to male rats, attenuated the passive avoidance (PA) retention deficit induced by the 5-hydroxytryptamine (HT)(1A) receptor agonist 8-hydroxy-2-(di-N-propylamino)tetraline (8-OH-DPAT) (0.2 mg/kg) The reduction in the postjunctional 5-HT(1A) receptor-mediated response after i.c.v. galanin was not associated with changes in the mRNA levels and agonist binding properties of cortical limbic 5-HT(1A) receptors, believed to be the target receptors mediating the PA deficit caused by 8-OH-DPAT. These results suggest that acute increases of galanin transmission in vivo even after 2 h can counteract limbic 5-HT(1A) receptor-mediated responses of relevance for affective disorders without significantly affecting gene expression and binding characteristics of cortical limbic 5-HT(1A) receptors.

The neuropeptide galanin as an in vivo modulator of brain 5-HT1A receptors : Possible relevance for affective disorders

The neuropeptide galanin is widely distributed throughout the central nervous system with multiple and diverse biological functions mediated by different receptor subtypes. In the rat, galanin-like immunoreactivity is expressed in a population of 5-hydroxytryptamine (5-HT, serotonin) neurons in the dorsal raphe with extensive projections to the forebrain areas, e.g., hippocampus. This review summarizes results from experimental studies in rodents showing that in vivo galanin is a potent modulator of brain 5-HT transmission, and in particular 5-HT1A receptor-mediated functions. Galanin, given intracerebroventricular (i.c.v.), was demonstrated to have strong inhibitory interactions with 5-HT1A receptor functions, particularly in the dorsal raphe but also in the hippocampus. Since pre- and postsynaptic 5-HT1A receptors in the dorsal raphe and hippocampus are implicated in the action of antidepressant drugs and in depressive disorders, it is suggested that galanin receptors may be an important target for development of novel antidepressant drugs. This view is supported by a recent study in the rat showing that the galanin antagonist M35, given i.c.v., could block the depression-like behavior in the forced swim test induced by galanin, while M35 produced an antidepressant-like effect on its own.

Estrogen Down-Regulates mRNA Encoding the Exocytotic Protein SNAP-25 in the Rat Pituitary Gland

Exocytosis is dependent on specific proteins that are located at the secretory granule membrane, in the cytoplasm or at the plasma membrane. The mRNA expression of synaptosomal‐associated protein of 25 kDa (SNAP‐25) isoforms SNAP‐25a and SNAP‐25b, vesicle associated membrane protein (VAMP) 2, mammalian homologue of unc‐18 (munc‐18) and Hrs‐2 was studied in the pituitary of ovariectomized rats after subcutaneous insertion of capsules containing estrogen or placebo using in situ hybridization. Estrogen treatment (0.25 mg estradiol) significantly decreased SNAP‐25a (32%; 10%) and SNAP‐25b (25%; 22%) mRNA levels in the anterior and intermediate lobes, respectively, whereas VAMP‐2, munc‐18 and Hrs‐2 mRNA levels remained unchanged. The results suggest that estrogen selectively regulates SNAP‐25 transcription in the pituitary gland, but leaves VAMP‐2, munc‐18 and Hrs‐2 mRNA levels unaffected.

5-HT1A receptor activation : short-term effects on the mRNA expression of the 5-HT1A receptor and galanin in the raphe nuclei

SYSTEMIC adminstration of the 5-HT1A receptor agonist 8-OH-2-(di-n-propylamino)-tetralin (8-OH-DPAT; 0.3 mg/kg, s.c.) was used to explore the effects of activation of 5-HT1A receptors on expression of mRNA coding for 5-HT1A receptor, tryptophan hydroxylase (TPH) and galanin in the ascending raphe nuclei. 8-OH-DPAT increased the hybridization signal of the 5-HT1A receptor by 105% in the dorsal raphe nucleus (B7) 30 min after the injection. No effects were seen at the later time points (2–8 h). In the median raphe nucleus (B8) and the B9 cell group in the medial lemniscus, 8-OH-DPAT induced a marked decrease in labeling 30 min after injection. At 8 h following 8-OH-DPAT injection, the effect had shifted to an increase in 5-HT1A receptor labeling by 68% in the B8 area. Importantly 8-OH-DPAT had no significant effects on the expression of mRNA coding for TPH and galanin. The results suggest an important and differential mechanism for the regulation of 5-HT1A receptor mRNA levels in the dorsal and median raphe nuclei. This regulation may be of importance for the differential control of the activity of the ascending 5-HT neurons, and hence for mood regulation. The results also indicate a dissociation between the effects mediated by 5-HT1A receptor functions and those regulating the coexisting peptide galanin in the dorsal raphe.