Neurotensin as Modulator of Glutamatergic Signalling: Relevance in Neurodegenerative Diseases

Rationale: Neurotensin (NT) is a tridecapeptide widely distributed in mammalian brain, where acts as a neurotransmitter or neuromodulator of classical neurotransmitters, mainly through the activation of its receptor NTS1. Several in vitro and in vivo studies have demonstrated the existence of close interactions between NT and dopamine (DA) systems both in limbic and striatal brain regions (Nemeroff CB., 1985; Binder EB., 2001; Caceda R., 2006). Because of the involvement of an over-activation of DA system in the development of neurological disorders such as schizophrenia, psychosis and dyskinesia, a strong attention was given to the study of complex interactions between NTS1 and D2 dopamine receptor, highlighting the existence of receptor-receptor interaction, potential target for developing new anti-schizophrenic drugs (Ferraro L., 2007). In addition, neurochemical and biochemical data indicate that NT plays a crucial role in regulating glutamatergic transmission, probably inducing an amplification of NMDA receptor signalling, even at threshold concentrations (lOnM) (Antonelli T., 2004). Results I: The neuromodulatory function of NT on glutamatergic signalling was studied in an in vitro model of primary cortical cultures, highlighting a dose-dependent effect (NT 0.1-300 nM) on glutamate release. In addition, NT show the ability to amplify the NMDA-induced (lOOnM) increase of glutamate release. The use of the NTS1 receptor antagonists, SR48692 (lOOnM) and the NMDA receptor antagonist MK-801 (l[iM), in combination with an effective concentration of NT, made possible to hypothesize that the mechanism involved could be an NTSl/NMDA receptor-receptor interaction (Antonelli T., 2004; Ferraro L., 2008) both at striatal and cortical level. It has been postulated that the accumulation of extracellular glutamate level and the consequent excessive activation of NMDA receptors (excitotoxic mechanism) contributes to neuronal death associated with chronic and acute neurodegenerative diseases (Olney JW., 1978). Since the data obtained to date suggest a NT-mediated strengthening on several glutamatergic functions in the central nervous system, our work was intended to deepen its possible involvement in glutamate-induced neurodegenerative mechanisms. The in vitro model of cerebral ischemia obtained by oxygen and glucose deprivation (OGD) showed a significant increase in extracellular levels of glutamate. In addition, significant alterations of biochemical and morphological parameters measured were observed. Increase the release of LDH, reduction of mitochondrial oxidative capacity (MTT levels), increased activity of caspase-3, increased number of apoptotic (fragmented) nuclei, increasead level of AN(+)/PI(-) immunoreactive cells and MAP-2 dendritic aggregations was measured 24 hours after the ischemic insult. The addition of NT (lOOnM) to the culture medium showed a significant increase in the OGD-induced changes, while cells pre-exposure to the NTS1 antagonist SR48692 (lOOnM) blocked the effect of both the neuropeptide and OGD exposure, alone or in combination. The results obtained with this in vitro model of cerebral ischemia, stress the involvement of NT activity in the eziopathogenesis of an acute neurodegenerative disease (Antonelli T., 2008). Results II: At basal ganglia level, NT induces an amplification of glutamate release, probably through a NTS1/D2 antagonistic interaction. This phenomenon could contribute to the degeneration of dopaminergic nigro-striatal neurons by the means of an excitotoxic mechanism, pathogenetic feature of Parkinson's disease (PD). In this contex, experiments were conducted with the in vivo microdialysis technique at striatal and cortical level, anatomical areas notoriously involved in PD. The results obtained again showed that a, potential, NTS1/NMDA receptor-receptor interaction induces a glutamatergic signalling amplification. The observed increase in glutamate extracellular levels induced by treatment with NMDA (100 and 500 uM) and NT (lO nM), showed once again to be partially blocked by treatment with NT antagonist SR48692 (Ferraro L., 2008). Given the potential neuroprotective role played SR48692, successive studies in an vivo model of PD achieved through unilateral lesion of the nigro-striatalpathway with the neurotoxin 6-idroxydopamine (6-OHDA) were done. Three experimental groups were tested for the turning rotation behaviour and by a challenge with NMDA lOOuM: lesioned rats, rats exposed only to vehicle and lesioned rats treated with the neurotensinergic antagonist. The animals exposed to SR48692 have shown a significant recovery for both the parameter of turning behaviour and responsiveness to pharmacological challenge with NMDA (Ferraro L., 2008). The results obtained can lead to the hypothesis that the use of selective NTS1 receptor antagonists, in combination with conventional drug treatments, could provide a new terapeutic approach for chronic and acute neurodegenerative diseases treatment, such as cerebral ischemia and Parkinson's disease.