12 Oct 2023
Job InformationOrganisation/Company
University of Trieste
Department
Department of Life Sciences
Research Field
Neurosciences » Neurobiology
Researcher Profile
First Stage Researcher (R1)
Country
Italy
Application Deadline
31 Dec 2023 - 12:00 (Europe/Rome)
Type of Contract
Temporary
Job Status
Full-time
Offer Starting Date
1 Feb 2024
Is the job funded through the EU Research Framework Programme?
Not funded by an EU programme
Is the Job related to staff position within a Research Infrastructure?
No
Offer DescriptionWe seek motivated applicants for a pre-doctoral / post-doctoral fellowship aimed at developing a novel chemo-optogenetic tool for drug-refractory epilepsy. The project will optimize ad-hoc probes that will sense extracellular acidic pH shifts associated with epileptic activity and optogenetically silence excitatory neurons to inhibit seizure generation. For the in vitro approach, the successful applicant will have a background in cellular / molecular neurobiology and molecular cloning, with some experience in culturing primary neurons and astroglia. For more information and to send your application, please contact Dr. Fabrizia Cesca at [email protected] by December 31 st, 2023. Please note that the starting date is flexible. Below a detailed abstract of the Research Project.
Epilepsy is a neurological disorder characterized by repeated seizures. Several therapeutic approaches are available but, unfortunately, around 30% of patients do not respond to medical therapies. In the last decade, optogenetics has emerged as a tool to both explore neuronal networks dynamics and to treat neurological conditions such as epilepsy. The optogenetic strategy is based on the expression, in precise brain areas, of light-sensitive proteins called opsins that are able to change the membrane potential upon wavelength-specific illumination, usually achieved using invasive optical fibers. Despite the many advantages of this technique, it still faces practical and translational challenges because of the difficulties of illuminating multiple and deep areas of the brain. In this scenario, the search of alternative light sources is an unmet need.
Luciferases are enzymes able to emit light upon addiction of their substrate coelenterazine and can be used to deliver endogenously generated light to opsins and modulate their action non-invasively. In this project, we aim to develop a closed-loop chemo-optogenetic nanomachine called pHIL (pH-sensitive inhibitory luminopsin) that senses the extracellular acidic pH shifts associated with epileptic activity and optogenetically silences excitatory neurons to inhibit seizure generation. In fact, seizure activity leads to an extracellular pH shifts toward acidosis, which in turn aggravates hyperexcitability by stimulating depolarizing acid-sensing ion channels. A triple chimeric probe pHIL has been designed in which a bioluminescent variant of Renilla luciferase (RLuc8) is coupled to a fluorescent pH sensor (EGFP mutant, called E2GFP) and the inhibitory opsin eNpHR3.0. In the proposed strategy, under acidic pH evoked by hyperexcitation, the fluorescence emission of E2GFP excited by endogenous RLuc8 UV-light will increase, in turn activating eNpHR3.0 to actuate a hyperpolarizing outward current that delays/silence epileptic activity. A first pHIL1.0 chimera has already been engineered and expressed in cell lines and primary hippocampal neurons. On the basis of promising preliminary data, we will: (i) optimize the chimera for optimal energy transfer between its building blocks; (ii) study the activation of the inhibitory opsin by the acidic pH in cell lines and primary neurons; (iii) express pHIL in the hippocampus and study pHIL responses to epileptic-like activity in cortico-hippocampal slices; (iv) test the efficacy of pHIL in vivo on drug-evoked tonic-clonic seizures and in experimental models of genetic epilepsy. We propose pHIL as a cell-autonomous close-loop nanomachine to counteract neuronal hyperexcitability and restore neural network homeostasis in drug-resistant epilepsy.
RequirementsResearch Field
Neurosciences » Neurobiology
Education Level
Master Degree or equivalent
Research Field
Neurosciences » Neurobiology
Education Level
PhD or equivalent
Number of offers available
1
Company/Institute
University of Trieste
Country
Italy
City
Trieste
Postal Code
34127
Street
Piazzale Europa, 1
Geofield
Where to applyCity
Trieste
Website
https: // www. units.it/
Street
Piazzale Europa, 1
Postal Code
34127
STATUS: EXPIRED