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Real-Time Electrochemical Measurements in the brain of living animals
 
March 4 - 8
João Laranjinha (laranjin@ci.uc.pt), Rui Barbosa

BEB Advanced Course 2013
CenMet Coimbra Labs

Real-time Electrochemical Measurements in the Brain of Living Animals

Goals: To provide a theoretical view coupled with experimental expertise on the approaches to study neurotransmitter dynamics in the brain in vivo (anesthetized and awake behaving rats), using state-of-the-art electrochemical methods.
This course is organized within the framework of the Coimbra Labs Division (CNC) of the Center for Microelectrode Technology, University of Kentucky at Lexington (Greg Gerhardt, Director).

Course outline
The course will consist on lectures by the morning and experimental sessions by the afternoon, using state-of-the-art technology for in vivo recordings. The experiments will be assisted by experts that have developed the technology for the in vivo approaches.

Organizers:
João Laranjinha (laranjin@ci.uc.pt)
Rui Barbosa (rbarbosa@ff.uc.pt)

Faculty List:
Greg Gerhardt (Kentucky University, Lexington, USA)
Martin Lundblad (Lund University, Sweden)
George Quintero (Kentucky University, Lexington, USA)
João Laranjinha (CNC)
Rui Barbosa (CNC)

Instructors:
Ana Ledo (Research Associate)
Cátia Lourenço (Post-Doctoral Fellow)
Ricardo Santos (Post-Doctoral Fellow)

COURSE OUTLINE

MONDAY, 4TH

9:00-10:30 (BEB Room)
Electrochemistry for Neuroscience. Rapid electrochemical measurements in biological system.
(Greg Gerhardt)
10:30-11:00 -  Break
11:00-12:30
Translational issues: From single cells to freely–moving animals.
(Greg Gerhardt)
12:30-14:00 - Lunch
14:00 – 17:00 (Location: Faculty of Pharmacy, Health Science Campus, Pólo 3)
Hands-on (Break into Groups)
Microelectrode sensing technology: practical aspects.
(Rui Barbosa)
Carbon fibers microelectrode fabrication: coatings, testing and calibration. Reference electrodes. Micropippetes …

TUESDAY, 5TH

9:00-10:30 (BEB room)
Studies of release and clearance of DA, NE and 5HT.
(Greg Gerhardt)
10:30-11:00 AM - Break
11:00-12:30
Dopamine dynamics. Coupling amperometry with Optogenetics.
(Martin Lundblad)
12:30-14:00 - Lunch
14:00 – 18:00 (Location: Group Redox Biology Labs)
Hands-on (Break into Groups)
Recordings of DA release and uptake in brain slices and anesthetized animals.
Recordings of NO, NE in brain slices.
Preliminary analysis

WEDNESDAY, 6TH

9:00-10:30 (BEB room)
Novel technologies. Ceramic-Based Microelectrode Arrays (MEAs). Glutamate and other neurochemicals.
(Greg Gerhardt)
10:30-11:00 - Break
11:00-12:30
Unorthodox neurochemicals and signaling: the case of nitric oxide
(João Laranjinha)
12:30-14:00 - Lunch
14:00 – 18:00 (Location: Group Redox Biology Labs)
Hands-on (Break into Groups)
Calibration of glucose and glutamate biosensors.  Amperometric measurements of glucose and glutamate in rats
Measurement of NO in brain slices and anesthetized animals.
Preliminary glucose and glutamate analysis 
Preliminary glucose and glutamate analysis 

THURSDAY, 7TH

9:00-10:30 (BEB room and Group Redox Biology Labs)
Lab-lectures: recordings in awake animals.
(George Quintero, Ana Ledo and Cátia Marques)
Brain surgery for freely moving experiments.
10:30-11:00 AM - Break
11:00-12:30
Data analysis
(George Quintero, Ricardo Santos, Cátia Marques and Ana Ledo)
12:30-14:00 - Lunch
14:30-15:15 (CNC auditorium, 2nd floor)
CNC weekly seminar
Development of Intraoperative Chemical Diagnostics Using Microelectrode Arrays for the Treatment of Neurological Disorders
(Greg Gerhardt)
15:30-18:30 (BEB room)

Students seminars
PowerPoint presentations of foure related papers on the above topics followed by discussion.   Each group (4 in total) will present a paper (a table will be provided).  Each presentation will be supplemented by a discussion lead by two groups who are advised to collude with the presenter beforehand with the aim of generating a good discussion.  The visiting experts will help with this discussion.

FRIDAY, 8TH

9:00-12:30 (Group Redox Biology Labs)
Experiments in awake animals
(George Quintero, Ana Ledo and Cátia Marques)
12:30-14:00 - Lunch
14:30-15:15 (CNC auditorium, 2nd floor)
CNC seminar
Early alterations in dopamine neurotransmission in progressive models of Parkinsons disease
(Martin Lundblad)
15:30-17:00 (BEB room)
Data analysis
Summing-up


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