Research Methods

1. in-vitro electrophysiology
2. Confocal imaging
3. Multiphoton imaging
4. Genetic manipulations
5. Biochemistry
6. Pharmacology
7. Behavior|

1. in-vitro electrophysiology
We record from slices from various brain regions. We do two types of recordings: extracellular recordings
and whole cell recordings. Each type of recording has its own unique setup. The whole-cell rig has also
live fluorescent capability for recognizing specific fluorescent structures (e.g. for axon bleb recordings).

2. Confocal imaging
In order to measure the length and assess the composition and density of the AIS, we reconstruct the
AIS structure by employing immunofluorescence and confocal imaging in fixed brain slices.

3. Multiphoton imaging
To investigate the altered calcium signaling as a result of the various manipulations that we induce,
we are employing multiphoton imaging with our interdepartmental multiphoton microscope.
We will use ratiometric imaging to measure calcium concentrations and dynamic calcium changes in
response to synaptic stimuli.

4. Genetic manipulations
Genetic manipulations that we employ are mainly in two ways: crossbreeding and viral transfections.
We use crossbreeding between transgenic mice, when we want to test the effects of one genetic change on
another mutation phenotype, while taking into consideration that the effect might be developmental and it
would be all over the brain. We use viral transfections when we want to introduce either overexpression of
a certain protein or knockdown a certain protein (by employing hsRNA viral vector) when we want to
induce a local and non-developmental change. We produce our viral vectors with the help of the
Center for Gene Manipulation in the Brain (CGMB) http://cgmb.haifa.ac.il/

5. Biochemistry
We employ basic biochemistry experiments with brain tissues like western blotting and
immunoprecipitations, in order to investigate proteins expression and protein-protein interactions.|

6. Pharmacology
We perform pharmacological experiments in slices, and examine the effects of various biochemical agents and
drugs either with electrophysiology or with biochemical assays.

7. Behavior
In our wish to understand the effects of our manipulations on the behaving animal we use many behavioral
paradigms. The most prevalent paradigms that we use are hippocampal dependent tasks like contextual fear
conditioning and Morris water maze. Other paradigms are employed when needed.