טכניון מכון טכנולוגי לישראל
הטכניון מכון טכנולוגי לישראל - בית הספר ללימודי מוסמכים  
M.Sc Thesis
M.Sc StudentOrbach Ady
SubjectThe Influence of the Voltage Dependant Anion Channel Gene
Expression over Cellular Functions Regulated by
the Peripheral Benzodiazepine Receptor
DepartmentDepartment of Medicine
Supervisor Professor Emeritus Moshe Gavish


Abstract

Peripheral benzodiazepine receptors are a trimeric complex of an 18kDa isoquinoline binding protein (IBP), a 32kDa voltage-dependent anion channel (VDAC), and a 30kDa adenine nucleotide transporter (ANT). They are located throughout the cell, especially in the mitochondria. PBRs have been associated with numerous cellular functions, such as, regulation of steroid biosynthesis, cell proliferation and apoptosis. The aim of this work was to shed some light on the significance of the VDAC1 subunit in the PBR complex and in PBR associated cellular functions.

Our project included the creation of plasmid vectors expressing the VDAC1 cDNA in both sense and anti-sense orientations, the transfection of C6 cells with the plasmids and testing the selected clones for several cellular functions. Resulting data are compared to data developed from a control group of cells that were transfected with the plasmid without any VDAC1 sequence.

We found an increase and a decrease in the whole cells’ binding and protein levels with the sense and anti-sense clones, respectively, vs. the control cells.

The VDAC1 antisense, surprisingly, was accompanied by an over expression of IBP.

Both clones proliferated faster than the control cells, while the sense clones proliferated fastest. In soft agar we found an increase in the number of medium and large colonies in sense clone and a trend towards increase in the large size colonies in antisense clone. We didn't find any significant differences in the apoptosis rate between the clones and the control, probably due to the fact that there was no significant change in the binding and protein levels in the mitochondrial fraction of the cells.

The conclusion is that VDAC1 alterations outside the mitochondria does affect the cells' proliferation rate but doesn't affect cellular apoptosis.