Title

Quantification of the Inhibition of Calcineurin by Protein Phosphatase Assay

Document Type

Oral Presentation

Location

SURC Ballroom A

Start Date

17-5-2012

End Date

17-5-2012

Abstract

The nervous system is full of neurons, cellular data processors. These neurons send out processes called axons to make connections with other neurons called synapses in regions far away from their starting point. Axonal development is a major subject of current biological research today. Dr. Daniel Selski, of the CWU Biology Department, has developed a project testing the effects of Calcineurin (CaN) on axonal development. Dr. Selski’s project entails using the immunosuppressant FK506 to inhibit CaN, using a chicken model, and measure the anatomical effects in the retina. However, our lab has not yet proven that FK506 significantly reduces the amount of active CaN within the chicken embryo. Our hypothesis is that FK506 is successfully inhibiting the target protein CaN. Testing and clearly demonstrating that FK506 inhibits CaN enzyme activity in our system will strengthen the interpretation of anatomical studies. This hypothesis is being tested by treating embryos on embryonic days 5-9 with FK506, then performing a protein phosphatase assay to determine how active CaN remains within retinal tissues. The assay data can be quantitated, using a visual wavelength spectrophotometer that measures the absorbance/amount of solution phosphorylated. The expected result is that treated retinas will show a measurable difference in CaN activity compared to control retinas. A significant difference in CaN activity between FK506 treated and control retinas would be evidence that our treatments with FK506 are inhibiting CaN.

Poster Number

19

Faculty Mentor(s)

Daniel Selski

Additional Mentoring Department

Biological Sciences

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May 17th, 8:30 AM May 17th, 11:00 AM

Quantification of the Inhibition of Calcineurin by Protein Phosphatase Assay

SURC Ballroom A

The nervous system is full of neurons, cellular data processors. These neurons send out processes called axons to make connections with other neurons called synapses in regions far away from their starting point. Axonal development is a major subject of current biological research today. Dr. Daniel Selski, of the CWU Biology Department, has developed a project testing the effects of Calcineurin (CaN) on axonal development. Dr. Selski’s project entails using the immunosuppressant FK506 to inhibit CaN, using a chicken model, and measure the anatomical effects in the retina. However, our lab has not yet proven that FK506 significantly reduces the amount of active CaN within the chicken embryo. Our hypothesis is that FK506 is successfully inhibiting the target protein CaN. Testing and clearly demonstrating that FK506 inhibits CaN enzyme activity in our system will strengthen the interpretation of anatomical studies. This hypothesis is being tested by treating embryos on embryonic days 5-9 with FK506, then performing a protein phosphatase assay to determine how active CaN remains within retinal tissues. The assay data can be quantitated, using a visual wavelength spectrophotometer that measures the absorbance/amount of solution phosphorylated. The expected result is that treated retinas will show a measurable difference in CaN activity compared to control retinas. A significant difference in CaN activity between FK506 treated and control retinas would be evidence that our treatments with FK506 are inhibiting CaN.