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Regulating RET receptor expression using targeted proteasomal degradation

Kavanagh, Patrick (2016) MOBT01 20161
Degree Projects in Molecular Biology
Popular Abstract
Parkinson Disease (PD) is a neurodegenerative disease affecting many neurons in the brain. Particularly nigrostriatal Dopamine (DA) neurons of the substantia nigra pars compacta (SNpc) appear to be the epicenter of the disease. Cell death of these neurons is believed to be the primary cause for motor defects and many of the symptoms seen in PD patients. Many experimental therapies look to prevent such cell death and subsequent disease. Leading this field is the Glial Derived Neurotropic Factor (GDNF) that has show much promise in animal models and clinical trials.

Ret is the primary receptor for GDNF in the DA neurons and increasing it may provide protection in PD similar to that seen in studies where exogenous GDNF is delivered or... (More)
Parkinson Disease (PD) is a neurodegenerative disease affecting many neurons in the brain. Particularly nigrostriatal Dopamine (DA) neurons of the substantia nigra pars compacta (SNpc) appear to be the epicenter of the disease. Cell death of these neurons is believed to be the primary cause for motor defects and many of the symptoms seen in PD patients. Many experimental therapies look to prevent such cell death and subsequent disease. Leading this field is the Glial Derived Neurotropic Factor (GDNF) that has show much promise in animal models and clinical trials.

Ret is the primary receptor for GDNF in the DA neurons and increasing it may provide protection in PD similar to that seen in studies where exogenous GDNF is delivered or expressed. This would be achieved by providing DA neurons with sufficient amounts of RET, allowing efficient activation of the required intracellular GDNF/Ret signalling pathways in target cells (Figure1).

Here we use Lentiviral Vectors to deliver the RET gene to DA neurons. We then use a destabilizing domain (DD) to provide sufficient regulations of the Ret protein. DD is conjugated to the Ret protein and is ubiqutinated marking the entire construct for degradation. This can be prevented by providing trimethoprim which binds the DD preventing its degradation and allowing protein activity.

Preventing 6OHDA lesion
Here we use a 6-hydroxydopamine (6OHDA) in rats to model PD and determine the effects of DDRET expression on motor symptoms and cell death. We see a failure of the DD conjugation to provide sufficient regulation of the Ret protein. However, there is apparent ability of Ret to provide protection to the required cell types when assessed by tyrosine hydroxylase (TH) a common marker for DA neurons.

Master’s Degree Project in Molecular Biology 60 credits 2016
Department of Biology, Lund University

Advisor: Luis Quintino
Advisors Lund University- Department of Experimental Medical Science (Less)
Please use this url to cite or link to this publication:
author
Kavanagh, Patrick
supervisor
organization
course
MOBT01 20161
year
type
H2 - Master's Degree (Two Years)
subject
language
English
id
8895229
date added to LUP
2016-11-18 11:26:08
date last changed
2016-11-18 11:40:25
@misc{8895229,
  author       = {Kavanagh, Patrick},
  language     = {eng},
  note         = {Student Paper},
  title        = {Regulating RET receptor expression using targeted proteasomal degradation},
  year         = {2016},
}