SSA: Elucidation of epigenomic and transcriptional mechanisms controlling stress-induced Fkbp5 gene expression in the hippocampus

Lead Research Organisation: University of Bristol
Department Name: Clinical Science at North Bristol

Abstract

This PhD studentship is an exciting project to obtain insight into how stress impacts on the brain at
the molecular level. Stress is part of everybody's life. It can become a health problem when a stressful
situation is extremely traumatic, when exposure to stress becomes chronic, and in cases where there
is an underlying genetic vulnerability. Recently, a genetic vulnerability (a so-called single nucleotide
polymorphism (SNP)) for developing major depression and posttraumatic stress disorder was
described for the gene Fkbp5 (Klengel et al. Nature Neurosci. 2013). The protein product of this gene,
Fkbp51, reduces the sensitivity of the glucocorticoid receptor for binding the stress hormones cortisol
or corticosterone, glucocorticoid hormones in man and rodents, respectively. Excessive stimulation of
the glucocorticoid receptor is thought to have detrimental effects on the brain; therefore, Fkbp51
plays an important role in keeping glucocorticoid receptor activity within healthy limits. Accordingly,
to maintain health and wellbeing appropriate expression of the Fkbp5 gene is of major importance.
Presently, however there is only limited information available about how the gene is regulated.
Glucocorticoid receptors are known to stimulate expression of the Fkbp5 gene and raise cellular levels
of Fkbp51 protein, thus resulting in reduction of receptor sensitivity to respond to stress hormone.
Recently, we discovered that glucocorticoid receptors bind to so-called glucocorticoid-response
elements within the Fkbp5 gene in the hippocampus after stress in vivo (Mifsud & Reul, Proc. Natl.
Acad. Sci. USA 2016).
The aim of this project is to elucidate how the activity of the Fkbp5 gene is regulated by glucocorticoid
receptors and epigenetic factors like histone modifications and DNA methylation status in vivo. You
will apply a range of state-of-the-art molecular techniques including chromatin immuno-precipitation
(ChIP), quantitative PCR, hnRNA and mRNA analysis and DNA methylation analysis, bioinformatics
analysis, radioimmunoassays as well as animal experimentation including surgeries. To obtain deeper
insight into the functional properties of particular segments of the Fkbp5 gene in vitro you will use the
front-line gene editing Crispr/Cas9 technology.
This project will be supervised by Professor Johannes Reul and Dr Oscar Cordero Llana at the University
of Bristol and Professor Jonathan Mill at the University of Exeter.

Publications

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Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/M009122/1 01/10/2015 30/09/2023
1942587 Studentship BB/M009122/1 18/09/2017 30/09/2021 Emily Price
 
Description SWBio DTP (BBSRC) In vivo skills award
Amount £5,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 07/2018 
End 09/2021
 
Title voluntary dosing to orally administer drugs/compounds to rodents in a stress-free manner 
Description For the voluntary dosing method, rodents (rat, mouse) are trained to drink a solution (diluted condensed milk) from a pipette in their home cage. After a few days, in our experience, 100% of all rats are compliant. On the experimental day, the (orally applicable) drug of choice is mixed into the diluted condensed milk and offered to the rats to drink. We have successfully applied this method for the stress-free delivery of mineralocorticoid and glucocorticoid receptor antagonists. The approach was successful as shown by measurable molecular and physiological endpoints. 
Type Of Material Improvements to research infrastructure 
Year Produced 2019 
Provided To Others? No  
Impact This method is an improvement regarding the welfare of the animal as restraint and injection is not required (Refinement). Furthermore, when conducting research to obtain insight into the effects of stress on the brain one doesn't want the delivery of a drug cause any stress as this would interfere with the outcome of the experiment. The voluntary dosing method prevents the generation of such non-specific stress for the animal. Therefore, the use of this method has considerably improved the quality of the experimental outcome in terms of effectiveness of the drug and robustness of the data. In terms of availability to others, the method doesn't require a specialist expertise. I think it is important to enhance awareness of this method and more researchers will apply it. 
 
Description Bristol Neuroscience festival 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact We had a stand at the Bristol Neuroscience Festival in June 2018 where schools from around the region as well as the general public could attend. We had an interactive game to learn about the stress response and the brain. We had a prize for the person who completed the game in the fastest time. As well as learning about the stress response we had several people doing their A-levels interested in a career in research and therefore asking our paths into what we are doing now.
Year(s) Of Engagement Activity 2018
 
Description School visit in Bristol 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact We went to a local school along with a member of the British neuroscience association. We attended different classes (year 2-5) throughout the day and tried to teach them different aspects of neuroscience by using interactive methods. They had a lot of interesting questions and already had a very good knowledge surrounding the topic areas.
Year(s) Of Engagement Activity 2018