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Changes in the expression and activity of mechanosensors with age and degeneration
This project will investigate whether and how the expression and activity of TRP Channels changes with age and degeneration in the human intervertebral disc. Methods: qPCR, immunohistochemistry and immunoblotting to investigate expression and caclium Imaging or patchclamping to measure activity.
Keywords: Intervertebral disc, mechanosensing, mechanotransduction, ion channel, TRP, PCR, immunohistochemistry, immunoblotting, gene expression, protein expression
Mechanical loading is an important external cue that is needed to maintain the functionality of mechanosensitive tissues, such as the intervertebral disc. The disc is subjected to strain and compression during daily activity and both are important in maintaining nutrition and tissue balance. The mechanisms of sensing and subsequently converting mechanical signals into electrical or chemical signals are called mechanosensing and mechanotransduction, respectively. Major players in mechanosensing/mechanotrandscuction are mechanically gated ion channels, such as TRP channels.
In the intervertebral disc, the mechanisms of mechanotransduction are not well established. In our own previous work, we were able to demonstrate that certain TRP channels are expressed in bovine and human intervertebral discs. As it has been described that disc cells respond differently to mechanical loading with increasing age/degeneration, it would be interesting to know whether the expression or activity of TRP channels changes during these processes.
Mechanical loading is an important external cue that is needed to maintain the functionality of mechanosensitive tissues, such as the intervertebral disc. The disc is subjected to strain and compression during daily activity and both are important in maintaining nutrition and tissue balance. The mechanisms of sensing and subsequently converting mechanical signals into electrical or chemical signals are called mechanosensing and mechanotransduction, respectively. Major players in mechanosensing/mechanotrandscuction are mechanically gated ion channels, such as TRP channels.
In the intervertebral disc, the mechanisms of mechanotransduction are not well established. In our own previous work, we were able to demonstrate that certain TRP channels are expressed in bovine and human intervertebral discs. As it has been described that disc cells respond differently to mechanical loading with increasing age/degeneration, it would be interesting to know whether the expression or activity of TRP channels changes during these processes.
The goal of this project is to analyze the gene and protein expression of TRP channel in human disc tissue (previously collected by our clinical partners) and to correlate the expression with the degree of degeneration, age, location of the tissue and disease status. Frozen tissue will be used to isolate RNA using a custom-made tissue pulverizer, reverse transcribed and subsequently used for gene expression analysis by qPCR. For protein expression analysis, frozen samples will be used for immunoblotting and formalin-fixed samples for immunohistochemistry.
Tasks: 70% Lab Work, 10% Method Development, 20% Documentation
The goal of this project is to analyze the gene and protein expression of TRP channel in human disc tissue (previously collected by our clinical partners) and to correlate the expression with the degree of degeneration, age, location of the tissue and disease status. Frozen tissue will be used to isolate RNA using a custom-made tissue pulverizer, reverse transcribed and subsequently used for gene expression analysis by qPCR. For protein expression analysis, frozen samples will be used for immunoblotting and formalin-fixed samples for immunohistochemistry.
Each year the IDEA League offers the students of its partner universities over 180 monthly grants for a short-term research exchange. In general, these grants are awarded based on academic merit. For more information visit http://idealeague.org/student-grant/