CHRAT Vacation Studentship - Summer 2013
Student: Ishani Barai
Supervisor(s): Dr Francesco Conti ( Dubowitz Neuromuscular Center)
Title of Project: The role of Integrins in the development of the neuromuscular system
Aim of Project: To investigate and identify the role of Integrin in regulating the development of the neuromuscular
junctions (NMJ)
Brief description of project and results. Please also comment on the value of your experience:
While the main focus of my placement was on the role of integrins, I initially spent time shadowing and gaining an
understanding of the various projects within the team. This provided me with a solid understanding of how the lab worked and
general hands on experience with the various laboratory techniques, including: immunohistochemistry, Western blotting,
plasmid isolation from bacterial cultures, cell culturing and working with electron and confocal laser scanning microscopes.
Integrins are a major class of adhesion proteins in various cell types, that mediate a connection between extracellular matrix
proteins, cytoskeletal and signalling proteins in the cytoplasm. Dr Conti’s group had identified defects in the neuromuscular
junction (NMJ), the synapse that allows the nervous system to communicate with the muscles and to trigger movement. In
particular, these studies had identified an integrin responsible for the correct assembly of active zones, the structures in the
nerve terminal that are essential for neurotransmitter release, and thus communication with the muscle. This integrin is also
responsible for organising the basal lamina, a layer of proteins between the nerve terminal and the muscle, which is important
for various synaptic functions. A third role of this is involved in regulating the correct number and patterning of the NMJs.
In light of these findings, my project aimed to look at whether other aspects of NMJ maturation were also under the control of
integrins. Using electron microscopy, I have helped determine the presence of defects in the basal lamina of mutant NMJs. Using
imaging techniques, Image J and GraphPad Prism I was able to assess that integrin mutant NMJs occupied a different area
compared to wild-type controls, suggesting defects in the maturation from immature junctions to larger pretzel-shaped NMJs. I
also prepared samples for electron microscopy, which will be used to evaluate defects in folding and in the assembly of the
synaptic basal lamina.
This work has the purpose to increase our understanding of the development of the neuromuscular system. Another implication
is that mutations in this protein may cause defects in neurotransmission within the muscle, potentially impairing movement and
result in muscle weakness. The results may be helpful in cases of myasthenic syndromes in patients, for which the genetic
diagnosis is yet unknown.
Having completed the project and produced some desirable results, I have come to understand the significance of patience. I
experienced several problems, including technological difficulties and human errors; however it was important to believe in the
science and work to optimise techniques. From a financial perspective, I also discovered the immense costs involved in
undertaking a study and the crucial reliance of research on grants and funding. The placement has given me an insight into the
challenges and rewards behind research, encouraging me to consider academic research alongside clinical medicine.
Having come into the project with very little lab experience, I am delighted to now have an understanding of numerous
laboratory techniques and have been able to carry them out independently. It has no doubt been an amazing experience and I
hope I can take a further part in the overall project.
I would like to give special thanks to CHRAT, ICH and the Dubowitz Neuromuscular Centre for providing such a fantastic
opportunity; especially Dr Francesco Conti and Jacob Ross, who took time out from their busy schedule to mentor, supervise and
help me understand the various aspects of the project. I would also like to thank everyone else who I worked with; it was truly a
valuable and enjoyable experience!