Check back on Monday for #6
Or, if you are feeling really keen, read about how Kevin actually did bring in the Mongol invasion of India in his PhD thesis and viva; here.
Two blog posts in the space of a month? You lucky readers you!
Recently, as part of the BBSRC DTP experience, I went on lab rotation to the wonderful Durham University, where I spent 4 days working with my third supervisor, the legendary Prof (uncle) Roy Quinlan. Roy and his former PhD student, and my new favourite former PhD student, Fred gave me the opportunity to learn how to extract a lens basement membrane from a cow eye (see videos below). While super squeamish at first, with all the blood and what not, it was amazing to get the chance to learn such a cool and useful technique from one of the leading lens scientists in the world! “But why is this such a cool technique?” I imagine I’d hear you ask if it was in fact possible for me to hear you through a computer screen as you read this blog.
The len’s basement membrane is quite a unique basement membrane, as it comprises only one cell type, in addition to being one of the thickest basement membranes in the body. This is particularly useful as we believe we can decellularise this basement membrane and use it to culture our Pierson syndrome cell models on, providing an ex vivo model of the basement membrane. Another cool aspect of using cow eyes for this model is that the animals were already being used for meat, so no part of the animal has went to waste!
After many, many goes, and the wonderful patience of the lovely Fred, I successfully managed to extract the basement membrane from the cow lens. I was also able to decellularise the lens surprisingly easily! I’d like to thank the lovely Quinlan lab for helping me out while I was in Durham and helping me take some really useful skills back to Liverpool, with special thanks to Roy (for organising the trip), Fred (for showing me the ways of cow dissection), and Alexia (for being super friendly to a weird scouser and showing me around, and also being camera woman for the videos above!).
Another Brucey Bonus point, I couldn’t have went to Durham on a more Christmassy weekend! I drove up in the middle of a snow blizzard, so Durham was a winter wonderland for the weekend I was there. While there, I purchased the newest member of the Hamill Lab at the Durham Christmas Markets. I know Derek the Reindeer will play a huge role in keeping me sane during my PhD!
So that was my weekend in Durham! It was a fabulous five days or so, and I look forward to many more days in the Quinlan lab!
Thanks for reading science homies!
Hi! I’m Conro Sugden (aka Conor), and I’ll start my PhD in the Hamill Lab in October investigating the mechanisms through which age and disease associated alveolar epithelial matrix changes drive development and progression of pulmonary fibrosis. This isn’t my first time being a part of the Hamill Lab; Dr. Hamill was my supervisor for 2 x 12 week projects during my MRes.
I studied my undergraduate degree in Biochemistry here in Liverpool, taking the chance to complete a sandwich placement at the University of Navarra in Pamplona, Spain (see below). Here I got my first taste of what it was like to be a research student, where I was part of a group developing novel plasmids to analyse Leishmania parasites.Following my undergraduate degree, I completed an MRes in Clinical Sciences, completing 3 x 12-week projects (2 of these projects, here and here, were supervised by Dr. Hamill!) in the Institute of Ageing and Chronic Disease, and this is where I fell in love with matrix biology. My work in the Hamill Lab so far has involved (with the help of Lee Troughton) using minigene constructs to investigate intron retention with alternative polyadenylation (IRPA) in the LAMA3, and then following up this research testing ways we can manipulate this ratio. The data from these projects has been exciting and encouraging, and I’m glad that I get to carry on being a part of this.
During my time in the Hamill Lab I’ve attended my first Burns night, an awesome ‘Assembly, Dynamics and Organisation of Filaments and Cellular Responses’ workshop at Durham University, and I’ve helped with the ‘Sunscreen Challenge’ at Meet the Scientists and other outreach events.I’m in the process of becoming a STEM ambassador, and I’m planning to bring the ‘Sunscreen Challenge’ to my old high school so that I can talk to the students about what it means to be a research student. Alongside my PhD research I’ll continue with the outreach activities, and I’ve also enrolled onto a Spanish language course (can’t let myself forget everything I learned when I was over there – I will be fluent one day!). Recently I wrote for The Biochemist blog about the link between inflammation and fibrosis, you can find that here.
Going forward, I couldn’t be happier to begin my PhD in the Hamill Lab. The atmosphere is great, as are the lab members. The research currently being carried out is extremely exciting, and I can’t wait to get stuck in and make my contribution to the field.
Thanks for reading,
It’s Valentina here, the first PhD student to have joined the Hamill’s lab. I am writing my first article on a very special day…on my way to Dundee to defend my thesis! It’s been a really cool adventure, full of incredible discoveries.
When I have started, the only available data on LaNts were related to the skin; after three years and many exciting experiments we now know the effects of LaNt a31 in the cornea…which may seem only a little organ of the body, but how amazing is it to be able to see the world?! We have loads of beautiful videos of live laminin’s deposition from corneal epithelial cells and we have established a LaNt a31 overexpression animal model, the first one ever!
Working in this lab has been fantastic and hopefully today I will be able to transmit my excitement to the examiners 🙂 If my work could open at least one more door to future projects and ideas, then I’ll be doubly proud.
Going on, the Hamill lab, laminins and LaNts are waiting to be completely undercovered 🙂 much more to follow on this exciting topic…
Soon to be (hopefully) Dr. Vale
Just back from another conference and Lee has added his recent poster to our growing collection. A quick rearrange and we have the wall of LaNt research!
Yes, we have gone too far, taking over a bunch of the poster boards on our floor but it is so cool seeing the work from the various members of my group coming together into a really big story. I recommend working from left to right as you look at them to get a picture from the gene to the protein to the whole tissue.
It starts at the transcript level with Lee’s poster showing LaNt regulation of laminin expression. Also including images of changing protein expression in squamous cell carcinoma from Valentina Barrera and Conro’s minigene analysis of critical residues involved in splicing regulation.
Next we move on to some of our cell biology work looking at LaNt a31 in the front of the eye where Vale B demonstrates differential distribution accacross the corneal epithelium and Lee and Valentina Iorio show knockdown and overexpression affecting cell migration/spreading
After that its into the molecular biology, with Vale I using fluorescently tagged LaNts and laminins to study their interplay in corneal epithelial cells in culture and also the effects LaNt overexpression has on junctional complex assembly and matrix organisation.
In Poster 4 Tobi and Vale B’s work brings us back to whole tissues, this time using limbal explants as a 3D model of corneal wound repair and includes our data demonstrating changes in LaNt distribution as wounds heal.
Finally Umar and Vale B show some quite preliminary work of LaNt roles in angiogenesis and the effect of overexpression/knockdown on HUVEC tube formation.
I will now be sending all new students to study all these posters before starting work in the group. Seems like an easy way to get up to speed!
My first post and its big news! The Biotechnology and Bioscience Research Council have decided to fund my application “Characterisation of LaNt regulation of basement membrane organisation in wound repair and angiogenesis.” Exciting times!
Below is the summary from the proposal. Please feel free to comment with ideas, suggestions, feedback, collaboration ideas etc.
Characterisation of LaNt regulation of basement membrane organisation in wound repair and angiogenesis In this work I propose to study the LaNt family of proteins which were recently identified and which I believe are important for processes such as wound repair, blood vessel growth and the spread of tumours. Through these studies, a deeper understanding of these processes will be obtained and this, in turn, may lead to identification of new treatment approaches for conditions such as chronic or slow-healing ulcers and cancer. The different tissues of the body are composed of defined combinations of specialised cell types and a mixture of proteins and sugars outside the cells, termed the extracellular matrix (ECM). Some of the cell types reside in and contribute to the production of this ECM, whereas others cover the outer (epithelial) and inner (endothelial) surfaces of regions of ECM as sheets of cells. Directly beneath these cell sheets, as well as surrounding nerves and muscles, there is an organised region of ECM termed basement membrane (BM). BMs provide the anchorage point for cells and are therefore important for stress resistance and structural integrity. In addition, BMs support the different behavioural requirements of a wide range of cell type at different times, including acting as the road upon which the skin cells migrate to close wounds. A major component of all BMs is the laminin family of proteins. Laminins assemble into cross shaped molecules that associate with one another to form a network. Formation of this network has been shown to involve a small region at the very end of the short arms of the laminin cross, which is termed a LN domain. The importance of this interaction is exemplified by a number of genetic diseases where specific defects in LN domains impact the laminin network and BM organisation resulting in skin blistering, eye defects, kidney failure or muscular dystrophy. However, despite this knowledge, the ways in which laminin networks form, how network organisation changes during different cellular processes and what drives those changes is yet to be fully understood. This project will focus on the LaNts which have been demonstrated to play a role in cell attachment and migration and which my preliminary data indicate is likely to be through regulating BM formation. Like the laminins, the LaNt (Laminin N-terminus) also contain a LN domain, this suggests that they can interact directly with laminins and modify the ways in which laminin networks are organised. Importantly, there are also tissue specific differences within the laminin family and these differences are likely to mean that the impact of the LaNts is cell type specific. This may also mean that LaNts play different roles during blood vessel growth or wound repair than during normal tissue function. In order to characterise the roles of LaNts in BM formation and the impact they have on cell behaviour and tissue function, this project will pursue three aims.
Together the data obtained from these studies will dramatically expand what is known about LaNts, about laminin network formation, about BM organisation and ultimately about wound repair, blood vessel growth and tumour progression. In the longer term, this may translate into identification of new strategies for therapy development .