Madeline Lancaster Profile picture
Oct 27, 2021 9 tweets 4 min read Read on X
I'm thrilled to share our latest published paper in @eLife
where we applied cryo-EM to brain organoids to look at ultrastructure of human axons with unprecedented resolution!

Check out the lovely cryo-CLEM clip below.
And a short 🧵 of what we found.

elifesciences.org/articles/70269
First off, we established a method to culture our air-liquid interface organoid cultures with EM grids to get outgrowth of axon bundles onto the grids. This enables capture of "clean" axons without dendrites like you normally get with cells in vitro.
Then, using correlative light and electron microscopy (CLEM) we could trace axon bundles and focus in on GFP labeled axons within bundles to explore their intracellular architecture.
We found that human growing axons are really unique! For example we know that microtubules are parallel and unidirectional in axons, and we could see that, but we even had the resolution to do subtomogram averaging and see individual protofilaments.
We also found some really interesting ER morphologies that seem to be unique to axons, with incredibly thin tubules almost completely lacking lumen, pointing to a primary role in lipid biosynthesis in this context, which makes sense given the huge surface area of axons.
Finally, we were surprised by the scarcity of ribosomes specifically in the axon shaft, a finding previously seen in more traditional EM, but now corroborated with cryo-EM which has the resolution to pick up even monosomes if they are there.
This has implications in terms of protein biogenesis, and suggests that local translation is not a major contributor along the length of the axon. Importantly, other neuronal processes (i.e. dendrites) had plenty of ribosomes, and we did not capture synapses or growth cones.
We hope that this big dataset of tomograms from human axons will be a useful resource for the community. Explore for yourself and access the full dataset on EMDB and EMPIAR (accession codes in the paper).
This was an awesome collaboration between my lab and @KukulskiWanda. It was the serendipitous result of our labs being next door, and many fruitful lunch breaks with co-first authors Patrick Hoffmann and Stefano Giandomenico. It helps to have friends that can do amazing science!!

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More from @mad_lancaster

Jan 21, 2022
New Lancaster lab publication out in @Nature! This was all @Dabrica’s idea and due to her ingenuity and hard work, with help from @ChiaradiaIlaria, @laupellegrini, and Alex Kalinka. Check it out and see our🧵below!

rdcu.be/cFi7f
Male and female brains differ in their total brain volume. They also show differential susceptibility for some neuropsychiatric disorders. We sought to explore the developmental origin of these differences by generating brain #organoids from male and female stem cell lines.
Adding to the cellular complement, we exposed organoids to sex steroids. Addition of androgens (testosterone and DHT) increased the numbers of basal progenitors and their proliferation while estrogen didn’t elicit an observable phenotype.
Read 9 tweets
Mar 24, 2021
📢New Lancaster Lab paper out now! Check it out, we've discovered a cool way evolution has played with cell shape to make our brains BIG! 🧵 cell.com/cell/fulltext/…
This is a question I've been interested in since starting my lab 6 years ago. And so this paper is a really big deal for me and the lab! So where to start...
We know that the human brain is about 3 times bigger than chimps' and gorillas' but why? How?
We can't (nor would we want to) do experiments on developing ape brains, so we approached this question by using brain organoids, little pea-sized replicas of early brain tissue. And when we made organoids from different apes, there was a clear difference in size!
Read 10 tweets
Aug 22, 2020
New preprint from the lab. We’ve joined the fight, and looked at tropism of the virus causing #COVID19 in the brain. Great collaboration with @AnnaAlbecka and Leo James group. Here’s a breakdown of what we find. 🧵

biorxiv.org/content/10.110…
We first look at expression of the viral receptors in human brain organoids and find not much expression, at least at the RNA level, in neural cells. BUT interestingly we find a lot of expression in the choroid plexus. So... what’s the choroid plexus you say?
The choroid plexus (ChP) is what makes your cerebrospinal fluid! It’s also a really important barrier that prevents things from entering the CSF from the blood. So it’s like a gatekeeper, protecting the brain from viruses, toxic compounds and immune cells and factors.
Read 11 tweets

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