Wait for the ride to Zurich, it's a gorgeous journey through vineyards! Unfortunately, there is no sunshine tomorrow...

Thanks Damien! But careful, not in the homecage, we still use an external setup. It's built to prioritize animal welfare and standardization, featuring a dark and safe environment, video recordings through infrared-permeable walls, built-in infrared light-source, slide-and-lock system, software...

By the way: To tackle this question, we have developed an automated pain and welfare monitoring system (combining "mouse grimace scale" and full-body pose-estimation with behavior flow analysis). Brilliant work by Oliver Sturman, the ETH 3R-Hub and our collaborator Katharina Hohlbaum. 💪

Notice that modest pain levels remain detectable with the grimace score for 24hrs after surgery. Neither meloxicam (5mg/kg) nor the combination of meloxicam(5mg/kg)+buprenorphine(0.1mg/kg) could eliminate these remaining pain levels. We detect strong side-effects from opioid treatment (hyperactive).

And saving the best for last: We integrate these results with previously published bulk and single-cell data from our lab and make these data freely accessible and searchable through an interactive app (ethz-ins.org/stressome2). This is a very large resource, we hope people like it - please share! 🙏

On a single-cell level, we reveal blunted chromatin accessibility changes after chronic stress for cAMP signaling, and a damped response that returns to baseline much faster for glucocorticoid receptor binding. This again points to two parallel mechanisms of habituation.

We replicate the profound damping of transcription on a single-cell, multi-omic level (24 biological samples), showing remarkable cell-type specificity in stress-induced transcriptional changes and diverse temporal dynamics. Estimating cellular activity we find that fewer neurons get activated.

Bioinformatic and experimental approaches (based on temporal dynamics and transcription factor analysis) identify two parallel, independent mechanisms of habituation: an early blunting due to cAMP signaling, a late blunting related to corticosterone signaling at the glucocorticoid receptor.

We find that the transcriptional stress response shows dramatic habituation in every single animal, leading to profound blunting of every stress induced gene. Habituation is already established after 10 days. There is no emergence of adaptive responses, and no change in baseline gene expression 😱!

First, we profiled the transcriptomic and chromatin accessibility response to an acute restraint stress challenge over time. Then we repeated this after 10 and 20 days of stress exposure (192 biological samples for transcriptomics, 32 for ATAC-seq).

Don't get too excited David... no noradrenaline data this time 😅🫣.

nope.

yes, we are all watching in disbelief, sorry ☹️. But thanks for the encouragement, Cate! We're about to dump a ton of data into the scientific aether, I just wasn't sure if anyone was still watching. May it provide some distraction from the madness...