Mixology

To try to make perfusion as humane as possible, we are getting help from vet Sarah Baert, who’s tackling this for us for her board certification with the American College of Animal Welfare. Sarah advises pre-medicating our mice with buprenorphine, and a previous study mixed powdered pills with Nutella to create an analgesic treat. But we don’t think we can get pills, just a liquid formulation. Would the liquid form mix with Nutella, or would it be like oil and water? I turned to Elena and her ancient arthritic cat Ginger for help: stealing a tiny bit of Ginger’s bupe, we tried mixing it with peanut butter (our Nutella proxy): a bit of ‘science in the kitchen’. And it worked! First problem solved, another 10 or so to go…

Thesis work, in and out of the lab

With the giant grant submitted to the university, I could finally pay attention again to my poor abandoned group. First I got their request forms in for returning (at least a bit) to the lab – very swiftly approved (thank goodness) by the wonderful Ryan. This was especially great for Michelle (top left), who needed to kill some more fish from a second cohort (for brain extraction) so that their ages and durations of differential housing could be the same as those from her first cohort.  With a pile of face masks and shields safely acquired, and a good lab protocol all figured out, she’s been working away in Fred Laberge’s lab most of the week.

And I also had a great meeting with Lindsey on Monday, on her parents’ lawn, about a monumental literature review she’s just wrapping up. It’s nice to be back!

The giant grant that’s dominated my life

Since February, but with growing pressure the over the last few weeks, my life has been subsumed by an NFRF Transformations proposal. This funding call aims to solve big problems using multidisciplinary teams, so myself and UK geographer Gail Davies have (with a team of about 25 people) have decided to solve the replication crisis (and other issues to do with animal use in research).

Below is a screenshot from my UoG OR-5 form, and yes those things that look like phone numbers really are totals: this 6-year project would cost $23.9 million!

 

The grant’s due in to the uni tomorrow AM, so it we spent yesterday getting feedback. This is last night’s lovely email from Jamie, which just about made my evening:

New email of the week

 

From the lovely Charlotte Winder:

***

 

So, what’s the work? A systematic review of how environmental enrichment affects rodent “models” of seven diseases that should be affected by stress (from cardiovascular disease to stroke). Our hypothesis is that conventional cages will make rats and mice more vulnerable to these health conditions.

And what’s the progress? Jess decided that for the first pre-screen of hits, which numbered some 10,000 papers, she was going to read 1,000 a day…. and she did it!! And her co-assessors, sister Alissa (who’s based in New Zealand, and scared them once by screaming during a meeting [there’d just been an earthquake]), and long time part-time research assistant Sonu Lidhar (who I’ve only ever communicated with by text; I call him “the mysterious Sonu” but really he’s just in Brampton) caught up with her a week or two later.  I just cannot wait to see what they find out!

Beautiful Balbs

Meet Coconut, Basma‘s last surviving adoptee from August 2018.  Like Emma’s Sugar (also still alive), Coconut became 3 years old in April!  Both were enriched housed from the moment they arrived in Guelph, which we strongly suspect helped their longevity. (Coconut’s looking physically great too. And Basma says yes this IS a recent photo, and no she has NOT had any work done).

In a slightly sorrier state (below) is this girl from our current cohort: victim of a barber this week. When barbering just involves hair removal, we don’t step in, but whisker removal’s different: mice use their whiskers like we use our fingers, so taking them away is quite the disability (and this mouse’s muzzle looks a bit sore, too).

With three cagemates, all fully whiskered, we don’t 100% know who the culprit was… but since only one was a C57, and in the last cohort all 11 of our past whisker-barbers were C57s (highly statistically significant), we know who it’s very very likely to have been. We’ve also learned by trying that while enriched-rearing greatly reduces this behaviour, moving barbers from standard to enriched housing does nothing to remedy it: by the time barbering appears it seems too late to change.

So, sadly that meant cervical dislocation for the C57: humane, but not something any of us want to see. When Covid’s over and we can get back to work again, we’re now determined to get to the bottom of this abnormal behaviour: why does standard housing turn some C57s into barbers? (And why are the poor old Balbs always the recipients, in our mixed strain set-ups?)  It’s really time to find out.

Zebra Fish ‘Chamber of Love’ pilots

Jaqui managed to get two trials of her mate choice tests in before we closed down research in the zebra fish lab: not enough data to run analyses obviously, but still a great way to learn how we can run things better when we get back going again. Below, a female  seems to like what she sees…

Linearity test – passed!!

Before everything shut down, Lindsey (L) just managed to complete her linearity test (a check that the amount of CO staining increases in a graded way, so that it can then be used to make quantitative assessments of CO activity). She was helped by Aileen (R), who finds she has a real penchant for lining up free-floating sections and gently gliding them onto a slide in the right order.

In the past Maria used smears (whole brain homogenates) of different thickness for this, but this time we used staining durations (doubly useful since this would also help narrow down the best incubation times).

So, sections from one brain were stained for 1, 2, 3, 4, 5 or 6 hours. By eye this turned out to look pretty promising … and then when actually tested: bingo!

CO staining protocol coming along

These brains get full marks for drama (though less for value). At 6 hours, the longest staining protocol Lindsey’s tried, these sections are too dark to be useful  – but superficially they look so cool!

Aileen and her mounted six whole brains on Friday, after Lindsey had subjected them to a range of incubation durations. This is to assess whether CO staining increases linearly with longer reaction times (and we hope it does, as we’ll know she has something she can validly quantify). Still some freezing artefacts and other problems to wrestle with, but this is all coming along very very nicely.

What’s the best treat for a mouse?

As we start planning a new judgment bias trial, hopefully more successfully this time, this time we’re playing the long game. For one, what treats do mice, even uninterested C57s, really like? (Since that might keep more of them more motivated for longer).

First Aileen and undergraduate research student Steph placed a very wide range of treats in their cages (imaginatively chosen to include freeze-dried chicken, any eggs and meal worms as well as the usual sweet treats), just to see what would regularly vanish quickly. Mice had to extract each treat from under some mesh tucked into a pill box compartment – below you can see a DBA busy foraging.

And the results are in: our mice have sweet teeth!  The appeal of savoury items was nothing compared to sugar sprinkles, popcorn and banana chips (though almond slivers got a look-in too).

Next up, an arena test: what each of our C57s select, when given a buffet in their very own rainbow coloured dispenser?

 

Email of the week

From Jamie. Yep, me him and Becky are designing our summer experiment again.