Assignment: The analysis and presentation of Promethion metabolic cage derived data
Task:
You are required to write up a mock-journal article that includes an introduction, methods, results, discussion and conclusion.
Introduction: A
ief background summary.
Methods: i.e. EchoMRI scan and Promethion metabolic cage.
Results: Using the given data you will be required to analyze, run statistics and produce graphs. Done
Discussion & conclusion: Talk about data with reference to literature and have strong, concise concluding remarks.
Required for submission:
· Word document of mock journal article
· Excel spreadsheet/(s) of your own data analysis, statistical tests and graphs.
Background:
4 Wild type control (C57
10) mice
4 mdx mice
6 day stay in Prometheon
EchoMRI pre and post for body composition
All groups provided with the same pre- and post-treatment echoMRI body composition data and Prometheon data.
All Groups will be required to analyze activity and body composition parameters:
· Energy expenditure and gas parameters
· Cage (non)Activity
· Body composition
All Groups will be required to make comparison of three items:
· Acclimatisation to the Prometheon cages
· Diurnal variation
· Dystrophic vs non-dystrophic (main comparison)
In vivo Analysis of Acclimatisation, Metabolism and Body Composition of wild type (C57
10) mice and mdx mice inside Prometheon cage system.
Abstract
Introduction: Additionally, we aimed to investigate
Methods:
Results:
Conclusion:
Introduction:
Materials & Methods:
Results:
Table 1: Average Energy expenditure of each group of mice during the 6-day period
WT (C57
10)
0.653
mdx
0.564
Fig. 1: Comparison of total energy expenditure of WT and mdx mice groups taken inside the prometheon cages with a total duration of 6 days
Table 2: averaged wheel running comparison for WT and mdx mice
Distance (m)
WT
7290.9
mdx
6465.6
Fig.2: comparison of the distance ran by each mice group using the running mill inside the prometheon cage.
Table 3: averaged pedestrian distance of each mice group inside the prometheon cage. Pedestrian distance refers to the activity of the mouse inside the cage (eg, wandering, walking around).
Distance (m)
WT
139.8
mdx
196.4
Fig 3: pedestrian distance travelled by each mouse group inside the prometheon cage, representing their activeness inside the cage. Measured during nocturnal hours.
Table 4: averaged non-activity of each WT and mdx group during nocturnal hours
x-beam reading
WT
34.97
mdx
42.08
Fig 4: graph showing the non-activity (lounging) of each mouse group inside prometheon cage
Table 5: measured average of duration of sleep (diurnal)
WT
87.14
mdx
80.77
Fig 5: comparison of WT and mdx mice and their sleep cycle
Table 6: Averaged gas parameters of WT and mdx mice inside the prometheon cage.
VO2
VCO2
mdx
XXXXXXXXXX
XXXXXXXXXX
WT
XXXXXXXXXX
XXXXXXXXXX
Figure 6: VO2 & VCO2 reading of each mouse group during the 6-day period
Fig 7: Fat percentage of WT and mdx mice before and after the 6-day duration inside the prometheon cage system
Fig 8: lean muscle mass measurements using echoMRI, comparing before and after the 6-day period inside the prometheon cages
Fig 9: water composition of both WT and mdx mice before and after the 6-day period inside the prometheon cage system
Fig 10: graph that shows the weight difference before and after the 6-day period in prometheon cage system
Discussion:
Conclusion:
Sleep (diurnal)
WT mdx XXXXXXXXXX XXXXXXXXXX
x beam
Gas Parameters
mdx VO2 VCO2 XXXXXXXXXX XXXXXXXXXX WT VO2 VCO2 XXXXXXXXXX XXXXXXXXXX
mL/min
WT vs mdx Energy Expenditure (Nocturnal)
WT (C57
10) mdx XXXXXXXXXX XXXXXXXXXX
x-beam reading
WT vs. mdx Wheel Meters
WT mdx XXXXXXXXXX XXXXXXXXXX
distance (m)
wt vs mdx ped distance
WT mdx XXXXXXXXXX XXXXXXXXXX
meters
Still
WT mdx XXXXXXXXXX XXXXXXXXXX
x beam
Week 4 – Workshop and Lab
in vivo Analysis of Physical Activity, Metabolism and Body Composition
Learning objectives
- Learn how to conduct live animal investigations (EchoMRI and Promethion
Metabolic System)
- The principle of both systems
- Determine primary outcomes of experimental design and inte
ogate the
differences obtained
o Mouse phenotypes (control strain versus mdx)
o Time-point measures
EchoMRITM: Body composition analysis
The EchoMRI™ Analyzers deliver precise body composition
measurements of fat, lean, free water, and total water
masses in live animals weighing up to 500, 700, 900 and
1100 grams. Scanning takes 0.5 – 3.2 minutes, depending on the precision options.
The live animals need no anesthesia and no special preparation before
measurement. EchoMRI™ Analyzers are exceedingly easy to operate. A training
period of one hour is sufficient for individuals without prior knowledge. Numeric
esults are stored and can be viewed again later at any time, as well as extracted
either into Excel and ASCII files or into Access database.
Figures above are from two different research papers from our lab. The lean and fat
mass index results are collated and finalised from raw EchoMRI data. Here, the
average values of the triplicate scans are taken and divided by the animal’s total
ody mass.
Promethion Metabolic Cage System
The Promethion cage system allows for in vivo measurement of many
physical activity and metabolic parameters. Promethion uses pull-mode air flow
generators, allowing the use of an unsealed “Home” cage environment for your
animal. Cage activity and monitoring functions, including running wheel, food hopper,
water bottle and scale, are built into the cage lid. You simply lift the lid to clean the
cage, leaving the animal in the security of its home cage. There is no handling of the
mouse, no unfamiliar new environment. This is a significant advancement over
conventional systems sealed cage systems, which can cause stress effects and
equire long acclimation times. In our studies, we typically house our mice for up to 1
week to allow for acclimitisation and familiarisation. Specifically, there are many
aspects of physical activity, behaviour and metabolism that we can elucidate from
this system including:
Energy expenditure (measured in kilocalories) during various levels
of physical activity
Wheel activity (distance, time, velocity)
Pedestrian meters (non-wheel activity)
Respiratory exchange ratios (RER)/substrate utilisation
Food and water intake
Sedentary/inactivity/sleep
Changes in body mass
Faecal and urine collection
Activity monitoring (location in cage)
With data recorded in real time, specific measures can be co
elated and contrasted
appropriately. For example: RER can be co
elated with running wheel intensity or
sedentary/sleep at that given time point – indicating substrate utilisation at that work
ate.
Mouse cage
The standard configuration model 3721 Mouse Cage comes with 2 cage tubs, cage
lid with 3 accessory holes, air flow manifold with filters and tubing, an acrylic block
and filter cover. Items include food/water hopper, body mass
acket and running
wheel capacities and are fixed in specific locations.
Total Activity Monitoring
The BXYZ Beam
eak Activity Monitor enables real time analysis of total activity with
a 0.25cm calculated centroid. Designed to be modular, the BXYZ can easily be used
stand alone, or synchronized with metabolic measurement and other elements of the
Promethion system. It is capable of ignoring
fixed objects, allowing flexibility to use
standard or customized cages without
interfering with activity monitoring. Using
Sable data analysis software, you’ll have
graphic representation of position within the
cage, calculation of total distance travelled, rearing information, and storage of raw
position vs. time data for other traceable, quantifiable analyses of activity level. The
lack ‘beam
eaks’ are displayed below and su
ound the cage for accurate
detection of raw positioning.
Wheel Activity Monitoring
The Promethion wheel monitoring system incorporates a durable, stainless steel
wheel into the cage for 180 or 360 degree monitoring
of voluntary wheel revolutions. Designed to integrate
with calorimetry, the wheel monitoring data can easily
e synchronized with RQ, VO2, beam
eak activity, or
any other parameter of the Promethion system. The in-
cage enrichment wheel provided with the Promethion
wheel monitoring system is designed not to restrict flow
around the wheel ensuring the most accurate and repeatable calorimetry data. The
system provides superior temporal resolution with real time monitoring of revolutions
per minute, permitting energy expenditure during running and resting periods to be
easily differentiated.
Food and Water Intake Monitoring
The Promethion food intake monitoring system features high precision sensors
capable of measuring real time food intake for mice and rats. The MM-1 Load Cell
with 3mg resolution can be used with the food hopper or any other Promethion mass
measurement device, allowing maximum flexibility in feeding such as varying the
location of the hopper within the cage and feeding liquid diets. The AC-2 Access
Control Module can be setup for paired feeding, yoked feeding, time and duration
limited feeding, and quantity limited feeding providing unmatched flexibility in
experimental design. The Promethion food intake monitoring system can be
incorporated into existing cage systems allowing the use of standard cages and
equiring no cage modifications reducing animal stress during acclimation. The
Promethion water intake monitoring system includes a standard water bottle and a
universal MM-1 Load Cell with a 3mg resolution. The MM-1 load cell can be used
with many different water bottle sizes and shapes allowing the use of existing
cleaning, sterilizing and filling supplies. The AC-2 Access Control Module allows
controlled access to liquid diets or water consumption. The system provides real time
monitoring, allowing for drip mitigation that differentiates between drinking bouts and
dripping events. Promethion also provides air flow through the cage keeping bedding
dry leading to healthier, happier animals. The Promethion water intake monitoring
system can be incorporated into existing cage systems allowing the use of standard
cages and requiring no cage modifications reducing animal stress during
acclimation.
Food hopper Water hoppe
Body Mass Monitoring
Promethion body mass monitors are in-cage
enrichment devices attached to a Promethion
universal MM-1 load cell. The body mass monitor
allows the real time recording of body mass when the
animal interacts with the device. The real time data
from the Promethion body mass monitor provides
automated, voluntary weighing of the animal – up to 800 mg – resulting in frequent
measurements of body mass and no handling stress on the animal during weighing.
Recording of body mass can be combined with food and water intake, and
synchronized with metabolic measurement, giving the investigator access to a more
complete picture of energy expenditure.
Data extraction and acquisitioning
ExpeData allows the user to explore acquired data in a rich graphic environment
where each parameter of the system has been
ecorded once per second enabling real time analysis
or user-selectable temporal resolution for comparison
to previous research. ExpeData automated analysis scripts can be run