Screening Protocol - General Phenotypes
1. At 11 weeks of age, body weight is recorded.
2. Under anesthesia (ketamine, xylazine, and acepromazine combination),
the following measurements are made: length of rat (nose to tip of tail) and
length of rat (nose to rump).
3. Blood pressure is measured under anesthesia (see Renal protocol), the
cardiac response to a dobutamine challenge is measured using
echocardiography (see Ultrasound protocol), and blood is collected for
standard clinical chemistry (see Biochemistry protocol).
4. Following the ultrasound protocol, the following measurements are made:
whole heart weight (g), left ventricle weight (g), right ventricle weight (g),
kidney weight (g).
5.
Preface: This protocol is written with the idea that all other protocols are operating
during this week, and that rat cohorts are overlapping from week to week.
Monday
Morning
Run CO2 challenge for movement/respiratory protocol
Remove rats from chambers once CO2 is over. Weigh rats and record
internal temperatures
For FHH and BN groups place rats on high salt diet as they are housed.
Shut down all Movement/respiratory protocol gases and computers
Transfer data from movement computers to server
Clean all cages in D101
Separate out rats for water consumption per Renal Protocol
Afternoon
Remove water bottles and nylabones from Movement chambers and clean
Remove wire rack and stand from Movement chambers and clean
Clean all Movement chambers
Setup clean Movement Chambers with bedding clean water bottles and
food
Setup water collection and start
Setup renal protocol urine collection racks and allow silicone spray to dry
Measure out water bottles for renal collection
Place rats in urine collection and inject 10mL of saline subcutaneous
Make vascular buffer
Tuesday
Morning
Spin urine collection
Anesthetize and shave rats 2 at a time
Nair the chests of the rats
Catheritize the jugular vein and carotid artery
Record 15min of anestitized blood pressure recording
Ultrasound Protocol
Blood Collection
Remove Kidney, Heart and aorta
Weight kidney and heart
Spin bloods
Pipette 1mL of urine from collection tubes into labeled vials
Clean and prepare lab space for the next day
Afternoon
Data Analysis (1-2 techs)
Vascular protocol (1 tech)
Record water collection values
Measure out water bottles for renal collection
Place rats in urine collection and inject 10mL of saline subcutaneous
Make vascular buffer
Wednesday
Morning
Spin urine collection
Anesthetize and shave rats 2 at a time
Nair the chests of the rats
Catheritize the jugular vein and carotid artery
Record 15min of anestitized blood pressure recording
Ultrasound Protocol
Blood Collection
Remove Kidney, Heart and aorta
Weight kidney and heart
Spin bloods
Pipette 1mL of urine from collection tubes into labeled vials
Clean and prepare lab space for the next day
Afternoon
Data Analysis (1-2 techs)
Vascular protocol (1 tech)
Record water collection values and finish collection
Measure out water bottles for renal collection
Place rats in urine collection and inject 10mL of saline subcutaneous
Make vascular buffer
Thursday
Morning
Spin urine collection
Anesthetize and shave rats 2 at a time
Nair the chests of the rats
Catheritize the jugular vein and carotid artery
Record 15min of anestitized blood pressure recording
Ultrasound Protocol
Blood Collection
Remove Kidney, Heart and aorta
Weight kidney and heart
Spin bloods
Pipette 1mL of urine from collection tubes into labeled vials
Clean and prepare lab space for the next day
Afternoon
Data Analysis (1-2 techs)
Vascular protocol (1 tech)
Breakdown urine collection rack and send to cage wash
Wash all nylabones and water bottles for renal protocol
Check pressures on movement chambers
Check to make sure movement protocol is ready for the next day
Friday
Morning
Data Analysis
Bring cohort into D101
Weight rats and record internal temperatures
Movement Protocol
Afternoon
Data Analysis
Check movement protocol periodically to make sure everything is
working properly
Additional Tasks
Order Animals
Animal Housing
Print Cage Cards, Group lists, Urine sample lists, DNA labels and Urine labels
ID all animals
Label Blood tubes, DNA labels, Urine labels and weight boats
Make vascular packets
Make Catheters
Order additional supplies