SUMMER STUDENT SUMMER STUDENT REPORTREPORT

葉明憲 National Tsing Hua UniversityLife Science 05Dr. Ying-Hue Lee Institute of Molecular BiologyAcademia Sinica R.O.C Date:2003/7/1~8/29

Project I:Mouse behaviorProject I:Mouse behavior

Project II:Mouse brain Project II:Mouse brain developmentdevelopment

Purpose: find special mutants Material:F3 EMS mouse

Selection type1. Rota rod2. Hole board3. Open-field4. Water maze

Result :

Project I mouse behaviorProject I mouse behavior

rotarodrotarod Major select mutants by its Major select mutants by its balance ability.balance ability.We control rotating speed andWe control rotating speed and accelerationacceleration timetime

Initial Initial speedspeed

Final Final speedspeed

Acceleration Acceleration timetime

practicpracticee 1rpm1rpm 10rpm10rpm 60sec60sec

Trail ATrail A 1rpm1rpm 20rpm20rpm 60sec60sec

Trail BTrail B 1rpm1rpm 20rpm20rpm 1sec1sec

Trail CTrail C 1rpm1rpm 60rpm60rpm 60sec60sec

Data analysisData analysisTrail A 1 to 20 rpm /60sec

trailA

048

121620

5 10 15 20rpm

numb

er of

mice

Average=18.29rpm16 mice up to 15rpm

It’s a easy job for mice.Still we can select the worst in balance ability.

TrailB di stri buti on

0

2

4

6

8

10

12

14

16

25 50 75 100 125 150 175 200sec

num

ber o

f mice

Trail B 1 to 20 rpm/ 1sec

Data analysisData analysis

High-speed motor coordination

The curve represent the division into two opposing extremessuccess or failure

Data analysisData analysisTrail C 1 to 60 rpm/60sec Select the best and the worst

TrailC di str i but i on

012345678

0 5 10 15 20 25 30 35 40 45 50 55 60rpm

num

ber o

f nice

Approach normal distribution.

Rotarod resultRotarod result

A secA sec A rpmA rpm B secB sec C rpmC rpm

worstworst1050410504 33.4533.45 10.5810.58 3.523.52 9.329.321050510505 39.1539.15 11.5711.57 2.512.51 9.049.04

excelleexcellentnt

1048510485 136.6136.622 2020 54.254.2 53.2253.22

88

1048610486 132.2132.277 2020 78.0278.02 58.1258.12

averaaveragege

102.8102.899 18.2918.29 100.9100.9

55 32.4832.48

back

Hole boardHole board

• A board has 4 X 4 holes arranged square in a plane

• Making the lighting dull enable the mice to take a look into the hole,and dark light make the mice Comfortable and in a leisurely manner. • We select by it curiosity and activity.

Data analysisData analysishole board distribution

02468

101214

20 30 40 50 60 70 80 90 100# of holes

# of

mice

The two trail correlation coefficient=0.405The data we obtain regard as activity reference.This result also could be compared with open-fieldIt should be more trails.

10496 have lowest frequency to look into the holeAlmost he just pass the hole and look the world outsideIf take off the plastic cover ,his activity raise a little

Besides, 10504 10507 don’t look into the holeit seems that they have no interest or they look clearly

Some doubtful mice!!Some doubtful mice!!

back

Open-field Open-field

As a mice,what would you do in a open field?

WT Find the exit 1.Run along two sides2.Diagonal direction

walk around and try to find out in the center1

WT

keeping looking for,try to jump out or give up

Lose his head. again and repeat many times

• Select by its courage

Data analysisData analysisThese two trails undergo on 7/25 and 8/26So we analysis by it track.

10504 have low activity both open field and hole board.And also he got a poor balance ability.

10485 10459 express a high number in both exp.We assume that his curiosity kills the cats.

HoleHole FieldField Rota Rota BB

Rota Rota CC

1050105044 45.9745.97 61.2361.23 3.52 s 3.52 s 9.3 s9.3 s

1048104855 117.3117.3 169.9169.9 54.2 s54.2 s 53.2 s53.2 s

TwoTwo trails(7.25 8/26) show a big trails(7.25 8/26) show a big change!! Why??change!! Why??

1. I calibrate the field in the different size.2. The wall around us turns white to green.3. The light is different.4. The mice is nerves.Above all,I think the first one is the main reason why I got the difference data.But the assume 2 maybe affect the mice feeling,we would design a new project to test whether it’strue or not.

Improvement :back

Water maze Water maze 3trails/day 10day3trails/day 10day

Mice are natural swimmers,and they are intelligent.

Put mice in a circular pool with radius 1.35meterMice will try to find a place to stand in the pool

This device want to select which is good or bad in Spatial learning and memory. Using a new staring point,he swims direct to the platform not try backtrack to old. (mouse will think out different strategy to find the plat)

Mouse learning behavior in Mouse learning behavior in water mazewater maze

As mice,what would you do in the pool?

1. find a place to stand. Path decide by what he used to do2. Set up a spatial map in his brain3. Knowing that platform would always be there4. Use all kinds of strategy to find it.5. Find three remarkable cues on the wall, and triangulate to find the platform. (spatial learning and memory )

Data analysisData analysis

trailtrail StartStartD0D0practipracticc

ee SESE

D1a~D4D1a~D4aa SESE

D4b~D6D4b~D6aa SWSW

D6b~D8D6b~D8aa NN

D8b~D9D8b~D9aa SESE

D9 D9 probeprobe

SE SE (no (no platforplatfor

m)m)

Total 26 trails

EW

S

N

1

23

4

SESW

What strategy they take?What strategy they take?

1.swim around at some distance form the pool

2.one way to destination

3.backtrack to previous starting location

4.guess the direction,and make a series of sweeps

5.looking at the cues,and triangulate platform

Two valueTwo value

1.no find the platform Q=(quad4%sc * quad4%path)^0.5 +(quad4%sc-quad4%path) selection for the worst spatial learning and memory

2. Find the platform S=(1-latent sc/average)*100+(1-path mt/average)*100 selection for the smart,but not necessary for spatial .

** Video track path is the most reliable method**

Remember that: What you design,what you get.

Probe testProbe test

There is no platform!!After 8 days train,we test the mice’s spatial learning.

1. direct to the previous platform2. looking forward something is here3. swim to the center and to quadrant4 repeatedly4. Stay here5. Ensure that the platform is not here. Swim out.

selectionselection

Probe test is a basic mean to analyze spatial learning.

back

10523104621045810487

1052310523

1046210462

1045810458

1048710487

Probe test is a basic mean to analyze spatial learning.

back

selectionselection

QuadQuad4 4

%sc%sc

QuanQuan4 4

%mt%mtpasspass0404

TimcTimct%04t%04

Laten Laten sc04sc04

1052105233 67.467.4 6262 1313 1212 5.55.5

1046104622 68.468.4 62.862.8 2222 18.918.9 11.311.3

1045104588 42.742.7 4242 1111 8.88.8 4.64.6

1048104877 20.320.3 20.120.1 44 3.33.3 1818

Result as followsResult as follows

1050410504 10505 lack balance ability. 10505 lack balance ability. 1048510485 good balance ability. good balance ability. 1050410504 is short of activity and is short of activity and

curiosity.curiosity. 1048510485 10459 have high activity and 10459 have high activity and

couragecourage

Next stepNext step

Assume that the phenotype is recessive aaUse wild type AA x aa F2 got all Aa F2 inbred Aa X Aa F3 got AA Aa aa 1 : 2 : 1 Then see if 1/4 offspring carry the same phenotype (aa)If yes abundant mice would use to analyze.

ThankThankss

Byebye~~