islet insulin production in diabetes model: spatial comparison of mouse strains
DESCRIPTION
islet insulin production in diabetes model: spatial comparison of mouse strains. mentor: Brian S. Yandell, UW-Biostat Trainee www.stat.wisc.edu/~yandell scientist: Donnie Stapleton, Attie Biochem Lab [email protected] objective of project: improve methods to distinguish "quality" - PowerPoint PPT PresentationTRANSCRIPT
![Page 1: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/1.jpg)
(C) 2005 BS Yandell UW Summer Research Program 1
islet insulin production in diabetes model:spatial comparison of mouse strains
• mentor: Brian S. Yandell, UW-Biostat Trainee www.stat.wisc.edu/~yandell
• scientist: Donnie Stapleton, Attie Biochem [email protected]
• objective of project:– improve methods to distinguish "quality"– of insulin production– between two strains of mice – based on 3-color images of islets in pancreas slices
![Page 2: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/2.jpg)
(C) 2005 BS Yandell UW Summer Research Program 2
why do this project?
• Knowledge/Skills needed– Basic statistics (t-test, ideally some nonparametrics)
– R programming (a la biostat training)
• Knowledge/Skills obtained– communication skills with scientist (lab biochemist)
– image analysis basics
– nonparametric density estimation (mechanics at least)
– experimental design considerations (nesting, balance, etc.)
![Page 3: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/3.jpg)
(C) 2005 BS Yandell UW Summer Research Program 3
project abstract• insulin is produced in the pancreas
– specifically the Islets of Langerhans by "beta" cells– (there are "alpha" cells as well.)
• scientists in biochemistry have imaged hundreds of islets– for two strains of obese mice
• BTBR.ob mimics type II (mature onset) diabetes mellitus• B6.ob appears healthy
• roughly a hundred islets per mouse, 3 mice per strain– can visually see differences in production of insulin in beta cells – have summary measurements on many islets to compare strains
• opportunity beyond data analysis– how to improve measurement process? – how to improve experimental design?
• goal: clearly infer differences in "quality" of insulin production– between BTBR.ob and B6.ob
• you can meet with the scientist and work with measurement system
![Page 4: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/4.jpg)
(C) 2005 BS Yandell UW Summer Research Program 4
of mice and men
• obese strains: BTBR.ob vs. B6.ob– obese BTBR mice mimic human diabetes
• high glucose, low insulin
– obese B6 mice appear non-diabetic
• congenic substrains A and B– mostly BTBR mouse genome– A congenic: insert small segment of B6– B congenic: essentially BTBR mice
![Page 5: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/5.jpg)
(C) 2005 BS Yandell UW Summer Research Program 5
glucose insulin
(courtesy AD Attie)
![Page 6: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/6.jpg)
(C) 2005 BS Yandell UW Summer Research Program 6
Type 2 Diabetes Mellitus
![Page 7: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/7.jpg)
(C) 2005 BS Yandell UW Summer Research Program 7
Insulin Requirement
from Unger & Orci FASEB J. (2001) 15,312
decompensation
![Page 8: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/8.jpg)
(C) 2005 BS Yandell UW Summer Research Program 8
islet degradation seriesA vs. B congenics
1 2 3 4 5healthy sick
![Page 9: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/9.jpg)
(C) 2005 BS Yandell UW Summer Research Program 9
comparison of 3 type A
and 2 type B congenic
mice
healthy sick
![Page 10: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/10.jpg)
(C) 2005 BS Yandell UW Summer Research Program 10
A and B congenic data
A congenics:Class.I Class.II Class.III Class.IV Class.V Total.IsletsA33-18 26 36 23 21 40 146A34-22 55 36 24 18 10 143A39-15 69 60 41 35 41 246
B congenics:Class.I Class.II Class.III Class.IV Class.V Total.IsletsA34-19 87 37 35 35 9 203A48-4 44 29 12 19 7 111
![Page 11: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/11.jpg)
(C) 2005 BS Yandell UW Summer Research Program 11
B6 Female&Male OB 14wk
![Page 12: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/12.jpg)
(C) 2005 BS Yandell UW Summer Research Program 12
Btbr Female& Male OB 14wk
![Page 13: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/13.jpg)
(C) 2005 BS Yandell UW Summer Research Program 13
Btbr Male & Female OB 14wks
![Page 14: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/14.jpg)
(C) 2005 BS Yandell UW Summer Research Program 14
total percent area for islets
0 50000 100000 150000
0.0
e+00
1.0
e-05
2.0
e-05
3.0
e-05
B6.1105
N = 21 Bandw idth = 6625
Den
sity
0 50000 100000 150000
0 e
+00
2 e
-05
B6.1169
N = 13 Bandw idth = 6111
Den
sity
0 50000 100000 150000
0 e
+00
2 e
-05
4 e
-05
6 e
-05
B6.1195
Den
sity
0 50000 100000 150000
0.0
e+00
1.0
e-05
2.0
e-05
BT.1419
N = 30 Bandw idth = 8943
Den
sity
0 50000 100000 150000
0.0
e+00
1.0
e-05
BT.1429
N = 34 Bandw idth = 1.246e+04
Den
sity
0 50000 100000 150000
0.0
e+00
1.0
e-05
2.0
e-05
BT.1507
Den
sity
12 13 14 15 16 17
0.00
0.10
0.20
0.30
B6.1105
N = 21 Bandw idth = 0.4719
Den
sity
12 13 14 15 16 17
0.0
0.1
0.2
0.3
0.4
B6.1169
N = 13 Bandw idth = 0.4596
Den
sity
12 13 14 15 16 17
0.0
0.2
0.4
0.6
B6.1195
Den
sity
12 13 14 15 16 17
0.00
0.10
0.20
0.30
BT.1419
N = 30 Bandw idth = 0.5141
Den
sity
12 13 14 15 16 17
0.00
0.05
0.10
0.15
0.20
0.25
BT.1429
N = 34 Bandw idth = 0.5427
Den
sity
12 13 14 15 16 17
0.00
0.10
0.20
BT.1507
Den
sity
log2 totalsraw totals
![Page 15: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/15.jpg)
(C) 2005 BS Yandell UW Summer Research Program 15
percent glu/som,
dapi, insulin
by mouse
(smooth histogram +
median)
0 20 40 60 80 100
0.00
0.04
0.08
0.12
B6.1105
0 20 40 60 80 100
0.00
0.02
0.04
0.06
0.08
B6.1169
0 20 40 60 80 100
0.00
0.02
0.04
0.06
0.08
B6.1195
0 20 40 60 80 100
0.00
0.02
0.04
0.06
0.08
0.10
BT.1419
0 20 40 60 80 100
0.00
0.02
0.04
0.06
BT.1429
0 20 40 60 80 100
0.00
0.02
0.04
0.06
0.08
0.10
BT.1507
![Page 16: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/16.jpg)
(C) 2005 BS Yandell UW Summer Research Program 16
sum of insulin,
glu/som, dapi
percents
60 80 100 120 140
0.00
00.
010
0.02
00.
030
B6.1105
N = 19 Bandw idth = 5.231
Den
sity
60 80 100 120 140
0.00
00.
005
0.01
00.
015
0.02
0
B6.1169
N = 13 Bandw idth = 8.969
Den
sity
60 80 100 120 140
0.00
0.01
0.02
0.03
0.04
B6.1195
Den
sity
60 80 100 120 140
0.00
00.
010
0.02
00.
030
BT.1419
N = 30 Bandw idth = 5.164
Den
sity
60 80 100 120 140
0.00
00.
010
0.02
0
BT.1429
N = 34 Bandw idth = 6.835
Den
sity
60 80 100 120 140
0.00
00.
010
0.02
00.
030
BT.1507
Den
sity
![Page 17: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/17.jpg)
(C) 2005 BS Yandell UW Summer Research Program 17
i
0 10 20 30 40 50 60 80 100 120 140
3050
7090
010
2030
4050
g
d
1020
3040
50
30 50 70 90
6080
100
120
140
10 20 30 40 50
tot
perc
ent
pairs
(co
rrel
atio
n) p
lot
![Page 18: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/18.jpg)
(C) 2005 BS Yandell UW Summer Research Program 18
inte
nsity
pairs
(co
rrel
atio
n) p
lot
i
600 620 640 660 680 700 720
700
750
800
850
900
600
620
640
660
680
700
720
g
700 750 800 850 900 1000 1200 1400 1600
1000
1200
1400
1600
d
![Page 19: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/19.jpg)
(C) 2005 BS Yandell UW Summer Research Program 19
calib
ratio
n st
udy
200 400 600 800 1000
1000
2000
3000
intensity
millisec
inte
nsity
200 400 600 800 1000
2000
040
000
6000
0
thresholded
millisec
thre
shol
ded
200 400 600 800 1000
2030
4050
6070
percent
millisec
perc
ent
200 400 600 800 1000
68
1012
14
signal.noise
millisec
sign
al.n
oise
![Page 20: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/20.jpg)
(C) 2005 BS Yandell UW Summer Research Program 20
90
80
70
60
50
40
30
140x103
12010080604020
Area (um^2)
B6 Btbr
Dot plo:t Insulin vs
Area of islet
![Page 21: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/21.jpg)
(C) 2005 BS Yandell UW Summer Research Program 21
50
40
30
20
10
140x103
12010080604020
Area (um^2)
B6 Btbr
Dot plot:
Glu/Som vs
Area of Islet
![Page 22: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/22.jpg)
(C) 2005 BS Yandell UW Summer Research Program 22
90
80
70
60
50
40
30
5040302010
Percent Glu/Som
B6 Btbr
Dot plot: Insulin vs
Glu/Som
![Page 23: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/23.jpg)
(C) 2005 BS Yandell UW Summer Research Program 23
Female ob/ob B6
![Page 24: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/24.jpg)
(C) 2005 BS Yandell UW Summer Research Program 24
Female ob/ob BTBR
![Page 25: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/25.jpg)
(C) 2005 BS Yandell UW Summer Research Program 25
Male ob/ob B6
![Page 26: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/26.jpg)
(C) 2005 BS Yandell UW Summer Research Program 26
Male ob/ob BTBR
![Page 27: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/27.jpg)
(C) 2005 BS Yandell UW Summer Research Program 27
Summary table for ob/ob B6 and Btbr islets
All animals ob/ob and 6 wk of age.#'s show mean of guassian fit for all islets analyzed.
Islet area Islet diameterStrain # islets (m2) (m) % Gluc&Som % DAPI % Insulin Total %
B6 F 54 12864 128 6 25 71 102Btbr F 104 14242 135 8 23 61 92
B6 M 107 13545 131 10 24 61 95Btbr M 106 11520 121 17 25 58 100
![Page 28: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/28.jpg)
(C) 2005 BS Yandell UW Summer Research Program 28
14 week Congenic Females
• Is there a difference between A & B congenics?
• Is there a difference between Btbr and the B congenic strain?
• How does the glucose and insulin values effect our comparisons?
![Page 29: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/29.jpg)
(C) 2005 BS Yandell UW Summer Research Program 29
Btbr Female OB 14wk
![Page 30: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/30.jpg)
(C) 2005 BS Yandell UW Summer Research Program 30
A33-18, Female OB A 14wk
![Page 31: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/31.jpg)
(C) 2005 BS Yandell UW Summer Research Program 31
A34-19, Female OB B 14wk
![Page 32: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/32.jpg)
(C) 2005 BS Yandell UW Summer Research Program 32
A34-22, Female OB A 14wk
![Page 33: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/33.jpg)
(C) 2005 BS Yandell UW Summer Research Program 33
A39-15, Female OB A 14wk
![Page 34: islet insulin production in diabetes model: spatial comparison of mouse strains](https://reader036.vdocument.in/reader036/viewer/2022062423/568146e8550346895db42074/html5/thumbnails/34.jpg)
(C) 2005 BS Yandell UW Summer Research Program 34
A48-4, Female OB B 14wk