1

XY

XX

DNA
DNA

DNA
DNA

kinetochore

DNA

G1

DNA
,G2
(primary constriction)

Metacentric

Submetacentric

Acrocentric
Telocentric

PHA
EBV

0.075M KCl

PB
Colcemide

Tissue culture
24-48h
1. PB Lymphocytes+PHA
2. Bon e marro w cells w/o PH A

Harvest

Hypoto nic treatment

0.075M K Cl
Fixation

Ethano l:A cetic acid 3:1

Spread on Slides

G-Banding
trypsin-Giemsa

FISH
DNA Probe

Karyotype

15 ,14 ,13

G D
G

NOR

22 21

RNA
(nucleolar organizer region)

A-G

(banding)

DAPI quinacrine
acridine orange

G
G

(Reverse)

quinacrine

Q
R

R
Ba OH

G
C

Ba OH

6
Yq
R
H

C
(nucleolar organizer region)

S
AgNO

NOR
R G

7
46,XY G

19

20

21

22
X

G
3
2

satellite
NOR
centromere
C-band

1
1 2

R-band
G-band
Telomere

20

1
11 1
2
1
2
1
22
3

21

High resolution banding

R G
methotrexate (MTX)

S
MTX
S

9
Tyler-Smith C, Willard HF. Mammalian chromosome structure. Current opinion in Genetics
and Development 3:390-397,1993
Craig JM, Bickmore WA. Chromosome bands-flavours to savour. BioEssay 15:349-354,1993
Gardiner K. Human genome organization. Current Opinion in Genetics and Development.
5:315-322, 1995

DNA
Y
NOR

C
Q

Yq 16 9

16qh+

Yq

Fragile sites

Xq27.3

16qh+

10
In Situ Hybridization (ISH)
DNA

FISH
(Fluoresence In Situ Hybridization)

FISH

DNA

DNA

FISH

FISH
R

Alu
unique sequences

(chronosome painting)

11
FISH

FISH

FISH

FISH

sat

BAC/YAC

PCP

WCP
Y

18
Centromere
specific

Unique
sequence

Partial
chromosome
paint

Whole
chromosome
paint

18

Interphase
FISH

FISH
M.R.Teixeira. Combined classical and molecular cytogenetic analysis of cancer . European
Journal of Cancer 38 :1580 1584, 2002

(TTAGGG)n

DNA
chromosomal breakage syndromes
Ataxia telangiectasia

Xeroderma pigmentosum

Fanconis anemia

Bloom syndrome

Fanconis anemia
12

Acentric
(lost)
(deletion)

del(5)(p13pter)
(cri du chat)

Interstitial deletion

Bloom syndrome

13

(Ring)

pericentric inversion

paracentric inversion

inv(3)(p12;q23)

14

reciprocal translocation

P
P
q

20 der(20)
q

der(2)
t(2;20)(p21;p23)

15
(Robertsonian)

45, XX,-14,-15,t (rob) (14q;15q)

16

DICENTRIC CHROMOSOME

ACENTRIC FRAGMENTS

ISO( )p

ISO( )q

17
illegitimate crossing-over

unequal crossing-over

unequal crossing-over

duplication

deletion

crossing-over

a'
b'
crossing-over within the inversion
c'
d'

a
b
c
d

d
c b

b
a b c d

18

de novo

45,X

40%
60%
30%
10%
10%
5%
5%

19

Down syndrome

47,XX or XY,+21

Pateau syndrome

47,XX or XY,+13

Edwards syndrome

47 XX or XY,+18

Down

(non-disjunction)

20
(non-disjunction)
II

DNA
dictyotene

I
I

21

Mother

Mat
Meiosis I

DNA

Father

Mat
Meiosis II

Mother

Mat
Meiosis I or II

Father

Pat
Meiosis II

22

I
II

I
II

23

24

25

Hassold T and Hunt P. To err (meiotically) is human: the genesis of human aneuploidy.
Nature review Genetics 2:280-291,2001
Reeves RH, et al. too much of a good thing: mechanism of gene action in Down syndrome.
Trends in Genetics 17:83-88,2001

RNA

____________________________________________________

26

21
14

balanced

unbalanced

unbalanced

zygotes

balanced
1/3

balanced
1/3

Tri 21
1/3

45,XY or XX, t(14q;21q)

q

46,XX or XY,-14,t(14q;21q)

Monosomy
21

Tri 14

Monosomy
14

Spontaneous abortion

27

46,XY or XX,t(A;21)

28

meiotic quadrivalent

Segregation 1:3

Gamete:
24 chromosome

Gamete:
22 chromosome

t (4;21) (q35;q22.2)

Parent
der(21) 21

der(4)

Offspring

Korenberg JR et al. Down syndrome phenotype: The consequences of chromosomal

imbalane. Proc. Nat. Acad. Sci. (USA) 91: 4997-5001, 1994

imbalane. Proc. Nat. Acad. Sci. (USA) 91: 4997-5001, 1994

29

obligatory
nondisjunction

microdeletions
contiguous
FISH

genes syndromes
contiguous genes syndromes
15q12

Angelman Prader-Willi
13q13
11p13

Anhiridia/Wilms tumor
17q15

Miller-Dieker

17q15

Miller-Dieker

30

15
13
12
11.2
11.1
11.1
11.2
12
13

cent

SNRPN

14
15
21.1
21.2
21.3
22.1
22.2
22.3
13

PML

24
25
26.1
26.2
26.3

q
SNRPN)

FISH
15q23

PML

Budarf ML, Emanuel BS. Progress in the autosomal segmental aneusomy syndromes (SASs):
single or multi-locus disorders? Human Molecular Genetics, 6:16571665,1997

(confined placental mosaicism-CPM)

31

Kalousek DK, Vekeman M. Confined placental mosaicism and genomic

imprinting. Clinical Obstetrics and Gynaecology 14:723-730,2000

uniparental disomy-UPD

nullisomic
gamete complementation

nondisjunction

(imprinting)

Hydatiform mole
DNA

Hydatiform mole

32

PWS

Gametes:

Ova:
disomy 14

Sperm:
nullisomic 14

Zygote: Normal karyotype

UPD 14

33

UPD
(microdeletion)

~70%

~30%

Robinson WP. Mechanisms leading to uniparental disomy and their clinical consequences.
BioEssays 22:452-459,2000

34

x
x
hydatiform mole
anaphase lag

unequal crossing-over

ambiguous genitalia