CHROMOSOMES

Courseoutl
ine:
A.CellDi
visi
on
B.Chromosomal St
ruct
ures
C.Chromosomal mutat
ions
D.Detecti
onofgenomeandchr omosomalmut
ati
ons
1.
Karyotyping
2.Fl
uorescenceInSit
uHybr
idi
zat
ion

CELLDI VISI
ON
CellDivi
si
on-Celldi
visi
onisthepr ocessbywhi chas inglecelldi
vides
i
ntot woormor edaught ercell
s,eachwi ththepot ent i
altogr ow,
Mei
osi
s
develop,andcarryoutspecif
icfunct i
ons.I
tisaf undament alprocess
i
nbi ol
ogyt hati
sessenti
alforthegr owth,repai
r,andr eproducti
onof
l
ivi
ngor ganisms
.

 Mei osi
si st hepr ocessbywhi chas ingl
ecelldi
videsintof our
daught ercel l
s,eachwi t
hhalft henumberofchromos omesof
thepar entcel l
,23i nsteadoft henor mal46.Meiosi
siscr i
ti
cal
fors exualr eproduction,asi tal l
owsf orthe producti
on of
gamet es .
Thes ear etheeggsands perm.
 Mi t
osisi sthepr ocessbywhi chas ingl
ecelldi
videsintot wo
i
dent icaldaught ercel l
s.Mitosisproducestwocel l
st hatar e
dupl i
cat es( 46chr omos omeseach)oft heori
ginalcell.This Mi
tos
is
kindofcel ldivisi
onoccur sthroughoutthebody,ex ceptint he
reproduct iveor gans.

STAGESOFMI
TOSI
S
 PROPHASE
During prophase,the chr
omatinint he nucleuscondens esi nto
vi
s i
blechromosomes ,whichbecomevi si
bleunderami cr
os cope.
Thecent r
osomes ,whichareres
ponsibl
eforor gani
zingt hes pindle
fi
bers,movet ooppos i
tepol
esoft hecell.Thenucl earenvel ope
breaks down,al l
owing the spi
ndlef i
berst o attach to t he
chromosomes .

 METAPHASE
Duringmetaphas e,t
hechromosomesl i
neupal ongtheequatorofthe
cel
l,cal
ledthemet aphas
eplat
e.Thes pindl
ef i
bers,whichar
eattached
tothecentromeresofeachchr
omos ome,ens urethatthechr
omos omes
areproperl
yali
gnedandcanbes epar
atedcor r
ectly.

 ANAPHASE
During thisst
age,t he si
sterchromati
ds ofeach repl
icated
chromosomes eparat
eandar epul
ledtowar
dsopposi
tepol esof
thecellbythespindl
efiber
s.Thisr
esul
tsint
heformati
onoft wo
i
dent i
calset
sofchromosomesatopposit
epol
esofthecell
.
  TELOPHASE
Duringtel
ophas e,thechromosomesr eachtheoppos i
tepol
esoft he
cel
l,andthenucl earenvelopebeginst oref
orm aroundeachsetof
chr
omos omes .Thechr omosomesbegint ouncoilandret
urntotheir
normal,di
ffusef orm.The s pi
ndl
ef i
bersdisas
semble,and the cel
l
beginst
opr epareforcytoki
nesi
s.

CATEGORI
ZATI
ONOFCHROMOSOMESBYTHELOCATI
ONOFCENTROMEREI
NTHECHROMOSOME

Thecentr
omer
er egioncontainsasetofhighl
yrepeti
ti
veal
phasatell
i
tesequences
,whichint
erf
erewit
h
chr
omosomecompact i
onsot hatt
hecentromer
eappearsasaconst
rict
ioninthemetaphas
echromosome.
Dependi
ngonthelocati
onofcentr
omere,
thechromosomecanbecl
assif
iedas;

 Metacentr
icchromosome’sarmsarer
elati
vel
yequalinlength
 onearm ofthesubmetacentri
cchr
omosomei sl
ongerthant heother.
 I
nacrocentri
cchromosome, onear
misextremelys
mal landt heotheri
sex
tremel
ylong.
 I
nthet el
ocentr
icchr
omos ome,onear
mi smissi
ngcompl et
ely.

CHROMOSOMALSTRUCTURES

Chromosomes-as tructurefoundinthenucl eusof

cel
lsthatcarr
iesgenet i
cinformat
ionint hef or
m of
DNA.
Chromosomals tr
uct
urer ef
erstotheor ganizati
onof
DNAandpr ot
einswithinourcells
,playingacr uci
al
rol
ei n gene regul
ation and mai nt
aining genet
ic
i
ntegri
ty.
 CHROMOSOMALMUTATI
ONS
Typesofchr
omos
omal
mut
ati
on:

A.Changei
nChromosomalStr
uctur
e/Chr
omosomal
Aberr
at i
on
B.
Changeinthenor
mal numberofChromos
ome-Aneupl
oidy

A.
Chromos omal Str
uct ure/Chromos omal Aberrat i
on
1.
DELETI ON-Los sofchr omos omal mat eri
al
2.I
NSERTI ON-Gai nofchr omos omal mat eri
al .Ar i
sef rom dupl icati
onofpar ti
cularregionswi t
hinthe
aff
ectedchr omos omeorf r
om f ragment sofot herchr omos omes .
*Del eti
onandI nsert
ioncaus esalteredbandi ngpat t
ernsandchangei ns i
ze
3.I
NVERSI ON-apar tofthechr omos omebr eaksof fandr ejoinsinther eversedi r
ect i
on(180-degreefl
ip)
 PERI CENTRICI NVERSI ON-t hecent r
omer ei sincludedi nt heinversionoft hechr omos ome.
 PARACENTRI CINVERSI ON-onl yinvolves equenceswi thinonear m oft hechr omos ome.
4.TRANSLOCATI ON-ar etheex changeofgenet icmat eri
al bet weendi f
ferentchr omos omes
 RECI PROCALTRANSLOCATI ON-par tsoft wochr omos omesex change;t hati s,eachchr omosome
breaks,andt hebr okenchr omos omesr eas sociateorr ecombi newi t
honeanot her.
 Us ually,aBALANCED TRANSLOCATI ON ( nor esult
si nnogai nnorl ossofchr omos omalmat eri
al,
sameamountofgenet i
cs equence. )
*Nophenot ypiceffectbuthasanef f
ectwhenger m cel lisnots ort
edpr operly.Unbalancedver si
on
canbehandedont otheof fs pri
ngr esulti
ngt ot oomuchormi ssi
ngs egment sofchr omos omes.
 ROBERTSONI ANTRANSLOCATI ON-Onl yi nvol vesacr ocent ri
cchr omos omes( 13,14,15,21,22)where
theyj ointoget hert o becomeonechr omos ome.Maybecomeunbal anced ( netgai n orlossof
chromos omal mat eri
alintheof fspri
ng)r esul ti
ngt odel etedorex tracopi esofchr omos ome.

OtherSt ruct
ural
Abnor
mal
i
ties
:
5.ISOCHROSOME-met acentr
ic chr omos ome
thatr esultsf rom t r
ansver s
es pli
tti
ng oft he
centromer e dur ing celldi vis
ion.Tr ansverse
spli
tt
ing caus est wo long armsort wo s hort
armst os epar ateintodaught ercel l
sinsteadof
normalchr omos omeswi thonel ongar m and
ones hortar m.Thear msofani s ochromos ome
aret hereforeequali nlengt handgenet icall
y
i
dent i
cal.
6.RINGCHROMOSOME-r esult
sfrom del eti
onof
genet i
cr egionsf rom endsoft hechr omos ome
andaj oiningoft heendst oform ar ing.

B.Aneuploi
dy
*Euploi
d-A cellorcel
lpopul
ati
on wi
th a nor
malcompl
ementof
chromosomes.

Syndr
omesr
esul
ti
ngf
rom Aneupl
oidy:

oPatauSyndr ome(TRISOMY1 3)
oEdwar ds’
sSyndr
ome( TRI
SOMY1 8)
oDownSyndr ome( TRI
SOMY21 /TRISOMYG)
oTurnerSyndrome( MONOSOMYX)
oKl
inefelt
erSyndr
ome( XXYi nmale)
oXYYSyndr omeinmal e
MUTATI
ONHI
STORY

1960-fi
rstchr
omosomemutati
ons
.PeterNowel
land Davi
d Hunger
for
d obser
ved an abnor
mal
l
ysmal
l
chromosome22inl
eukemi
acel
l
s,whichwasl
abel
edt
he“Phi
ladel
phi
a”chr
omosome.

J
anetRowl
ey-us
ingchr
omos
omebandi
ng,
tumorcel
l
snotonl
ylos
tgenet
icmat
eri
al,
butt
heyex
changedi
t.

1972-shef i
rstdescr
ibedt hetransl
ocat
ionbetweenchr omos omes8and21( 8;
21)inpat i
entswit
hacut e
myeloblasti
cleukemia.ThePhi l
adelphi
achromos omewast her esul
tofar eci
procalexchangebet ween
chromos ome9andchr omos ome22. (
14;
18)t
ransl
ocationinf ol
li
cul
arlymphomaandt het(
15;17)tr
ans
locati
on
i
nacut epromyelocyt
icleukemia.Thi
swasalsothef i
rstevidencethatcancerhadagenet i
cbas is

Det
ect
ionofgenomeandchr
omos
omal
mut
ati
ons
1.Karyotyping
Karyotype-di rectobs ervati
onofacompl ete
setofmet aphas e chr omos omes str
ucture
arranged by s i
ze i n a cel l(largestt o
smallest)
.
- Kar yotype i s t he nor mal
nomencl atureus edt odes cr
ibethenormalor
abnor mal, cons tit
utional or acqui red
chromos omalcompl ementofani ndi
vidual
,
ti
ssue,orcel ll
i
ne.
*Kar yotypeofnor malhumanhas23pai rsof
chromos omes( 22aut os omaland 1pai rof
sexchr omos omes –XX-f emale,XY-mal e).

TheMet
hodst
oPr
epar
etheKar
yot
ype
CONVENTI ONALCYTOGENETI CS
Convent i
onalcyt ogenet i
ct echniquesareusedr outinel
yt odeterminenumer icalchromos omalabnor mal i
ti
es
orst r
uct uralrearrangement s,mainlytr
anslocations,inanygi vencel ltype.Forex ampl e,itcanbeus edfor
detect i
ng genomi c aber r
ations,incl
uding bot h gains and l oss es ofs egment s oft he genome and
rearrangement swi t
hinandbet weenchr omos omes .Ther esoluti
onofs tandar dcyt ogenet i
cst echniques
remai nedl i
mited, however ,wi t
hacountofappr oximately400– 500bandsperhapl oi
dgenome.
Typically,cell
sar ear r
es t
edatt hemetaphas estagebycol cemi d.Subs equent l
y, cel
lsarefixedandpl acedon
as l
idef oll
owedbys taini
ngwi ththechemicaldyeGi emsa,afterpr oteolyti
cenzyme( t
ryps i
n)treatment .The
Giems adyegener atesdis t
inctchromosome- specif
icpatternscalledG- bands .Fig.13.
3repr esentsanex ampl e
ofthehumans et.TheGi ems adyecanmaps t
ructuralaberrati
onscl ear l
yandi sacommonl yus eds tai
ning
met hodf oranal yzingchr omos ome.
Speci
menus ed:
Reagent sus ed: 1.
Blood-1heparinizedgr eentop,
1.
Phytohemaggl utinin- is added t o a cel l 5-7ml(
chil
d-3ml,infant-2-2.
5ml)
cult
urewhen pr epar i
ngcel lsforkaryotyping.
2.Bonemarrow-Gr eent op,2-
2.5m
The pur pos e of t he phyt ohemagglutinin
3.Amniot
icfl
uid-18- 20ml
tr
eatmenti st os timul atemi tosi
sint hecell
s.
2.
Colcemi d/Colchi cine-Col cemidi sa mi t
otic
i
nhibit
or. Atypeofdr ugthatbl ockscellgrowt h
bys t
oppingmi tos is(celldivisi
on).
3.
Potassium Chl oride(KCl orpot assi
um salt
)-I
fa
cell
isplacedi nahypot oni csoluti
on,bl
oodcel l
growsins ize. I
ti sus edt ohel pproducedi st
inct
bandingpat t erns .
4.Gi
ems a-stain
5.Methanol -
forf ixation
BandingPat
ter
ns
Techni
que Pr ocedur e Bandi
ngpatter
n
Gbanding •Pr oteolysi
swi t
htrypsi
n Darkband:
A-Tri
chregi
on
•stai
ningwi thGiemsadye Li
ghtband:
G-Crichr
egi
on
Rbanding •Heatdenat ure(87°Cfor10 Darkband:
G-Crichr
egi
on
min) Li
ghtband:
A-Trichr
egi
on
•stai
ningwi thGiemsadye
Qbanding •St ai
ningwi th Darkband:
A-Tr
ichregi
on
Quinac r
inefluores
centdye Li
ghtband:
G-Cri
chregion
Cbanding •Denat ure wi th Bar i
um Darkband:
Centromere
hydroxide
•stai
ningwi thgiemsadye

2.
Fluor
escenceI
nSi
tuHybr
idi
zat
ion

FISH wor ksbyat t

achi ngaDNA pr obet oas pecifi
c
chr omos omalDNA s equence.Thi sDNA pr obehasa
fl
uor escentmol eculeat t
ac hedt oi tthathel psi nt he
trac i
ng.FISHispr ettys i
mpl e,scienti
stsprepar eaDNA
probewi thaknowns equenceofni trogenbas es.This
DNApr obeisthenat tachedt oaf l
uorescentmol ecule
thatcancr eatelumines cence. Theyputt hispr obei na
mi croscopicglasss l
idet hatal readyhaschr omos ome
stuck t oi t.The pr obe t hen at taches i tselft oa
compl ement arys equenceont hechr omos ome.Si nce
theDNApr obehasaf l
uorescentdyeat tachedt oit,i
t
becomeseas ytol ocat ewher e,int hechr omos ome,i s
thepr obeat tached.Thi stellsthel ocati
onofas pec i
fi
c
DNAs equence.

CHROMOSOMALDI
SORDERS

Resul
tsofkaryot
ypinganalysesareexpr
essedasthenumberofchr
omos
omespernucl
eus(
nor
mal
is46)
,the
typeofsexchromos omes(normalisXXorXY),f
oll
owedbyany
geneti
cabnormalit
iesobserved.

 46,
XX,del
(7)(
q13)-denot
esadelet
ioni
nthel
ongar
mq
ofchr
omos ome7atr egi
on1
,band3.

 46,XY,t
(5;1
7)(
p1 3.
3;p1
3) - denotes a tr
ansl
ocat
ion t
betweent hes hortarmspofchromosomes5and17and
region 1,band 3,s ubband 3,and regi
on 1
,band 3,
respecti
vely.

 47,
XXY-Kl
i
nefel
ters
yndr
omei
scaus
edbyanex
traX
chr
omos
omeinmales.

 47,XX+21-karyot
ypeofafemalewit
hDownsyndrome
(t
ri
s omy21
)res
ulti
ngf
rom anex
trachromos
ome21.
 AUTOSOMALCHROMOSOMESABNORMALI
TIES

 Tr
isomy21-Forthetr
isomyofaut osomalchr
omos omes,thefi
rsthuman chromos
omaldi
sor
der
di
scover
edandthemostimpor
tantoneist
heful
ltr
isomyofchromosome21(
+21).

 Tri
somy1 8-t hepr
esenceofthr
eecopiesofchromosome18,el
ici
tsEdwar
ds yndr
ome.Edwar
ds
syndromeischar
act
eri
zedbyl
owbi r
thweight
,anabnor
mall
ysmall
head,andheart
,ki
dney,
andl
ung
defects
.

 Tri
somy1 3-thepresenceofthreecopi esofchr
omos ome13,causesPat
ausyndr
ome.Childr
enwith
Patausyndromewill
oftenhavecl eftl
i
psandpalates
,extr
afinger
sortoes
,hear
tdefect
s,s
everebr
ain
abnormali
ti
es,andmalfor
medi nternalorgans
.

 Tri
somy8-War kanySyndrome.Babi
esbor
nwi t
hWarkanysyndr
ometypi
cal
lyhaveacl ef
tpal
ate,
di
sti
ncti
vefaci
alfeatur
es,heartdef
ects
,malf
ormedj
oint
s,abnor
malormiss
ingkneecaps,andan
abnormall
ycur
veds pi
ne(scol
i
osis
).

 Prader
-Wil
l
iSyndrome-Pr
ader
-Wi
l
lis
yndr
omer
esul
tsf
rom t
heabs
enceornonex
pres
sionofagr
oup
ofgenesonchromosome1
5.

SEXCHROMOSOMEABNORMALI
TIES

 Tur
nerSyndr
ome-Turnersyndr
ome(45,X)i
sanex
ampl
eofmonos
omy,i
nwhi
chagi
rli
sbor
nwi
th
onl
yonesexchr
omos
ome, anXchromosome.

 Kl
i
nef
elt
erSyndr
ome-r
esul
tedf
rom anaddi
ti
onal
Xchr
omos
ome.

 Tri
pleXSyndrome(Tris
omyX)-Somef emalesar
ebor nwithtri
pleXsyndrome( tri
somyX)i nwhich
thereisanextraXchr omosome.Thet ri
somyisnotas s
ociat
edwi thphys i
calfeaturesandoften
causesnomedicalsymptoms.As mal
lpropor
ti
onofthosewithtr
ipl
eXs yndromewi llhavemenstr
ual
probl
emsorlearni
ngdisabil
i
ties
,butthevastmajor
it
ydevelopnor mal
lyandwi thoutimpairmentof
anys or
t.

Ref
erences
:

Buki
ngham,L.(
2019).Mol
ecul
arDi
agnos
tics Fundament
als
,Met
hods
,andCl
i
nicalAppl
i
cat
ions
,3r
d
edi
ti
on,1
79pp.F.A.Davi
sCompany.I
SBN9780803699540

ChangHui
Shen(
201
9).
Diagnos
ticMol
ecul
arBi
ology,
331
pp.
Els
evi
erI
nc.
ISBN978-
0-1
2-802823-
0