Dr Shilpa Goyal

Overview, mendelian disorders and pattern

of inheritance
Class 1

Hereditary
Familial
Congenital

congenital means born with

not all genetic diseases are congenital

Standard Pedigree Symbols

A functional unit that is regulated by

transcription and encodes a product, either a
protein or RNA
There are about 30,000 genes in the human
genome (2% code for protein)
A single gene can generate multiple spliced
mRNA products which are translated into
proteins and are subject to complex
posttranslational modification

defined as a permanent change in the DNA

Origin
germ cells transmitted to progeny
somatic cells cancer and some congenital
malformations

Types of mutation

Chromosomal mutation structural changes within

the chromosome translocations, deletions, etc
Genome mutation loss or gain of whole
chromosomes: monosomy and trisomy
Gene mutation alterations at the level of the gene

The Genetic Code

Result from substitution of a single base in the

DNA
Coding portion of gene
Missense result in substitution of one amino
acid for another in the coded protein
conservative function not affected
nonconservative function altered
Nonsense
stop codon results in truncated protein
Noncoding portion of gene
promoter and enhancer regions
posttranslational processing defective splicing

0 Thalassemia: Point Mutation Leading To

Premature Chain Termination

Downloaded from: Robbins & Cotran Pathologic Basis of Disease (on 18 July 2005 09:03 PM)
2005 Elsevier

Deletion of multiple of 3 bases

Frameshift mutation

genetic code is altered distal to the mutation

usually leads to stop codon

Blood Group O: Single-base deletion at

the ABO locus

Downloaded from: Robbins & Cotran Pathologic Basis of Disease (on 18 July 2005 09:03 PM)
2005 Elsevier

Tay-Sachs Disease: Four-base Insertion In

The Hexosaminidase A Gene

Downloaded from: Robbins & Cotran Pathologic Basis of Disease (on 18 July 2005 09:03 PM)
2005 Elsevier

Three-base Deletion In The Common

Cystic Fibrosis (CF) Allele

Downloaded from: Robbins & Cotran Pathologic Basis of Disease (on 18 July 2005 09:03 PM)
2005 Elsevier

INTERFERE with protein synthesis

SUPPRESS transcription, DNARNA
PRODUCE abnormal mRNA
DEFECTS carried over into TRANSLATION
ABNORMAL proteins WITHOUT impairing
syntheses

Classificationofgeneticdisorders
SingleGeneDisorders
Mutationsinsinglegenes
Male
Multifactorialdiseases
Variantsingenes

+environment

Chromosomedisorders
Chromosomalimbalance

All are caused by mutations in single genes of

large effect

Transmission patterns of single gene disorders

Dominant
Heterozygotes withonecopy ofthealteredgene
areaffected

Recessive
Homozygotes withtwocopies ofthealteredgeneare
affected

Xlinkedrecessive
Maleswithonecopy ofthealteredgeneonthe
Xchromosomeareaffected
Male

Disease occurs when only one allele at

given gene locus is present
Penetrance

proportion of patients who have the gene who

express the trait (expressed as %)

Expressivity

degree to which trait is expressed e.g

neurofibromatosis cases

Role of new mutations

1) BOTH SEXES INVOLVED

2) 2) GENERATIONS

NOT SKIPPED

HUNTINGTON DISEASE
NEUROFIBROMATOSIS
MYOTONIC DYSTROPHY
TUBEROUS SCLEROSIS
POLYCYSTIC KIDNEY
HEREDITARY SPHEROCYTOSIS
VON WILLEBRAND DISEASE
MARFAN SYNDROME
EHLERS-DANLOS SYNDROMES (some)
OSTEOGENESIS IMPERFECTA
ACHONDROPLASIA
FAMILIAL HYPERCHOLESTEROLEMIA
ACUTE INTERMITTENT PORPHYRIA

Reduced production of gene product or inactive

protein.
If Enzyme protein: not manifested usually
enzymes are usually present in excess
heterozygotes have half of normal enzyme level

Protein involved in pathways with feedback

inhibition:

LDL Receptor Protein in familial hypercholesteremia

One subunit of a multimeric protein

e. g. collagen (trimeric molecule)
dominant negative

Less common than loss of function

mutation
Endow normal protein with toxic properties
Nearly always autosomal dominant
e. g. Huntingtons disease

Disease occurs only when both alleles at

given gene locus are present
Parents are usually normal
Nearly all inborn errors of metabolism are
recessive

Expression of the disorder more uniform

than with dominant diseases
Complete penetrance is common
Onset is frequently early in life
New mutations may occur but are rarely
detected

GENERATIONS SKIPPED

CF
PKU
GALACTOSEMIA
HOMOCYSTINURIA
LYSOSOMAL STORAGE
-1 ANTITRYPSIN
WILSON DISEASE
HEMOCHROMATOSIS
GLYCOGEN STORAGE
DISEASES

HgbS
THALASSEMIAS
[Link]
EHLERSDANLOS(some)
ALKAPTONURIA
[Link]
FRIEDREICHATAXIA
SPINALMUSCULARATROPHY

Both alleles of a gene pair are fully

expressed in the heterozygote
Blood group antigens
Histocompatibility antigens

Nearly all X-linked disorders are recessive

Dominant and recessive apply only to the
female males are hemizygous
Absence of father-son transmission
All daughters of affected male are obligate
carriers

DUCHENNE MUSCULAR DYSTROPHY

HEMOPHILIA , A and B
G6PD DEFICIENCY
AGAMMAGLOBULINEMIA
WISKOTT-ALDRICH SYNDROME
DIABETES INSIPIDUS
LESCH-NYHAN SYNDROME
FRAGILE-X SYNDROME

1) MALES ONLY, sons of affected males are OK

2) GENERATION SKIPPING DOESNT MATTER

Thank you

Mechanism and examples of Single gene

disorders
Class 2

1. Enzyme defects and their consequences

eg. Lysosomal storage disorders

2. Defects in membrane receptors and transport

systems
eg. Familial hypercholesterolemia

3. Alterations in structure, function, or quantity of

nonenenzyme proteins
eg. Hemoglobinopathies,Thalassemias, Marfans

4.

Mutations resulting in unusual reactions to drugs

eg. Glucose -6-phoshpate dehydrogenase (G6PD), Cytochrome P450

enzymes

Accumulation of the substrate

Metabolic block and decreased amount of the
product ( lack of feedback inhibition)
Failure to inactivate a tissue damaging
substance
Lysosomal storage disorders
Lesch-Nyhan Syndrome: deficiency of HGPRT- gout
1- antitrypsin deficiency
neutrophil elastase inactivation is deficient
unchecked activity - lung and liver damage

Defects in membrane receptors

and transport systems
Familial hypercholesterolemia
Cystic fibrosis

Hemoglobinopathies

sickle cell disease abnormal -chain

Thalassemias

decreased synthesis or chains of hemoglobin

Abnormal Structural Proteins

collagen Ehlers-Danlos syndrome

elastin Marfans syndrome

Muscular dystrophies

Glucose -6-phoshpate dehydrogenase (G6PD)

G6PD activity - protects RBCs from oxidative stress
drugs that block G6PD (e.g. primaquine) can cause
severe hemolysis in patients who lack this enzyme

Cytochrome P450 enzymes

used by the liver to metabolize many drugs

changes in CYP enzyme levels affect drug
metabolism

Defects in structural proteins

Marfan syndrome is a disorder of the

connective tissues of the body, manifested
principally by changes in the skeleton, eyes,
and cardiovascular system
Inheritance:
70% to 85% - Autosomal dominant
Sporadic - new mutations

Defect in extracellular glycoprotein fibrillin-1,

which forms a scaffolding for deposition of
elastin fibers
mutations in FBN1 gene (chromo 15q21) abnormal protein
this abnormal protein disrupts assembly of
microfibrils
Microfibrils are most abundant in aorta, ligaments,
and ciliary zonules(support lens)

Marfans Disease - Clinical

Skeletal most
striking feature
Exceptionally tall
Pectus excavatum or
pigeon chest deformity
Scoliosis
Joint laxity
Arachnodactyly
Ratio of the upper
segment to the lower
segment of body 2
SDs below mean

Marfans
Cardiovascular
dilatation and aneurysms of ascending aorta
floppy mitral valve

Ocular
Bilateral ectopia lentis

Clinical

skeletal changes
ectopia lentis
aortic aneurysm (ultrasound or x-ray)

Histology

cystic medial necrosis of aorta (autopsy)

dissecting aortic aneurysm most common cause
of death

Genetic and molecular

problematic because there are 500 distinct

mutations
detection of fibrillin defects in cultured skin
fibroblasts and DNA analysis of the gene by RFLP.

Defect in collagen synthesis or structure

Type I and III collagen affected
Tissues rich in collagen skin, ligaments and
joints
Hyperextensible, fragile skin
Hypermobile joints, predisposition to
dislocation
Serious internal complications

Defects in receptor proteins

Possibly the most frequent Mendelian disorder,

with a gene frequency of 1:500
Mutation of the gene encoding the low density
lipoprotein (LDL) receptor
Heterozygotes

2-3 x elevation of serum cholesterol

tendon xanthomas and premature atherosclerosis in early
adulthood

Homozygotes

5-6x elevation of serum cholesterol

tendon xanthomas and premature atherosclerosis develop
earlier
may have myocardial infarction by age 20 years

Syn of VLDL by liver into bloodstream.

LPL of adipose tissue capillaries cleaves it into
IDL (less TG, high chol esters content)
50% IDL taken up by liver by LDL receptors
and recycled into VLDL
Rest of IDL converted into LDL (chol rich)
LDL receptors in liver n systemic take up LDL
through coated pits and convert it into chol
used for memb syn
Neg feedback for more chol syn

Genetics
Chromo 19, many mutations
5 classes
Class 1: no syn of receptor protein
Class II: receptor protein cannot be transported from
ER to golgi
Class III: LDL binding domain defective
Class !V: failure to internalize the protein by coated
pits
Class V: acid dependent dissociation of receptor and
bound LDL fails

Disorders associated with defect in

enzymes

Galactosemia: galactose-1phosphate uridyltransferase

Albinism: tyrosinase

Lesch-Nyhan
Syndrome

Downloaded from: Robbins & Cotran Pathologic Basis of Disease (on 18 July 2005 09:03 PM)
2005 Elsevier

Tissue- where most of material to be degraded is

present
Location- where degradation normally occurs
Gaucher Disease, Type I
glucocerebroside in cell membranes of senescent
leukocytes and erythrocytes
reticuloendothelial cells of spleen, bone marrow

Tay-Sachs Disease

GM2 ganglioside
neurons of central nervous system

Lysosomes contain acid hydrolases that catabolize

the breakdown of complex molecules
Lysosomes may contain
substances from cellular organelles (autophagy)
bacteria and other exogenous material (heterophagy)

Lysosomal storage diseases result from the lack of

any protein essential for their function
lack of lysosomal enzyme
dysfunctional enzyme
defective post-translational processing of enzyme

Downloaded from: Robbins & Cotran Pathologic Basis of Disease (on 18 July 2005 09:03 PM)
2005 Elsevier

Tay-Sachs mc (GM2 gangliosidosis)

Hexosaminidase A deficiency

GANGLIOSIDES are ACCUMULATED

Ashkenazi Jews (1/30 are carriers)
CNS neurons a site of accumulation
Deterioration of mental and physical abilities
CHERRY RED spot in Macula

MANY types, but the metachromatic

leukodystrophies (CNS), Krabbe, Fabry, Gaucher,
and Niemann-Pick (A and B) mc
SULFATIDES, CEREBROSIDES, SPHINGOMYELIN are
the accumulations

Most common storage disease

Autosomal recessive
Deficient enzyme is glucocerebrosidase
cleaves glucose from ceramide
glucocerebrosides accumulate

Glucocerebrosides are derived from

catabolism of lipids in cell membranes of
senescent white and red blood cells
Accumulate in macrophages of bone marrow,
liver, spleen and lymph nodes
symptoms appear in adulthood
splenic enlargement
bone marrow involvement
type I disease does not cause neurological disease
and is compatible with long life

Gaucher cell - crumpled paper

cytoplasm, upto 100um size.

Morphology

Gaucher cell is characteristic

Glucocerebrosidase assay

diagnostic of homozygous disease

heterozygote values overlap with normal

Genetic

presence of 150 alleles complicates genetic

diagnosis

TYPES A, B, C
SPHINGOMYELIN BUILDUP
Sphingomyelinase (ASM), is the missing enzyme
MASSIVE SPLENOMEGALY
OFTEN FATAL in EARLY LIFE, CNS inv, ORGANOMEGALY
Affected cells upto 90um size, foamy cytoplasm
EM- Zebra bodies

HURLER/HUNTER, for I and II, respectively

DERMATAN sulfate, HEPARAN sulfate
buildup, respectively

coarse facial features

clouding of the cornea
joint stiffness
mental retardation
URINARY EXCRETION of SULFATES COMMON

OTHER LYSOSOMAL STORAGE

DIS.

FUCOSIDOSIS
MANNOSIDOSIS
ASPARTYLGLYCOSAMINURIA
WOLMAN (CHOL., TRIGLYCERIDES)
ACID PHOSPHATE DEFICIENCY
(PHOS. ESTERS)

ALKAPTONURIA
NOT a LYSOSOMAL ENZYME DISEASE
FIRST inborn error of metabolism TO BE
DESCRIBED
HOMOGENTISIC ACID accumulates
HOMOGENTISIC ACID OXIDASE

BLACK URINE
BLACK NAILS (OCHRONOSIS), SKIN
BLACK JOINT CARTILAGE (SEVERE ARTHRITIS)

MANY TYPES (at least 13)

Type 2 (Pompe), von Gierke, McArdle, mc

Storage sites:

Liver, Striated Muscle (Skel + Ht)

Defects in proteins that regulate cell growth

Example of a defect in a protein affecting cell

growth Tumor suppressor gene
Autosomal dominant
Type I
relatively common, 1 in 3000
50% of cases lack positive family history and are
new mutations
penetrance is 100%, but expresivity is very variable

Type II

less common and not discussed further here

Gene for neurofibromatosis type 1 (NF1)

Chromosome 17q11.2
Encodes for a protein (neurofibromin) which
down-regulates the RAS signal transduction
pathway
NF-1 belongs to a family of tumor suppressor
genes

Multiple neurofibromas dispersed anywhere

in the body
cutaneous
subcutaneous
plexiform diagnostic for NF-1

Multiple pigmented skin lesions, including

caf au lait spots
Pigmented iris nodules called Lisch nodules

Caf Au Lait Spots

Lisch Nodules

Plexiform Neurofibroma

Diagnostic Criteria for Neurofibromatosis Type 1

Korf, B. R. et al. N Engl J Med 2005;352:1800-1808

Until recently no tests were available

NF-1 is a large gene with 60 exons

gene has high mutation rate
hundreds of mutations have been reported and
almost no two families share the same mutation

Specialized methods are now available

sequencing of entire gene

protein truncation analysis neurofibromin

Thank you

Multifactorial inheritance and

Chromosomal disorders
Class 3

Multi-FACTORIAL, not just multi-GENIC SOIL

theory
Common phenotypic expressions governed by
multifactorial inheritance

Hair color
Eye color
Skin color
Height
Intelligence
Diabetes, type II

Expression determined by NUMBER of genes

Overall 5% chance of 1st degree relatives having it
Identical twins >>>5%, but WAY less than 100%

Goldman: Cecil Textbook of Medicine, 22nd ed.

 Cleft

lip, palate
Congenital heart disease
Coronary heart disease
Hypertension
Gout
Diabetes
Pyloric stenosis
MANY, MANY, MANY, MANY
MORE..

Chromosome mutation structural changes

within the chromosome
deletion
inversion
translocation

Genome mutation loss or gain of whole

chromosomes: monosomy and trisomy
sex chromosomes
autosomes

Types Of Chromosomal
Rearrangements.

Downloaded from: Robbins & Cotran Pathologic Basis of Disease (on 18 July 2005 09:03 PM)
2005 Elsevier

Most common chromosomal disorder

Affects 1 in 750 newborns overall, but is
related to maternal age
1 in 1550 live births of mothers > 20 years
1 in 25 live births of mothers > 45 years

Usually results from meiotic nondisjunciton

of chromosome 21
4% result from Robertsonian translocation
of chromosome 21 to another chromosome
1% result from mitotic nondisjunction of
chromosome 21 during early
embryogenesis: mosaics

95%peoplehavethreeseparatecopiesofchromosome21
trisomy21

4%havetheextracopyofchromosome21becauseofa
Robertsoniantranslocation

Nondisjunction

1%havemosaicism withnormalandtrisomy21celllines(and
usuallyhavemuchmilderfeaturesbecauseofthepresenceofthe
normalcells);occurspostzygotically

Nondisjunction

Clinical Features of Down Syndrome

Downloaded from: Robbins & Cotran Pathologic Basis of Disease (on 18 July 2005 09:03 PM)
2005 Elsevier

Amniocentesis

most common modality

performed at 15-17 weeks gestation

Chorionic Villus Sampling (CVS)

second most common

performed at 10-12 weeks gestation

Percutaneous umbilical blood sampling (PUBS)

performed in second and third trimesters
usually prompted by ultrasound abnormalities of
fetus

Edwardssyndrome (trisomy18)
1in3000births
multiplemalformations(especiallyheart,kidneys)
clenchedhandswithoverlappingfingers

Patausyndrome (trisomy13)
1in5000births
multiplemalformations
affectsmidlinestructuresparticularly:
incompletelobationofbrain;cleftlip;congenitalheartdisease

Bothsyndromeshaveaverypoorprognosis:majorityofbabiesdyinginfirstfew
[Link](veryunusual)thereisseverementalretardation.

Patau syndrome - also known as trisomy

13 and trisomy D.
Affects about 1 in 12,000 live births.
More than 80% of infants with Patau
syndrome die within their first year of life.

The Simian line, or an

abnormal palm pattern
that is usually a symptom
of Patau syndrome.

Cayden Phipps: 3A - Abrams

92

First observed by Thomas Bartholin in

1657.
However, the actual genetic and
chromosomal aspects were discovered by
Dr. Klaus Patau in 1960, hence the name
Patau syndrome.

The cells have three copies of chromosome 13 instead of

the normal two, as well as extra material from the extra
chromosome attached to another chromosome, resulting
in changes.
Most cases occur as random events during the formation
of gametes - An error in meiosis.

Cayden Phipps: 3A - Abrams

94

A small percentage of cases occur when only

some of the bodys cells have an extra copy of
chromosome 13, resulting in a mixed
population of cells with differing numbers of
chromosomes. This is called Mosaic Patau.

A baby with a cleft palate, a common

abnormality of Patau syndrome.

Cayden Phipps: 3A - Abrams

95

Nervous system problems:

Mental and motor disabilities similar to that of

autism
Microcephaly, or a less rounded brain resulting in
more of an egg-shaped skull
Eye structure defects:

Microphthalmia, or crossed eyes (may involve one eye

or both)
Cataracts
Sensory Nystagmus, or involuntart twitching of the eye
Optic nerve hypoplasia, or the underdevelopment of the
optic nerve

Sensory nystagmus
96

Muscular and skin problems:

Polydactyly, or extra fingers/toes

Low-down ears
Prominent heels and deformed feet, called rockerbottom feet
Strange palm patterns, commonly called the Simian
line
Overlapping of the fingers over thumb
Cleft palate

Polydactyly

The Simian line

Rocker-bottom feet

Vascular Problems:

Kidney problems
Heart defects such as ventricular septal defect

Kidney Problem

The disease shown

right is called Polycystic
kidney disease (PKD).
This is a disorder in
which clumps of cysts
develop within your
kidneys. Cysts are small
round sacs containing
water-like fluid.

Since most infants with Patau syndrome die

within the first year of life, special
management/procedures are necessary to
fix defects to allow the child to survive for
as long as possible

DiGeorge Syndrome,
Velo(soft palate)Cardio(heart)Facial(face) Synd
Conotruncal anomaly face syndrome
Congenital Thymic Aplasia,
mnemonic C-A-T-C-H:
Cardiac Abnormality (especially Fallot's
Tetralogy)
Abnormal facies
Thymic aplasia
Cleft palate
Hypocalcemia

Because of a DELETION, this cannot be

detected by standard karyotyping and
needs FISH

Cytogenetic abnormalities involving sex

chromosome

Klinefeltersyndrome
47,XXY
1in1000males
Infertility(atrophictestesdonotproducesperm)
Poorlydeveloped2ndysexualcharacteristicsinsome(lackoftestosterone)
Tall

Turnersyndrome
45,X
1in5000females
99%arelostspontaneouslyinpregnancy
Shortstature
Primaryamenorrhoea(ovariesinvolutebeforebirth)
Congenitalheartdisease(coarctationofaorta)20%

Problems related to sexual development and

fertility
Discovered at time of puberty
Retardation related to the number of X
chromosomes
If you have at least ONE Y chromosome, you
are male

Imbalances of X-chromosomes are better

tolerated than those of autosomes
Lyonization Mary Lyon

during 16th day of embryonic life one Xchromosome in females is randomly inactivated
inactivation persists in all subsequent cells

Increased number of X-chromosomes in

either males or females lead to mental
retardation

 Hypogonadism

#1

found at puberty

cause of male
infertility

NO

retardation unless more Xs

47, XXY
82% of the time
L----O----N----G legs, atrophic
testes, small penis

A male hypogonadism that occurs when there

are two or more X-chromosomes and one or
more Y-chromosomes
Incidence is 1 in 500 male births
Usually (82% of cases) 47,XXY

maternal (60%) or paternal (40%) nondisjunction

during meiotic divisions

15% are mosaics, usually 46,XY/47,XXY

Testicular abnormality does not develop

before puberty

seminiferous tubules are atrophic resulting in

reduced spermatogenesis, infertility, small firm
testes, and increased FSH
testosterone levels are reduced
impotence and increased LH
lack of secondary male sexual characteristics

Mental retardation is unusual but IQ may be

below normal
Mosaics are less severely affected

45, X is the proper designation

Mosaics common
Often, the WHOLE chromosome is not missing,
but just part
NECK WEBBING
EDEMA of HAND DORSUM
CONGENITAL HEART DEFECTS most FEARED

Results from complete or partial monosomy

of the X-chromosome in females
Most common sex chromosome abnormality
in females, incidence 1 in 1000 live births
Classical cytogenetics
45,X (57%)
structural abnormalities of X-chromosomes (14%)
mosaics (29%)

GENETIC SEX is determined by the PRESENCE or

ABSENCE of a Y chromosome
TRUE HERMAPHRODITE: OVARIES AND TESTES,
often on opposite sides (VERY RARE)
PSEUDO-HERMAPHRODITE:
MALE: TESTES with female characteristics (Y-)
FEMALE: OVARIES with male characteristics (XX)

Thank you

Other genetic disorders and diagnosis

Class 4

Triplet repeats

Fragile X (CGG)
Others: ataxias, myotonic dystrophy

Mitochondrial Mutations: (maternal)

(LEBER HEREDITARY OPTIC NEUROPATHY)

Genomic IMPRINTING:
paternal allele, contradicts Mendel)

Gonadal MOSAICISM:

cells)

(Inactivation of maternal or

(only gametes have mutated

Fragile-X syndrome
Triplet expansion consists of repeating
sequences of 3 nucleotides, usually including
guanine (G) and cytosine (C) and may occur in
coding or noncoding regions of the gene
Triplets undergo expansion during
gametogeneis
Above a certain threshold of repeats, function is
impaired and disease results
Currently comprise 20 diseases

Affects males
Long face
Large mandible
Large everted ears
Large testicles (90%)
Some ptsHyperextensible joints, high arched palate,
mitral valve prolapse

Second most common cause of mental

retardation (after Downs syndrome)
Mutation is present in an untranslated portion
of the Familial Mental Retardation Gene (FMR1)
Loss of function mutation

Fragile-X

Cytogenetic abnormality
appears as a constriction
in the long arm of Xchromosome

Downloaded from: Robbins & Cotran Pathologic Basis of Disease (on 18 July 2005 09:03 PM)
2005 Elsevier

Male carrier

detected by pedigree analysis and genetic tests

20% are clinically and cytogenetically normal

Daughters are obligate carriers

50% are affected (i.e. retarded)

transmit disease to grandsons of male carrier

Risk of phenotypic effects

depends on position in pedigree

Sherman paradox

Anticipation

defect worsens with each successive generation

Fragile-X Pedigree

Downloaded from: Robbins & Cotran Pathologic Basis of Disease (on 18 July 2005 09:03 PM)
2005 Elsevier

Cytogenetics
PCR based molecular methods of repeats in
FMR-1 gene
normal 10 -55 repeats
premutation 55-230 repeats in carrier state
mutation 230 - 4000 in full clinical syndrome

Affected females

unfavorable lyonization
Shermans paradox: likelihood of MR is much higher
in grandsons than in brothers of transmitting males

Autosomal trinucleotide repeat disorder

Expansion occurs during spermatogenesis
Expansion occurs in exons coding the highly
conserved cytoplasmic protein huntingtin
Toxic gain in function produces
movement disorder
dementia

 Mitochondria have their own DNA, encoding for

proteins involved mainly in oxidative metabolism
These genes are derived from the ovum, since the
sperm has little cytoplasm and few mitochondria
Inheritance is via the mother

Leber optic atrophy

mitochondrially inherited (mother to all offspring)
degeneration of retinal ganglion cells (RGCs) and
their axons
Progressive loss of bilateral vision; blindness
Cardiac defects
Minor neurological manifestations
affects predominantly young adult males.

is a genetic phenomenon by which

certain genes are expressed in a PARENT-OFORIGIN specific manner.
Selective inactivation of either maternal or paternal
allele before fertilization
Transmitted to all somatic cells

Prader-Willi syndrome

del of band q12 on chromo15 (paternal)

Mental retardation, short stature, hypotonia,
obesity, small hands and feet, hypogonadism

Angleman syndrome

del of same chromo region from mother

MR, ataxic gait, seizures, inappropriate laughter
happy puppets

Applications:
Detection of inherited mutations
Detection of acquired mutations
Accurate diagnosis and classification of
neoplasms
Diagnosis of infectious diseases
Determination of identity, transplantation,
paternal testing, forensic medicine

Advanced maternal age >34yrs

A previous child with a chromosomal
abnormality
A carrier parent of X-linked genetic disorder
A carrier parent of translocation or inversion

Multiple congenital abnormalities

Unexplained mental/developmental
retardation
Suspected aneuploidy (downs)
Suspected unbalanced autosome (imprinting)
Suspected sex chromosome abnormality
Suspected fragile X syndrome
Infertility (to rule out sex chromo abn)
Multiple spontaneous abortions (both
partners)

Cytogenetic analysis: Karyotyping

Molecular analysis

Arrest of mitosis in metaphase by colchicine

followed by staining of chromosomes
In interphase cells, the genetic material is
dispersed and chromosomes are not visible
Identification of each chromo on pattern of
distinctive light and dark bands
Giemsa stain G banding
Described as: 47,XY, +21 (Downs)
Total no of chromo, sex chromo, abnormality
Short arm p
Long arm q

 A,B,C,D,E,F,G depends on chromosome length

A longest
G shortest

ARMREGIONBANDSub-BAND,
numbering from the centromere progressing
distally

Banded karyotype of X chromosome

Routine banded karyotype detects

Abnormal number of chromosomes
Large duplications and deletions
Balanced rearrangements (translocations,
inversions)

F.I.S.H.

(gene probes)

greatly enhances G-banding

Fluorescent InSitu
Hybridization
Uses fluorescent
labelled DNA
fragments, ~10,000
base pairs, to bind
(or not bind) to its
complement

FISH
fluorescent in situ hybridization: (FISH) A technique used to identify
the presence of specific chromosomes or chromosomal regions
through hybridization (attachment) of fluorescently-labeled DNA
probes to denatured chromosomal DNA.
Step 1. Preparation of probe. A probe is a fluorescently-labeled segment of
DNA complementary to a chromosomal region of interest.

Step 2. Hybridization. Denatured chromosomes fixed on a microscope slide are

exposed to the fluorescently-labeled probe. Hybridization (attachment) occurs between
the probe and complementary (i.e., matching)
chromosomal DNA.

FISH
Step 3. Visualization. Following hybridization, the slide is examined
under a microscope using fluorescent lighting. Fluorescent signals
indicate the presence of complementary chromosomal DNA; absence
of fluorescent signals indicate absence of complementary
chromosomal DNA.

Green signal =
Normal control
Pink signal =
Chromosome region of interest

Normal control:
Two green signals
Two pink signals

Patient with deletion:

Two green signals
One pink signal

FISH detects small (submicroscopic)

chromosome
Deletions
Duplications
Translocations
Is

applicable to interphase cells

TRIPLE CHROMOSOME
#20

A DELETION in
CHROMOSOME #22

SPECTRAL
KARYOTYPING

Diagnosis of single gene disorders

by recombinant DNA
Direct gene diagnosis
Indirect DNA diagnosis: linkage analysis

Polymerase Chain Reaction

Direct gene diagnosis

1. Some mutations alter certain restriction sites
on DNA .eg mutations in factor V
- 2 primers which bind to 3 and 5 ends of a
normal allele are produced.
- DNA between the primers is amplified
- Then digested by Mn11 enzyme
- Mutant gene- 2 products
- Normal gene 3 products

Direct gene diagnosis

2. Allele specific oligonucleotide hybridization
- Mutation does not alter cutting site of enz
- Control and pt DNA amplified using primers that flank
mutation site
- Each sample applied on a filter paper as a dot
- 2 oligonucleotides which have at their centre single
base by which normal and mutant genes differ are
produced and radiolabelled
- Allowed to hybridize with control & pt DNA
- Normal pts- strong signal
- Hetrozygotes faint signal
- Homozygotes no signal

Direct gene diagnosis

Mutations affecting length of DNA
- 2 primers that flank the region affected by
trinucleotide repeats (in fragile X) amplify the
intervening sequences
- Differential migration of amplified DNA
products on a gel according to size

Indirect DNA diagnosis: linkage

analysis
When information about gene sequencing is
lacking in a disease
To differentiate abnormal chromosome from
normal based on polymorphisms (variations)
in DNA sequences
- Site polymorphism
- Length polymorphism

Indirect DNA diagnosis: linkage analysis

Site polymorphism
Restriction fragment length polymorphisms
- Single base pair changes can produce or
abolish sites for restriction enzymes
- Length of DNA fragments produced after
digestion with certain enzymes is altered
- DNA fragments of different length detected by
southern blot

Indirect DNA diagnosis: linkage analysis

Length polymorphism
Human DNA has short repetitive sequences
of non coding DNA
The number of repeats varies in individuals
Microsatellite repeats- less than 1kb, 2-6bp
Mini satellite repeats- 1-3kb, 15-70bp

MOLECULAR DX by DNA PROBES

BIRTH DEFECTS, PRE- or POST- NATAL

TUMOR CELLS
CLASSIFICATIONS of TUMORS
IDENTIFICATION of PATHOGENS
DONOR COMPATIBILITY
PATERNITY
FORENSIC

H&E tissue
structures
ImmunoAntigen
Proteins
GENES that
MAKE those
PROTEINS

Thank you