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Journal of


Short communications

Springer-Verlag 1988
J Neurol (1988) 235 : 485-486

Familial Alzheimer's disease (FAD):

co-segregation between alleles at the D 2 1 S l l D N A marker
and the FAD gene in a particular pedigree
F. David 1, F. Clerget 2, and G. Lucote I
1Laboratory of Melocular Genetics, INTS, 6 rue Alexandre Cabanel, F-75015 Paris 15~me, France
ZLaboratory of Epidemiology Genetics, INSERM U. 155, Chateau de Longchamp, Bois de Boulogne, F-75016 Paris 166me, France

Summary. Segregation studies of Alzheimer's disease (AD) gene

and a cloned D N A probe (D21Sll), which detects an EcoRI
restriction fragment length polymorphism for a sequence located in the medial part of the long arm of chromosome 21,
are reported in a large pedigree, in which AD is transmitted as
an autosomal dominant mendelian trait. In this pedigree, the
A D gene co-segregation with one of the alternative alleles at
the probe raises the possibility of using such a marker for presymptomatic diagnosis of individuals at risk for the disease.
Key words: Familial Alzheimer's disease - Polymorphic chromosome 21 D N A probe - Presymptomatic diagnosis - At-risk

Alzheimer's disease (AD) is a degenerative disorder of the
central nervous system, characterized by progressive impairment of memory and other intellectual functions beginning in
middle to late adult life. The primary cause of AD remains unknown, although both genetic and environmental factors have
been considered; several large families have been reported
that display autosomal dominant transmission of the disorder,
and consequently the familial form of A D (FAD) observed in
these pedigrees is clearly caused by a genetic defect. Recently,
it has been shown that this genetic defect causing FAD maps
on chromosome 21 [8]. The hypothesis of an eventual linkage
between the F A D gene and several polymorphic probes located on the long arm of chromosome 21 has been investigated in the recent past in a large family, in which the FAD is
transmitted as an autosomal dominant mendelian trait [2-4]
but without any evidence of tight linkage between the gene
and the chromosome 21 markers studied. In the present paper
we report, in the same family, co-segregation between a polymorphic allele of probe D21Sll [9] and the FAD gene, suggesting that such a linked D N A marker, might be used for presymptomatic diagnosis in this particular pedigree.

Subjects and methods

The F A D family studied is a five-generation pedigree shown
in Fig. 1. It is a large kindred of Italian descent [5], representOffprint requests to: G. Lucotte

ing the Italian and the French branches, where the diagnosis of
F A D for deceased family members of generations I and II was
documented by examination of medical records and questioning of the actual members, by neuropathological examination
of post-mortem brain tissue for III8 and III13, and in living
members with standard clinical criteria or psychometric classical tests (for III10, III1, III6 and I I I l l ) . Carriers of the FAD
gene generally do not develop symptoms until the fourth decade
of life, and consequently the genetic status of asymptomatic
individuals remains uncertain. In this family, however, there
is a well-defined period during which symptoms usually appear (III2 and III4, more advanced in age, do not have the
disease). One individual in this pedigree (IV12) describes
subtle clinical symptoms (notably loss of memory), which are
sufficient to establish a practical definitive diagnosis.
The genotype of each available member of the pedigree
was ascertained by digesting 20 gg genomic D N A with EcoRI
and TaqI restriction enzymes. The resultant D N A fragments
were resolved to size by horizontal agarose gel electrophoresis
in TBE buffer, and these denatured fragments were then transferred to a nylon filter support (Zeatabind-AMF/Cuno) by
Southern blotting.

/ A=2.
B :1.9



Fig. 1. Part of the familial Alzheimer's (FAD) pedigree studied. Symbols used: square, male; circle, female; solid symbol, affected with
FAD; slashed symbol, deceased. Pedigree and clinical data from the
pedigree were collected by other members of our research group, who
were blind to the results of the molecular genetic study. Individual
genotypes at the locus D21Sll are indicated above each symbol. To
preserve confidentiality, not all of the individuals in the pedigree (and
their birth order) are shown


Table 1. Lod scores for familial Alzheimer's disease (FAD) with DNA marker D21Sll. The lod score value at 0 = 0 is low (because of allelic frequencies in the standard panel population), but positive. Lod scores were calculated using a LIPED program corrected for age of onset (a gene
frequency of 0.000I was assumed for FAD defect). Peak lod score (el was obtained for + 1.02 at 0 = 0.00
Recombination fractions (0)
Lod scores

+ 1.02













D N A p r o b e D 2 1 S 1 1 ( p l a s m i d p P W 2 3 6 B) was labelled with

[~,_32p[ a d e n o s i n e t r i p h o s p h a t e ( A m e r s h a m ) by r a n d o m oligonucleotide priming, and t h e n hybridized to the filter for 48 h
at 65C in 6 s t a n d a r d citrate of s o d i u m (SSC), 1 D e n h a r d t ' s solution. 0.5% sodium dodecyl sulphate, a n d sonicated a n d d e n a t u r e d s a l m o n testis D N A (100[ag/ml). T h e
filters were w a s h e d in 1 SSC at 65C, a n d exposed to Xray film ( K o d a k X A R - 5 ) with a D u p o n t C r o n e x intensifying
screen at - 7 0 C for 2 - 3 days.

Results and discussion

Samples restricted with EcoRI a n d TaqI were e x a m i n e d using
p r o b e D21S11. With EcoRI (Fig. 2) a c o n s t a n t b a n d was observed at 0.9 kb, and two p o l y m o r p h i c b a n d s at 2.9 (allele A )
and 1.9 kb (allele B); with TaqI the variable b a n d s were at 5.5
or 4 - 1 . 5 k b [9]. In o u r family, as previously o b s e r v e d [1, 6],
t h e r e was a c o m p l e t e c o n c o r d a n c e in the findings with the
p r e s e n c e of the EcoRI and TaqI restriction sites a n d cons e q u e n t l y t h e r e is a strong linkage disequilibrium b e t w e e n the
p r e s e n c e or a b s e n c e of these two p o l y m o r p h i s m s .
Figure 1 shows that in o u r p e d i g r e e III10 is informative,
b e i n g h e t e r o z y g o u s A B ; c o n s e q u e n t l y D 2 1 S l l alleles can b e
t r a c k e d in the family to study co-segregation b e t w e e n these
two alleles and F A D gene. The b r o t h e r of III10, III8, is h o m o zygous B; III2 a n d III4, m o r e a d v a n c e d in age, are h o m o zygous A. C o n s e q u e n t l y , in the third g e n e r a t i o n of the pedigree, F A D allele co-segregates with allele B of the D 2 1 S l l
p r o b e . In the f o u r t h g e n e r a t i o n I V I Z at the b e g i n n i n g of the
disease, is h e t e r o z y g o u s A B ; the B allele of IV12 was inherited from his ( d e d u c e d ) m o t h e r A B . In the right part of the
pedigree, the B allele of III8, III10 and III13 was consequently
i n h e r i t e d f r o m the (also d e d u c e d ) g r a n d f a t h e r II6. U n d e r this
hypothesis, all individuals h a v i n g i n h e r i t e d the B allele from
diseased p a r e n t s (like I V I 0 , I V I 6 a n d V3) are at high risk of
d e v e l o p i n g A D in the future. T a b l e 1 shows the likelihood of
linkage to F A D at various r e c o m b i n a t i o n fractions relative to




Testing a D N A m a r k e r such as D 2 1 S l l , located in the

21q21 region of c h r o m o s o m e 21 [6, 9], should also p e r m i t a
b e t t e r definition of the location of the F A D gene a n d facilitate
future a t t e m p t s to isolate a n d characterize the gene affected.
F o r the time being, the existence of such a D N A m a r k e r ,
tightly linked to the F A D gene, raises the possibility of its use
for p r e s y m p t o m a t i c diagnosis for at-risk individuals. H o w e v e r ,
before these individuals can b e offered a n i n f o r m e d choice, it
will be necessary to confirm the results o b t a i n e d with additional p r o b e s from the same c h r o m o s o m a l region, and to define r e c o m b i n a t i o n frequencies b e t w e e n these m a r k e r s a n d
the F A D gene.

Acknowledgements'. This work was supported by grants from INSERM

(contract number 868027; Director: Denise Salmon), France-Alzheimer, and the MRES. We are grateful to Dr. Paul C. Watkins (Integrated Genetics) for the gift of recombinant pPW236B plasmid.

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Fig. 2. Variable EcoRI bands detected by probe D21Sll. A constant

band is present at 0.9 kb and variable bands at 2.9 (A allele) or 1.9 kb
(B allele)

Received March 10, 1988 / Received in revised form June 10, 1988 /
Accepted June 20, 1988