Hereditas 90: 59-83 (1979)

Satellite association in human metaphases.

A comparative study of normal individuals,
patients with Down syndrome and their parents
ALF HANSSON
Institute of Genetics, Lund, Sweden and The John F . Kennedy Institute,
Department of Medical Genetics, GlostruplCopenhagen, Denmark

HANSSON. A. 1979. Satellite association in human metaphases. A comparative study of normal

individuals. patients with Down syndrome and their parents. - Hereditas 90: 59-83. Lund, Sweden.
ISSN 0018-0661. Received September 27, 1978
Satellite association (SA) was studied in lymphocyte cultures from normal individuals, patients with
Down syndrome (DS) and parents, totally 455 individuals and 22,750 metaphases. SA tendency was low
in children under 10 years both in controls and DS patients, increased at the age of 10 to 19 years,
decreased in the controls aged 20 to 29 years, and again slightly increased in individuals over 30 years. A
drastical decrease was found in females over 70 years. Sex differences in the SA patterns were few,
except for the parents of DS,the mothers having a higher SA tendency, especially for chromosome 21,
also compared with the control individuals. This increase was more prominent in mothers aged 20 to 29.
The fathers of DS also, but not to the same extent, had higher SA tendency of chromosome 21,
compared to the controls. The occurrence of paternal non-disjunction may reflect this tendency.
A mathematical model for associations with two acrocentrics showed, strikingly, that in the DS
patients associations involving chromosome 21 were less frequent than expected.
The results support the evidence that high SA tendency increases the risk of non-disjunction. The
human SA patterns are, however, polymorphic with complex background.
Alf Hansson, Institute of Genetics. University of Lund,S-223 62 Lund. Sweden

Associations between the acrocentric chromo-
somes are very obvious in human metaphases.
The phenomenon, called satellite association (SA),
was first reported by FERCUSON-SMITH and HAND-
MAKER (1961), HARNDEN (1961) and OHNOet al.
(1961) and observed in mitotic metaphases. The
phenomenon of SA was also reported in meiosis
by FERCUSON-SMITH (1964).
Nonrandomness of the SA pattern
The question of non-random participation of the
acrocentric chromosomes in SA was discussed by
many authors even before the banding techniques
made it possible to identify the individual chromo-
some types within the D and G groups. A higher
tendency of the G group chromosomes to be
involved in SA has been reported by e.g. ZANG and
BACK(1966) and COHEN and SHAW (1967). On the
basis of autoradiographic identification, other au-
thors reported a random association pattern of the
D group chromosomes (SHAWet al. 1969;
NAKAGOME 1969; and CUEVAS-SOSA. 1970). Later it
has been shown however, by analysis of SA on
autoradiographically labelled or banded chromo-
somes that individuals with non-random SA pat-
terns are frequently found (PATIL and LUBS1971;
COOKE1971, 1972a; and NAKAGOME 1973). Fur-
thermore, by means of analysis of variance, JACOBS
et al. (1976) found that there was a highly signifi-
cant heterogeneity among individuals for the SA
tendencies of all the acrocentrics. These results
confirmed earlier observations indicating a stable
and specific association pattern in individuals
studied for SA (ZANGand BACK1967, 1968;
ROSENKRANZ and FLECK 1969; and HANSSON 1970a).
Influence of the age factor
The relation between age and SA has been dis-
cussed by many authors and their results have so-
metimes been rather contradictory. PROKOFIEVA-
BELCOVSKAYA et al. (1966, 1968) found that the
number of cells with SA, the number of associa-
tion complexes per cell and the number of
acrocentrics per SA complex increased with age
60 A.HANSSON
up to 40 years (n = 26) and then decreased again in
individuals over 53 years ( n = 12). Some of these
results Were supported by ARAGATZUNI (1967), who
studied six individuals aged 80 to 86 years.
BOGOMAZOV and DOROSHENKO (1968) studied indi-
viduals in the age range of 4 to 3 I years and found
the highest number of cells with SA and most SA
complexes per cell in four individuals aged 19
years. The other individuals studied were in the
age ranges of 4 to 9 years and 22 to 3 1 years (n = 4
and n = 14, resp.). The sexes were pooled in all
these studies.
GOODMAN et al. (1969) studied the SA tendency
counted as mean number of association com-
plexes per cell in females of three age groups:
newborn, 19-21 years and 67-93 years. They
found the highest number of SA complexes per
cell in the old age group and the lowest number of
complexes in the group between 19 and 2 1 years.
The most systematic and detailed report on the
age-related variation of SA was published by
COOKE (1972b). A sample of 123 normal females
was split into 12 different age groups from “0-5
years” to “over 55 years”. The highest number of
associations was found in age group 16-20 years
followed by a gradual fall to age 41-45. Above that
age, the results tended to fluctuate.
MATTEVIand SALZANO (1975) studied two age
groups: 11-13 years and 62-69 years of both
females and males (30 individuals of each sex in
both age groups). They found significantly more
cells with DD and DG associations in the younger
age group. In the pooled age groups the only sex
differences found was a higher number of cells
with DG associations in females. However, the
number of chromosomes involved in the SA com-
plexes did not vary significantly, neither with age
nor with sex.
L l E M et al. (1977) studied different aspects of SA
in 167 normal individuals and reported a low SA
tendency in newborns and in individuals older
than 50 years and an increased tendency in the age
ranges up to 20 years in males and 25 in females.
Also females had a comparatively higher associa-
tion tendency than males in most age groups.
Variations of the nucleolus organizing region
of the acrocentrics
Differences in the SA tendency were also found
between individual variants of homologous
acrocentrics (BAUCHINGER and SCHMID 1970; RoccHi
et al. 1971; GIGLIANI et al. 1972; DE CAPOA et al.
1973; S C H M l D et d. 1974; ZANKL and ZANG1974;
Herediras 90 (1979)
PHiLLiPs 1975; and HAYATA et al. 1977). The rela-
tionships between the tendency of SA and the
nucleolar organizing region (NOR) of the human
acrocentrics have been discussed from three dif-
ferent angles of approach:
( I ) The length of the nucleolar constriction and
double satellites. Several reports indicated that
chromosomes with increased length of the con-
striction (with or without double satellites) have
higher SA tendencies than chromosomes with
shorter or no visible nucleolar constriction
BR0GGER 1969; BAUCHINCER and SCHMlD 1969;
RoccHlet al. 1971; GimANiet al. 1972; D E C A P O A ~ ~
ai. 1972; ScHMio et al. 1974; and ZANKL and ZANG
1974). ORYE (1974) studied this relation in the G
group chromosomes and arranged the chromo-
somes in five groups according to the length of the
constriction. The SA tendency increased with the
length of the NOR but only up to a moderate
length. Very long nucleolar constrictions were
negatively correlated with the SA tendency.
(2) Variation in the number of genes for rRNA.
EVANS et al. (1974) studied in situ hybridization
and found that acrocentrics with large amounts of
rDNA were not preferentially involved in SA.
WARBURTON et al. (1976), on the other hand, re-
ported a positive correlation between the SA
tendency of an acrocentric chromosome and its
rDNA content. However, certain individual
chromosomes were clear exceptions.
(3) The size of N-band. HAYATA et al. (1977)
studied the SA tendency and the N-band of indi-
vidual acrocentrics (identified by means of Q-
band) after silver staining. They compared the
homologues of the individual acrocentrics and
found that the homologue with larger N-band was
involved in association complexes more often
than that with smaller N-band. These results were
confirmed by MILLER et al. (1977) who also com-
pared them with the reported (WARBURTON et al.
1976) correlation between the amount of rDNA
and SA. This comparison indicated that SA was
correlated rather with the size of N-band than with
the amount of rDNA and that, therefore, silver-
stained N -bands indicate function rather than
structure of the NOR’S.
Under the assumption that the SA tendency
reflects the capacity of the satellite stalk, the
association pattern should be a property
characteristic of any given acrocentric chromo-
some. This may explain the findings of HANSSON
(1970b, 1971) that, in the SA tendency of non-
banded D and G group chromosomes and the D/G
ratio of associated chromosomes, siblings of
Hereditas 90 (1979) SATELLITE ASSOCIATION I N H U M A N METAPHASES 61

monozygotic twin pairs showed a better con- cell division, the risk of non-disjunction is most
cordance than dizygotic twin siblings. PHILLIPS likely increased. The risk may be further aggra-
(1975) reported that the association pattern of a vated by the thin fibres occasionally visible be-
child usually was the result of the combination of tween the satellite regions of the chromosomes in
the specific SA tendencies of the acrocentrics SA’s. These connectives are found by light
received from the parents. Any exceptions found microscopy in some SA’s, and HENDERSON (1973),
should reflect the functional variability of the by means of hybridization of 3H-labeled rRNA to
NOR’s. There is some evidence of such a regula- human chromosomes, found silver grains between
tion of rDNA genes. MARSHALL et al. (1975) satellite regions without visible fibres, indicating
studied mouse-human hybrid cells, and those cells subvisible strands.
which retained mouse chromosomes and selec-
tively lost human chromosomes produced only
mouse 28s rRNA even when human acrocentric SA in relation to Down syndrome
chromosomes were present. CROCEet al. (1977) Several papers have reported evidence of an in-
studied other mouse-human hybrid cells, which creased SA tendency in mothers of Down
segregated mouse chromosomes, and only human syndrome (DS) children (HANSSON and MIKKELSEN
28s rRNA was produced. In the same manner, 1974; MATTEI1974; LEEDHAM. personal communica-
MILLERet al. (1976a, b) studied mouse-human tions). On the other hand, COOKE and CURTIS (1974)
hybrid cells after N-banding (silver staining) and and TAYSI (1975) were not able to confirm these
found that only mouse chromosomes showed N- results. HANSSON and MIKKELSEN (1976, 1978)
bands in cells segregating human chromosomes, studied QM variants of chromosomes 21 and SA
and only human chromosomes showed N-bands in in order to examine both the origin of the extra
cells segregating mouse chromosomes. HUBBELLchromosome 21 in DS and the significance of SA
and Hsu (1977) studied N-bands in neoplastic and on non-disjunction. Maternal non-disjunction was
normal human cells after silver staining. In spite of reported in 19 and paternal non-disjunction in 7
the fact that the tumor cells had between 11 and 18 families. The idea that high SA tendency may
acrocentric chromosomes, only 6-9 of them influence the risk of non-disjunction was strongly
showed N-bands, thus, about the same number as supported by the fact that the SA tendency of
in the normal cells. These results indicate that chromosome 21 of the parents with non-disjunc-
there exists a precise genetic regulation of the ac- tion was highly significantly increased compared
tivity of the NOR’s, which, in its turn, will in- with a control group.
fluence the phenomenon of SA.

Influence of nucleoli
It is well known that some human acrocentrics are
Materials and methods
preferentially involved in Robertsonian transloca- The SA patterns were studied in normal individu-
tions and non-disjunction, and, therefore, SA has als, parents of DS and patients with DS. The
received much attention because of its possible numbers of the different individuals studied were:
relation to these abnormalities. Most likely, SA’s
Female controls 142
and fusions of nucleoli are different manifestations
Male controls 124
of the same phenomenon, and acrocentrics be-
Mothers of DS 63
longing to one association complex will organize a
Fathers of DS 55
common nucleolus. Consequently, two frequently
Female DS 35
associated acrocentrics are more likely to be in-
Male DS 36
volved in translocations with each other. MIK-
KELSEN et al. (1975) thus found a significant in- The material, totally 455 individuals, included 40
crease of 13-21 (and 13-15) associations in a families, in which the parents and the child with
mother (with a normal chromosome constitution) DS were studied. One of these families had three
of a t(13q21q) carrier. Studies of fluorescent var- children, two of which had DS and the third
iants indicated that the translocation originated (chromosome constitution unknown) died with
from the mother. serious malformations two days old (Fig. 1). In
If sticky material from a large nucleolus or- another family, patients with DS were found in
ganized by several acrocentrics persists through two generations (Fig. 2).
62 A.HANSSON Hereditas 90 ( I 979)

Table I. The mean AI's f standard errors of the 68 h in Parker 199 medium with 20% fetal calf
acrocentrics in cells cultured 48 h (A) and 68 h (B). serum in the other.
Statistical t-test of the types of cultivation. n: The analyses for SA were performed on G -
number of individuals studied in each sample. banded chromosomes. Silver-staining according
The degrees of significance are denoted by to GOODPASTURE and BLOOM(1975) after identifica-
*=0.05>P>0.01, **=O.Ol>P>O.OOl, tion of the chromosomes by fluorescence (Q-
*** =P < 0.001 band) was also carried out in some of the individu-
als studied.
Individuals Chromo- Type of cultivation t The criteria for evaluating SA were the same as
studied some
No. A B in HANSSON (1970a). The tendencies of the indi-
vidual acrocentrics to be involved in SA were
Female 13 0.38f0.020 0.34f0.031 1.08
indicated in the association index (A1 = the
controls 14 0.37f 0.017 0.35f 0.023 0.67 number of associating chromosomes of a specific
(A: n=99) I5 0.34f0.017 0.31 k0.025 0.99 type divided by the total number of this type).
(B: n =43) 21 0.48 k 0.019 0.45 f 0.028 0.89 The calculations of the expected number of
22 0.35 f 0.014 0.32k 0.027 0.99
different association complexes with two chromo-
Male 13 0.32f0.011 0.31f0.027 0.34 somes in Figs 9-11 were made according to the
controls 14 0.33f0.012 0.32f0.030 0.31 formula presented in HANSSON (1975).
(A: n=93) 15 0.28 f 0.012 0.27 f 0.023 0.39 The weighted mean values were calculated ac-
(B: n = 3 I ) 21 0.34f0.015 0.33f0.033 0.28
22 0.31 f0.016 0.29f0.030 0.59
cording to the formula:
I -
c-xi
Mothers 13 0.33f0.020 0.36f0.016 -1.17
of DS 14 0.35 f 0.018 0.36f 0.028 -0.30 X,=m?
(A :n =43)
(B: n =20)
15
21
0.31f0.019 0.34f0.033 -0.79
0.38f0.027 0.44fO.044 -1.16
z- I
m9
22 0.30f0.018 0.35f0.030 -1.43
X,: the weighted mean
Fathers 13 0.36f0.017 0.30f0.028 1.83 Xi: the original means from which the weighted
of DS 14 0.33k0.018 0.32k0.030 0.29
(A: n =37) I5 0.33 f 0.020 0.29 f 0.037 0.95 mean was calculated
(B: n = / 8 ) 21 0.38 f 0.021 0.34f 0.047 0.78 mi: the standard errors of the original means
22 0.32 f 0.023 0.30f 0.035 0.48

Female DS 13 0.32f 0.017 0.29f0.025 0.99

(A: n = 2 / ) 14 0.31 f 0.014 0.30f 0.018 0.44
(B: n = / 4 ) I5
21
0.31f0.017
0.35 k 0.015
0.28f0.031
0.35 k 0.025
0.85
0.12
Results
22 0.30f 0.016 0.35 f 0.028 -1.55 Comparison between the two methods of cultivation
Male DS 13 0.32f 0.020 0.27 f 0.012 2.14; The material of the two laboratories are compared
(A: n=20) 14 0.32 f 0.018 0.30f 0.016 0.83 in Table 1 by means of the average AX'S for the
(8:n = / 6 ) I5 0.29f0.014 0.24f0.013 2.62;
21 0.39f0.022 0.34f0.016 1.84
different groups of individuals studied, one
22 0.37 k 0.025 0.28 f 0.021 2.79.; laboratory corresponding to type of cultivation A,
the other to B. In most cases, cells cultured for 48
h (A) had somewhat higher AI's than those
cultured for 68 h (B). However, only three
significant differences were found, all in the male
The normal material was composed of vol- DS. On the other hand, all the AI's of the mothers
untary healthy individuals from Iceland, of DS were higher in the cells cultured for 68
Copenhagen and the south of Sweden. The h. After all, most differences found between
Icelanders were originally involved in a family the laboratories were small and may well be due to
study for structurally variant chromosomes (QM). chance. Therefore, in the continued calculations
All analyses were arranged as blind studies, 50 and considerations, the material of the two
metaphases from each individual being analysed
laboratories were pooled in order to attain an
for SA, totally 22,750 metaphases. The cells were increase in the number of individuals in each
cultured in two laboratories according to standard
group studied.
whole-blood techniques: 48 h in Mc Coy medium
with 20% human serum in the one laboratory, and
 Hereditas 90 (1979) SATELLITE ASSOCIATION I N H U M A N METAPHASES 63

Table 2. The mean AI’s f standard errors in normal individuals. The material has been divided into age
groups. n: number of individuals studied in each age group

Normal Chromo- Age in years

individ- some
uals No. <I 1-9 10-19 20-29 30-39 40-49 50-59 60-69 B 70 Total

Female (n=19) (n=7) (n=10) (n=24) (n=19) (n=23) (n=17) (n=16) (n=7) (n=/42)
13 0.3OfO.019 0.30f0.036 0.38f0.024 0.30f0.020 0.34f0.022 0.33f0.019 0.35f0.018 0.35f0.040 0.33 f0.w0.32f0.012
14 0.30f0.023 0.32f0.030 0.36f0.023 0.31f0.022 0.32f0.021 0.32f0.021 0.35f0.026 0.34f0.022 0.36f0.062 0.33f0.008
15 0.27f0.021 0.31f0.040 0.31f0.038 0.25f0.017 0.30f0.023 0.30f0.023 0.26f0.022 0.35f0.024 0.34f0.066 0.28fo.01 I
21 0.31f0.022 0.33f0.043 0.38f0.027 0.30f0.024 0.34f0.027 0.34f0.027 0.38f0.028 0.42f0.029 0.30f0.026 0.34f0.013
22 0.28f0.028 0.29f0.023 0.36f0.050 0.24f0.019 0.36f0.028 0.36f0.028 0.32f0.026 0.37f0.025 0.39 f0.060 0.31f0.016
Male (n=7) (n=7) (n=6) (n=13) (n=18) (n=14) (n=/2) (n=32) (n=/5) (n=124)
13 0.26f0.039 0.30f0.019 0.36f0.041 0.31f0.023 0.39f0.025 0.35f0.022 0.32f0.035 0.35f0.020 0.36f0.032 0.33f0.013
14 0.31f0.046 0.31f0.034 0.35f0.036 0.32f0.017 0.31f0.027 0.36f0.032 0.36f0.033 0.34f0.017 0.35 f0.036 0.33f0.008
I5 0.29f0.023 0.25f0.029 0.29f0.041 0.27f0.025 0.32f0.033 0.36f0.024 0.34f0.038 0.37f0.019 0.41 f0.036 0.32f0.017
21 0.37f0.043 0.29f0.023 0.32f0.053 0.28f0.037 0.39f0.034 0.30f0.034 0.34f0.045 0.44f0.017 0.42 f0.031 0.37f0.018
22 0.30f0.024 0.22f0.024 0.28f0.073 0.22f0.024 0.37f0.033 0.31f0.033 0.39f0.032 0.36f0.019 0.37f0.032 0.31f0.021

AI, sex and age - Each acrocentric separately

Controls. - The individuals were also grouped
according to age. The different mean AI’s and the
standard errors of normal female and male indi-
viduals are given in Table 2. In the lowest age
group of the controls (under 1 year), all the
children were below 1 month of age. The AI’s can
differ very much between separate individuals,
which may influence the SA patterns if only a few
individuals are studied. All the same, some trends
were found. Thus, the highest A1 was most fre-
quently found for chromosome 21 (both female
and male) and the lowest for chromosomes 15
(female) and 22 (male). Statistical analysis con-
cerning sex differences in the SA tendency were
also performed. The five different mean AI’s in
each age group were tested with t-test. The only
significant differences (P< 0.05) were found for
chromosome 15 in the age group 40-49 years, and
for chromosome 21 in the age group 70 years and
above. In both cases, the AI’s of the males were
higher. However, in spite ‘of the fact that the
differences were non-significant, all the AI’s of the
females were higher than the corresponding in-
dexes of the males in the age groups between I and
19 years.

Table 3. The mean AI’s ? standard errors of parents of DS. The

material has been divided into age groups. n: number of individuals
studied in each age group
Parents Chromo- Age in years
of DS some
No. 20-29 30-39 P 40 Total

Mothers (n = 2 6 ) ( n =26) (n=ll) (n =63)

13 0.34 f 0.023 0.39f 0.022 0.37 f 0.040 0.37 f 0.015
14 0.35 f 0.015 0.38 f 0.022 0.37 f 0.035 0.37 f 0.012
15 0.32f 0.018 0.33 f 0.022 0.32 f 0.030 0.33 f 0.013
21 0.51 ? 0.020 0.44 f 0.02 1 0.51 f 0.038 0.48f 0.014
22 0.36f 0.019 0.32 f 0.023 0.36 f 0.026 0.34f0.013

Fathers (n = 2 1 ) (n = 19) (n = 1 5 ) (n =55)

13 0.34 f 0.024 0.35 f 0.025 0.35 f 0.03 I 0.35f 0.015
14 0.36f 0.018 0.38 f 0.022 0.33 f 0.024 0.36f 0.012
I5 0.34_+0.020 0.29f0.018 0.342 0.029 0.32f0.019
21 0.43 f 0.029 0.36 f 0.03 1 0.41 fO.037 0.40f0.019
22 0.32k 0.024 0.31 f 0.028 0.33 f0.021 0.32f0.014
64 A. HANSSON Hereditas 90 (1979)

Table 4 . The AI’s of the members of the families presented in Fig. 1 (family A) and Fig. 2 (family B)
Chromosome Family A Family B
NO.
1: I 1:2 I1:2 II:3 I: I I:2 11: I I1:2 Ill: I

13 0.37 0.41 0.28 0.37 0.3 I 0.29 0.32 0.33 0.35

14 0.31 0.43 0.39 0.42 0.27 0.38 0.32 0.32 0.2 I
I5 0.30 0.44 0.21 0.29 0.27 0.44 0.35 0.31 0.23
21 0.56 0.56 0.47 0.37 0.54 0.40 0.32 0.55 0.27
22 0.57 0.50 0.29 0.46 0.30 0.32 0.28 0.37 0.20

Parents of DS. - The parents of DS were grouped
into three age groups (20-29, 30-39 and 3 4 0
years). The mean AI’s of the five acrocentrics are
given in Table 3. The differences both between the
sexes and between the age groups were very small
except for chromosome 21. In all age groups, the
Of DS had higher Of chromosome 21
than the but the increase was O n l y
significant in the age range of 20 to 29 years
(p<0*05) and when the groups were ’‘eld
(P< 0.01).
The different AI’s found in the families with two
children with DS, as presented in Fig. I and 2 are
given in Table 4. In family A (Fig. l ) , both parents
(aged 56 and 61 years) had highly increased A1 of
chromosome 21. However the analyses of SA
were performed when their children were 27 and
22 years old. In family B (Fig. 2), the grandmother
(I: I ) and the mother (II:2) of a newborn child with
DS (1II:I) had increased A1 of chromosome 21.
The grandparents had also got a child with DS.
Both the grandparents and the parents were young
when their children with DS were born (26, 27, 21
and 22 years old, respectively).
with Ds. - The AI,s of the patients with
DS are recorded in Table 5. Most patients were
below two months of age and only one female and
one male were over 30 years (31 and 44, respec-
tively). Irrespective of sex, the highest A1 was
most frequently found for chromosome 21 and the
lowest for chromosome IS. A few differences in
the association patterns were found between the
sexes, e.g. for chromosome 21 in the age groups
above 10 years, but due to the high standard errors
of these mean AI’s, none of the differences were
significant.
Parents of DS versus controls. - Statistical
analyses were also performed between the parents
of DS and the control individuals in the same age
group. In order to get the same age distribution in
the control groups as in the parents of DS aged 40
years and more, the control groups were compos-
ed of individuals aged 40-49 years and of indivi-

I I

II 3

t Ill t
0 46,XX
0 46,XY 0 46,XX
0 47,XX,+21 0 46,XY
rn 47,XY,+21 0 47,XX,+21
Boy, with multiple malformations
Fig. I . Pedigree of a family with two children with Down Fig. 2. Pedigree of a family with Down syndrome in two
syndrome. subsequent generations.
Hereditos 90 (1979) SATELLITEASSOCIATIONI N HUMAN METAPHASES 65

Table 5 . The mean AI's k standard errors of the patients with DS. The material has been divided into
age groups. n: number of individuals studied in each age group
Patients Chromosome Age in years
with DS No.
<I 1-9 10-19 P 20 Total

Female (n = 16) (n = 8 ) (n=7) (n = 4 ) (n = 3 5 )

13 0.30f0.018 0.2920.034 0.28k0.024 0.39f0.064 0.31f0.014

14 0.30f0.015 0.30f0.027 0.32f0.025 0.36k0.039 0.31f0.011
I5 0.25f0.018 0.27f0.044 0.25f0.025 0.38+0.063 0.30f0.016
21 0.34f 0.018 0.35 f 0.032 0.36f 0.034 0.39f 0.053 0.35 f 0.014
22 0.29 f 0.027 0.32 f 0.025 0.35 f 0.039 0.35 f 0.044 0.32 f 0.016

Male (n =23) (n = 4 ) (n = 5 ) (n = 4 ) (n = 3 6 )
13 0.29f0.018 0.27f0.029 0.32f0.022 0.31f0.028 0.29f0.013
14 0.30f 0.015 0.24 f 0.026 0.36f 0.019 0.35 f 0.034 0.31 f 0.012
I5 0.26f0.013 0.25f0.045 0.29k0.010 0.26k0.053 0.26f0.011
21 0.35f0.017 0.33f0.046 0.46f0.047 0.32f0.021 0.36f0.015
22 0.31f0.024 0.36f0.047 0.35k0.046 0.32f0.029 0.32f0.018

duals from the age groups 50-69 years, randomly
selected (by computer). The AI's of those control
groups are presented in Appendix 1. It is obvious
that the parents of DS aged 2&29 years have the
highest AI's compared with the controls (Table 6).
The increase was most prominent in the mothers
and for chromosome 21. Chromosomes 15 and 22
also have significantly increased AI's in this age
group, but not to the same extent, at least for the
mothers. In the higher age groups, the only
significant differences between the parents of DS
and the controls were found for the increased A1
of chromosome 21 in the mothers of DS. The
different age groups within each sex were also
pooled and the AI's of all the parents (Table 3)
were compared with the controls (Appendix 1). In
the mothers of DS, all the acrocentrics had
significantly increased AI's, the increase being
most prominent for chromosome 21 (Table 6). In
the fathers of DS only chromosome 21 had a
significantly increased association tendency.

Table 6 . Statistical 1-test of the mean AI's in the Table 7 . Statistical t-test of the mean AI's in the
parents of DS and the control individuals. The patients with DS and the control individuals.
table gives the r-values, and the degrees of The table gives the r-values, and the degrees of
significance are denoted as in Table 1. A negative significance are denoted as in Table 1. A negative
t-value indicates that the mean A1 is higher in the 1-value indicates that the mean A1 is higher in the
controls controls
Individuals Chrome Age in years Individ- Chrome Age in years
compared some uals some
No. 20-29 30-39 240 Total compared No. <1 1-9 10-19 320 Total

Mothers of 13 1.48 1.69 0.77 2.38' Female 13 0.30 -0.12 -2.81' 1.26 -0.62
DSI 14 1.50 1.99 1.30 3.00'; DSl 14 0.07 -0.60 -1.23 0.97 -0.71
Female 15 2.82'' 1.09 1.28 3.08'' Female 15 -0.55 -0.74 -1.32 1.71 -0.89
controls 21 6.77.'' 2.72.' 3.91'. 7.78*** controls 21 1.06 0.21 -0.35 1.67 0.32
22 4.16". -0.83 1.79 2.26* 22 0.28 1.12 -0.17 1.11 0.96

Fathers of 13 0.84 -0.90 0.53 0.15 Male 13 0.67 -0.81 -0.78 0.03 -0.62
DSI 14 1.64 2.01 -0.99 1.20 DSI 14 -0.14 -1.67 0.34 0.78 -0.90
Male 15 2.28* -0.77 -0.37 -0.08 Male I5 -1.22 -0.17 0.09 -0.59 -0.50
controls 21 3.01'' -0.57 1.66 2.32* controls 21 -0.58 0.68 1.92 0.55 1.74
22 2.98** -1.33 -0.29 -0.09 22 0.18 2.56. 0.83 2.04 3.04''

5
66 A. HANSSON Hereditas 90 (1979)

Patients with DS versus controls. - Statistical

analyses were also performed between the pa-
tients with DS and the control individuals (sex and
age matched) and are presented in Table 7. The
control groups both for DS patients aged 20 years
and above and for the total material of DS were
composed of individuals randomly selected as
described above in connection with Table 6. The
AI's of these control groups are given in Appendix
2. The female DS in the age range of 10 to 19 years
had consistently lower AI's than the control indi-
viduals in the same age, and the female DS over 20
years had indexes higher than their controls.
However, the only significant differences found
were the increased A1 of chromosome 22 of the
male DS (aged 1-9 years, and total) and the low A1
of chromosome 13 in female DS aged 10-19 years.

-
AI, sex and age Acrocentrics combined
Weighted means of the At's of the five acrocentrics
were calculated for each group, according to the
formula presented above, and plotted in Fig.
3a-5a. In the parents of DS only chromosomes 13,
14, 15 and 22 were included because of the high
values of the index of chromosome 21. Weighted
means for the average number of SA complexes
per cell were also calculated for each age group
and plotted in Fig. 3b-5b. Furthermore, the mean
numbers of acrocentrics per SA complex were
C calculated and plotted in Fig. 3c-5c.
Controls. - In the controls (Fig. 3) the pattern was
largely the same in both sexes. The lowest as-
sociation tendencies (Fig. 3a) and numbers of SA
complexes per cell (Fig. 3b) were found in
newborns, children up to 9 years and individuals
in the age range of 20 to 29 years, whereas the
highest values were mostly found in individuals in
the age ranges of 10 to 19 years and above 30
years. However, the mean A1 and, to some ex-
t tent, also the mean number of SA complexes per
cell of females aged 70 years and above were
t lower than in the males. The difference was
significant for the mean A1 (P<O.O5). The mean
z' Ii
2.rI number of acrocentrics per SA complex was low-
est in newborns, females aged 40-49 and 70 and
.I 1-0 I040 2040 30-30 4040 5060 6 0 4 0 170
above (Fig. 3c). The most prominent sex differ-
A 9e ences were also found in these age groups.
Fig. 3 a-c. Different aspects of satellite association in normal
individuals; a: the mean association indexes (fstandard errors) Parents o f D S . - In all the age groups the mothers
of all the acrocentrics; b: the mean numbers ( 5 standard errors) of DS had both higher mean A1 and more SA
of association complexes per cell; c: the mean numbers complexes per cell than the fathers, and the differ-
(+standard errors) of acrocentrics per association complex. ences increased with age (Fig. 4a and b). Another
obvious trend may be seen in Fig. 4c: the size of
Hereditas 90 (1979) SATELLITE ASSOCIATION I N H U M A N METAPHASES 67

the SA complexes increased with age in the moth-

ers of DS but decreased in the fathers.
Patients wirh DS. - The patients with DS are
38 l a
.36
presented in Fig. 5 . In the female DS aged 20 years
and above, the mean A1 (Fig. 5a), the number of
SA complexes per cell (Fig. 5b) and the mean
'
3

.32
it
number of acrocentrices per SA complex (Fig. 5c)
were higher than in the lower age groups. In the 30
male DS, this was the case even in the patients in
the age range of 10 to 19 years at least for the latter
-< .28

two parameters. Still, probably due to the low .26

number of individuals studied and the high
standard errors of the means, the only significant I I
sex differences were the increased mean A1 of
the females aged 20 years and above (P< 0.05) and
the high number of acrocentrics per SA complex
in males aged 10 to 19 years (P< 0.001).
Parents of DS versus controls. - The results in
Fig. 3-5 were also submitted to a systematical
statistical comparison. As the mean AI's of the
parents of DS did not include chromosome 21, the
statistical comparisons between the parents and
the controls were performed on other indexes than Y
those plotted in Fig. 3a, and calculated merely 0
L 1.2
from chromosomes 13, 14, 15 and 22. These new a
indexes of the controls were only slightly lower n
than the original. Both the young (20-29 years)
5 11

and the old ( 3 4 0 years) mothers of DS had Z

significantly increased indexes (P< 0.01 and
< 0.05, respectively) compared with the controls C C
(Fig. 3a and 4a). Furthermore, the young parents
had also significantly more SA complexes per cell
than the controls (Fig. 3b and 4b; mothers: U r t
+
v)

'I f
P < 0.001, fathers: P < 0.05). On the other hand, \ 2.4
II)
the older parents had significantly increased
numbers of chromosomes per SA complex (Fig. ;
3c and 4c: mothers aged 30-39 and 3 4 0 years: 510
P < 0.01 and fathers aged 30-39 years: P< 0.05). E 2.3
e
x
Patients with DS versus controls. - The patients u
with DS were statistically compared with the Y
0
controls. The mean A1 (all acrocentrics) of the 2.2
0
n
Fig. 4 a-c. Different aspects of satellite association in parents of s
Z
patients with Down syndrome; a: the mean association indexes
2.1
(fstandard errors) of chromosomes 13, 14, 15, 22; b: the mean
numbers (fstandard errors) of association complexes per cell;
a1 1.9 10-19 120
c: the mean numbers (+ standrad errors) of acrocentrics per
association complex. A9=
Fig. 5 a-c. Different aspects of satellite association in patients
with Down syndrome; a: the mean association indexes Fig. 4. Fig. 5.
(fstandard errors) of all the acrocentrics; b: the mean numbers
(fstandrad errors) of association complexes per cell; c: the
mean numbers (-+standarderrors) of acrocentrics per association
complex.
68 A.HANSSON Hereditas 90 ( 1 979)

Femole
Male

13-13 14-14 15-15 13-14 13-15 14-15 21-21 22-22 21-22 13-21 13-22 1 4 - 2 1 14-22 15-21 15-22

Type of asrociolion

Fig. 6. The mean numbers (kstandard errors) of different satellite association pairs found in 50
metaphases of the normal individuals. All sizes of association complexes were included, but the
complexes with more than two chromosomes were divided into association pairs possible
(for details see text).

1
18. 1 1 1 Fernole

16

14

% 12

ul
0
2 10
a
c 8
c

$ 6
-B
0
U
n
5
Z

13-13 14-14 15-15 13-14 13-15 14-15 21-21 22-22 21-22 1 3 - 2 1 13-22 1 4 - 2 1 14-22 15-21 15-22

T y p e of ossociation

Fig. 7. The mean numbers (k standard errors) of different satellite association pairs found in
50 metaphases of parents of patients with Down syndrom. All sizes of association complexes
were included. but complexes with more than two chromosomes were divided into association
pairs possible (for details see text).
Hereditas 90 (1979) SATELLITE ASSOCIATION IN H U M A N METAPHASES 69

Fernole
Male

13-13 14-l4 15-15 13-U 13-15 14-15 21-21 22-22 21-22 13-21 1312 14-21 14-22 15-21 15-12

Type of ortociation

Fig. 8. The mean numbers (?standard errors) of different satellite association pairs found in
50 metaphases of patients with Down syndrom. AU sizes of association complexes were
included, but complexes with more than two chromosomes were divided into association pairs
possible (for details see text).

female DS aged 1&19 years was significantly

lower, and the mean A1 of the female DS aged 20
years and more was significantly higher than the
corresponding indexes of their control groups
(Fig. 3a and 5a). These mean AI's of the female
DS agreed much better with the indexes of the
control groups with 10 years younger individuals.
However, to a lesser extent, these mean AI's also
well agreed with the indexes of the controls aged
10 years more than the patients, which may sup-
port the idea of premature ageing of the patients
with DS. The mean AI's of the female DS up to 9
years age and of all the age groups with male DS
were close to the similar inhexes of the controls.
Furthermore, most differences between the con-
trols and the patients were rather small for the
mean number of SA complexes per cell and the
mean number of acrocentrics per complex (Fig.
3b, 3c, 5b and 5c). Only one highly significant
difference was found (P<O.OOI), the number of
acrocentrics per SA complex being higher in
newborn female DS.
Analysis of SA complexes
Most SA complexes were composed of only two
chromosomes. In Fig. 6 8 , all association com-
plexes are presented irrespective of size, but
complexes with more than two chromosomes
were divided into "association pairs". Thus, an
association complex with the constitution:a-b-c
was divided into a-b, a-c and b-c association
pairs. In this manner, it was possible to study
whether some chromosomes were involved in the
same SA complexes more often than others and
thus could be assumed to organize a common
nucleolus more often. The numbers of association
pairs in the figures represent the means of all the
individuals studied of each type. The correspond-
ing results of the different age groups are pre-
sented in Appendix 3-5.
Two homologous chromosomes can only form
one type of association, two non-homologous
chromosomes may form four different types, and
two non-homologous chromosomes, one of which
70 A.HANSSON Hereditas W (1979)

Table 8. Statistical t-test of the sex factor in the Table 9. Statistical t-test of the incidence of
incidence of different association pairs of acro- association pairs of acrocentric chromosomes in
centric chromosomes in controlh, parents of DS parents of DS and controls (age groups pooled).
and patients with DS (Fig. 6-8, respectively). Negative t-values indicate higher frequencies in
Negative t-values indicate higher frequencies in the controls. The degrees of significance are
males. The degrees of significance are denoted denoted as in Table 1
as in Table 1
Association Individuals compared
pairs
Association Individuals studied Mothers Fathers Female Male
pairs of DS/ ofDS/ DSI DS/
Controls Parents Patients Female Male Female Male
of DS with DS controls controls controls controls

13-13 1.09 0.46 0.00 13-13 -0.74 -0.49 -1.34 -1.53

14-14 I .03 -1.86 0.97 14-14 -0.30 0.43 -0.34 -0.34
15-15 -1.41 0.69 -0.35 15-15 -1.48 -1.76 -1.12 -2.47'
13-14 0.73 0.23 -I .23 13-14 2.76'' 2.17' -1.50 -0.32
13-15 -0.51 0.43 0.07 13-15 2.09' 0.78 -1.35 -1.26
14-15 - I . I4 0.26 -0.11 14-15 2.95" 1.05 -1.90 -3.82***

2 1-2 I -0.51 0.87 -0.19 2 1-2 I 3.06" 1.88 5.31*** 6.64'"

22-22 1.50 -0.82 - I .37 22-22 -2.53' -0.72 -2.44. 0.27
2 1-22 -0.43 1.40 -1.03 2 1-22 4.54'" 2.76'' 3.99'" 5.Ol"'

13-21 -0.63 2.92'' 1.14 13-21 6.17"' 1.94 2.43' 3.20"

13-22 0.40 0.98 0.65 13-22 I .55 0.33 -1.11 0.14
14-21 -1.30 1.59 -1.19 14-21 5.78.'. 2.83.. 3.36" 3.33''
14-22 I .49 -0.65 0.00 14-22 0.57 1.23 -1.24 -0.11
15-2 I -2.94" 1.28 0.92 15-21 5.57''' 1.35 2.16' I .56
15-22 -1.57 0.90 -0.22 15-22 1.21 -0.08 -1.24 -0.56

is trisomic may form six different types of associa-
tions (for explanation see HANSSON 1975). These
principles are well illustrated by the lengths of the
bars in the diagrams of Fig. 6-8, in which the
incidences of SA pairs in the controls, the parents
of DS and the patients with DS are presented,
respectively. It should be noted that in the control
individuals, the parents of DS, as well as the
patients with DS, the most frequent association
pair was 21-22. Also the other pairs involving
chromosome 21 and the pair 13-14 were very
frequent.
Injluence of rhe sex factor. - Statistical analyses
of the possible influence of the sex factor were
performed on the results diagrammatically pre-
sented in Fig. 6-8 (Table 8). Very little sex differ-
ences were found. In the controls the only
significantly deviating value was the high number
of 15-21 associations of the males. In addition, all
other types of association pairs with chromosome
21 were somewhat more frequent in the males
than in the females, but the trend was probably
negligible. In the parents of DS, all types of
association pairs with chromosome 2 1 involved
were found more frequently in the mothers than in
the fathers. However, only the increase of 13-21
associations was significant. In the patients with
DS, no significant differences were found.
Statistical analyses between the sexes were also
performed in the different age groups, but only
few differences were significant and may be
caused by chance.
Parents of DS versus controls. - In Table 9, the
numbers of different association pairs of the par-
ents of DS and of the patients with DS were
statistically compared with those of the control
individuals. All types of association pairs with
chromosome 2 1 were significantly more frequent
in the mothers of DS than in the female controls.
Also, some other pairs (13-14, 13-15 and 14-15)
were more frequent in the mothers of DS but not
to the same extent. In contrast, there were
significantly more 22-22 associations in the con-
trols, whereas all other pairs with chromosome 22
were more frequent in the mothers of DS. The
number of association pairs were also studied for
separate age groups. The most prominent differ-
ences between the mothers of DS and the controls
Hereditas 90 (1979)
1 13-13 14-14 15-15 13-14 13-15 14-15
$ *$ 3" *"
* *
Fig. 9. The mean differences (k stadard errors) between the observed and the expected numbers of association complexes with two
chromosomes found in normal individuals. A positive difference implies that the observed number of association complexes was
more frequent than expected. The information under the diagram indicates if the differences deviated significantly from 0. The
dcgreesofsignificanceare denotedby *=0.05>P>0.01,*+=0.01>P>O.o01, ***=P<O.o01.
were found for the age groups,above 30 years
(Appendix 6).
In the fathers of DS, the number of 21-22, 14-21
and 13-14 association pairs were significantly in-
creased compared with the male controls (but not
to the same extent). The other association pairs
with chromosome 21 were also more frequent in
the fathers of DS although not significantly so
(Table 9). For the individual age groups, only few
significant differences were found between the
fathers of DS and the controls (Appendix 6).
Patients with DS versus controls. - The patients
with DS were also compared with their controls.
Irrespective of sex, most association pairs with
chromosome 2 1 were significantly more frequent
in the patients. In contrast, all other types of
association pairs but two in the males were more
frequent in the controls (Table 9). The same
phenomenon was found in the different age
groups, but because of the small number of indi-
viduals in some of the groups, the results were
somewhat more influenced by chance (Appendix
7).
Incidence of different types of associations,
observed versus expected
A high number of associations between two
acrocentrics can result when either these chromo-
somes associate frequently with any acrocentric
or when these chromosomes have a specific
tendency to associate with each other. The ex-
pected number of SA complexes with two
SATELLITE ASSOCIATION I N H U M A N METAPHASES 71
2l-21 22-22 21-22 13-21 13-22 14-21 14-22 15-21 15-22
** * *
*
chromosomes was calculated by means of the
formula presented in HANSSON (1975). Since, be-
cause of the great variety of such complexes, only
few larger SA complexes were found of each type,
it was decided to limit the calculations to associa-
tions with just two chromosomes. The mean dif-
ferences between the number of association com-
plexes observed and those expected on the basis
of the formula are plotted in Fig. 9-1 1. Positive
differences imply that the observed number of
association complexes is higher than expected. In
case this increase is statistically significant, it may
be assumed that there is a specific tendency of the
chromosomes involved to associate with each
other.
Controls. - Irrespective of sex, all homologous
associations between the D group chromosomes
(DD associations) were fewer than expected in the
controls (Fig. 9). However, associations between
13 and 14 and between 13 and 15 were more fre-
quent than expected. The homologous GG pairs
occurred in about the expected frequency, but
pairs of 21 and 22 were significantly more frequent
than expected. Most types of DG associations
were less numerous than expected, but in general,
the trend was very weak, and only 13-21 and
13-22 associations of the females were signifi-
cantly fewer than expected.
Parents of DS. - In the parents of DS, the
deviations from the expected number of associa-
tion complexes were more marked than in the
controls (Fig. 10). In spite of this, the same
general association pattern prevailed as in the
72 A.HANSSON Heredifas 90 (1979)

t a Female
Male

1 13-13 H-14 15-15 13-14 13-15 14-15 21-21 22-22 21-22 13-21 13-22 14-21 1 4 1 2 15-21 1 5 4 2

Type of association
* * * * $2 3 * * **
3* * +** 3 * ** *
+* +* **
Fig. 10. The mean differences (fstandard errors) between the observed and the expected numbers of association complexes with
two chromosomes found in parents of patients with Down syndrome. A positive difference implies that the observed number of
association complexes was more frequent than expected. The information under the diagram indicates if the differences deviated
significantly from 0. The degrees of significance are denoted by * =O.OS >P>O.OI, ** =0.01 >P>O.Ool, *** =P<O.OOI.

controls, i.e. fewer homologous and more non-
homologous associations than expected. The
homologous associations, both for G group
chromosomes and for D group chromosomes were
less numerous than expected in both sexes. A
significantly increased number of 2 1-22 associa-
tions was noticed for the mothers of DS and the
fathers as well, the increase being most prominent
in the mothers. The number of DG associations
was significantly lower than expected in three
cases (14-22 in the mothers, 13-21 and 14-22 in
the fathers) and higher in one (14-2 I in the fath-
ers). The only signifcant difference between the
sexes was found in 14-15 associations, where the
number of complexes observed was higher than
expected in the mothers of DS but not in the
fathers.
Patients with DS. - All types of non-homologous
DD associations were found significantly more
often than expected in the patients of DS irrespec-
tive of sex (Fig. 11). However, the most striking
characteristic pattern was that all types of com-
plexes with chromosome 21, except the 21-22
pair, were much more infrequent than expected.
This pattern was most obvious for 21-21 associa-
tions. As was the case for the control groups and
to some extent also the parents of DS, there was a
very good agreement between the sexes of the
patients with DS.
Influence of age. - The differences between the
observed and the expected numbers of SA com-
plexes with two chromosomes were also studied
in the different age groups. These results are
presented in Appendix 8-10. There were no com-
mon age dependent patterns, and the few varia-
tions in the association patterns of the different
age groups may well be caused by chance.
Discussion
The great variability in the SA pattern among
human individuals has caused much discussion of
the grounds for and the significance of the SA
phenomenon. The fact that the amount of silver-
stained material in the NOR’S and the SA
tendency of the acrocentrics are strongly corre-
lated (MILLERet al. 1977), indicates that the SA
pattern might illustrate functional properties of the
individual acrocentrics.
Influence of time of cultivation and
methodology factors
Several authors (NANKIN 1970; ZHDANOVA 1972;
MATTEVI and SALZANO 1975) have reported a higher
SA tendency in lymphocytes cultured for 48 h than
in those cultured for 72 h. These findings would be
easily explained, under the assumption that the
cells have a higher SA tendency in the first mitosis
after the stimulation with phytohemagglutinin
(PHA) than in the second. If this is true the
agreement between the results of the two
laboratories in the present experiments (Table 1)
Hereditas 90 (1979)
40
2.0
::
c
0.0
!!
+e
g -2.0
s
3 -4.0
-6.0
13-13 U-14 15-15 1344 13-15 14-15
Type of association
* *** *** ** ** **
**
Fig. I I . The mean differences (?standard errors) between the observed and the expected numbers of association complexes with
two chromosomes found in patients with Down syndrome. A positive difference implies that the observed number of association
complexes was more frequent than expected. The information under the diagram indicates if the differences deviated significantly
from0.Thedegreesof significancearedenoted by *=0.05>P>0.01,
would mean that the same cell generation after
PHA-stimulation was studied although the times
of cultivation were different, 48 h and 68 h. This
explanation was also supported by our observa-
tion that in the 68 h culture only few metaphases
were found after 48 h.
JACOBSet al. (1976) found no significant differ-
ences in SA patterns between duplicate cultures
set up from the same blood sample or between
different slides from the same culture. They also
studied the influence of different observers on SA
in the cells analysed, and also here they were
unable to demonstrate any significant differences.
Growth factors, age and SA
COOKE (1972b) has suggested that some growth
factors, sex hormones or thyroxin, may influence
the SA tendency. This problem was investigated
by NILSSON et al. (1975) who reported an increased
association tendency in patients with hyper-
thyroidism. Such factors may underlie some of
the variation among the different age groups in
this material.
SATELLITE ASSOCIATION IN HUMAN METAPHASES 73
0 Fornolo
m Mob
ff
21-21 2*22 21-22 13-21 13-22 14-21 14-22 15-11 15-22
$* 3* * ** ** * * ** * * ** **
**=0.01>P>O.o01, ***=P<O.o01.
Several contradictory observations on the influ-
ence of age on SA have been reported. Many of
the reports, however, are difficult to compare
because the parameters recorded were not the
same. Aspects like the frequency of cells with SA,
the number of association complexes per cell, the
number of acrocentrics per SA complex and/or
the number of acrocentrics involved in association
per cell were discussed in the different reports.
Also, in some materials, the sexes have been
pooled.
Many reports have indicated a low association
tendency in newborns and in children below 10
years’ age and then an increase in the teens, which
is in very good agreement with our results. This
pattern may, at least partly be due to sex hormonal
influence. The drastical decrease in SA tendency
in individuals between 20 and 29 years found in
the present study has also been reported in
females by GOODMAN et al. (1969) and to some
extent also by COOKE(1972b) and LIEMet al.
(1977). The slight increase of the SA tendencies
we found in age groups above 30 years is well in
accord with certain other reports, e.g. GOODMAN et
74 A.HANSSON Herediras 90 ( I 979)

al. (1969) who compared females in the age ranges more than 29 years (especially the mothers) had
of67-93 years and 19-21 years, and COOKE(1972b) larger SA complexes than the controls. This is a
who compared females in the age ranges of 3 6 4 0 factor that may also increase the risk of nbn-dis-
years and above 55 years. COOKE, however, also junction, since the higher number of acrocentrics
found a slight decrease in association tendency in will organize larger nucleoli, which are likely to
females in the age range of 41 to 54 years. Most produce more persistent nucleolar material than
studies of SA in individuals of high age deal with small nucleoli. Thus the difference in SA between
females and report a decrease in the SA tendency, the controls and the parents has different dimen-
just as we found in females aged 70 or more but sions in different age groups. This finding may, at
not in the somewhat younger age groups in the least partly, explain the somewhat contradictory
present study. The differences among the age reports which have been published concerning the
groups ranging from 30 to 70 years have been connection between a high SA tendency and non-
small in most studies. In view of the fact that the disjunction. Age factors also, like the decrease in
SA pattern can differ considerably among indi- the chiasma frequency just mentioned, may con-
viduals irrespective of age and sex, these fluctua- tribute to non-disjunction in old parents of DS.
tions are most likely due to chance. The low SA Furthermore, the analyses of SA in the parents
tendency of newborns and young children might be aged more than 40 years, and in a few parents aged
related to the observation of several authors that 30 to 39 years, were performed more than one
lymphocytes cultured for 72 h also show low year, in one family even more than 20 years, after
degree of SA. Therefore, the age of the lympho- the birth of the child with DS. The SA patterns of
cytes and/or the time since the previous cell these parents may have changed during this time.
division must be of significance. The finding of chromosomal variants, e.g. by
means of fluorescence microscopy has indicated
that the extra chromosome 21 in DS is of paternal
Increased A1 and non-disjunction
origin in 25 to 30 % of the cases (MIKKELSEN et al.
The strongly significant increase of the A1 of 1976; MAGENIS et al. 1977; HANSSON and MIKKELSEN
chromosome 21 in mothers of DS indicated that 1978). These observations may reflect the in-
the SA tendency will influence the risk of non-dis- creased SA tendency observed in the fathers of
junction. This idea is further supported by the DS .
results in the families with two DS patients (Fig. 1 PHILLIPS (1975) found that the association pat-
and 2), in whom the SA tendency of chromosome tern of children usually was the result of a com-
21 was highly increased. In addition, the high A1 bination of the patterns of their parents. In our
of chromosme 21 will increase the indexes of the material, however, the A1 of chromosome 21 in
other acrocentrics involved in the same associa- the children with DS was lower than expected
tion complex. However, other factors must also from the high indexes of the parents of DS. The
influence non-disjunction. For instance, non-dis- differences between the parents of DS and the
junction in mothers at late fertile age may be controls were much greater than between the pa-
caused by a decline in the number of chiasmata. tients with DS and their controls (Table 6 and 7).
The results of STENE et al. (1977) indicate that These results may support the idea that a gene-
males aged over 55 years run a higher risk that dose mechanism underlies the phenomenon of
their newborn children will have DS. LUBS(1976) SA, as has been suggested and discussed in an
reported that females aged under 20 years also run earlier paper (HANSSON 1975). That discussion was
an increased risk having a child with DS, actually based on the findings that the intact homologues
about the same risk as females between 35 and 40 of the acrocentrics taking part in Robertsonian
years. In these young mothers, sex hormonal translocation were involved in SA more fre-
factors may contribute to the high values of mean quently than expected.
AI, of mean numbers of SA complexes per cell
and of the size of SA complexes found in this age
group (Table 2 and Fig. 3). Different SA complexes of all sizes
The most conspicuous increase of the A1 of Chromosomes frequently involved in the same
chromosome 21 was found in mothers of DS aged association complex should have a higher
20 to 29 years. The increase in the higher age tendency to form Robertsonian translocations
groups, too, was significant h t not to the same with each other than with chromosomes which are
extent. However, the parents of DS when aged rarely found in the same SA complex. JACOBS et al.
Hereditas 90 (1979)
(1974) analysed the distribution of acrocentrics
involved in Robertsonian translocations in man
and demonstrated an excess of t(DqDq) and a
deficiency of t(DqGq) and t(GqGq). Furthermore,
the involvement of acrocentrics in each group was
non-random. Thus, there was an excess of
t( 13q14q) as compared with other translocations
among D group chromosomes. Similar investiga-
tions have shown that t(14q21q) was the most
frequent translocation involving D and G group
chromosomes (ROWLEYand PERCAMENT 1969;
COHEN1971; HECHT and KIMEIERLINC 1971; MIK-
KELSEN 1973 and HAMERTON et al. 1975). In all types
of individuals studied, the most frequent DD as-
sociation was between chromosome 13 and 14,
and the most frequent DG association was be-
tween chromosome 14 and 21 (Fig. 6 8 ) . These
results are in agreement with the non-random
involvement of acrocentrics in Robertsonian
translocations. On the other hand, association
pairs 13-21 were frequently found, even though
the t(13q21q) is a rare type of translocation be-
tween chromosome 21 and a D group chromo-
some. This shows that other factors besides SA
must be involved.
SA complexes with two chromosomes involved
and preferential association
MAITEI(1974) reported a similar excess of 21-22
association pairs in both controls and parents of
DS as found in our material. Also the other types
of association pairs were found mostly in the same
frequencies in both our materials and in that of
MATIXI.JACOBSet al. (1976) found, however, no
significant evidence for preferential association be-
tween any acrocentric when applying to their data
a mathematical model for pairwise associations.
All the results plotted in Fig. 9-11 are mean
values, and according to the construction of the
formula, the sum of all these means equals 0.
Thus, the occurrence of high positive or negativ
deviations from the expected number for a certain
association type will influence the values of the
other associations. This means that small non-
significant differences are most likely negligible.
According to the results of the control individu-
als in Fig. 9, chromosome 13 had a specific
tendency to associate with chromosome 14 and
15. Likewise, chromosome 21 and 22 also had a
specific tendency to associate with each other.
Associations between homologous chromosomes
were found less frequently (the D group) or to the
same extent as expected (the G group) according
SATELLITE ASSOCIATION IN HUMAN METAPHASES 75
to the formula above. Thus, there was no
tendency to “somatic pairing” of the short arms
of these chromosomes but these findings might
mean that the NOR and/or other parts of the short
arms of the acrocentrics produce different sub-
stances quantitatively and/or qualitatively of im-
portance for SA. There were no specific trends in
the association of the different types of DG
chromosomes except that associations 13-21 and
13-22 were few in the females. The high number of
13-21 association pairs in Fig. 6 must, therefore,
depend on factors other than a specific tendency
to association between these chromosomes, e.g.
high AI’s of the chromosomes in question. It may
also be a matter of influence that the results in Fig.
6 include SA complexes with more than two
chromosomes, divided into association pairs.
In the parents of DS (Fig. lo), the association
pattern found was about the same as in the con-
trols, but the mean differences were more prom-
inent. In the patients with DS (Fig. 11). however,
quite another kind of association is found. The
very low number of observed associations involv-
ing chromosome 21 (especially 21-21) strongly
supported the idea that a gene-dose compensation
mechanism might be active in the formation of
SA.
Conclusion
Our results illustrate the high complexity of the
phenomenon of SA. Genetic as well as non-genet-
ic factors must be of importance. Moreover, the
association patterns may reflect the activity and
the regulation of the NOR’S. The results indicate
that SA play an important role in non-disjunction.
Still, at least in individuals at advanced reproduc-
tive age, other factors also must be involved in
the chain of events leading to the formation of
aneuploid gametes.
Acknowledgments. - The author wishes to express his gratitude
to Dr. Margareta Mikkelsen for having generously supplied a
considerable part of the material and for valuable discussions.
He would also like to express sincere thanks to Professor Albert
Levan and Dr Gunnar Ising for constructive criticism and to
Mrs. Hanne Poulsen and Mrs. Birthe Jespersen for technical
assistance. This work has been supported by grants from the
Danish Medical Research Council, The Danish National Board
of Social Welfare, Department of Mental Retardation, and from
“Prenatalforskningsnamnden” and the Nilsson-Ehle Foundation
in Sweden.
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78 A.HANSSON Hereditas 90 (1979)

Appendix I . The mean AI's f standard errors of Appendix 2 . The mean AI's f standard errors of
control groups constructed for statistical com- control groups constructed for statistical com-
parisons with DS parents aged 2 4 0 years, and parisons with DS patients aged 3 2 0 years, and
total number. The mean AI's k standard errors of total number. The mean AI's f standard errors of
the control groups for the younger DS parents are the control groups for the younger DS patients are
presented in Table 2. n: number of individuals presented in Table 2. n: number of individuals
Control Chromosome Age in years Control Chromosome Age in years
individuals No. individuals No.
P 40 total B 20 total

Female (n =29) (n = 72) Female (n = 3 0 ) (n =66)

13 0.34f0.015 0.32f0.011 13 0.30f0.017 0.32f0.012
14 0.32f0.019 0.32f0.012 14 0.31 f 0.018 0.32f0.012
I5 0.28 f 0.016 0.28f 0.01 I 15 0.27 f 0.017 0.29f0.013
21 0.33+0.021 0.32f0.014 21 0.29 f.0.020 0.34f 0.015
22 0.30f 0.021 0.30f 0.014 22 0.30f.0.019 0.29f0.017

Male (n = 18) (n =45) Male (n = 16) (n =36)

13 0.33 f.0.018 0.34f0.013 13 0.31 fO.021 0.31 f0.018
14 0.36+0.021 0.34f0.014 14 0.32 f 0.020 0.33 f 0.021
15 0.35 f 0.020 0.33 f 0.015 I5 0.30+0.027 0.27 f 0.014
21 0.33 f 0.027 0.34 f 0.019 21 0.30+ 0.030 0.31 f.0.020
22 0.34+ 0.023 0.32f0.017 22 0.24 2 0.022 0.25 f.0.016
Hereditos 90 (1979) SATELLITE ASSOCIATION IN HUMAN METAPHASES 79

Appendix 3. The numbers of different association pairs found in 50 metaphases of each control
individual. The means f standard errors were calculated for the different age groups. The numbers of
individuals studied in each group are given in Table 2
Control Ass. Age in years
individ. pair
<I 1-9 10-19 20-29 30-39 4&49 50-59 60-69 b 70

Female 13-13 1.4f0.35 3.1f0.81 4.7f0.56 3.6f0.91 2.8f0.41 2.3f 0.40 3.1f0.54 2.8f0.68 2.0f 0.50
14-14 2.1 f 0.33 2.550.78 4.8f0.91 3.3f0.65 2.1f0.36 2.1 f0.41 3.2 f 0.59 2.6f 0.55 3.7f 1.04
15-15 0.9f 0.25 2.5f0.93 1.7f0.33 2.5f 1.20 2.4fO.51 1.9f 0.40 I .8 f 0.45 3.3 f 0.59 2.7 f 0.78
13-14 10.4f 1.04 17.1+ 1.60 15.2f2.37 16.5f2.31 10.6f1.24 11.65 1.03 13.6f1.25 11.8f1.38. 14.7 f 2.66
13-15 8.8f 0.93 12.6f 2.29 14.8f2.60 13.8f2.40 10.6f 1.51 10.4f 1.22 8.7f 1.35 12.3f 1.63 14.8f3.51
14-15 8.7 f 0.99 10.8f 1.52 10.7f 1.28 13.3f2.38 9.2f 1.32 10.0f 1.08 9.3f 1.48 10.9f 1.59 l5.7f3.57
21-21 2.8 f 0.76 3.Ok0.65 5.52 1.82 3.0f 1.24 3.850.58 3.3f 0.56 4.0f 0.75 4.5 f 0.78 2.0 f 0.47
22-22 2. I f 0.53 3 . l f 1 . 4 1 4.7f1.26 2.5f1.00 3.2f0.55 2.7f 0.41 2.5f0.61 3.9f0.58 5.Of I .07
2 1-22 9.7 f I .26 13.8f 2.77 17.2f4.58 1O.Of3.80 12.0f 1.67 11.9f 1.50 13.45 1.67 14.3f 1.25 11.4f 1.71
13-21 9.7f 1.16 13.8f 1.10 15.Of3.56 12.4f2.43 11.0f1.51 10.2f 0.95 12.7f 1.31 l5.lf2.44 10.6f 1.42
13-22 8.0f 1.07 12.4f2.54 9.7f2.14 9.6f2.03 12.5+1.15 8.6f 1.14 10.4f 1.34 I l . l f 1.22 13.8f 2.20
14-21 8.6f 0.90 14.6f 1.91 14.3f2.97 10.5f2.90 10.8fO.84 12.0f 1.23 12.9f 1.60 12.6f 1.63 10.8 f I .22
14-22 8.2f 1.04 9.6% 1.49 13.3f2.33 8 . 9 f 1.67 ll.lfO.98 10. I f 1.05 10.6f 1.19 13.1f 1.64 15.4f 2.86
15-21 8.6f 1.16 11.32 1.81 14.3f 1.31 11.3f2.83 8 . 8 t 1.23 8.8f0.87 9.2f0.97 11.6f 1.86 10.2f 1.06
15-22 7.9+ 1.06 8.4f 1.70 12.0f 1.86 9.6f 1.76 9.8f 1.41 8.4f 1.15 8.6f 1.12 13.3f 1.82 17.0f 3.39
Male 13-13 1.4 t 0.69 3.6f0.87 2.8f 1.16 2.6f 0.82 3.3 f 0.59 2.1f0.29 1.5f0.47 2.6? 0.41 2.2 f 0.58
14-14 2.6f 0.84 4.0f0.63 3.4f 1.54 2.3f 1.01 I.lf0.35 2.8t0.77 3.5f0.49 2.0f 0.32 2.1 f 0.66
15-15 2 . l f 1.18 1.4f0.51 2.0f0.00 2.4 f 0.63 0 . 8 f 0.25 1.9f0.37 3.0f0.59 3.3 f 0.44 3.0f0.66
13-14 6.lf 1.62 14.6f 1.94 15.4f2.06 14.6f 3.60 12.9+ 1.61 14.8f2.02 12.8f 1.85 11.5 f 0.77 9.6f 2.27
13-15 7.9f 1.52 12.4f2.68 11.6f 1.60 12.6 f 2.47 8.6f 1.67 13.9f 1.26 12.5f2.13 12.5f 1.41 11.2f 1.66
14-15 1O.Of 2.29 11.4fO.60 10.4f2.20 11.6f 1.59 8.0f 2.67 14.2f 1.75 11.2f 1.24 10.7 f I .OO 12.8 f 2.09
21-21
22-22
4.4 f I .04 3.4f 2.23 2.2f0.73 3.4f 1.12 3.9 f 0.72 2.2f0.78 2.8f0.76 4.5 f 0.44 4.5 0.93 +
2.0f 0.38 2.6f0.93 1.2f0.73 1.8 f 0.68 2.3 f 0.45 2.0f 0.63 3.5 f 0.59 3.4f0.50 1.7f0.56
21-22 12.4f 0.75 11.4f4.91 8.4f2.25 8.8f 2.09 12.5f 3.46 10.5f2.18 I3.SfZ.qO 15.6f0.92 13.8 f I .95
13-21 7.6f0.90 11.6f2.06 11.4t2.38 I1.9f 2.23 12.9f 1.72 11.0f 1.79 11.3f 1.77 13.6f 1.04 13.2f 1.95
13-22 6.9f0.91 8.8f2.18 16.6f4.43 8.3 f 2.72 13.8f 2.72 9.8f0.93 11.32 1.24 10.2 f 1.08 10.92 0.89
14-21 12.0f 1.98 15.2f 3.77 13.4+ 2.01 12.6f 1.78 11.6f 2.26 12.4f 1.62 11.9f 1.45 12.7f 1.15 12.8+ 1.53
14-22 9.1 f2.01 1O.Of 1.33 8.4f4.03 7.5f 1.44 7.1f1.62 11.0f2.00 13.7f 1.84 9.8 f 0.79 9.12 1.10
15-21 I l . l f 1.56 11.8f3.10 8.622.62 13.5 f 3.26 8 . 5 f 1.80 10.2f 1.25 11.5f2.38 14.5f 1.11 13.6f 2.13
15-22 8.4 f 0.87 12.2f 2.82 9.2f 3.44 7.1 f 2.39 9.9f 2.50 10.9f 1.57 14.4f 1.87 12.4f 1.05 11.1f1.31
80 A.HANSSON Hereditas 90 (1979)

Appendix 4 . The numbers of different association pairs found in 50 metaphases of each DS parent.
The means k standard errors were calculated for the different age groups. The numbers of individuals
studied in each group are given in Table 3
Ass. Mothers aged in years Fathers aged in years
pair
20-29 30-39 B 40 20-29 30-39 L 40

13-13 2.0 f 0.36 3. I f 0.58 l.0f0.55 I .8 f 0.44 2.6f 0.72 2.4 f 0.44
14-14 1 . 5 f 0.26 2.9+ 0.42 2.4f 1.44 2.8f 0.38 3.1 f 0.63 2.5 f 0.50
15-15 1.5 f 0.17 2.0f 0.29 I .2 f 0.58 1.6f0.37 1.Of 0.24 2.1 f 0.52
13-14 13.0f 1.14 18.5f 1.53 14.2+ 3.15 13.9f 1.14 l6.5f 2.01 14.5 f 1.93
13-15 12.35 1.11 15.2f 1.35 1 O . O f 1.76 13.4f 1.17 13.0f 1.48 12.5 f 1.98
14-15 l2.7f 0.74 15.5f 1.87 12.0f3.19 13.5f 1.18 12.8f 1.54 13.5 f 1.73

21-21 5.7f0.57 3.9f 0.50 6.6f 2.20 5.1 f 0.76 3. I f 0.68 4. I f 0.96
22-22 2.3 f 0.32 I .3 f 0.22 1.8 f 0.92 2.6f 0.45 2. I f 0.49 1.7 f 0.42
21-22 17.2 f I .36 16.4f 1.40 23.0f 3.56 16.6f 1.57 14.6f 2.29 15.2 f 1.75

13-21 16.5f 1.17 18.7f 1.50 21.4f4.01 14.65 1.39 13.5f 1.83 13.7f 1.59
13-22 11.1f 1.19 12.7f 1.31 12.222.22 11.65 1.22 9.9f 1.11 11.3f 1.79
14-21 17.5f 1.06 17.0f 1.34 25.8f4.99 16.7f 1.56 15.6f 1.45 13.9f 1.88
14-22 10.7f0.73 10.5 f 0.93 11.62 3. I4 11.3 f 0.99 11.6f 1.50 10.9f 1.44
15-21 14.8f 1.01 14.6f 1.33 17.0f4.04 14.5f 1.23 11.4f 1.03 12.4f 1.69
15-22 9.8f 0.86 9.6f 0.87 10.8 f 2.76 11.72 1.08 10.2f 1.10 10.3f 1.11

Appendix 5 . The numbers of different association pairs found in 50 metaphases of each DS patient.
The means k standard errors were calculated for the different age groups. The numbers of individuals
studied in each group are given in Table 5
Ass. Female DS aged in years Male DS aged in years
pair
<I 1-9 10-19 > 20 <I 1-9 10-19 120

13-13 l.6f 0.24 2. I f 0.46 2.3 f 0.75 1.5 f 0.29 1.4 f 0.38 1.5t0.29 2.8f 1.07 2.8f 1.32
14-14 I .9 f 0.40 2.6f 0.81 2.3 f 0.56 2.8f 0.63 1.3 f 0.29 2.3f 0.95 2.8 f 0.86 3.6f 1.17
15-15 0.9f 0.26 I. I f 0.34 0.9f 0.30 1.5f0.87 0.8f0.18 1.5f0.29 1.4 f 0.51 2.2f 1.20
13-14 10.5f 0.97 9.4f 1.72 8.5 f 1.44 l l . 5 f 1.55 10.6 f 0.97 1O.Of 1.78 12.2 f 2.60 14.6f 2.49
13-15 8 . 8 f 1.58 7.0f 2.37 5.5 f 0.85 14.5 f 2.33 8.3 f 0.91 4.8f 1.44 8.8f 1.32 9.0f 3.35
14-15 8. I f 0.93 6.9f2.21 6.5 f 1.32 10.5f 0.96 7.7f 0.55 5.3f 1 . 1 1 9.2 f 0.66 9.8f 3.15

21-21 7.7f 0.94 8.0f 2.60 10.3f 1.99 9.3f 2.75 7.3 f 0.65 6.8+ 1.75 13.4f2.38 9.5f 2.90
22-22 1.3f0.29 1.6f 0.53 1.8f0.56 2.8f 0.25 1.9f 0.34 2.8f 1.31 2.0f 0.89 1.3f0.25
2 1-22 16.3f 1.41 22.4 f 3. I I 18.5 f 2.37 1 8 . 8 t 2.06 19.2f I .70 19.0 f 4.92 24.8 f 4.76 17.3 f 2.75

13-2 I 14.6f 1.48 14.9f 2.48 15.3 f 2.80 20.0f 2.68 12.5f 1.11 10.5 ?r I. 19 19.4f 1.86 16.5f 1.55
13-22 7.5f0.98 10.0f 1.72 9.6f 1.76 1 I . O f 3.58 7.4 f I .04 7 . 0 f 0.71 10.2f 1.28 10.3 f 1.49
14-21 13.7f0.89 16.I f 2.74 16.0f 1.24 17.3f 3.52 1 5 . O f 0.89 14.0f 1.29 24.4f 4.27 18.3f 1.80
14-22 7.42 0.95 7.6f 1.29 10.0f 1.07 ll.5f2.90 6.8 f 0.58 8.5 f 2.18 11.8f 2.96 12.5f 1.26
15-21 12.2f 1.03 15.3f 3.01 13.5f 1.10 15.3 k 3.25 I I . O f 0.68 14.3 t 5.20 18.4f 2.09 12.3f 1.65
15-22 7. I f 0.84 8.7f2.10 9.0f 1.58 8.3 f 1.44 7.8 f 0.94 8.Of 1.87 8.8f 1.07 7.5f 1.19
Hereditas 90 (1979) SATELLITE ASSOCIATION IN HUMAN METAPHASES 81

Appendix 6 . Statistical analysis (t-test) of the numbers of association pairs in the DS parents and the
controls. The table gives the r-values, and the degrees of significance are denoted by * =O.OS >P>O.Ol,
** = 0.01 > P> 0,001, *** = P < 0.001. A negative t-value indicates that the number of association pairs
found is higher in the controls
Ass. Mothers of DWfemale controls aged in years Fathers of DS/male controls aged in years
pair
20-29 30-39 240 20-29 30-39 840

13-13 -1.64 0.42 -2.28. -0.86 -0.75 0.87

14-14 -2.57. I .64 0.13 0.46 2.78" -1.18
15-15 -0.83 -0.68 -1.20 -1.10 0.58 -1.64
13-14 -1.36 4.01'" 0.88 -0.19 1.40 0.30
13-15 -0.57 2.27' -0.44 0.29 1.98 -0.22
14-15 -0.24 2.75'' 0.57 0.96 1.56 0.05

21-21 1.98 0.13 1.so I .26 -0.81 1.34

22-22 -0.19 -3.21" -0.80 0.98 -0.30 -1.77
21-22 1.78 2.02' 3.081' 2.98'. 0.51 1.57

13-21 1.52 3.62*'* 2.77'' I .03 0.24 1.39

13-22 0.64 0.11 1.13 1.11 -1.33 0.51
14-21 2.27' 3.92*** 2.941' 1.73 I .49 0.78
14-22 0.99 -0.28 0.46 2.17' 2.04' -0.50
15-21 1.16 3.20'. 2.14' 0.29 1.40 0.38
15-22 0.10 -0.12 0.81 I .75 0.11 -1.47

Appendix 7. Statistical analysis (t-test) of the numbers of association pairs in DS patients and the
controls. The table gives the t-values, and the degrees of significance are denoted as in Appendix 6.
A negative r-value indicates that the number of association pairs found is higher in the controls
Ass. Female DWfemale controls aged in years Male DS/male controls aged in years
pairs
<I 1-9 10-19 P 20 <I 1-9 10-19 3 20

13-13 0.47 -1.07 -2.56' -2.32' 0.00 -2.29' 0.00 0.07

14-14 4.B 0.09 -2.34' -0.48 -1.46 -1.49 -0.34 1.01
15-15 0.00 -1.41 -1.79 -0.64 -1.09 0.17 -1.18 0.07
13-14 0.07 -3.28" -2.42' -1.58 2.38' -1.75 4% 0.I5
13-15 0.00 -1.70 -3.40' * 0.54 0.23 -2.50. -1.35 -0.65
14-15 -0.44 -1.45 -2.288 -0.79 -0.98 4.83+" -0.52 -0.23

21-21 4.05." 1.87 1.78 2.07' 2.36' I .20 4.50'' I .95

22-22 -1.32 -1.00 -2.10 0.20 -0.20 0.12 0.70 -0.92
21-22 3.49" 2.06 0.25 1.86 3.66" 1.09 3.11' 1.24

13-21 2.61' 0.41 0.07 2.29' 3.43'' -0.46 2.65' I .67

13-22 -0.34 -0.73 -0.04 0.22 0.36 0.09 -1.39 0.55
14-21 4
.
03
' 0.45 0.53 1.51 1.38 -0.30 2.33' 2.49'
14-22 -0.57 -1.01 -1.29 0.48 -1.10 -0.59 0.68 2.67'
15-21 2.32. 1.14 -0.47 1.20 -0.06 0.41 2.92' 0.00
15-22 -0.59 0.1 I -1.23 -0.63 -0.47 -1.24 -0.11 -0.20

6
82 A.HANSSON Hrreditas 90 (1979)

Appendix 8. The differences between the observed and the expected numbers of SA complexes with
two chromosomes as found in the control individuals. The mean differences f standard errors were
calculated for each age group. A positive difference implies that the observed number of association
complexes were more frequent than expected. The numbers of individuals studied in each group are
given in Table 2

Control Ass. Age in years

individ. pair
<I 1-9 10-19 20-29 30-39 40-49 50-59 60-69 a70

Female 13-13 -0.5t0.18 1.5f0.29 O.OfO.39 -0.lf0.31 - 0 . l f 0 . 2 3 0.2fo.30 - 0 . l f 0 . 2 4 -0.3f0.26 -0.5fo.30

14-14 -0.3f0.14 1.520.56 -0.5f0.30 0.3f0.28 -0.1f0.20 -0.5f0.17 - 0 . I f 0 . 2 2 -0.1+0.28 -0.3+0.3.(
15-15 -0.5fO.IS -0.4f0.21 -0.2f0.35 -0.6f0.19 0.4f0.31 -0.5f0.14 0.0f0.14 O.Of0.24 -0.6f0.34
13-14 0.8f0.49 -0.7f0.83 1.0f0.42 I.0f0.46 0.2f0.27 0.7f0.44 1.6f0.44 -0.IfO.40 I.Of0.69
13-15 0.7f0.44 I.2f0.79 -0.3f0.93 0.0f0.31 0.4f0.38 1.0f0.54 4 . 2 f 0 . 4 2 l . l f 0 . 5 0 0.5fo.74
14-15 0.2f0.42 -1.2f0.98 0.250.94 0.3f0.63 -0.3f0.41 0.4f0.37 -0.lf0.61 -0.8fO.33 0.2f0.43
21-21 -0.3f0.30 2.Ofl.08 -0.2f0.34 -0.1f0.14 0.2-10.28 0.3f0.26 0.4f0.54 0 . I f 0 . 4 0 0.1fO.28
22-22 -0.3f0.20 0.5f0.43 -0.3f0.33 0.3f0.23 0.1f0.16 0.1f0.20 -0.2f0.27 -0.1+0.20 0.5i-O.53
21-22 1.3f0.51 -0.3f0.51 0.9f0.75 0.8f0.65 0.2f0.43 1.3f0.39 1.0f0.47 0.2fO.44 0.4f0.75
13-21 - 0 . l f 0 . 3 9 -1.9f0.77 -1.Of0.48 0.2f0.49 -0.4f0.39 -1.OfO.35 -0.4fo.54 0.1fo.42 0.2fo.47
13-22 -0.5f0.32 -2.Of0.53 l.OfO.86 -0.9f0.42 0.3f0.41 -1.0f0.34 -0.9f0.37 -0.4f0.42 -1.Ofo.72
14-21 -0.3f0.37 1.2f0.93 0.8f0.69 -1.3f0.40 0.3f0.42 -0.4k0.43 -0.4f0.40 0.5f0.53 -0.1f0.35
14-22 - 0 . l f 0 . 4 9 O.Jf1.03 -1.lfO.63 -0.4f0.32 -0.3fO.35 O.Of0.39 -0.8f0.38 0.4f0.44 -0.5fo.77
15-21 O.Of0.53 -0.4f0.65 -0.2f0.66 0.5f0.35 -0.6f0.42 -0.lfO.36 -0.7fO.38 -0.5f0.70 -0.2fO.65
15-22 O.OfO.45 0.6f0.68 0.3t0.47 0.0f0.40 -0.2f0.38 -0.5f0.33 0.7f0.49 -0.1f0.44 0.8fO.84
Male 13-13
-0.4 f 0.23
14-14
-0.4 f 0.18
15-15
0.1 f 0.21
13-14
-0.6f 0.54
13-15
0.3 f 0.40
14-15 l.Sf0.59
21-21
0.Of 0.36
22-22
-0. If 0.31
2 1-22
0 . 9 f 0.58
13-21
0.9f 0.65
13-22
0.2 f 0.52
14-21
0.Of 0.52
14-22 -0.3 f 0.47
15-21
-1.3f0.28
15-22
- 1 . 1 f 0.56
Hereditas 90 (1979) SATELLITEASSOCIATION I N H U M A N METAPHASES 83

Appendix 9. The differences between the observed and the expected numbers of SA complexes with two
chromosomes as found in the DS parents. The mean differences k standard errors were calculated for
each age group. A positive difference implies that the observed number of association complexes was
more frequent than expected. The numbers of DS parents studied in each group are given in Table 3

Ass. Mothers aged in years Fathers aged in years

pair
2&29 30-39 3 40 20-29 30-39 a 40

13-13 -0.4 f 0. I6 -0.9f0.14 -0.8 f 0.30 -0.6f0.17 -0.5f 0.19 -0.4f0.11

14-14 -0.7fO.l I -0.3 f 0.22 - 0 . 1 f 0.44 -0.4 f 0.23 -0.3 f 0.25 -0.2f 0.35
15-15 -0.6f0.16 -0.4f0.15 -0.3f 0.13 -0.7 f 0.20 -0.7f 0.1 I 0.2k0.29
13-14 0.4+ 0.39 I .7 f 0.40 -0.3 f 0.73 0.9 f 0.45 0. I f 0.29 0. I ? 0.52
13-15 0.6f 0.32 0 . 2 f 0.34 0.6 f 0.58 0.7f 0.41 I .4 f 0.4 I -0.2f 0.32
14-15 0.7 f 0.26 0.5 f 0.36 0.3 f 0.56 0.2f 0.33 -0.4 f 0.32 -0.3 f 0.47

21-21 0.3 f 0.34 -0. I f 0.36 -0.62 0.46 -0.2 f 0.20 -0.4 f 0.22 -0. I f 0.48
22-22 -0.3 f 0.19 -0.6f 0.12 -0.7f0.14 0.0 f 0.23 -0.3f0.21 -1.2fO.12
21-22 0.8 f 0.49 1.7 f 0.41 l.2f0.66 0.4f 0.43 0.9 f 0.63 I. I f 0.81

13-21 -0.7 f 0.34 0. I f 0.36 0.6f 0.56 -0.2 f 0.32 -0.4f 0.42 0.4 f 0.63
13-22 0.3 f 0.38 - 0 . 1 f 0.36 0.4f 0.57 -0.2f0.21 -0.4f 0.38 0.3 f 0.65
14-21 0.5 f 0.33 -0.4f 0.37 1.3f 0.83 0.5 f 0.39 I.If0.44 0.2 f 0.62
14-22 -0.5 f 0.3 I -1.Of 0.30 -1.lf0.28 -0.8 f 0.30 -0. I f 0.42 0.2 f 0.52
15-2 1 -0.2 f 0.40 -0.3 f 0.45 - I . O f 0.73 0. I f 0.40 0. I f 0.41 -0.7 f 0.47
15-22 -0.4 f 0.30 0.3 f 0.36 0.3 f 0.48 0.5 f 0.25 0.2f 0.42 0.6 f 0.45

Appendix 10. The differences between the observed and the expected numbers of SA complexes with
two chromosomes as found in the DS patients. The mean differences k standard errors were calculated
for each age group. A positive difference implies that the observed number of association complexes
was more frequent than expected. The numbers of parents studied in each group dre given in Table 5

Ass. Female DS aged in years Male DS aged in years

pairs
<I 1-19 10-19 2 20 <I 1-9 10-19 320

13-13 0.2f0.18 0.2 f 0.30 0.4 f 0.44 -0.8f 0.37 -0.2fO.IS 0.4 f 0.67 0.3 f 0.38 -0. I f 0.42
14-14 0.3f 0.27 0.8f0.53 0.3 f 0.30 -0.3f 0.27 -0.2f 0. I9 0.2 f 0.43 0.4f 0.29 0.5 f 0.55
15-15 -0.3f0.15 -0. I f 0.24 -0.4 f 0. I4 -0.2 f 0.2 I -0. I f 0.15 0.3 f 0.24 -0.4 f 0.24 0.2f0.25
13-14 1 . 1 f 0.30 0.7f 0.65 2.2f0.67 0.9f 0.35 1.8 f 0.41 2.2f0.31 0.8f 0.86 1.1f 1.05
13-15 I . I f 0.37 0 . 6 f 0.77 - 0 . 1 f 0.76 2.8f 1.14 l.5f0.43 0. I f 0.43 0.2f0.41 -0. I 50.65
14-15 1.4f 0.31 O.Of 0.34 -0. I f 0.54 0. I f 0.67 0.9f 0.3 I -0.7+ 0.57 0.6f 0.88 I .O f 0.71

21-21 -3.2f0.32 -3.8f0.83 -2.8 f 0.90 -2.2 f 0.31 -3.3 f 0.28 -2.7f 1.89 -3.3 f 0.72 -2.3f 1.32
22-22 -0. I f 0. I5 -0.7f 0.21 -0.6f 0. I8 0.5f 0.38 -0. I f 0.19 -0.4f 0.48 -0.3 f 0.25 -0.3f 0.07
2 1-22 +
I .O 0.48 1.1f 1.26 -0.8f 1.16 -1.2f 0.95 I . I f 0.46 0.3 f 0.73 -0.2f 0.43 -0.5k 1.41

13-21 -0.8f0.40 -0.9f 0.56 -1.6f 1.05 -0.8 f 0.45 -1.2f 0.31 -2.4f 0.31 -0.9f0.82 -1. I f 0.78
13-22 0.4 k 0.37 1.2f0.57 0.8 f 0.52 0.4f 0.30 0.2 f 0.38 1.1fO.83 l.lf0.46 l.7f 0.90
14-21 -1.2f0.29 -0.8 f I .06 -0.8f 1.05 -0. I f 0.75 0. I f 0.33 -0.8f 0.25 -1.Of 1.24 - I . I f 0.68
14-22 0.5 f 0.40 0.4 f 0.54 0.72 0.88 1.3f 1.08 -0.2f 0.22 0.3 f 0.98 0.82 1.07 0.7 f 0.79
15-21 -1.lfO.42 0.2 f 0.72 -0.2 f 0.64 -1.3 f 0.82 -1.5f0.44 0.2f 2.35 0.1 f 0.74 -0.3 f 0.67
15-22 0.9? 0.33 I. I f 0.52 3.0f 1.02 0.6f 0.81 1 . O f 0.5 I 1.9f0.95 l . 4 f 0.29 0.9 f 0.68