CYTOGENETIC P R A C T I C E IN A MENTAL R E T A R D A T I O N
CLINIC*
F. R. SERGOVICH, Ph.D.
While cytogenetics has been a long
established area of biological investiga­
tion, relatively little effort has been
devoted in the past to an elucidation of
the role that chromosomes play in hu­
man developmental errors. With the
evolution of new cytological techniques
and concepts within the last decade, a
variety of human syndromes have been
found to have a chromosome aberration
as their etiology. This of course is of
general medical interest but one could
question the significance of this field in
psychiatric practice in general and for
those actively engaged in mental retard­
ation work in particular. This question
can partially be elucidated by a consider­
ation of frequencies of chromosome
anomalies in various human populations,
Recent work by Dr. D. H . Carr at
the University of Western Ontario
Medical School, on the frequency of
chromosome abnormalities in spontane­
ous abortion, indicates that these me­
chanisms account for a significant pro­
portion of foetal loss. Approximately
20% of recovered foetuses or embryonic
sacs showed some form of chromosome
abnormality. Generally, these abnormal­
ities were similar to those found in viable
infants, although somewhat more bizarre
forms were occasionally encountered.
Buccal smear surveys on newborn popu­
lations in Berne (1), Edinburgh (7) and
Winnipeg (8), indicate that 2/1,000
males are chromatin positive (i.e., have
at least one extra sex chromosome), that
0.37/1,000 females lack a sex chromo­
some and about 1.8/1,000 females possess
three X chromosomes rather than two.
"Presented at the Canadian Psychiatric Association
Annual Meeting, Halifax, N.S., June 18, 1965.
'Director, Cytogenetic Department, Children's Psy­
chiatric Research Institute and Lecturer, Department
of Anatomy, University of Western Ontario, London,
Canada.
Canad. Psychiat. Ass. J. Vol. 12 (1967)
35
1
Since on the average about 1/600 live-
born infants are mongoloid and they
clearly possess a chromosome abnormal­
ity, one can estimate that chromosome
anomalies can be found in the newborn
with a frequency in the order of 0.6%.
The true frequency of abnormalities in
newborns may be somewhat higher since
these estimates are primarily based on
buccal smear tests and not on direct
chromosome studies. It is clear that a
controlled study of the chromosomes in
a large population of newborns is neces­
sary to determine this frequency pre­
cisely.
When one compares the frequency of
sex chromosome anomalies in institutions
for the mentally retarded, a marked in­
crease over that of the general popula­
tion is observed. Maclean et al (6) found
1.07% chromatin positive males and
0.24% double positive females (i.e., pos­
sessing three X chromosomes instead of
the normal pair). Similarly, the fre­
quency of sex chromatin positive males
among male sexual psychopaths (9)
tends to approach that of the institution­
alized mental retardate, i.e., approximate­
ly a ten-fold increase over the newborn
population. Exact frequency figures of
all types of chromosome abnormalities
(i.e., sex chromosome plus autosomes)
are unknown and must await a chromo­
some survey of a large population of
mental retardates. However, a reasonable
estimate can still be made. A cross sec­
tion of a given large institution such as
we have in Ontario indicates that be­
tween 10 and 15% of the patients are
affected with mongolism. It is a well-
established fact that this disease is caused
by the presence of an extra small chro­
mosome. Another 1 or 2% of patients
have a sex chromosome abnormality,
36 CANADIAN PSYCHIATRIC ASSOCIATION JOURNAL Vol. 12, No. 1

while possibly another 1 or 2% may degree of mental and physical impair­

have a different autosomal anomaly. ment to warrant structured care.
Based on these estimates we can see that This discussion will be primarily con­
as many as 15 to 20% of institutionalized cerned with the cytogenetic methods
mental defectives have a demonstrable and principles which are currently in use
chromosome abnormality. at The Children's Psychiatric Research
Data obtained from animal and plant Institute in London, Ontario. This labor­
cytogenetics suggests that gross chromo­ atory has a threefold function — diag­
somal changes such as we are able to nosis, genetic counselling and basic re­
detect in mental retardates are relatively search. An attempt at this time will be
infrequent. Small changes, such as in­ made to illustrate how the integration
versions of whole segments, as well as of relatively straightforward techniques
duplications and deletions of small seg­ can be used to obtain data which are
ments, appear to be the more frequent meaningful from both a practical and
aberrations that have been found to have theoretical point of view.
occurred naturally. In clinical cytogene­
The use of sex chromatin determina­
tics our technical methods are still too
tions will be discussed first because of
crude to detect these types of variations
the simplicity in technical procedure as
except in the most highly favourable
well as the ease in interpretation of re­
circumstances. For example, if there
sults. This procedure is used routinely
were a small duplication on an area of
on all patients coming to our clinic. A
a chromosome that is normally very
scraping of buccal mucosa cells is smear­
small, it would appear quite striking to
ed on a slide, fixed in alcohol, stained
an experienced observer. Similarly, if a
with a nuclear dye, then mounted in
relatively small chromosome was deleted
permanent medium. Cells from a normal
for a segment it would produce a mark­
male and a normal female processed in
ed distortion of the chromosome com­
this manner are shown in Figure 1.
pared to the normal member of the pair.
'iVIale nuclei' (1 a) and 'Female nuclei'
On the other hand, loss of a segment of
( l b ) are readily distinguishable in this
similar size in a large chromosome or an
type of preparation. Sex chromatin
insertion of a similar segment into this
(arrow) is the large, darkly stained body
chromosome could easily be overlooked
which is closely applied to the nuclear
because of normal variability in chromo­
membrane and can readily be noticed
some size (even between homologues)
in the female cells, while the male nuclei
during the different stages of mitosis. It
are relatively vesicular and homogenous.
therefore is conceivable that for each
The number of these highly characteris­
type of chromosome abnormality which
tic bodies is directly related to the num­
can be detected in an affected person,
ber and type of sex chromosomes pre­
there may be many more which escape
sent in the karyotype. A normal male
notice because of technical inadequacy,
karyotype is shown in Figure 2. Twenty-
and it seems reasonable to assume that
two pairs of autosomes are present as
the figure of 15 to 20% given for the
well as a single X and a single Y chromo­
frequency of chromosome aberration in
some. The paired chromosomes are
a retarded population is a minimal figure
arranged in order of decreasing lengths
and the true frequency may be higher.
as well as into groups, the latter group­
At the present time, a chromosome
ings being formulated on the basis of
abnormality of one form or another
ease of identification as well as general
appears to be the most frequent recog­
similarity. The important feature in this
nizable etiological factor in the causa­
illustration is that a normal male has one
tion of mental retardation — particularly
large X chromosome and one small Y
in those individuals with a severe enough
chromosome and shows no sex chroma-
February 1967 CYTOCKN K T I C PRACTICE 37

FIGURE 1 (a) Chromatin negative buccal smear nucleus. The nucleus is relatively vesi­
cular with no significant chromatin masses, (b) Chromatin positive nucleus from a normal
female. Sex chromatin (arrow) is an intensely staining mass approximately lu in diameter
that is applied to the inner surface of the nuclear membrane. The presence of one mass
indicates that two X chromosomes are in the chromosome complement.

tin in interphase nuclei of buccal smears.
In contrast, the next Figure (Fig. 3)
shows a chromosome array of a normal
female. A l l the autosomes, (i.e., non-sex
chromosomes) are morphologically sim­
ilar to that of the male except that two
large X chromosomes are present and
interphase nuclei of buccal smears show
a single sex chromatin body. A simple
rule can be formulated from a study of
correlations between the number of sex
chromatin bodies in interphase nuclei
with the number of sex chromosomes
present in the chromosome complement.
This rule, by the way, has withstood all
testing in a number of conditions so far.
We can say that the Y chromosome does
not contribute to sex chromatin forma­
tion and that a single sex chromatin mass
results from the presence of two X
1 instances, however, sex chromatin does
chromosomes. I f three X chromosomes
were present, there would be two sex
chromatin bodies; if there were four X
chromosomes present, there would be
three sex chromatin bodies, and so on.
The next Figure (Fig. 4) illustrates this
point. This is a buccal smear from a boy
suffering froin a variant of Klinefelter's
syndrome. Three sex chromatin masses
can be seen in many cell nuclei. On the
basis of the rule governing the relation­
ship between the number of sex chroma­
tin bodies and the number of X chromo­
somes, one could predict that he would
have four X chromosomes plus a Y
chromosome. The next illustration (Fig.
5) shows the peripheral leukocyte chro­
mosome complement from this patient.
As can be seen in the lower right-hand
corner of the array, four X chromosomes
are present, one more than the number
of sex chromatin masses in his buccal
smear. In general, then, this relatively
simple technique can be used with a
high degree of success for early diagnosis
(even perinatally) of Klinefelter's syn­
drome, as well as other forms of sex
chromosome abnormality. In certain
not tell the complete story. While it
accurately determines the correct num­
ber of X chromosomes present, it pro­
vides no information on the Y chromo­
some. T o obtain this, one must resort to
examination of chromosme spreads in
dividing cells rather than the interphase
characteristics of the resting cell. Briefly,
this is done by drawing a small sample
38 CANADIAN PSYCHIATRIC ASSOCIATION JOURNAL Vol. 12, No, 1

il U
C

U Kn h u u 10 n
D E

4A IA >*
13 14 15
I I 11 i i
17 18

it it
19 20
** 21
Aft
mm
22
•'
XY
FIGURE 2. Normal male chromosome complement. Twenty-two autosomes are arranged
in order of decreasing lengths and ordered into groups which are based on ease of identifica­
tion or similar morphology. In addition, a sex chromosome complex of X Y is present and
placed at the lower right corner of the array.

of heparinized peripheral blood, treating chromatin body in his buccal smear. On

this with a plant extract to selectively
precipitate erythrocytes and placing the
supernatant containing the leukocytes
into sterile culture medium. A mitotic
stimulant is added and three days later
cell division is stopped at the metaphase
stage with colchecine. After swelling,
fixing and squashing the cells on a slide,
the chromosomes are stained and then
examined and photographed. Enlarge­
ments are then made and the chromo­
somes cut out and paired.
Figure 6 shows the chromosomes from
a Klinefelter who showed only one sex
the basis of sex chromatin one would
expect that his karyotype would be 44
autosomes plus an X X Y sex chromosome
complement. However, the leukocyte
preparation indicated that he in fact had
48 chromosomes, the second extra ele­
ment being a Y chromosome. This
strongly implies that instead of an error
occurring during germ cell maturation,
as is likely the situation in most cases of
Klinefelter's syndrome, the chromosomal
error probably occurred during the first
cleavage divisions of embryonic life.
Another situation where the sex chro-
February 1967 CYTOGENETIC PRACTICE 39

.B.

tillII 12

II I I I I
13 14 15 16 17 18

19 20 21 22
FIGURE 3. Normal female chromosome complement. Autosome pairs are similar to that
of the normal male, however, a sex chromosome complex of X X replaces X Y .

matin test does not give a clear-cut
diagnosis is in those instances of chro­
matin negative buccal smears associated
with a female phenotype. In most cases,
this sex chromatin pattern would indi­
cate that the patient has only 45 chromo­
somes, the missing element being the
other sex chromosome. However, parti­
cularly in children, it may become neces­
sary to distinguish between gonadal dys­
genesis (or Turner's syndrome) and the
syndrome of testicular feminization.
The latter condition is presumably a
genetic disturbance characterized by the
presence of a normal female phenotype
and external genitalia, absence of the
uterus and the presence of testes some­
where in the inguinal canal. These in­
dividuals possess a male chromosome
complement with X Y chromosomes and
this karyotype can readily be verified
by leukocyte culture. Since the abnormal
testes found in these girls often become
malignant, it is of more than academic
interest to determine the chromosome
complement at the earliest opportunity.
Another instance where peripheral
leukocyte culture is routinely used in
40 CANADIAN PSYCHIATRIC ASSOCIATION JOURNAL Vol. 12, No. 1

ents were less than 35 years of age at

mm
FIGURE 4. Buccal smear from a Klinefelter
syndrome variant. Three sex chromatin masses
are present (arrows).
our laboratory is in mongolism where it
is of considerable value in assisting phy­
sicians in genetic counselling. A l l mon­
goloids are karyotyped, and if the par­
the birth of the patient their chromo­
some complements are likewise studied.
While most cases of mongolism are like­
ly due to the inheritance of a 'free' extra
chromosome from the mother and the
frequency of this sporadic event in­
creases with maternal age, occasionally
a young parent will be shown to be a
'carrier' for a translocation. Figure 7
illustrates the chromosome complement
from a regular 'trisomic' mongoloid.
There are 22 pairs of autosomes in this
karyotype, but in the 21-22 or G
group, an extra member is present
(arrow). In a translocation mongol,
however, such as seen in Fig. 8, the extra
member is not free but is attached to
another chromosome. In this case the
chromosomes involved being those in
the 13-15 or D group. Here, instead

.B.

U11
U M « IX Ift XI XI 8 10 12

i\>w\ (fc«p"S» J k X J l i $ ilife

13 14 15 16 17 18

h i '
19 20
m
21 22
AIM*XXXXY
FIGURE 5. Peripheral leukocyte chromosomes of a male patient with three sex chromatin
masses in his buccal smear. Four X chromosomes are present in addition to the Y ,
February 1967 CYTOGEN KTIC PRACTICE
.A.„
XS HI 81 AH ]IS KH tx 8 9
13 14 15
jt 11
19
m m
20 21
.6.
FIGURE 6. Peripheral leukocyte chromosomes from single sex chromatin positive Kline­
felter's syndrome. T w o Y chromosomes are present in addition to two X chromosomes.
of three pairs of V-shaped chromosomes
being present, there are only five, and
instead of five chromosomes in the 21-
22 or G group, there are onlv four.
However, an extra member is present
which resembles those in the C or 6-
12 group and has been paired in the
D group (arrow). This chromosomal
situation is interpreted as representing a
fusion of a G chromosome with a D
chromosome, i.e., a translocation. I f a
parent is a carrier for this abnormal
chromosome, i.e., the fusion chromo­
some is present as well as a loss of a
member of the D and G groups within
a complement of 45 chromosomes, she
has a very high risk of producing more
abnormal children as well as carriers.
These carriers, in turn, have a high risk
(theoretically as much as one in three)
41
10 II 12
I
16
I I
f i l l
17
* A
• • • •
18
22
M. XXYY
4X
of producing additional mongoloid chil­
dren, as well as carriers themselves. We
can thus see that in families such as these
it is possible that a number of mongols
could be found scattered through many
generations of the family. Generally,
this carrier state is found in young
parents of mongoloids since the etiology
of mongolism in these cases is indepen­
dent of the ageing effect usually attri­
butable as the cause of this disease. Be­
cause most women, including carriers,
tend to have their children at younger
ages, it is in the younger age group of
parents of mongoloids where this type
of chromosomal lesion is most likely to
be discovered.
From the data that is presently avail­
able on the offspring of a mongoloid
patient herself, we can conclude that
42 CANADIAN PSYCHIATRIC ASSOCIATION JOURNAL Vol. 12, No. 1

.B.

SH i^x ^X 4^ 8ji Art

8 10 II 12
.D- -E.

A A A * I * jfliJJi ^d^ti i K J ^
13 14 15 16 17 ~ 18

.6.

19 20
Jfefc
21
rfl^ JPb* ^Kfc
22
StX Y
FIGURE 7. 'Regular' trisomic mongolism chromosomes. A n extra 'free' chromosome
(arrow) designated as number 21 is present in excess of the normal complement.

normal and mongoloid children are
produced in approximately equal pro­
portions. This fits well with theoretical
expectation. Similarly, parents who have
had a mongoloid child at an early age,
or who have had more than one mongol,
have occasionally been found to be
'mosaics' for the extra chromosome, i.e.,
some of their cells contain 47 chromo­
somes. Because the abnormal cells are
few in number and need not be present
in all tissues, presumably there has been
minimal damage exerted on the carrier
himself. However, from what is known
about the offspring of mongoloids, the
risk of recurrence of the anomaly in
these mosaic parents is presumably in­
creased over that expected by chance.
For some time our laboratory has been
actively screening young parents, at­
tempting to discover translocation car­
riers and mosaicism, primarily for gene­
tic counselling purposes. This function
of our laboratory is a highly important
one, particularly from a preventative
point of view, since one can predict
with a high degree of certainty the risk
the parent faces of producing an ab­
normal child, and the family can be
spared the psychological and sociologi­
cal shock that accompanies this disease.
It is clear that the use of relatively
simple procedures can be of significant
value for early diagnosis and genetic
counselling in mental retardation, al­
though occasionally one must resort to
more sophisticated and difficult techni­
ques in order to accomplish these aims.
In particular, the skin culturing techni­
que is of great value, although there are
a number of disadvantages in using this
for routine analysis. Basically, what is
involved is that a small sample of skin,
usually from the forearm, is minced into
February 1967 CYTOGENETIC PRACTICE 43

3-

111II ft It it
8 9 10 12

i t f < l#
13 14
i t if
16 17
it
18

i i « ftf,
19 20 21 22
FIGURE 8. D / G (13-15/21) translocation-mongolism karyotype. Extra chromosome is
present attached to a medium sized chromosome (arrow).

1 mm. square particles and placed in method, it is still not insurmountably

special culturing tubes. The latter con­
tain a mixture of human A B serum and
chick embryo extract. The medium is
changed frequently and after good
growth is obtained the slides are pre­
pared in the usual manner.
The most serious drawback of this
procedure is that considerable effort
must be expended in changing media
and explants. Also, four to eight weeks
of growth are needed before results are
obtained. While this technique is more
delicate technically than the leukocyte
difficult for use in a routine laboratory,
and the results that can be obtained are
highly rewarding. Figure 9 depicts the
karyotype of a young lad who showed
a normal chromosome complement in
both leukocyte and direct bone marrow
preparations. The boy himself showed a
number of congenital anomalies asso­
ciated with a distinct facies. Physical
examination when he was four years
nine months showed that he was small
(below the third percentile) in height,
was below the third percentile in weight
44 CANADIAN PSYCHIATRIC ASSOCIATION JOURNAL Vol. 12, No. 1

11 K U ft i i

M ^X IX 11 11 11 11 10 II 12

0(\ m § i
13 14 15 16 17 18

<7

20 21 22 XY

FIGURE 9. Skin fibroblast chromosomes. An extra chromosome is present in the C

(6-12) group (arrow). This is likely autosome number 6, 7 or 8 since no sex chromatin was
present in the skin culture or buccal smear nuclei.

and was below the third percentile in
head circumference. A convergent stra­
bismus was present, as was a downward
slant to the eyes and deformed ear lobes.
His neck was short with some' slight
webbing and some limitation of neck
movement. A l l his fingers were short
and stubby and a single flexion crease
was present on the fifth digit rather than
the usual two. The large toe was de­
formed and there was a limited move­
ment of his feet and hands. An intra­
venous pyelogram showed a rotated
right kidney and a spina bifida was
present at lumbar three to five. The
posterior neural arches were also missing
in the sacrum and could not be seen in
the lower dorsal vertebrae. Pelvic X-rays
showed splitting of the body of L-4.
This represents a congenital anomaly
due to failure of segmentation. There
was some mottling of the hone texture
of the metacarpals, phalanges- and el­
bows. The boy was mildly retarded with
an I.Q. of 62 on the Kuhlman scale, but
it is uncertain whether this is a result
of an apparent deafness or as a result of
his chromosome anomaly.
The pertinent features in the karyo­
type of this individual are illustrated in
Figure 9. These are metaphase chromo­
somes from skin cultured from the fore­
arm. A l l the chromosome groups are
within the normal limits of variability
except for the C (6-12) group. A n extra
chromosome is present here and has been
1'cbruary 1967 C Y T O G E N E T I C PRACTICE
placed apart from the others (arrow).
After pairing the chromosomes to the
best of our ability in many metaphase
plates, the extra chromosome appears to
be one of the largest, probably a mem­
ber of pair 6, 7 or 8. Since the X chro­
mosome has the same morphology, it
may be argued that this is an X chromo­
some and the boy is affected with Kline­
felter's syndrome. However, because of
the wide variety of congenital anomalies
which are present and because he is sex
chromatin negative in skin cultures
which showed the extra chromosome,
this interpretation is unlikely. Lympho­
cyte and bone marrow show a normal
chromosome complement, so this patient
is a mosaic and the lesion probably arose
during the course of embryonic devel­
opment.
Another procedure that requires a
somewhat greater degree of sophistica­
tion, although still within the capability
of a diagnostic laboratory, is the use of
strip-film autoradiography. The method
is identical to the leukocyte culturing
technique except that in the final six
hours of culture a radioactive isotope of
thymidine is added to the dividing cells.
Since thymidine only becomes incorpor­
ated into desoxyribose nucleic acid,
which is only found in chromosomes,
we have a method which is useful in
identifying them. This procedure hinges
on the fact that different autosomes re­
plicate D N A at slightly different times
during cell division, although the patern­
al and maternal homologues presumably
replicate exactly at the same time and
in exactly the same location. This situa­
tion is somewhat more complex in the
case of sex chromosomes, but rules have
been established which account for
their differences. Figure 10 illustrates
how it is possible to identify morphologi­
cal variations using strip-film autoradio­
graphy. This is the metaphase comple­
ment from another boy who presents a
wide variety of congenital anomalies
associated with severe mental defect. A l l
chromosome pairs appear normal except
45
in the sex chromosome complex. An
experienced human cytogenetist would
suspect that in this case the X chromo­
some may be abnormal because it re­
sembles those in the B (4-5) group.
Normally, the centromeric constriction
is placed somewhat more medially than
in this chromosome. However, as this
appearance is not striking to the casual
observer, other methods must be found
to clearly establish that an abnormality
has actually occurred. Radioactive label­
ling unequivocally shows the type of
lesion we are dealing with in this case.
Before we can illustrate this, it is
necessary to establish the labelling pat­
terns that occur in so called 'cold X ' .
The next karyotype (Fig. 11) is an
autoradiograph of the labelling pattern
in mongolism. The upper rows of this
karyotype represent chromosomes over
which blackened silver grains lie. These
are areas in which radioactive material
was incorporated into the chromosome,
the radiation exposing a radio-sensitive
emulsion. Below these are the identical
chromosomes from which the emulsion
has been removed in order that they
can be recognized and paired. For the
present it is only necessary to note that
the 'cold X ' tends to label at the centre
rather than at the distal ends as illustrated
by the blackened silver grains in the
central portion of the X chromosome
(arrow). As a matter of interest, it can
be noted that in the G group the three
members that have been paired as chro­
mosome 21 tend to have the same inten­
sity of labelled silver grains overlying
the chromosome, whereas the other two
acrocentric members of the group tend
to have a decreased amount of radio­
activity. Returning to the karyotypes of
the patient with the question of how
autoradiography can be useful in identi­
fying the chromosome abnormality, if
in fact it really does exist, the next figure
(Fig. 12) represents the X and the B
(4-5) groups from four metaphase plates
from cultures of the patient. It can be
easily seen that the labelling pattern of
 46 CANADIAN PSYCHIATRIC ASSOCIATION JOURNAL
Sf is it a u si u
M it **
13 14 15
^»
19
j i
20
FIGURE 10. Abnormal X chromosome. This chromosome (arrow) resembles those in
the B (4-5) group whereas a normal X possesses a more median constriction.
the two 4-5 pairs are different from
each other, but that of the two bomo-
logues of each pair are roughly similar
to each other. The chromosome that re­
sembles these in morphology and that is
presumably the X , has a strikingly dif­
ferent labelling pattern. Not only is this
pattern different from the 4-5 group,
but it also differs completely from a
normal X . Here the labelling pattern is
more intense at the distal portions of the
chromosomes, whereas if we recall the
previous illustration (Fig. 11) a normal
'cold X ' tends to label at the centre.
The most likely interpretation of this
autoradiographic pattern is that a 'peri­
Vol. 12, No. 1
10 ii
U ft) fit
16 17 18
21 22
Si
X Y
centric' inversion has occurred. A dia­
grammatic representation of the mechan­
ism whereby this phenomenon can oc­
cur is shown on the next figure (Fig.
13). The top strand represents a chromo­
some with the hypothetical gene se­
quence A B C D , etc. A break could occur
in this strand and if fusion occurred at
the intersection where the chromosome
has looped, the sequence can be changed
from ABCD centromere E F G , etc. to
K D centromere E F , etc. The total length
of the chromosome remains unchanged
but there is a shift in the position of the
centromere associated with a linear
change in gene sequence. Thus, a por-
February 1967 CYTOGEN E T I C PRACTICE 47

Di if

BA %% St « I I ii to

&4 *.* jwfr

13 14
XX 1 4 i
16 17 18
l

19 20 21 XY

FIGURE 11. Autoradiographic labelling pattern in 'regular' trisomic mongolism. Karyo­

type arranged in double rows according to size and groups in the usual way. The lower
rows represent a complement which contains radioactive material within the chromosomes,
but the cell had not been exposed to a radiosensitive emulsion. The upper rows represent
the identical chromosomes from which the supporting medium has been removed and re­
placed with emulsion. Blackened, silver grains lying over the chromosomes give a rough
measure of the concentration and locality of incorporated D N A precursors.. The X chromo­
some appears to incorporate label into the central portion under the conditions used rather
than at the ends.

tion of the material that in this case was
labelling at the centre, can now be seen
near the distal portion of the chromo­
some in a manner similar to that shown
by the abnormal X chromosome in
Figure 12. Peripheral leukocyte cultures
on the parents and siblings of this pa­
tient showed a normal appearing X
chromosome so it appears likely that the
congenital anomalies and mental defect
48 CANADIAN PSYCHIATRIC ASSOCIATION JOURNAL Vol. 12, No. 1

1?
\
FIGURE 12. Array representing the X and B (4-5) groups as well as autoradiographic
patterns of these chromosomes. The B group shows a distinct difference in labelling pattern
from the abnormal X which tends to show highest radioactivity in the distal regions rather
than in the centre.

have occurred in this patient as the result
of this inversion of chromosomal mater­
ial. In other words, the anomalies may
be due to a 'position effect' caused by
the adjacent positioning of chromosomal
material not normally so occurring.
In the final section of this discussion
we will attempt to indicate how the use
of relatively straightforward techniques,
such as the peripheral leukocyte method,
can be used to throw new light on the
etiology of psychiatric disorders.
The etiology of schizophrenia, infan­
tile or adult, is still unknown and has
been the subject of a number of highly
controversial discussions. Some statisti­
cal evidence hints that genetic factors
may play a role in some families, yet
environmental influences have a recog­
nized impact upon developing personal­
ity. It would thus appear possible that
several different etiological mechanisms
may produce similar or identical schizo­
phrenic symptomatology. One of these
mechanisms may be due to minor
chromosome aberrations. There is con­
siderable evidence that schizophrenic
disorders in men affected with Klinefel­
ter's syndrome are elevated above that
expected in the normal population. Like­
wise, buccal smear surveys of institu­
tionalized schizophrenics tend to show
an elevated frequency of Klinefelter's
syndrome over that of the general popu-
February 1967 CYTOGENETIC PRACTICE

B C D E F G H

K D E F G H I J C B A

FIGURE 13. Diagrammatic representation of a pericentric inversion. After formation of a

loop and subsequent breakage and fusion a shift in the position of the centromere occurs.
In this instance a portion of the material that was in the central region (white area) has
relocated to a more distal position. Associated with this is a change in gene order from
A B C D centromere E F G H I J K t o K D centromere E F G H I J C B / 4 .

lation. These suggestions, plus the results
of Biesele, Schmid and Lawlis ( 2 ) as
well as Turner and Jennings (10), who
found chromosome aberrations in three
children with schizophrenic disorders,
have prompted us to make a retrospec­
tive study of the chromosomes in chil­
dren with this disorder.
Twenty children who had previously
been diagnosed as falling under the cri­
teria of pure functional childhood schizo­
phrenia (5) were studied by the psychia­
trists at our Institute. Chromosome
aberrations were found in two. The next
figure (Fig. 14) illustrates the lesion
found in one patient. There is the normal
number of 46 chromosomes in the com­
plement and all groups appear to be
within the normal limits of variability
except in the G (21-22) group. One
chromosome here is found to have a
short arm that is strikingly different
from the other members of the group,
exhibiting a metacentric rather than
acrocentric appearance. Whether this
represents a duplication of a portion of
chromosome in the short arm or whether
the chromosome has lost its normal short
arm and been replaced by a whole seg­
ment (i.e., translocation), cannot clearly
be established from our preparations.
There are suggestions that an insertion
so CANADIAN PSYCHIATRIC ASSOCIATION JOURNAL Vol. 12, No. 1

ft I I If 11 I I
II II II At I I I I I I
6 7 8 9 10 II 12

*• l i I ft * l l i •
13 44 15 16 17 |8

.G.

19 20 21 22 , X X

FIGURE 14. Karyotype in Childhood Schizophrenia. A member of the G (21-22) group

(arrow) has a metacentric appearance likely due to the presence of an enlarged short arm.

of material has occurred because the
distal portion of the chromosome has
some of the characteristics of satellited
regions since there is generally a vari­
ability in relative size of the region and
in its staining capacity. This type of
chromosome variation has been described
in a number of individuals. In some
instances the individual exhibited men­
tal retardation associated with physical
abnormalities; in others no obvious mal­
formations were present. The significant
point of the karyotype within the con­
text of our discussion is that it is a very
rare phenomenon and does represent a
chromosome complement with an excess
of material over the norm, even though
the expression of the abnormality is as
yet unknown.
Figure 15 illustrates the abnormal
karyotype found in the second patient
suffering from childhood schizophrenia.
In this case a portion of what we have
chosen to call chromosome number 11
has translocated onto the long arm of a
member of chromosome pair 8. The
chromosome complement is presumably
'balanced' since there is neither a detec­
table excess nor significant deficiency of
chromosomal material. In this particular
family, a sibling, the father and paternal
grandmother carried the identical trans-
February 1967 CYTOGENETIC PRACTICE
i I I If It li
B 7 8 9
JD.
li l i I I
13 14 15
19 20 21
FIGURE 15. Karyotype in Childhood Schizophrenia. Patient is a carrier for a transloca­
tion of material from chromosome number 11 to chromosome number 8.
location. A l l of these individuals are
physically and psychologically normal.
Further family studies are in progress
and we hope to complete these within
a relatively short time.
Only one study of chromosome vari­
ability in a random population is present­
ly available. Court-Brown et al (4)
found that 2.9% of 207 men showed
some form of chromosomal variation
and 1.3% of 231 women. Most of these
were due to hereditary variations in the
length of the Y chromosome, which ac­
counts for the higher frequency in men.
Variations in the short arm of the D
SI
3.
10
II 11
16 17 18
JS-
22 XY
group chromosomes as well as in the
total length of number 16 made up the
majority of the 'marker' chromosomes.
However, in two instances, one in a
male and. one in a female, a structural
rearrangement had occurred between
two chromosomes in a manner similar to
our second patient, although not invol­
ving the particular exchange chromo­
somes for which he was a carrier. A
third individual in their series possessed
a structurally abnormal Y chromosome.
The remainder possessed G (21-22)
group chromosomes which included a
member who was similar in morphology
to that of our first patient.
 52 CANADIAN PSYCHIATRIC ASSOCIATION
It is difficult to comment on the signi­
ficance of the present findings in a
meaningful way, since we do not have
a random series of individuals karyo­
typed in our laboratory. On the other
hand, Court-Brown et al found only
eight individuals out of 438 who possess­
ed either abnormal G group chromo­
somes or structural rearrangements.
Pooling our data with that of Biesele,
Schmid and Lawlis (2), Book, Nichtern
and Gruenberg (3) and Turner and
Jennings (10) we find that 4 in 41 child­
hood schizophrenics possess a chromo­
some aberration — a fivefold increase
over Court-Brown et al's' random series.
It may thus be that structural chromo­
some abnormalities may predispose an
individual to psychological defects, but
the expression of these is perhaps modi­
fied or regulated by other constitutional
factors.
Summary
Chromosome abnormalities are known
to be associated with a wide variety of
psychiatric disorders varying from sev­
ere mental retardation to aberrant social
behaviour. Principles and methods used
in contemporary cytogenetic laborator­
ies are discussed and an attempt is made
to illustrate how the integration of these
techniques can be of considerable value
for diagnostic and genetic counselling
purposes.
References
1, Bergemann, E . : Qeschlechtschromatinbest-
stimnmngen am Nmgebornen. Schweiz.
med. Wschr., 91: 292-294, 1961.
2, Biesele, J. J . , Schmid, W . and Lawlis, M.
G.: Mentally retarded schizoid twin girls
with 47 chromosomes. Lancet, 1: 403-405,
1962.
3, Book, J. A., Nichtern, S. and Gruenberg,
E , : Cytogenetical investigations in child­
hood schizophrenia. Acta Psychiat. Scand.,
39: 309-323, 1963.
JOURNAL Vol. 12, No. 1
4. Court-Brown, W . M., Jacobs, P. and
Brunton, iVL: Chromosome studies on ran­
domly chosen men and women. Lancet, 2:
561-562, 1965.
5. Creak, M., Chairman.: Schizophrenic syn­
dromes in childhood; progress report of a
working party. Brit. med. J . , 2 : 889-890,
1961.
6. Maclean, N., Mitchell, J . M., Harnden,
D. G., Williams, J., Jacobs, P. A., Buckton,
K. A., Baikie, A. G., Court-Brown, W . iM.,
McBride, J. A., Strong, J . A., Close, H . G .
and Jones, D. C : A survey of sex-chromo­
some abnormalities among 4,514 mental
defectives. Lancet, 1: 293-296, 1962.
7. Maclean, N., Harnden, D. G., Court-
Brown, W . M., Bond, J . and Mantle, D. J.:
Sex-chromosome abnormalities in newborn
babies, ibid, 1: 286-290, 1964.
8. Moore, K . L . : Sex reversal in newborn
babies, ibid, 1: 217-219, 1959.
9. Mosier, H . D., Scott, L . W . and Dingman,
H . F.: Sexually deviant behaviour in Kline­
felter's syndrome. J . Pediat. 57: 479-483,
1960.
10. Turner, B. and Jennings, A. N.: Trisomy
for chromosome 22. Lancet, 2: 49-50, 1961.
Acknowledgements
The writer gratefully acknowledges
the financial assistance of the Medical
Research Council for certain phases of
research discussed in this manuscript.
Resume
On sait que les anomalies des chromo­
somes sont le corollaire de toute une
gamme de troubles mentaux, allant de
Parrieration mentale grave jusqu'au com­
portement social aberrant. L'auteur
traite des principes et des methodes uti­
lises dans les laboratoires modernes de
cytogenetique et il cherche a illustrer de
quelle facon l'integration de ces techni­
ques peut se reveler fort utile aux fins
du diagnostic et des conseils en matiere
de genetique.