Genetica, Vol.
50, 2: 99-104, 1979
CYTOGENETICS OF THE SOUTH-AMERICAN AKODON RODENTS (CRICETIDAE) VI
POLYMORPHISMIN AKODON DOLORES (THOMAS)*
N.O. BIANCHI, Susana MERANI & Martha LIZARRALDE
Institute Multidisciplinario de Biologfa Celular (IMBICE) C.C. 403, 1900 La Plata, Argentina
ReceivedDecember4, 1978/AcceptedJanuary 16, 1979
The cytogenetic analysis of 68 specimens of Akodon
dolores (Rodentia: Cricetidae) collected in three dif-
ferent localities of the central region of Argentina
showed the presence of chromosomal polymorphisrns
of the Robertsonian type in chromosomes 1, 3, 4 and
5. The variability in chromosomes 3, 4 and 5 was
correlated with the geographical area of collection.
Animals with low chromosome numbers and two
biarmed chromosomes in pairs 3, 4 and 5 were mainly
found in the southern most area of collection (simple
homozygous). Conversely, in the northern most local-
ity forms with an increased frequency of acroeentrics
and high modal numbers (double homozygous) pre-
dominated. The central part showed the highest con.
centration of heterozygous specimens. It is speculated
that Robertsonian rearrangements of chromosomes 3,
4 and 5 originated in the central area and that DH
and SH animals migrated from there to the North and
South respectively.
The polymorphism of chromosome 1 was not cor-
related with the geographical areas. In the three local-
ities surveyed the frequency of double homozygous
animals was very low while the frequency of heter-
ozygous and simple homozygous specimens was high.
It is assumed that the high frequency of heterozygous
animals is maintained through positive selection. The
possible correlation between the chromosome 1 poly-
morphism and the infection with virus Junin (ethio.
logical agent of the haemorrhagic fever in Argentina) is
discussed.
* This work was supported by grants from 'Consejo Nacio-
nal de InvesfigacionesCientificas y T6~micas', the 'Comisi6n
de Investigaciones Cientificas de la Provincia de Buenos
Aires', and the Organization of American States.
Introduction
Chromosomal variability plays a fundamental role in
the evolution of mammals. Robertsonian rearrange-
ments, inversions and/or translocations give rise to
different structural modifications of chromosomes
which may follow different fates according to their
effect on the genetic balance of the population in-
volved. Those producing decreased fitness of bearers
are eliminated by natural selection, whereas some
others are incorporated at different rates depending
upon their ecological adaptive value. Thus, in a cer-
tain way, chromosomal rearrangements represent ex-
periments of evolution. Numerous species with an un-
stable chromosomal complement show evidence of
these rearrangements which often give rise to poly-
morphic chromosome pairs.
It has been previously reported that several species
of Akodon (Rodentia, Cricetidae) exhibit remarkable
chromosomal polymorphisms (Bianchi et al. 1969,
Kiblisky et al. 1977). Moreover, there is evidence
showing the presence of a considerable autosomal
variability in Akodon dolores. Therefore the aim of
this paper is to describe in detail this variability and
to analyse its distribution in three different popula-
tions of Akodon dolores.
Material and methods
A total of 68 specimens of Akodon dolores were col-
lected in three different localities of the Argentine
central region (Laguna Larga and Dean Furies, Pro-
vince of Cordoba and La Carrera, Province of Cata-
marca) (Fig. 1).
99
 Cytogenetic studies in bone marrow and spleen
were done according to standard procedures (Bianchi,
1966). Moreover, chromosomal analysis of some spec-
imens was performed with the microcharnber tech-
nique (Bianchi et al., 1972). Chromosome spreads ob-
tained from animals belonging to each population
were treated with trypsin to induce G bands (Wang
& Fedoroff 1972). Chromosome identification was
based on chromosomal morphology and G-banding
patterns. All the specimens included in this paper
were deposited in the collections of the 'Museo de
Ciencias Naturales de La Plata' and the 'IMBICE'.

Results

At the present time it is not known whether the

oL.C. Robertsonian rearrangements to be described below
oD.F. arose by centric fusion or fission of the Akodon dolo-
O! res chromosomes. However, for the sake of simplicity
it will be assumed that all the karyotypes found in
the species derived from a basic karyotype having the
lowest chromosome number and the highest number
of biarmed elements. The chromosome number of
this karyotype was 34. Pairs 1-5 and 16 are meta-
centrics and can be easily identified by their sizes and
centromere location. Pairs 6 to 15 are acrocentric
with a progressively decreasing size; therefore, the
identification of these chromosomes is only tentative.
The X and Y chromosomes are both acrocentrics; the
X being one of the largest and the Y the smallest
acrocentric chromosome of the complement
1000 (Table 1).
0 500
t , 1 The distribution of G-bands in the basic karyotype
km of A. dolores is shown in Figure 2. This idiogram is a
Fig. 1. Geographical areas of collection. L.L.: Laguna Larga, composite representation of all the G-bands observed
D.F.: Dean Funes, L.C.: La Carrera. in 10 well banded karyotypes; accordingly, it in-

Table 1
Chromosome length and arm indices of Akodon dolores

Chromosome Length Arm index Chromosome Length Arm index

(*) 1 18.15% + 0.84 1.23 9 3.41% + 0.30

(*) 1 a 9.90% + 0.72 8.05 10 3.37% + 0.28
(,) 1 b 8.47% + 0.68 3.8 11 3.26% ± 0.20
2 13.06% + 0.68 1.33 12 3.10% + 0.32
3 10.37% + 0.72 1.44 13 3.04% ± 0.20
4 9.54% + 0.70 1:24 14 2.66% ± 0.20
5 8.38% + 0.60 1.09 15 2.29% + 0.38
6 4.74% + 0.20 16 1.52% + 0.30 1.01
7 4.25% + 0.40 X 5.72% + 0.6
8 3.42% + 0.30 Y 2.78% + 0.3

(*) Length of chromosome 1 is valid for animals with 42 chromosomes.

Length of la and l b chromosomes is valid for animals with 43 chromosomes.

100
b!
4 lb
4 ~
3~
+
2-
1--
o-
1
L i
2-
3~
4 3 4 5 6 7 8 9 10
4*-
2
o~
1 la
.f.
2~
3- 11 12 13 14 15 16 X Y
Fig. 2. Pattern of G-bands in the idiogram of A. dolores. Chromosomes are divided in segments comprising 1% of the haploid set
(scale at the left of each row of chromosomes). Black, dotted and empty regions represent dark, dull and bright bands, respectively.

dicates the average location of each one of the bands
detected.
The analysis of the G-banding pattern allowed a
correct identification of all chromosomes in the com-
plement. Moreover, it also made it feasible to de-
termine that the chromosome arms in A. dolores have
an exact correspondence with the chromosome arms
ofA. molinae (Bianchi et al. 1973). The only differ-
ences found between the idiograms of the two species
are due to improvements in the induction of G-band-
ing; some regions which were originally represented as
a single band are now known to be formed by two or
three bands in A. molinae as well as in A. dolores.
Modifications to the basic karyotype of A. dolores
were produced by Robertsonian rearrangements in-
volving the chromosomes 3, 4 and 5 and a Robert-
sonian rearrangement plus two probable pericentric
inversions in the case of chromosomes 1. The poly-
morphisms of each chromosomal type produced three
varieties of animals: (a) specimens with two biarmed
homologues, simple homozygous (SH) animals; - (b)
specimens with only one biarmed chromosome and
two acrocentric (or two subterminal in the case of
chromosome 1) homologues to the arms of the meta-
centric chromosome (the homology was determined
by G-banding, Fig. 3), these specimens were identi-
fied as heterozygous (Ht); - (c) animals in which the
metacentric pairs were replaced by two pairs of acro-
centric (or subterminal) chromosomes; these animals
were named double homozygous (DH). Chromosome
numbers in A. dolores varied continuously from 34 to
40 according to the combination of polymorphisms
in the different chromosome types involved. The
system of karyotype construction and nomenclature
employed in A. dolores is similar to that previously
used in A. molinae (Bianchi et al. 1973); that is: (a)
the acrocentric (or subterminal)chromosomes homo-
logous to a metacentric element were placed in the
karyotype with the centromeres facing each other
and in the same location as the biarmed chromosomes
(Fig. 3) and numbered according to their position in
the karyotype, and the chromosomes pairing with the
long and short arm of a metacentric element were
respectively identified with the letters 'a' or 'b'
(Fig. 3); - (b) karyotypes were identified by the rood-
Fig. 3. G-banding pattern in Htl, Ht3, Ht4 and Ht5 chromo-
some groups.

101
Table 2
Distribution of chromosome polymorphisms of Akodon dolores in three regions of Central Argentina

Locality Number of animals Chromosome Polymorphisms Modal FN (x)

d 9 ..Number
Chr. 1 Chr. 3 Chr. 4 Chr. 5

SH 10 24 24 18
Laguna larga 13 11 Ht 12 - - 8 34-36 44
DH 2 - - -
SH 14 5 7 9
Dean funes 9 13 Ht 7 14 14 8 35-38 44
DH 1 3 1 5
SH 11 - - -
La Carrera 10 12 Ht 11 12 14 2 39-40 44
DH - 10 8 20

(x) The small arms of chromosomes la and lb were not counted.

al chromosome number, and by the chromosomes types
which appear as SH, Ht or DH; for instance, a male
karyotype heterozygous for chromosomes 1 and 4,
simple homozygote for chromosome 3 and double
homozygote for chromosomes 5 is identified as 38;
Ht 1, SH3, Ht4, DH5/XY. The table 2 shows the total
of animals studied in each locality, the frequency of
polymorphisms observed and the distribution of these
polymorphisms in the geographical areas.
As mentioned earlier it is not known whether the
Robertsonian rearrangements detected derived
through centromere fusion or breakage. However, the
evidence accumulated in A. molinae seems to indicate
that the polymorphism of the pair 1 in this species
arose via centromeric breakage plus two pericentric
inversions. (Bianchi et al. 1973, Bianehi et al. in pre-
paration). Since the pair 1 ofA. molinae andA. dolo-
res are alike (shared chromosomes, Bianchi & Bian-
chi 1969) it seems reasonable to assume a similar
mechanism to explain the origin of the polymorphism
of chromosomes 1 in A. dolores.
Discussion
The finding of animals heterozygous for one to four
Robertsonian rearrangements poses the problem of
the eventual segregational impairment in the fertility
of bearers. The studies of Bruere & Mill (1971) and
Bruere et al. (1972) apparently show no fertility de-
crease in sheep having single or double structural
heterozygosity of the Robertsonian type. On the
other hand, the presence of meiotic irregularities with
the appearance of gametic and zygotic aneusomy has
been shown in bulls and mice heterozygous for fusion
metacentric chromosomes. Ford & Evans (1972)
and Gropp et al. (1974) have been able to show that
different types of single Robertsonian metacentric
heterozygotes in mice produce variable rates of meio-
tic non-disjunction ranging from 4-to 26% of un-
balanced metaphase II figures. Both groups of authors
also remark that the non-disjunction rate is consider-
ably higher in female than male gametogenesis. The
result of these meiotic irregularities is the appearance
of aneuploid fetuses which are usually aborted at vari-
able stages of development.
The frequency of abnormal meiotic segregation in
A. dolores exhibiting "different combinations of
Robertsonian rearrangements is under analysis in our
laboratory at the present time. The data are still pre-
liminary and no conclusions can be made at present.
The information obtained with A. molinae, a spe-
cies which has the same type of chromosome 1 poly-
morphism as A. dolores, seems to indicate some
fertility impairment existing in heterozygous ani-
mals. However, the surprising observation was made
that the frequency of DH specimens in the crosses of
Ht x Ht was very low indeed and that these animals
were highly infertile (Bianchi et al., in preparation).
The very low frequency of DH 1 specimens in the

102
three wild populations of Akodon analysed in this
report (Table 1) probably indicates the existence of a
similar impairment for the appearance of these
animals in A. dolores. On the other hand, the usual
Finding of DH 3, 4 and 5 specimens (Table l) suggests
that these types of chromosome complements do not
produce remarkable fertility impairment in the bear-
ers.
Table 2 shows that, with the exception of chromo-
some l, all the other polymorphisms are correlated
with the geographic area. In the Laguna Larga collec-
tion there exists a clear predominance of SH forms
with no DH animals, whilst the northern locality of
La Carrera exhibits no SH specimens and a pre-
dominance of DH animals. The lack of SH in La Car-
rera, a panmictic population formed by Ht and DH
animals, is rather striking and it emphasizes the high
adaptive value of SH forms to southern and of DH to
northern regions of the collecting area. To a certain
extent our findings are similar to the results obtained
by Yosida et al. (1971, 1974) in rats. These authors
were able to demonstrate a different geographical dis-
tribution of rats differing in their karyotypes by two
Robertsonian rearrangements. Moreover Yosida &
Tsuchiya (1969) and Yonda et al. (1971) were also
able to show that temperature variation may produce
remarkable changes in the frequency of hetero-and
homozygous animals for a pericentric inversion of
chromosome 1. Laguna Large and La Carrera differ in
several environmental conditions such as winter tem-
perature, altitude and vegetation. However, up to the
present moment no correlation between these envi-
ronmental variations and the karyotype of the
rodents has been established.
Dean Funes, which is approximately located in the
middle part of the collecting area seems to be a region
of hybridization between SH and DH forms. Accord-
ingly, it is possible to speculate that Dean Furies may
have been the geographical region at which Robert-
sonian rearrangements for chromosomes 3, 4 and 5
originated and that SH and DH animals migrated
from there to the South and North respectively.
The behavior of chromosome 1 has been studied
in A. molinae. The data existing in this species seem
to indicate that chromosomes la and lb arose by
centric breakage of chromosome 1 and pericentric in-
version of the two new elements formed (Bianchi et
el. 1973). Moreover, there also exists information
indicating that DH1 specimens of A. molinae are
rare with considerable infertility. Since chromo-
some 1 of A. molinae and A. dolores is similar it
seems valid to extrapolate to the latter species the
conclusions obtained with the former. Therefore,
taking into account that, in the three localities
surveyed, there is a low number of DH1 and a high
frequency of Htl and SHI A. dolores, it seems
reasonable to assume that the maintainance of Htl
specimens is favored by selection. Which selective ad-
vantage is playing a role is not known at the present
time. It is, however, interesting that Akodon is one of
the genera known to be the reservoir of the Junin
virus, the aetiological agent of haemorrhagic fever in
Argentina. Previous studies performed in Calomys
(Rodentia, Cn'cetidae) apparently show that the in-
fection with Machupo virus (a virus closely related to
the Junin virus) can produce variability in the fertility
rates of different strains of these rodents. (Report
469 Comisi6n de Investigaciones Cientfficas de la Pro-
vincia de Buenos Aires). Accordingly, the relationship
between viral infection, chromosome 1 poly-
morphism and fertility of Akodon is now under
analysis in our laboratory.
The authors wish to thank Mr. Omar Molina for his
technical assistance.
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104