Heredifas 109: lS9-lhN (1988)

Reciprocal translocation in four boars producing decreased litter size

INGEMAR GUSTAVSSON

Department of Animal Breeding and Genetics, Swedish University of Agricultural Sciences,

Uppsaln, Swtieti

(iL,sI A \ ssoh. I . 14x8. Reciprociil ti-aiislo~iitioiiin tour hoar\ producing decruascd litter size. ~ Heredifas
I O Y : I S Y - l 6 X . Lund. Swcilcn. ISSN OUIX-06hl. Received November 27, IY87

Four hoar\ with ii normal semen picture. normal sexual hehaviour and normal incidences of repeat breeding
and m;iltormation\ hut giving rise to considerably decreased litter size were found t o he translocation car-
rier\. 7 w j ol them had ;I tranalocation involving chrom(i\rimc I . rcp( Ip+.14q-) a n d rcp(lq-;17q+). re-
\pc.ctivcly; both were presumptivc new mutations. T h e other 2 boars, littermates. h a d the same reciprocal
. . ' rcp(Sq-;8q+). which was presumably inherited from their d a m . All t h r e e translocationscon-
' .Iocdticin.
t r .in5
cerned uncqual-\ized chromosome segments and five of the breakages preceding t h e translocations occurred
i n KRA positive h m d s c l m c to the centromeres and telomcrcs. Mitotic and meiotic chromcisomc morphol-
ogy w a \ Inve\tlgated

lii,g(wiur Gir.sfav.sson,Departrniwf of Animal Rrerdirig arid Genetics. Swrdish Unrver.sity of Agricultural

S r . i r r i w \ . S-7.56 07 Uppsul[i 7, Swrdm

Chromosome aberrations as fertility-reducing fac-
tors have been reported in several domestic animals
(GLISIAVSSON 1980). but have only rarely been con-
sidered in breeding programs. In the pig, aberra-
tions, most often reciprocal translocations, have
been described sporadically in boars producing de-
creased litter size (for review, see P o r ~ s c u1982).
Over the few years there have been reports indicat-
ing that reciprocal translocations are probably more
common i n pigs than was earlier believed (Gus-
TAVSSON et al. 1983) and sometimes attain economi-
cally important proportions (GCISTAVSSON and SE r -
I E K G K E1984).
N Some time ago, three new reciprocal
translocations were found in boars having a normal
sexual behaviour and a normal semen picture yet
giving rise to considerably reduced litter size. Be-
cause of the latter fact, the boars had been taken out
of service. Since chromosomal aberrations are still
only rarely found in domestic animals, the mitotic
and meiotic appearances of the translocations are
described here and their origin and possibilities of
persisting in the population are briefly discussed.
Materials and methods
The boars were 15-16 months old and of the
Swedish Yorkshire breed. Sexual bchaviour and
semen picture were normal, there werc no indic;i-
tions of an increascd incidence of repeat breeding or
malformations, but litter size was considerably re-
duced. All 4 boars had been used on farms produc-
ing pigs for meat. Case 1 . This boar demonstrated
an average of 7.5 (range 2-1 1 ) piglets calculated on
39 litters. The average litter size of the farm had ear-
lier been 11.5. There was no repeat breeding and
the number of malformations, according to the
farmer, was very low. The dam was available for
cytogenetic investigation but the sire, which in 104
litters produced a mean of 11.7 piglets, had been
culled before the invzstigation. Case 2. Boar no. 2
had in 18 litters produced an average of 7.6 (range
4-13) piglets on a farm which had earlier had an av-
erage of 12.7piglets per litter. There was one repeat
breeding and the incidence of malformations was
5 %. None o f the parents was alive, but the dam had
farrowed 6 litters with an average of 10.1 (range 8-
12) and the sire had produced 10.6 as an average in
22 littcrs. Case3. This boar had been mated with 75-
100 sows and gilts on different farms. The records of
litter size kept by the owner of the boar were veryin-
complete, but the average litter size was 4.7 (range
2-6) in 18 matings on 5 different farms randomly
chosen. Case 4 . Boar no. 4 was a littermate of no. 3,
reported almost simultaneously from another area
o f Swedcn. I t had been mated with 37 females on
one single farm, giving 6.7 (range 3-11) as an aver-
age litter size. The numbers of repeat breeding, and
stillborn and malformed piglets were 3 and 12 %,
respectively. The sire in cases 3 and 4 was available
160 I GLISIAVSSON Hereditas 109 (1988)

for cytogcnetic studies but the dam had been culled both conventionally stained and banded karyotypes
before the investigation. She had farrowed 6 litters, (Fig. 1 and 4a). The RBA banded chromosomes
with an average of 7.8 (range 4-10) piglets. (Fig. 1) showed that the larger part of chromosome
Peripheral blood lymphocyte cultures were set up 14 had been translocated to the p arm of chromo-
at repeated times in the conventional way from the some 1, and by applying the G T G technique (Fig.
4 boars and available parents. RPMI 1640 contain- 421) it was found that the breakages had occurred in
ing glutamine plus 30 Yo fetal calf serum and the most distal negative (KBA-positive) band of
pokeweed (Grand Island) were used as culture chromosorne arm Ip and in the proximal part of
medium and mitogen, respectively. After approxi- chromosome 14 (Fig. 1 and 4a). However, the
mately 3 days, colchicine (I00 pg/ml) was added breakage in l p had not occurred in the terminal
half an hour before harvest and to some of the cul- THA-positive band (Fig. Sa). Application of the
tures to be studied by the RBA technique (see QFQ, CBCi and Ag-l gave no complementary in-
below), S-bromodeoxyuridine (BrdU) in a concen- formation about the translocation. According to the
tration of 200 pg/ml was added 8 hours before har- standard proposed (Committee for the Standard-
vest. Some other cultures to be used for high resolu- ized Karyotype of the Domestic Pig 1988), the
tion GTG studies were treated with ethidium translocation can be designated rcp( 1;14)(p25;q15).
bromide (10 pglml) one hour before harvest At metaphase I of meiosis, the translocation
(IKEUCHI and SAsAKi 1979). The lymphocytes were chromosomes formed, in 17 investigated cells, a
then treated with a 0.075 M KCI solution for 15 min, large chain bivalent with an interstitial chiasma
fixed in methano1:acetic acid (3: 1) and air-dried on (Fig. 6a).
cold wet slides. Besides conventional staining with The dam of the boar had normal chromosomes.
Giemsa, preparations were stained according to the
following banding techniques: QFQ (CASPEKSSON et
Case 2
al. 1971), RBA (DUTKILLAUX et al. 1973), T H A
(DLI.1RILI-AUX 1973; GL~STAVSSON IY83), GTG (SEAB- The conventional stainings demonstrated a fore-
K K H T 1971), CBG (SUMNEK 1972) and Ag-l (BLOOM shortened q arm in one chromosome of pair 1 and a
and GOODPAST~JRF 1976). Some of the preparations somewhat extended small telocentric chromosome
were also sequentially studied with QFQ-RBA (Fig. 2 and 4b). The banded preparations showed
(Dl:TKIl.LAIJX 1975) and QFQ-CBG (RUBENS-Eli5et that the latter was chromosome no. 17. By applying
al. 1978) stainings. the GTG technique (Fig. 4b) it could be established
After taking samples of the testicles at slaughter, that the breakages had occurred in GTG-negative
male meiosis of the 4 boars was studied according to (RBA-positive) bands in the central part of the q
the technique described by PATHAK et al. (1976) as arm of chromosome 1 just distal to the interstitial
applied by KING(1981). The preparations were THA-positive band (Fig. S b), and close to the cen-
either stained conventionally with acetic orcein tromere of chromosome 17. No centromeric
(2 Yo) or banded according to the CBG (SUMNEKheterochromatin, demonstrated by the CBG and
1972) and the T H A (DUTRII.L.AUX 1973) technique. T H A (Fig. Sb) techniques, had been transferred to
The nomenclature is the one used in human lq-, however. 'The QFQ and Ag-l techniques gave
cytogenetics and now also to some extent adopted no complementary information about the transloca-
for use in domestic animals. The chromosomes were tion. According to the international standard, the
arranged according to the pig standard (Proceed- translocation can be described as rcp( 1;17)(q21;
ings of the First International Conference for the q l l ) .
Standardization of Banded Karyotypes o f Domestic This translocation showcd quadrivalents of vary-
Animals 1980) and the aberrations designated ac- ing morphology at the metaphase I stage of meiosis.
cording to the banding system agreed to (Comniit- In 21 cells analysed, 7 quadrivalents could be clas-
tee for the Standardized Karyotypc of the Domestic sified as ring-shaped and 14 as chain-shaped (Fig.
Pig 1988). 6b).

Results Cases 3 and 4

Case 1
A deviating karyotype due to a reciprocal transloca-
tion, rcp(lp+;l4q-) could easily be identified in
In conventionally stained preparations from the
probands, one small one-armed chromosome, i n
addition to the six normal one-armed ones, could be
identified. O n closer examination, this chromo-
Hereditac 109 (1980)
Fig. 1. RBA-banded karyotypc of the boar carrying the rcp(lp+;l4q-). Bar indicates 10prn.
some had, however, a small distinct second arm.
There was also a large submetacentric chromo-
some, the second largest submetacentric one ac-
cording to size, which had n o homologue. By apply-
ing the RBA (Fig. 3) and the GTG (Fig. 4c)
techniques, the long arm of the small chromosome
dcmonstrated the same patterns as the p arm of
chromosome 5 . and the distal half of the q arm ofthc
large submetacentric chromosome corresponded in
pattern t o Sq. The remaining part of the large
TRANSLOCATIONSA N D DECREASED L I ~ E SIZE
R IN PIGS 161
chromosome corresponded in patterns to chromo-
some 8 (Fig. 1 and4c). The deviations could be attri-
buted to a translocation between chromosomes 5
and 8 . Apparently, most of Sq had been translo-
catcd t o chromosome 8 after a break in the proximal
KBA negative (GTG positive) band. Applicationof
the T H A technique (Fig. Sc) demonstrated that the
centromere o f chromosome S was not involved i n
the translocation. The break in chromosome 8 had
occurred in thc most distal RBA positive (GTG
162 I . GUSTAVSSON
Fig. 2. RBA-banded karyotype of the boar carrying the rcp(lq-;17q+). Bar indicates 10 p n
negative) band of the q arm. The chromosome ma-
tcrial of the terminal segment o f chromosome 8
could, howcvcr, not be identified by available
techniques in chromosome 5q-. These observations
wcrc supported by studies on thc QFQ and THA
(Fig. Sc) patterns, while thc Ag-I technique sup-
ported the identification of chromosome 8. The
CRG patterns gave no further information about
the translocation. According to thc landmark sys-
Herrditas 109 (1988)
tem proposed, the translocation can be designated
rcp(5;8)(q12;q27).
Eighty-eight (74 ‘L) of 119 metaphase cells inves-
tigated in case 4 demonstrated a ring-shaped quadri-
vdent, while the remainder (31 or 26 YO)had a
chain-shaped quadrivalent. Corresponding figures
in case 3 werc 20 (8.5 ‘YO) and 5 (IS [YO) out of 34 cells
investigated. With the aid o f the T H A technique,
the details o f the quadrivalent could be elucidated

Fig. 3. RBA-karyotype o f one o f the boars carrying the rcp(5q-;8q+)
(Fig. 6c, d). The chain-shaped configuration was ap-
parently due to the absence of a chiasma between
5q- and 8q.
The sire had normal karyotype.
Discussion
By somatic chromosome analysis it was possible
with available banding techniques to establish that
TRANSLOCATIONS A N D DECREASED LITTER SIZE IN PIGS 163
Bar indicates 10 p m
two of the translocations were reciprocal. The for-
mation of a ring-shaped quadrivalent in the first
meiotic metaphase of rcp(5q-;8q+) demonstrated
conclusively that there had been a reciprocal ex-
change of segments in this translocation too.
Chromosome 1 has earlier been implicated in
spontaneous reciprocal translocations in the pig
(LOCNISKAR et al. 1976; FORSIER et al. 1981; G U S -
I'AVSSON et al. 1983), as also have chromosome 5
( P o r t x x et al. 1983),chromosomc 8 (GusrAvssoNet
164 I CilJSI'AVSSON Hereditas 109 (1988)

Fig. 4a-c. GTG-banded translocation chromosomes of three cells of each translocation with schematic figures
indicating breakage and rejoining points (arrowheads) in the normal and the translocated chromosomes, re-
spectively. of (a) rcp( lp+;14q-). (b) rcp( lq-;17q+) and (c) rcp(5q-;8qi-). Bar indicates 10 p m .

Fig. 5 a-c. THA-banding pattern\ of (a) rcp( I p t ;14q-).
(b) rcp(lq-;17q+), and (c) rcp(5q-;Xq+). a All thcposi-
tive terminal band of 1 p had been translocated. b The
break in chromosome I prcccding thc translocation had
apparently occurred distally to the interstitial positive
TRANSLOCATIONS A N D DECREASED LITTER SIZE I N PIGS 165
al. 1983). chromosome 14 (HAGELTORN et al. 1976;
MADAK al.~1978;
~ PoPEsmand LEGAULT 1979), and
chromosome 17 (Poiwcrr and BOSCHEK 1986). These
observations agree closely with the hypothesis of a
non-random distribution of reciprocal transloca-
tions in the pig ( F m s a n d STRANZINGER 1982) with 1
and 14 being the chromosomes most often involved.
Five of the chromosome breakages preceding the
rejoining of chromosomes in the present transloca-
tions occurred close to the centromeres and telo-
meres, presumably similar to the non-random pat-
tern earlier observed in man (Yci et al. 1978). All
breakages except the one in chromosome 5 occur-
red in RBA-positive bands, which appear to be pre-
ferentially involved in spontaneous reciprocal trans-
locations (NAKAGOME and CHIYO 1976). It is quite
possible, however, that with increased resolution
the breakage site of 5q will prove to be in an inter-
band between two dark bands resolved from the
proximal dark G T G band. In man, an excess of
breakages in a single chromosome band has been
described (e.g.. FORD^^ al. 1981), and in the pig
there are similar indications of non-random pat-
terns. In the chromosome arm Ip, breakages pre-
ceding two spontaneous (LoCNisKARet al. 1976; Gus-
TAVSSON et al. 1083) and one induced (FRIES and
SrRANziNciEK 1982) reciprocal translocation have
been observed in the proximal segment distributed
over three bands. Similar indications concern the
most proximal band of chromosome arm 1Iq, which
has been involved in two translocations. In one of
the present translocations, rcp(5q- ;8q+), it was
interesting to note that the breakage site of 8q, in-
volving exactly the same band, was described earlier
(G~JSTAVSSON et al. 1983) as being involved in a
rcp( Ip- ;8q+). When studying induced aberra-
tions, FRIES and STRANZINGER (1982) even observed
that three of four breakages preceding rearrange-
ments of chromosome 2, had occurred in the same
band. This does not mean, however, that recurrent
breakage in the same band also affects exactly the
same site. The recurrent breakage in the same band
of chromosome arm l l q referred to above appa-
rently affected different sites, as in one of them-
but not in the other-the translocation of chromo-
some material also included a transfer of CBG- and
T H A b:und and n o centromcric hetcrochroimtin had hccn
transferred from chromosome 17. c N o ccntromeric
hctcrochromntin had been transferred from chromosome
5 t o chromsome X q + . Bar indicates I 0 pin.
166 I . GUS’I’AVSSON Hereditas 109 (1988)

Fig. 6 a-d. Diakincsis-metaphasc 1 o f the translocations: chain-shaped quadriv:ilcnt o f (a)

rcp( lp+;14q-), (b) rcp(lq-;17q+), (d) rcp(k-;Xq+) and ringshaped quadrtvalent of (c)
rcp(5q- ;Xq+). The chromosomes havc been stained conventionally (a, b) and according t o
the THA technique (c, d). The quadrivalcnts are indicated by arrows, and the bar indicates
IOpm.

THA-positive material (GWIAVSSON and SETTEK-
( i 1 l t . N 19x4). The preponderance o f breakages affect-
ing certain chromosomes, chromosome segments,
and even chromosome bands. has often been discus-
sed and different theories have been proposed ( e . g . ,
JA(.oiISCt al. 1974; Yuet 211. 1978), but there isstill no
conclusive cxplanation o f the c;iusc(s) o f this non-
random pattern.
Unfortunately, the meiotic techniquc is not yet
satisfactorily devclopcd f o r pig chromosomes (KIN(;
198I), and conclusive results about the outcome ot
disjunction must therefore await studies on early
embryos. Since there arc no such observations, we
can only speculate about the mode of chromosome
segregation. The meiotic technique, which includes
;I prolonged hypotonic treatment giving chromo-
somes ii fuzzy ;ippearance makes detailed studies
difficult. I n the present work, the CBG and THA
techniques wcrc both used ;IS far as possible, be-
c;iusc, depending o n hase composition o f the cen-
Hrredirus 1OY i1YHXJ
tromeres (LINet al. 1982). the centromeres o f the
two-armed and the one-armed chromosomes arc
stained weakly and strongly, respectively, by the
CBG technique. On applying the T H A technique.
the condition is quite the reverse; centromeres of
two-armed chromosomcs stain brightly but centro-
meres of one-armed chromosomes stain darkly
( G L ~ S T A 1983).
VSS~~
Although the rcp(5q- ; X q + ) concerned unequal-
sized chromosome segments. the fkrmation prefer-
entially of a ring-shaped quadrivalent in the first
meiotic metaphase would result preferentially in a
2:2 alternate and adjacent I disjunction. according
to experience in man ( J A I H ~etKal. I 1980). I n the
rcp(lq-;17q+), the occurrence of a chain-formed
quadrivalent, such as observed in most first meta-
phases. probably increases the incidence of adja-
cent-2 segregation ( H A Z I I . K I1971).
~\ The observa-
tion in the latter translocation that the smallest
chromosome could be iissociatcd with two different
chromosomes probably means that sometimes no
chiasma will form. leaving 17q as a univalent.
Rarely, observations o f something like a precocious
disjunction or absence of chiasma have indicated ii
possible 3.1 disjunction. The importance of a short
interstitial segment, besides thc extreme disparity in
the length of chromosomes involved, for the occur-
rence of a 3:1 disjunction has been emphasized by
LINIENBAIIM and BOOKOW (1975). Since there is an
inhibition of chiasma formation in the vicinity of the
breakage point, a short inter\titial segment should
predispose to a I l l + I configuration, giving inter-
change trisomy and tertiary mono- and trisomy in
the zygote. The quadrivalent ofthe remaining trans-
location, rcp(lp+;l4q-). had a fairly symmetrical
appearance due to thc interstitial chiasma. I t is
interesting that the translocation fulfils most of the
criteria set up in man (Jtu.t3b.Ki et al. 1980) for a high
incidence of 3: 1 disjunction; the total length of arms
not involved in the translocation plus interstitial
segments situated between the centromeres and the
breakpoints differs at least with a mean of 1:3, the
overall length of translocated segments differs with
at least the same ratio, and one of the chromosomes
is acrocentric and most often behaving as a univa-
lent. A III+I configuration was not observed in the
rcp(lp+;l4q-), but this probably does not exclude
the occurrence of a high frequency of 3:l disjunc-
tion.
From experience with translocations in pigs (e.g.,
KINGet al. 1981) and some other mammals such as
mice (e.g., S m w t ~ e al.
t l97l), we can assume that
chromosomally unbalanced segregation products
arc formed and develop into unbalanced sper-
1KANSl.OCATIONS A N D D E C R t A S E D LITTER SIZE I N PIGS 167
matozoa which arc capable of fertilizing. The oc-
currence o f such chromosomally unbalanced zy-
gotes, presumdiy becoming eliminated, would
explain the reduction in litter size observed. Since
the translocated segments wcre fairly large (DANIEL.
1979) and also rich in RBA-positive material-vari-
ation of which is not readily tolerated (KoRENijrx; et
al. 1978)-it is quite conceivable that the unba-
lanced zygotes were eliminated early in embryonic
life, although comparison o f two other transloca-
tions, rcp(13q-;14q+) and the rcp(llq+;lSq-)
did not, however. indicate such relationships in the
pig ( K I N G al.
C ~1981). There wasnoincreasein num-
hers of stillborn and/or malformed piglets in the
offspring o f the boars presented here, which agrees
with observations in the other pig translocations
(AKEiSS0N:Ind Hrww(’soh 1972; K I N G Ct al. 1981;GLJS-
I’AVSSON and Si:rwKmtN 1984). I t must he assumed
that the offspring either had a normal karyotype or
were balanced carriers of the translocation con-
cerned.
The mean litter sizes in cases 1 and 2 were 7.5 and
7.6, respectivcly, which. compared with the mean
for the farm, when a normal boar was used, means
corresponding reductions of 35 ‘%I and 40 ‘YO. The
dam of case I was normal and since the surviving pa-
rents of the 2 boars had produced large litters, it is
quite conceivable to assume that both transloca-
tions were new mutations.
The rcp(5q- ;8q+) was probably maternally de-
rived, with the dam being a carrier of the transloca-
tion, since recurrence o f the same mutation in the
gametes or derivation of two fertilizing sper-
matozoa from the same mutant stemline cell of an
individual are improbabilities. The low average lit-
ter size of 7.8 for this dam, estimated on 6 litters (7,
10, 4, 9, 7 piglets in litter order), also supports this
assumption. T o become approved as a breeding
dam in Sweden the least number of pigletsshould be
8 as an average of the litters farrowed and this dam
was therefore periodically approved for breeding
purposes. She originated from a litter consisting of
11 piglets, but according to the owner of the farm,
the grand dam had also demonstrated fairly small
litters (8.1 as an average of 6 litters, range 5-12) and
it is therefore quite possible that the translocation
had been inherited maternally for at least two gen-
erations. It is evident that, in the females, random
factors can easily affect mean litter size because of
the very restricted number of litters on which esti-
mations are made. thus rendering the artificial
selection for increased litter size inoperative. This is
probably one important way in which some translo-
cations escape detection and elimination. The litter
rizes of the proband boars wcrc 4.7 and 6 . 7 , rcspec-
tively. N o rcliahle figures f o r the cxpected normal
littcr size wcrc available for case 3, however, since it
had bccn used o n scveral farms. T h e average value
for case 4, compared with the normal mean for thc
farm, gives a reduction o f 33 '!h. I t is not known if
this information, available for liltcr size, indicates
true diffcrencc between the sexes.