J Appl Genet 51(3), 2010, pp.

309–317

Original article

Chromosome rearrangements and survival of androgenetic

rainbow trout (Oncorhynchus mykiss)

K. Ocalewicz1, S. Dobosz2, H. Kuzminski2, J. Nowosad1, K. Goryczko2

1
Department of Ichthyology, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland
2
Department of Salmonid Research, Inland Fisheries Institute in Olsztyn, Zukowo, Poland

Abstract. The purpose of this work was to quantify the impact of spontaneous and X-radiation-induced chromo-
some rearrangements on survival rate of androgenetic rainbow trout (Oncorhynchus mykiss). Various doses of X
irradiation (50, 150, 250, 350 Gy) were used for inactivation of nuclear DNA in oocytes. After the irradiation,
eggs were inseminated with normal sperm from 4 males derived from a strain characterized by Robertsonian rear-
rangements and length polymorphism of the Y chromosome. The haploid zygotes were exposed to a high hydro-
static pressure (7000 psi) to duplicate the paternal DNA. Neither Robertsonian chromosome polymorphism nor
the Y chromosome morphology impaired the viability of the androgenetic embryos and alevins. Moreover, sur-
vival of eyed embryos of the androgenetic rainbow trout increased significantly with increasing doses of oocyte
X irradiation. After 6 months of rearing, only specimens from the 250 and 350 Gy variants survived. The number
of fingerlings with remnants of the maternal genome in the forms of chromosome fragments was higher in the
250 Gy group. Intraindividual variation of chromosome fragment number was observed, and some individuals
exhibited haploid/diploid mosaicism and body malformations. Individuals irradiated with less than 250 Gy died,
presumably because of the conflict between intact paternally derived chromosomes and the residues of maternal
genome in the form of chromosome fragments.

Keywords: androgenesis, chromosome fragments, sex chromosomes, survival rate, X irradiation.

Introduction eggs. After fertilization, haploid zygotes are

Androgenesis is a reproductive process in which
diploid offspring inherits only paternal nuclear
DNA. Although natural (spontaneous) andro-
genesis is observed in some plant and animal spe-
cies (McKone and Halpern 2003), paternal
chromosome inheritance can be induced inten-
tionally in fish (Pandian and Koteeswaran 1998;
Devlin and Nagahama 2002; Komen and
Thorgaard 2007).
Artificial androgenesis includes 3 steps: (1) in-
activation of nuclear DNA in oocytes; (2) fertiliza-
tion with untreated or cryopreserved sperm; and
(3) diploidization of paternal chromosomes. The
irradiation with UV, gamma rays or X rays is ap-
plied to destroy the nuclear genome of irradiated
exposed to a thermal or high-pressure shock, sup-
pressing the first mitotic division of the zygote.
When diploidization is not applied, androgenetic
fish start developing but they usually die at the
eyed-stage of embryogenesis (Komen and
Thorgaard 2007).
Studies leading to obtain viable and fertile
androgenetic progenies are advanced in rainbow
trout (Oncorhynchus mykiss). Uniform, homozy-
gous androgenetic rainbow trout are used for ge-
netic mapping and genome sequencing studies
(Young et al. 1998; Nichols et al. 2003), detection
of markers for quantitative trait loci (Martinez
et al. 2005), and examination of physiological ef-
fects of mitochondrial DNA variation (Brown and
Thorgaard 2002). Moreover, androgenetic rain-

Received: October 6, 2009. Revised: January 7, 2010. Accepted May 4, 2010.

Correspondence: K. Ocalewicz, Department of Ichthyology, University of Warmia and Mazury in Olsztyn, Oczapowskiego 5,
10–957 Olsztyn, Poland; e-mail: con@uwm.edu.pl
310 K. Ocalewicz et al.
bow trout lines have been used in studies concern-
ing the evolution of sex chromosomes, the process
of sex determination, and differentiation (Felip
et al. 2004; Ocalewicz et al. 2007). Androgenesis
has also been applied to recover the nuclear
genomic information from cryopreserved rainbow
trout spermatozoa (Babiak et al. 2002).
Interspecies androgenesis (thought to be a useful
method in restoration of extinct species) has been
induced and androgenetic development of rain-
bow trout in eggs of Yellowstone cutthroat trout
(Oncorhynchus clarkii bouvieri Richardson) re-
ported (Brown and Thorgaard 2002).
Rainbow trout shows spontaneous chromo-
some rearrangements/aberrations regarding the
number of sets of chromosomes and the number or
structure of individual chromosomes. Natural
rainbow trout triploids have been described by
several authors (Thorgaard and Gall 1979;
Flajshans and Rab 1987; Ocalewicz and Dobosz
2009). Robertsonian polymorphism resulting in
diploid chromosome number ranging from 58 to
64, and a constant chromosome arm number (FN)
equal to 104, are widely observed among speci-
mens from various rainbow trout populations and
strains (Thorgaard 1983; Ocalewicz 2002). Addi-
tionally, length polymorphisms of the Y and X
chromosomes have been described in this species
(Thorgaard 1977; Ocalewicz and Dobosz 2009).
On the other hand, rainbow trout chromosome re-
arrangements may appear in the course of incom-
plete inactivation of paternal or maternal nuclear
genome during induction of gynogenesis
(Chourrout and Quillet 1982) and androgenesis
(Parsons and Thorgaard 1985). Residues of irradi-
ated nuclear DNA in the form of chromosome
fragments can destabilize a gynogenetic or
androgenetic genome by their uncontrolled incor-
poration into the intake chromosomes. The resi-
dues can also provoke chromosome rearrangements
or disturb cell divisions and may interfere with em-
bryonic development of gynogenetic or
androgenetic organisms.
The primary goal of this study was to use
androgenesis for evaluating the influence of spon-
taneous as well as induced chromosome rear-
rangements on the survival rate of androgenetic
rainbow trout.
Materials and methods
Fish and gamete collection
Gametes were collected in April 2005 from
broodstocks of rainbow trout from the Rutki strain
kept at the Department of Salmonid Research, In-
land Fisheries Institute in Olsztyn, Rutki, Poland.
The strain had been established based on 5 founder
strains (Oleœnica and Jastarnia from Poland;
Saitama from Japan; A13 and Donaldson from the
USA, imported from Finland), as described by
Dobosz et al. (1992). The Rutki strain is polymor-
phic for Robertsonian fusions, and the diploid
chromosome number in fish from the strain ranges
from 2n = 59 to 2n = 62 (Ocalewicz 2002). More-
over, 2 different morphological forms of the Y
chromosome are observed in rainbow trout from
the Rutki strain. The longer form, YL
(undistinguishable from the X chromosome), and
the shorter form, YS (not X-like, similar to
acrocentric autosomes) differ in length of the short
arm (p). Moreover, YS lacks a pericentromeric
cluster of AT-rich chromatin and 5S rDNA se-
quences (Ocalewicz et al. 2007). Oocytes were
collected from six 5-year-old females. The eggs
were pooled and kept in a plastic container on ice
before further treatment. The 4 rainbow trout
males used in the experiment were randomly cho-
sen and stripped just before insemination.
Induction of androgenetic development
In order to destroy maternal nuclear DNA, pooled
oocytes were transported to the Clinic of Radio-
therapy, Medical School of Gdansk, Poland,
where they were exposed to X-ray radiation by us-
ing a medical linear accelerator (Clinac 2300).
Some oocytes were not subjected to irradiation
and left as controls. The eggs were irradiated with
50 Gy, 150 Gy, 250 Gy or 350 Gy. During and af-
ter irradiation, the oocytes were covered with
black foil to avoid photo reactivation of maternal
nuclear DNA. After irradiation, the oocytes were
transported back to the Department of Salmonid
Research, Rutki. Oocytes from each irradiation
variant, as well as untreated oocytes, were further
divided into 4 batches and inseminated independ-
ently with semen collected from the 4 males.
High-pressure shock (48 265 kPa for 4 min) was
applied 350 min after fertilization to some of the
inseminated eggs, to double the haploid chromo-
somes set in the zygote during their first mitotic di-
vision (Chourrout 1984; Parsons and Thorgaard
1985). Fertilized irradiated eggs that were not sub-
jected to the high-pressure shock, as well as un-
treated fertilized eggs, were left to develop as
androgenetic haploid controls and normal diploid
controls, respectively. The irradiated and fertilized
batches subjected to the high-pressure shock con-
tained about 2100 eggs each, while haploid
 Androgenesis in rainbow trout
androgenetic and normal diploid control batches
contained about 500 eggs each.
Survivability of androgenetic rainbow trout
The experimental and control groups were incu-
bated in 2 separate replicates under routine pro-
gram conditions performed at the Department of
Salmonid Research, Rutki. Survival rates of
androgenetic rainbow trout embryos and hatched
larvae obtained due to the irradiation with 50 Gy,
150 Gy, 250 Gy or 350 Gy, as well as controls,
were analyzed during fish ontogeny. The fertiliza-
tion rate of androgenetic haploid, diploid and un-
treated embryos was calculated 48 h after egg
insemination. Embryo survival was assessed at the
eyed-stage, 6 months after hatching, as the number
of eyed embryos divided by the number of fertil-
ized eggs. Survival rates of haploid and diploid
androgenetic eyed embryos were compared (hap-
loid/diploid assay) to evaluate the impact of the
high-pressure shock on survival rate of
androgenetic rainbow trout.
Cytogenetic screening of fish
After 6 months of rearing, 27 out of 55
androgenetic rainbow trout (offspring of 4 fathers)
were chosen for the karyological analysis:
11 specimens from variant 250 Gy, and 16 speci-
mens from variant 350 Gy. As a control, 16 normal
untreated rainbow trout specimens were analysed.
The remaining progenies have been kept for fur-
ther breeding experiments. The morphological de-
velopment of the fish was examined. The
phenotypic sex of the studied specimens was de-
termined by histological examination of the go-
nads after fish dissection (wet squash of gonadal
tissue under a light microscope).
Somatic metaphase plates were prepared from
head kidney cells of studied fish, using conven-
tional air-drying technique (Rab and Roth 1988).
Chromosomes were stained in buffered Giemsa
(10%, 7 min) for visualization. For better charac-
terization, metaphase spreads were stained with
DAPI. Three drops of antifade solution
Vectashield, containing DAPI (1.5 µg mL–1)
(Vector, Burlingame, USA) were dropped onto a
slide, and the preparation was covered with a
coverslip.
Statistical analysis
The survival rate (p) was subjected to arcsin root
transformation (T = arcsin p) (Babiak 1998).
The transformed data showed a normal distribu-
311
tion, and the variances were homogenous
(Cochran’s and Bartlett’s test). Analysis of vari-
ance (ANOVA) was made for transformed sur-
vival rate data in various experimental groups, and
significance of the differences between groups
was assessed with the Duncan multiple range test.
Two levels of significance (significant P £ 0.05;
highly significant P £ 0.01) were designated to
verify the hypotheses.
Results
Survival rates of haploid and diploid
androgenetic rainbow trout embryos during
embryogenesis
No significant differences in survival rates be-
tween androgenetic haploid and diploid offspring
of 4 males were observed (P > 0.05). However, the
post-hoc Duncan test revealed that the survival
rates of haploid and diploid androgenetic embryos
observed from the time of fertilization to the stage
of eyed embryos increased significantly with in-
creasing doses of ionizing radiation applied during
the maternal DNA inactivation (P < 0.05) (Ta-
ble 1). The mean survival rates of haploid
androgenotes at this stage of embryogenesis were
3.0%, 18.6%, 24.4%, and 28.9% in irradiation
variants 50 Gy, 150 Gy, 250 Gy, and 350 Gy, re-
spectively (percentage was calculated in relation
the to fertilization rate) (Table 1). For diploid
androgenetic individuals, the mean survival rates
of androgenetic offspring in the same irradiation
variants were 0.78%; 4.30%; 7.52%, and 10.6%,
respectively (Table 1). The comparison of surviv-
ability between haploid and diploid androgenetic
rainbow trout from the same irradiation variants at
the stage of eyed embryos was applied to evaluate
the impact of the diploidization factor, namely the
high-pressure shock, on the survival of
androgenetic embryos during embryogenesis. Ap-
parently, the pressure shock decreased signifi-
cantly the survival rate of the diploid androgenetic
offspring in each irradiation variant (P < 0.001)
(Table 1).
Survival rates of androgenetic rainbow trout at
hatching
The survival of haploid and diploid androgenetic
embryos after the eyed stage decreased dramati-
cally. No haploids from any irradiation variant and
no diploid fish from the 50 Gy variant hatched
(Table 2). For further statistical analysis, all
androgenetic offspring in the 150 Gy, 250 Gy, and
312 K. Ocalewicz et al.

Table 1. Survival rates of haploid and diploid androgenetic rainbow trout (Oncorhynchus mykiss) eyed embryos –
progenies of 4 males – after X-ray irradiation (0–350 Gy)
Father Survival (%, mean ± SD) of androgenetic eyed embryos
no. 50 Gy 150 Gy 250 Gy 350 Gy Control
1n 2n 1n 2n 1n 2n 1n 2n 1n 2n
2.24 ± 0.32 ± 15.3 ± 2.62 ± 22.3 ± 7.37 ± 25.8 ± 6.86 ± 16.4 ± 92.4 ±
1
0.25 0.45 0.78 0.90 1.05 0.08 0.18 0.77 9.67 2.24
2.06 ± 0.79 ± 15.0 ± 4.17 ± 22.6 ± 6.33 ± 24.9 ± 10.4 ± 16.1 ± 94.0 ±
2
0.58 0.17 1.10 0.71 4.14 3.04 4.78 1.54 9.83 1.07
4.02 ± 0.99 ± 19.6 ± 4.49 ± 25.9 ± 8.78 ± 28.4 ± 12.6 ± 19.5 ± 96.1 ±
3
0.31 0.08 4.89 0.15 3.95 0.64 1.55 3.96 10.4 0.28
3.85 ± 1.00 ± 24.5 ± 5.92 ± 26.9 ± 7.61 ± 36.4 ± 12.3 ± 22.9 ± 92.5 ±
4
1.82 0.05 0.90 0.86 6.97 1.94 0.54 0.27 13.0 3.33
Overall 3.04 ± 0.78 ± 18.6 ± 4.30 ± 24.4 ± 7.52 ± 28.9 ± 10.6 ± 18.7 ± 93.8 ±
mean 1.21 0.35 4.55 1.37 4.05 1.67 5.20 2.94 10.6 2.25

350 Gy variants were studied. Survival rate of
hatched larvae was statistically the highest in the
350 Gy irradiation variant (P < 0.01) (Table 2).
After 6 months of rearing, only 11 specimens sur-
vived from the 250 Gy variant and 44 specimens
from the 350 Gy variant (Table 2).
250 Gy variant was greater than in the 350 Gy
group, and ranged from 1 to 4, whereas up to 3
fragments were observed in the fish from the
350 Gy group (Figure 2). Also intraindividual
variation of the chromosome fragment number
was observed (Table 3). No chromosome frag-

Table 2. Number of viable specimens of diploid androgenetic rainbow trout

(Oncorhynchus mykiss) hatchlings and fingerlings
No. of viable specimens
Father no.
50 Gy 150 Gy 250 Gy 350 Gy
hatch. fing. hatch. fing. hatch. fing. hatch. fing.
1 0 0 5 0 26 2 27 10
2 0 0 7 0 15 1 31 11
3 0 0 2 0 38 3 38 11
4 0 0 6 0 22 5 50 12

Chromosomal analysis ments were observed in the karyotypes of un-

Modal chromosome number of androgenetic off-
spring of male 1 was 60. Androgenetic progenies
of male 2 were characterized by diploid chromo-
some numbers of 58 and 60. Metaphase spreads
consisting of 62 chromosomes were found in
androgenetic offspring of male 3. Androgenetic
progenies of male 4 had 58, 60 and 62 chromo-
somes. Two specimens from the 350 Gy variant
exhibited co-existence of haploid and diploid cell
populations among the studied head kidney cells.
In one specimen, the quality of metaphase spreads
was very poor and we were not able to provide any
cytogenetic characteristics (Figure 1, Table 3).
Chromosome fragments of various size were
detected in 9 out of 11 (~82 %) and 6 out of 16
(~38 %) specimens from the 250 Gy and 350 Gy
groups of androgenotes, respectively. None of the
chromosome fragments was bigger than the small-
est intact rainbow trout chromosomes. The num-
ber of chromosome fragments in fish from the
treated rainbow trout individuals.
Sex genotypes and sex ratio among androgenotes
Phenotypic sex was determined in all
cytogenetically studied specimens. Phenotypic fe-
males and males were found among the examined
androgenetic offspring of males 2 and 4 (Table 3).
Only female offspring of male 1 (2 individuals)
and only male offspring of male 3 (3 individuals)
were examined in this regard. Male progenies of
males 2 and 3 showed YSYS sex genotypes. Male
offspring of male 4 had YLYL sex chromosomes.
All androgenetic females exhibited XX sex geno-
types (Table 3).
Examination of fish morphology
Six androgenetic rainbow trout (one from the
250 Gy variant and 5 from the 350 Gy variant)
showed body malformations, namely shortened
 Androgenesis in rainbow trout 313

Figure 1. Metaphase spreads with (a) 29 chromosomes (haploid cell) and (b) 58 chromosomes (diploid cell) from
androgenetic mosaic (haploid/diploid) rainbow trout female. Arrows indicate X chromosomes

Figure 2. Residues of maternal DNA in the form of (a) 1, (b) 2, (c) 3, and (d) 4 chromosome fragments (arrowheads) in
androgenetic rainbow trout: (a) female, 2n = 60, XX; (b) male, 2n = 60, YSYS; (c) male, 2n = 60, YLYL; and (d) male, 2n =
62, YSYS. Yellow arrows point to X chromosomes, while white arrows indicate YL chromosomes

body and higher width of the body, compared to chromosome fragments, and one of them was a
their androgenetic siblings and fish from the con- mosaic of haploid/diploid cells.
trol group (Table 3). Two morphologically under-
developed specimens were carriers of
314 K. Ocalewicz et al.

Table 3. Cytogenetic analysis, gonadal sex, and body shape of androgenetic progenies of rainbow trout
(Oncorhynchus mykiss)
Number Irradiation Father no. Body shape Gonadal sex No. of chromosomes Sex genotype No. of chromosome
dose(Gy) fragments
1 250 1 normal female 60 XX 0–1
2 250 1 normal female 60 XX 0–1
3 250 2 normal female 60 XX 0?
4 250 3 normal male 62 Y SY S 0–4
5 250 3 normal male 62 Y SY S 0–4
6 250 3 abnormal male ND ND ND
7 250 4 normal male 60 YLYL 0–2
8 250 4 normal male 62 YLYL 0–3
9 250 4 normal female 60 XX 0–1
10 250 4 normal male 60 YLYL 0–4
11 250 4 normal male 60 YLYL 0–3
12 350 2 abnormal male 58 Y SY S 0?
13 350 2 abnormal male 60 Y SY S 0–2
14 350 2 abnormal male 30/60 Y SY S 0–2
15 350 2 normal female 58 XX 0?
16 350 2 normal female 29/58 XX 0–1
17 350 2 normal female 58 XX 0?
18 350 4 abnormal female 60 XX 0?
19 350 4 normal male 60 YLYL 0?
20 350 4 normal female 60 XX 0?
21 350 4 normal female 58 XX 0?
22 350 4 normal female 60 XX 0?
23 350 4 normal male 60 YLYL 0?
24 350 4 normal male 60 YLYL 0–1
25 350 4 normal male 60 YLYL 0–3
26 350 4 normal female 60 XX 0–2
27 350 4 abnormal female 60 XX 0?

Discussion (Phillips et al. 1999). Our results confirm the ear-

Results of karyological analysis of the
androgenetic rainbow trout progenies suggested
that their fathers were characterized by various
chromosome numbers, resulting from the
Robertsonian fusions and fissions. This is in
agreement with previously published data con-
cerning the Rutki strain, whose chromosome num-
ber varies from 2n = 59 to 2n = 62 (Ocalewicz
2002). Although animals that are heterozygous for
Robertsonian rearrangements might show
subfertility caused by the aberrant pairing and
malsegregation during meiosis, we did not ob-
serve any significant differences in survival rates
between androgenetic offspring of the 4 males ob-
served from the time of fertilization to the stage of
eyed embryos. Similarly, no significant difference
in viability of pink salmon (Oncorhynchus
gorbusha) offspring from crosses between speci-
mens showing different cytotypes related to chro-
mosome number and structure caused by
chromosome fission and inversion was found
lier suggestion that heterozygotes for simple
Robertsonian rearrangements resulting from
centric fusions or fissions do not suffer from any
reduction in fitness (Phillips and Kapuscinski
1988).
In the males from the studied rainbow trout
strain, 2 morphological forms of the Y chromo-
some are observed. Sex genotypes of the male
androgenetic progenies indicated that males 2 and
3 had a short form of the Y chromosome (YS),
while male 4 had its longer form (YL). In the mam-
malian species, the Y chromosome rearrange-
ments may be responsible for the loss of key locus
or loci or abnormality during pairing and segrega-
tion with the X chromosome and thus may cause
infertility or reduce fertility in males (Affara
2003). In the present study we may have eluci-
dated the interaction between rainbow trout Y
chromosome morphology and reproductive effi-
ciency of males with YL and YS chromosomes.
The lack of significant differences in survivability
between androgenetic and normal offspring of
4 males used as sperm donors in the experiment
 Androgenesis in rainbow trout
suggested the lack of such an association. Presum-
ably, the partial deletion of the Y chromosome
p-arm, which is considered as a rearrangement
leading to the formation of the YS chromosome
morph (Ocalewicz et al. 2007), did not comprise
any locus or loci whose absence might affect fit-
ness of the males with the XYS sex genotype. On
the other hand, partial interstitial translocation or
deletion of the rainbow trout X chromosome
p-arm region adjacent to the 5S rDNA locus and
the telomeric region or a terminal deletion adja-
cent to the 5S rDNA sequences may be lethal in
the females homozygous for this rearrangement
(Ocalewicz and Dobosz 2009).
Decrease of the survival rates of androgenetic
rainbow trout paralleled with decreasing doses of
radiation applied for the inactivation of maternal
nuclear DNA is known as the Hertwig effect and
might have its basis in deleterious chromosome re-
arrangements and disturbances of the cell divi-
sions caused by the residues of maternal nuclear
genome that endured the X ray-enucleation. Previ-
ously, the Hertwig effect has been observed when
increasing doses of UV radiation were applied to
inactivate the tiger barb (Puntius tetrazona
Bleeker) oocyte nuclear DNA (Kirankumar and
Pandian 2003). A too little dose of radiation ap-
plied for the inactivation of nuclear DNA in sperm
decreased the survival rate of the gynogenetic sea
bass (Carillo et al. 1995; Felip et al. 1999).
The increased number of the chromosome
fragment carriers and increasing number of the
fragments with decreasing radiation doses ob-
served among androgenetic rainbow trout individ-
uals were consistent with those for the loach
(Misgurnus anguillicaudatus Cantor) (Fujimoto
et al. 2007).
Androgenetic rainbow trout fingerlings with
residues of the maternal genome contained popu-
lations of cells with various numbers of chromo-
some fragments. As the kinetochore-lacking
chromosome fragments have been gradually lost
during the cell cycle (Fujiwara et al. 1997), 2 or
more cell fractions showing different genotypes
could have originated as a result of uneven elimi-
nation of the fragments from the zygote after its
first mitotic division. In some cases, inheritance of
the fragments was stabilized during fish ontogeny
(Ocalewicz et al. 2004), although its mechanism
remained unclear.
The presence of the chromosome fragments
apart from the diploid set of chromosomes was
considered to trigger the germ cell development
and cause irregular ovarian development
315
in gynogenetic females (Krisfalusi et al. 2000). On
the other hand, adult androgenetic rainbow trout
with gamma-radiation-induced chromosome frag-
ments were morphologically normal and fertile
(Ocalewicz et al. 2004).
Our analysis of survival of androgenetic hap-
loid and diploid rainbow trout showed that
high-pressure treatment decreases significantly
the survivability of androgenetic rainbow trout.
Several-fold higher mortality of diploid
androgenotes in comparison to their haploid sib-
lings observed during early stages of development
mirrored the level of deleterious effect of the
high-pressure shock. Apart from the desta-
bilization of the microtubules, the high-pressure
shock may induce deformations of other cell
organelles (Parkkiren et al. 1993; Crenshaw et al.
1996) or provide viable mosaic haploid/diploid in-
dividuals (Zhang and Onozato 2004). Among
cytogenetically studied androgenetic rainbow
trout, 2 fish exhibited populations of haploid and
diploid cells (Figure 1, Table 2). Mosaic fish can
suffer from numerous body malformations
(Tanaka et al. 2003) and indeed one mosaic speci-
men in our experiment exhibited disturbances in
body growth and development (Table 3).
Conclusions
Spontaneous chromosome rearrangements, like
Robertsonian chromosome polymorphism, did not
impair fertility in the rainbow trout males from the
Rutki strain. Additionally, progenies of males ex-
hibiting the Y chromosome length polymorphism
showed similar survival rates during the early de-
velopment. This may be interpreted as the lack of
association between the Y chromosome morphol-
ogy and reproductive performance in this species.
On the other hand, both partial inactivation of the
maternal nuclear genome and high-pressure shock
significantly decreased the survival rate of
androgenetic rainbow trout. The androgenetic
rainbow trout individuals irradiated with less than
250 Gy died.
Acknowledgement. We thank Joanna Nowaczyk,
Bodr University College, Norway, for her help in the
early stages of the experiment, and Prof. Miros³aw
£uczyñski, University of Warmia and Mazury in
Olsztyn, Poland, for critical reading of the manu-
script. This work was supported by the University of
Warmia and Mazury in Olsztyn (project no.
0804.0809).
316 K. Ocalewicz et al.
REFERENCES
Affara NA, 2003. The role of the Y chromosome in
male infertility. Reprod Med Rev 11: 67–85.
Babiak I, 1998. Experimental design and statistical
analysis in research on fish reproduction. Archiv
Polish Fish 6: 5–34.
Babiak I, Dobosz S, Goryczko K, Kuzminski H,
Brzuzan P, Ciesielski S, 2002. Androgenesis in
rainbow trout using cryopreserved spermatozoa: the
effect of processing and biological factors.
Theriogenology 57: 1229–1249.
Brown KH, Thorgaard GH, 2002. Mitochondrial and
nuclear inheritance in an androgenetic line of rain-
bow trout, Oncorhynchus mykiss. Aquaculture 204:
323–335.
Carillo M, Zanuy S, Blazquez M, Ramos J, Piferer F,
Donaldson EM, 1995. Sex control and ploidy ma-
nipulation in sea bass. In: Environmental impacts of
aquatic biotechnology. Paris: OECD: 125–143.
Chourrout D, 1984. Pressure-induced retention of sec-
ond polar and suppression of first cleavage in rain-
bow trout: production of all triploids, all tetraploids,
and heterozygous and homozygous dip-
loid gynogenesis. Aquaculture 36: 111–126.
Chourrout D, Quillet E, 1982. Induced gynogenesis in
rainbow trout: sex and survival of progenies: pro-
duction of all triploid populations. Theor Appl
Genet 63: 201–205.
Crenshaw HC, Allen JA, Skeen V, Harris A,
Salmon ED, 1996. Hydrostatic pressure has differ-
ent effects on the assembly of tubulin, actin, myosin
II, vinculin, talin, vimentin, and cytokeratin in
mammalian tissue cells. Exp Cell Res 227:
285–297.
Devlin RH, Nagahama Y, 2002. Sex determination and
sex differentiation in fish: an overview of genetic,
physiological, and environmental influences.
Aquaculture 208: 191–364.
Dobosz S, Goryczko K, Olech W, Zyczynski A,
Friedrich F, Kohlman K, 1992. Incomplete diallele
cross of rainbow trout strains imported to Poland.
Fortschritte der Fischerewissenschaft 10: 75–82.
Felip A, Fujiwara A, Young WP, Wheeler PA,
Noakes M, Phillips R, Thorgaard GH, 2004. Poly-
morphism and differentiation of rainbow trout Y
chromosomes. Genome 47: 1105–1113.
Felip A, Piferrer F, Carillo M, Zanuy S, 1999. The rela-
tionship between the effects of UV light and ther-
mal shock on gametes and the viability of early
developmental stages in a marine teleost fish, the
sea bass (Dicentrarchus labrax L.). Heredity 83:
387–397.
Flajshans M, Rab P, 1987. A finding of the triploid rain-
bow trout, Kamloops form (Salmo gairdneri
Kamloops). Zivoc Vyr 32: 279–282.
Fujimoto T, Sakao S, Yamha E, Arai K, 2007. Evalua-
tion of different doses of UV irradiation to loach
eggs for genetic inactivation of the maternal ge-
nome. J Exp Zool 307: 449–462.
Fujiwara A, Abe S, Yamaha E, Yamazaki F,
Yoshida MC, 1997. Uniparental chromosome elim-
ination in the early embryogenesis of the inviable
salmonid hybrids between masu salmon female and
rainbow trout male. Chromosoma 106: 44–52.
Kirankumar S, Pandian TJ, 2003. Production of
androgenetic tiger barb, Puntius tetrazona.
Aquaculture 228: 37–51.
Komen H, Thorgaard GA, 2007. Andro-
genesis, gynogenesis and the production of clones in
fishes. Aquaculture 269: 150–173.
Krisfalusi M, Wheeler PA, Thorgaard GH, Cloud JG,
2000. Gonadal morphology of female diploid
gynogenetic and triploid rainbow trout. J Exp Zool
286: 505–512.
Martinez V, Thorgaard GH, Robinson B, Sillanpaa MI,
2005. An application of Bayesian QTL mapping to
early development in double haploid lines of rain-
bow trout including environmental effects. Genetics
Res 86: 209–221.
McKone MJ, Halpern SL, 2003. The evolution of
androgenesis. Am Nat 161: 641–656.
Nichols KM, Young WP, Danzmann RG, Robison BD,
Rexroad C, Noakes M, et al. 2003. A consolidated
linkage map for rainbow trout (Oncorhynchus
mykiss). Anim Genet 34: 102–115.
Ocalewicz K, 2002. Cytogenetic markers for X chro-
mosome in karyotype of rainbow trout from Rutki
strain. Folia biologica (Krakow) 50: 10–14.
Ocalewicz K, Babiak I, Dobosz S, Nowaczyk J,
Goryczko K, 2004. The stability of telomereless
chromosome fragments in adult androgenetic rain-
bow trout. J Exp Biol 207: 2229–2236.
Ocalewicz K, Babiak I, Kasprzycka B, Dobosz S,
Kuzminski H, Goryczko K, 2007. Occurrence of
two forms of Y chromosome in rainbow trout
(Oncorhynchus mykiss) males from Rutki strain.
Aquaculture 270: 546–551.
Ocalewicz K, Dobosz S, 2009. Karyotype variation in
the albino rainbow trout (Oncorhynchus mykiss
(Walbaum)). Genome 52: 347–352.
Pandian TJ, Koteeswaran R, 1998. Ploidy induction
and sex control in fish. Hydrobiologia 384:
167–243.
Parkkiren JJ, Lammi MJ, Pelttari A, Helminen HJ,
Tammi M, Virtanen I, 1993. Altered Golgi appara-
tus in hydrostatically loaded cartilage chondrocytes.
Ann Rheum Dis 52: 192–198.
Parsons JE, Thorgaard GH, 1985. Production of
androgenetic diploid rainbow trout. J Hered 76:
177–181.
Phillips RB, Kapuscinski AR, 1988. High frequency of
translocation heterozygotes in odd year populations
of pink salmon (Oncorhynchus gorbuscha).
Cytogenet Cell Genet 48: 178–182.
Phillips RB, Matsuoka MP, Smoker WW, Gharett AJ,
1999. Inheritance of a chromosomal polymorphism
in odd-year pink salmon from southeastern Alaska.
Genome 42: 816–820.
Rab P, Roth P, 1988. Cold-blooded vertebrates.
In: Balicek P, Forejt J, Rubes J, eds. Methods of
chromosome analysis. Brno, Cytogeneticka sekce,
Ceskoslovenske biologicke spolecnosti pri CSAV,
Czech Republic: 115–124.
 Androgenesis in rainbow trout
Tanaka M, Kimura S, Fujimoto T, Sakao S,
Yamaha E, Arai K, 2003. Spontaneous mosaicism
in normally fertilized and gynogenetically induced
progeny of the kokanee salmon Oncorhynchus
nerka. Fish Sci 69: 176–180.
Thorgaard GH, 1977. Heteromorphic sex chromo-
somes in male rainbow trout. Science 196:
900–902.
Thorgaard GH, 1983. Chromosomal differences among
rainbow trout populations. Copeia 1983: 650–662.
317
Thorgaard GH, Gall GAE, 1979. Adult triploids in a
rainbow trout family. Genetics 93: 961–973.
Young WP, Wheeler PA, Coryell VH, Keim P,
Thorgaard GH, 1998. A detailed linkage map of
rainbow trout produced using doubled haploids.
Genetics 148: 839–850.
Zhang X, Onozato H, 2004. Hydrostatic pressure treat-
ment during the first mitosis does not suppress the
first cleavage but the second one. Aquaculture 240:
101–113.