Comp. Biochem. Physiol. Vol. 118A, No. 1, pp. 183–190, 1997 ISSN 0300-9629/97/$17.

00
Copyright  1997 Elsevier Science Inc. PII S0300-9629(96)00415-X

Effects of Accelerated Photoperiod Regimes on the

Reproductive Cycle of the Female Rainbow Trout:
I—Seasonal Variations of Plasma Lipids Correlated
with Vitellogenesis
E. Bon,1,2 G. Corraze,3 S. Kaushik,3 and F. Le Menn1
1
Laboratoire de Biologie de la Reproduction des Poisson, U.A. INRA, Uuniversité Bordeaux I, Avenue des,
Facultés, F-33405 Talence Cedex, France; 2 Salmonidés d’ Aquitaine, Route de Taller, F-40260 Castets, France;
and 3 Laboratoire de Nutrition des Poissons, Unité Mixte INRA-IFREMER, Station d’ Hydrobiologie, F-64310
Saint Pée sur Nivelle, France

ABSTRACT. Female rainbow trout were exposed to a simulated natural photoperiod (control group N) and
to two accelerated photoperiod regimes (S9 and S6 groups). Early spawning was achieved in both the S9 and
S6 groups, coupled, however, with a reduction of mean egg size. To investigate this reduction of egg size, some
morphometric parameters, as well as some plasma parameters such as lipids and vitellogenin levels, were regularly
measured. Regardless of the photoperiod regime, a preferential utilization of lipids for energy purposes was ob-
served during previtellogenesis and Type I-vitellogenesis, whereas during Type II-vitellogenesis, mobilized lipids
were preferentially used for lipoproteins and notably for vitellogenin synthesis. Different patterns of the visceroso-
matic index, triacylglycerols, and non-esterified fatty acids were observed during previtellogenesis and Type I-
vitellogenesis, whereas-similar patterns of all paremeters were observed during Type II-vitellogenesis. From these
results it may be concluded that previtellogenesis and Type I-vitellogenesis are photosensitive periods, whereas
Type II-vitellogenesis is probably under the control of an endogenous biological rhythm synchronized by the
photoperiod. Our findings also demonstrate that the reduction of egg size could be explained neither by lower
plasma TG levels nor by lower vitellogenin (Vtg) levels, since both were much higher during Type II-vitellogen-
esis. comp biochem physiol 118A;1:183–190, 1997.  1997 Elsevier Science Inc.

KEYWORDS. Biological clock, endogenous rhythm, photoperiod, plasma lipids, rainbow trout, reproductive
cycle, vitellogenin

INTRODUCTION existing broodstocks, but also an extension of the availabil-

ity of fertilized eggs at any time of the year. Thus, in the
In rainbow trout, gonadal maturation is characteristically
last decade, photoperiodic manipulations of the reproduc-
an annual event with gonadal growth commencing in
tive cycle have been developed on commercial farms to
spring, in preparation for spawning in the autumn or winter
overcome the seasonal aspect of spawning (8). Many experi-
months (9). Most salmonids, including rainbow trout, ap-
ments performed on rainbow trout have demonstrated that
pear to rely on the seasonally changing cycle of daylength
exposure of fish to advanced or compressed seasonal light
for the timing of gonadal recrudescence, maturation and
cycles induces precocious gonadal development, whereas
spawning (9,12,13). In wild stock, this dependence on light
delayed or extended seasonal photoperiods result in later
is of considerable adaptative significance. Indeed, it has
spawning (8,12,14). However, advancing the time of
been demonstrated that the annual change in photoperiods
spawning generally produces smaller egg sizes, which are not
continuously induces an autonomous endogenous circan-
acceptable for general sale, whereas neither the egg quality
nual clock (12,13). This ensures the production of offspring
nor the potential growth of the fry produced are altered
at a time of the year when their chances of survival are
(7,9).
optimal (9,12,13).
In teleosts, as in other oviparous vertebrates, vitellogen-
The recent increase in the retail market for trout requires
esis is a crucial period in the female reproductive cycle,
not only an optimisation of the reproductive potential of
characterized by a pronounced growth of oocytes. Much of
this growth results from the uptake of specific proteins, pre-
Address reprint requests to: E. Bon, Laboratoire de Biologie de la Reproduc- dominantly a hepatically-derived yolk lipoprotein precur-
tion des Poissons, U.A INRA, Université Bordeaux I, Avenue Des Fa-
cultés, F-33405 Talence Cedex, France. sor: vitellogenin (24,36). Also, fish exhibit seasonal varia-
Received 12 March 1996; accepted 10 October 1996. tions in their lipid metabolism especially related to the
184
reproductive process (3,17,38,40). The primary aim of the
present study was to determine whether egg size reduction
was due to an alteration of lipid metabolism and vitello-
genin synthesis during vitellogenesis. Consequently, some
morphometric parameters and some plasma parameters were
measured regularly in rainbow trout reared under normal
and accelerated photoperiod regimes. In this study, morpho-
metric parameters (gonadosomatic, hepatosomatic, and
viscerosomatic indices) were used as indirect indicators of
the physiological stage of lipid metabolism and vitellogen-
esis, whereas plasma parameters (triacylglygerols, non-ester-
ified fatty acids, total cholesterol, and vitellogenin) were
used as direct indicators.
MATERIALS AND METHODS
Fish and Photoperiod Conditions
Three hundred sixty 2-year-old female rainbow trout (On-
corhynchus mykiss) with an average body weight of 1.46 6
0.16 kg were randomly selected after their first spawning in
December 1991. They were divided into three groups of 120
fish, and 10 per group were individually identified by a num-
bered tag attached through the base of the dorsal fin. Fish
were reared under natural temperature conditions (8.5 6
0.5°C) in 4m3 covered tanks continuously supplied with
spring water. They were fed twice a day with a pelleted
broodstock diet (Trouvit, Trouw France) containing 45%
crude protein and 9% crude fat, distributed at a rate of 1%
body-weight per day over the first 3 months following
spawning, then 0.7% over the next 2 months, and finally
0.3% up to the next spawning.
All groups were maintained under artificial photoperiod
in tanks illuminated by a 75-W bulb providing approxi-
mately 950 lux at the surface of the water. Daylength was
automatically controlled by 24-hr electronic time clocks,
and the switch-off time was manually adjusted each week
according to the prescribed schedule (Fig. 1). In the control
FIG. 1. Photoperiod regimes. N: Photoperiod regime simu-
lating natural conditions (control group); S9 and S6: Accel-
erated photoperiod regimes (treated groups).
E. Bon et al.
group (N), fish were subjected to the photoperiod regime
they would normally receive in their natural environment.
The spawning period of controls was expected to occur in
late November. In the two other groups (S9 and S6), fish
were subjected to accelerated photoperiod regimes leading
to earlier spawning. Accelerated photoperiod regimes were
obtained by an abrupt increase in daily light from the previ-
ous 8L: 16D to 16L : 8D in December, immediately after
spawning in the S6 group, and in February, 2 months after
spawning in the S9 group. Afterwards, daylight was progres-
sively reduced from 16L: 8D to 8L: 16D, exactly as in the
control group (Fig. 1).
Biological Samples
Twice a month until spawning, the 10 tagged fish from each
group were anesthetized with 2-phenoxyethanol (0.03%),
weighed, and blood sampled. The blood was collected from
the caudal sinus into heparinized microtubes immediately
refregerated on ice. Blood samples were centrifuged (2000
g, 10 min, 4°C) and the plasma was collected, frozen on dry
ice, and stored at 220°C until use. At spawning, the tagged
fish were individually stripped, and the diameter of 20 to
30 eggs per female was immediately measured under a bin-
ocular microscope to determine mean egg size.
Also, twice a month until spawning, a further 10 non-
tagged fish per group were randomly selected, anesthetized,
weighed, and killed. The gonads, the liver, and all the adi-
pose tissue associated with caeca and viscera were removed
from the body cavity and weighed to determine respectively,
the gonadosomatic index (GSI), the hepatosomatic index
(HSI) and the viscerosomatic index (VSI). Despite a 4 day
fast, their digestive tracts still contained undigested food,
which was removed and the VSI was calculated. GSI, HSI,
and VSI were calculated as organ to body-weight ratios.
Analyses
The plasma concentrations of triacylglycerols (TG), non-
esterified fatty acids (NEFA), and total cholesterol (CHOL)
were evaluated colorimetrically using commercially avail-
able enzymatic kits, respectively: Triacylglycerol N Wako
and NEFA C Wako (Unipath, France), and Cholesterol
CHOD-PAP (Boehringer Mannheim, France). Plasma sam-
ples were diluted in NaCl 9 g/l prior to assay to avoid any
protein precipitation. The results were expressed in milli-
grams per deciliter of plasma (mg/dl).
The plasma concentrations of vitellogenin (Vtg) were de-
termined by microassay using an ELISA for rainbow trout
Vtg developed by Bon et al.(6). Vtg was coated on microti-
tration plates and competed with free Vtg (plasma samples
or standards) for the binding on a specific antibody directed
against Vtg. The antigen-antibody complexes formed were
detected by the peroxidase-anti-peroxidase (PAP) method.
Absorbances were measured at 492 nm with a Titertek EIA
Plasma Lipids and Vitellogenesis 185

microplate reader. The results were expressed in milligrams

per milliliter of plasma (mg/ml).

Mode of Data Expression

All data is expressed as mean 6 standard error (m 6 SEM).
Comparison of means were made using non-parametric tests
(30).

RESULTS
Morphometric Parameters
GONADOSOMATIC INDEX (GSI: FIGS 2 AND 3a) Based on
the seasonal profile of GSI, the reproductive cycle of the
control group (N) was divided into physiological phases: a
very slow development phase (VSD) from December to
May during which GSI remained at a minimum (around
0.5%); a slow development phase (SD) from May to mid-
September during which GSI increased slowly but steadily
from 0.5% to 6%; and a rapid development phase (RD) from
mid-September to November during which GSI rose
sharply, reaching a maximum value of 16% in late Novem-
ber when fish entered the spawning period. The fish
spawned over a 6-week period starting in early November.
Immediately after the spawning, during the post-ovulatory
phase (Pov) in December, GSI decreased abruptly and re-
turned to basal values. Regardless of the photoperiod re-
gime, the seasonal profile of GSI showed the same pattern
(Fig. 3a). In the S9 group, fish began to spawn in late August
and continued over a 4-week period to mid-September. In

FIG. 3. Morphometric parameters. Effects of accelerated

photoperiod regimes on the seasonal profile of (a) gonadoso-
matic index (GSI), (b) hepatosomatic index (HSI), and
(c) viscerosomatic index (VSI). The time of spawning is indi-
cated by ‘‘O.’’

FIG. 2. Gonad growth, spawning time and egg size. Effects
of accelerated photoperiod regimes on the seasonal profile
of GSI, on the time of spawning and on egg size (n 5 20
per group). The time of spawning is indicated by ‘‘S.’’ The
reproductive cycle is divided into four physiological phases:
VSD 5 very slow ovarian development; SD 5 slow ovarian
development; RD 5 rapid ovarian development; and Pov 5
post-ovulatory period.
the S6 group, the first fish began to spawn in late May, and
the last, 2 weeks after, in early June (Fig. 2). However, a
significant decrease in egg size was observed at spawning in
the S9 and S6 groups (Fig. 2). Compared to the controls
(4.7 6 0.06 mm in diameter), the egg size was reduced by
0.3 mm in the S9 group and by 0.9 mm in the S6 group,
representing, respectively, a loss of 6 and 19%. A similar
decrease in GSI values was also observed at the end of RD
when females entered the spawning period, with, respec-
tively, a loss of 10 and 27% of the ovarian mass. The dura-
tion of RD and Pov remained unchanged in both the S9
and S6 groups. However, the duration of VSD and/or SD
186 E. Bon et al.

was significantly modified. In the S9 group, the duration of

VSD was 35% shorter than in the controls, whereas SD
remained nearly the same. In the S6 group, the duration of
VSD was 56% shorter and early SD 46% shorter than in
the controls.
HEPATOSOMATIC INDEX (HSI; FIG. 3b). In all groups, HSI
varied little during VSD and mid-SD, exhibiting values be-
tween 1.3 and 1.7%. HSI began to increase significantly in
late SD, 4 months before spawning, reaching maximum val-
ues at the very end of SD that is the beginning of RD, 2
months before spawning. The main difference between the
accelerated groups and the controls was the maximum val-
ues of HSI which were 9.5 and 33.9% higher in S9 and S6,
respectively.
VISCEROSOMATIC INDEX (VSI; FIG. 3c). In controls, during
VSD, a significant increase in VSI (123.9%) was observed
(peak A), immediately followed by a great decrease
(226.9%). The minimum values were observed at the very
end of VSD. During SD, a slight increase (116%) was ob-
served (peak B) rapidly, followed by a decrease (232%).
Finally, at spawning, in November, VSI values exhibited a
total loss of over two-thirds of the initial values. Under the
accelerated photoperiod regimes, no significant differences
between groups were observed at any time throughout the
reproductive cycle for both maximum and minimum VSI
values. However, compared to controls, the duration of peak
A was 2 months shorter in the S9 group, and there was no
peak A in the S6 group. Peak B was not affected by the
photoperiod regimes either in terms of duration or in maxi-
mum values. During Pov, a rapid increase in VSI was ob-
served in both the control and the S9 groups, whereas in
the S6 group, VSI continued to decrease.

Plasma Parameters
TRIACYLGLYCEROLS (TG; FIG. 4a) AND NON-ESTERIFIED
FATTY ACIDS (NEFA, FIG. 4b). During gonadal maturation,
transient increases in plasma TG and NEFA levels were ob-
served. In controls, TG levels ranged from 350 to 900 mg/
dl, whereas NEFA levels ranged from 15 to 50 mg/dl. At
the beginning of VSD, a significant decline in TG and
NEFA levels was observed, followed by a rapid rise in TG
and NEFA levels (the beginning of peak A), leading to high
values in March for TG (620 mg/dl) and in early April for
NEFA (50 mg/dl). This increase was followed by a progres-
sive fall which ended at the limit between VSD and SD, as
concerns TG, and in mid-SD as concerns NEFA. During
early SD, TG levels showed constant values (400 mg/dl)
and began to increase in July (the beginning of peak B),
reaching a maximal value of 900 mg/dl in September, 2
months before spawning, between SD and RD. Thereafter,
TG levels gradually decreased until the spawning period
(late November), reaching basal values during Pov. NEFA
FIG. 4. Plasma lipids. Effects of accelerated photoperiod re-
gimes on the seasonal profile of circulating (a) triglycerides
(TG), (b) non-esterified fatty acids (NEFA), and (c) total
cholesterol (CHOL). The time of spawning is indicated by
‘‘O.’’
variations paralleled those of TG with a time-lag of 2 weeks.
Under the accelerated photoperiod regimes, TG peak A ex-
hibited a lower amplitude and a shorter duration in the S9
group and was clearly absent in the S6 group. Regardless of
the photoperiod regime, TG peak B appeared 2 months be-
fore spawning, between SD and RD. The amplitude of TG
peak B, in the S9 group was similar to controls (900 mg/
dl), but much higher in the S6 group (1200 mg/dl). Com-
pared to controls, the NEFA peak A remained unchanged
in the S9 group, whereas it was absent in the S6 group. The
NEFA-peak B amplitude was similar in all groups. However,
its duration was clearly shorter in both the S9 and S6
groups.
Plasma Lipids and Vitellogenesis
FIG. 5. Plasma vitellogenin. Effects of accelerated photope-
riod regimes on the seasonal profile of plasma vitellogenin
(Vtg). The time of spawning is indicated by ‘‘O.’’
TOTAL CHOLESTEROL (CHOL; FIG. 4c). In all groups, the
seasonal profile of CHOL showed similar fluctuations with a
major peak in early-VSD (around 550 mg/dl), a progressive
decrease during VSD and mid-SD, constant values until the
end of SD (around 300 mg/dl), and a slight increase during
Pov. However, several minor differences were observed be-
tween groups with respect to the minimum levels reached.
In controls, the lowest CHOL levels occurred 1 month be-
fore spawning, during RD, and in the S9 and the S6 groups,
3 months before spawning, in late SD. The slight increase
in CHOL levels observed at spawning in the control group
occurred 2 months earlier than in the S9 and S6 groups.
Finally, compared to controls and to the S9 group, CHOL
levels were higher in the S6 group at spawning and during
Pov.
VITELLOGENIN (Vtg; FIG. 5). In controls, Vtg levels gradu-
ally decreased during early VSD. The lowest Vtg levels (less
than 1 mg/ml) were measured in mid-VSD and the first
significant increase in late VSD. After a slow but gradual
increase from 2.4 to 10 mg/ml in early SD, Vtg levels rapidly
increased up to 35 mg/ml between SD and RD. During RD,
Vtg levels increased once again up to a maximum of 60 mg/
ml, which was reached 2 weeks before spawning. Vtg levels
remained high (around 50 mg/ml) until spawning in late
November, and started to decrease gradually just afterwards.
Elevated Vtg levels (around 20 mg/ml) were still present 1
month after spawning, during Pov. Under the accelerated
photoperiod regimes, no significant differences with respect
to the seasonal profile of Vtg were observed between groups.
The main difference was in the maximum values reached
by Vtg levels. Indeed, at the of RD, Vtg levels were 39%
higher in the S9 group (83 mg/ml) and 96% higher in the
187
S6 group (116 mg/ml) than in the control group N (60 mg/
ml).
DISCUSSION
The abrupt increase in daylight from 8L: 16D to 16L : 8D,
we applied at the beginning of the reproductive cycle in the
S9 and S6 groups, produced earlier spawning, i.e., 3 and 6
months respectively, compared to the control group N.
These photoperiod changes had the additional effect of syn-
chronizing the time of spawning of individual fish. Thus,
the period of time between the spawning of the first and
the last fish was 6 weeks in control, which is similar to rain-
bow trout maintained under natural conditions (9), but
only 4 weeks in the S9 group, and 2 weeks in the S6 group.
However, one of the undesired effects of photoperiodic al-
terations on spawning time was a change in egg size. Indeed,
in the S9 and S6 groups, advancing spawning time produced
smaller eggs compared to controls. These observations are
in complete accordance with previous studies performed on
rainbow trout by other authors (7,12,13). Measurements of
circulating levels of lipids (TG, CHOL, and NEFA) and
Vtg, as well as measurements of morphometric parameter
values (GSI, HSI, and VSI), provided a better understand-
ing of the physiological origin of the reduction in egg size.
(a) Early Stages of Gonadal Development
(Previtellogenesis and Type I Vitellogenesis)
In controls, during early VSD (March–May), a rapid in-
crease in VSI was observed simultaneously with a drop in
TG and NEFA plasma levels, suggesting a rapid recovery of
the perigastric adipose tissue (PAT). This depletion in
plasma TG levels, as well as NEFA levels, is probably related
to a storage of plasma lipids in the PAT. Indeed, lipids in
rainbow trout are stored primarily as TG in the visceral fat
(PAT), dark muscle, and to a lesser extent in the liver (32).
Lipid stores acquired during this period could be related to
the intensive food uptake which generally occurs just after
the time of spawning. Regarding plasma CHOL levels, the
slight increase detected immediately after spawning could
be related to the gonadal resorption, as some authors have
suggested in other teleost species (10,16,40). The relatively
high levels of Vtg (30–50 mg/ml) observed during early-
VSD cannot be related to the lipoprotein biosynthetic ac-
tivity of the liver since the HSI remained constant. These
high Vtg levels could be due to the clearance of the re-
maining Vtg from the blood but could also be explained by
the presence of ovarian yolk proteins massively released into
the bloodstream in the course of the follicular atresia (2).
During late-VSD (March–May), a subsequent decrease
in VSI was observed simultaneously with a transient in-
crease of TG and NEFA levels (peak A). In teleost species,
mobilization of lipids from tissue reserves such as the PAT
188
or the carcass has been commonly reported during sexual
maturation (4,27). However, this lipidic flux was not di-
rected towards gonads or the liver for storage since neither
GSI nor HSI fluctuated. Consequently, the progressive de-
crease of TG and NEFA levels that occurred during late
VSD suggests a preferential utilization of mobilized visceral-
lipids for energy purposes and for structural proliferation in
ovaries. During this period, a transient increase of CHOL
levels was observed, immediately followed by a progressive
decrease. This could be explained partly by the mobilization
of CHOL from the adipose tissue and its storage in the liver
for bile formation (19,32). The decrease in CHOL level
could also be explained by the possible utilization of CHOL
as a substrate for steroid biosynthesis (3,38), and notably
for 17β-estradiol production which occurs at a low rate dur-
ing this period (15). This could explain the slight increase
in Vtg levels we observed during late VSD since Vtg is syn-
thesized by the liver under the effect of 17β-estradiol (34).
However, GSI remained at a minimum (around 0.5%), sug-
gesting that gonad development is quiescent, and that this
period, referred to as VSD, probably corresponds to previtel-
logenesis.
During early SD (May–July), a transient increase in VSI
was observed, immediately followed by a depletion (peak
B). This decrease in VSI probably reflects a mobilization of
mesenteric fat stores to support the energy costs incurred
by gonad development (21,31). Indeed, for the first time,
GSI exhibited a slight but significant increase. Even though
a major part of the mobilized TG is still required during
early SD to supply energy demands, a substantial portion is
directed towards the liver for the biosynthesis of lipopro-
teins, and also incorporated into developing ovaries as ovar-
ian recrudescence creates a large demand for fatty acids
(23,27). Lipid inclusions, consisting largely of TG, have
been observed in growing oocytes during this period (39).
They generally result from the uptake and the lipolysis of
circulating lipoproteins such as VLDL, very low density li-
poprotein, or VHDL, very high density lipoprotein, which
are the main form of transport of mobilized visceral lipids
(5,26,32,38). According to these observations, early SD
probably corresponds to Type I-vitellogenesis (22), also re-
ferred to as endogenous vitellogenesis (35) or the cortical
alveolus stage (37).
Under the accelerated photoperiod regimes (the S9 and
S6 groups), the duration of previtellogenesis (VSD) and
Type I-vitellogenesis (early SD) were clearly reduced. Dur-
ing these periods, although oocyte development is minor in
terms of weight, it is physiologically important in terms of
setting up organites involved in future synthetic and endo-
cytotic activities. Our results show that compared to con-
trols, the S9 and S6 groups did not reconstitute their initial
PAT mass, as evidenced by the alteration and, finally, the
suppression of VSI peak A. This deficiency in the recovery
of PAT mass probably reflects a deficiency in mesenteric fat
reserves since similar alterations of plasma TG and NEFA
E. Bon et al.
profiles were also observed. In contrast, plasma CHOL re-
mained unchanged. Usually, mobilization of lipids from
PAT is accomplished by the activation of lipolytic enzymes
under hormonal regulation and notably under estradiol reg-
ulation (3,11,20,32). Recent studies have demonstrated
that modifications in photoperiod regimes may produce
clear differences in the patterns of secretion of this steroid
hormone (12,13). It is proposed that such hormonal differ-
ences might alter the hormonal regulation of lipogenetic
and/or lipolytic enzymes, preventing any storage and/or
mobilization of the mesenteric fat during the early stages of
gonad development in both the S9 and S6 groups. These
results clearly demonstrate that during previtellogenesis and
Type I-vitellogenesis the photoperiod is mainly responsible
for the modulation of the biochemical processes implicated
in reproduction.
(b) Late Stages of Gonad Development
(Type II Vitellogenesis)
During late SD (July–September), the simultaneous decline
in VSI values, in TG, and in NEFA plasma levels, as well
as the concomitant increase in HSI levels, probably reflect
the intensification of the hepatic biosynthesis of Vtg, the
main egg yolk precursor. Indeed, the seasonal profile of Vtg
clearly shows that Vtg levels increased significantly, reach-
ing 35 mg/ml at the end of late SD. During this period, the
slow but steady increase in GSI from 1.3 to 6% probably
reflects the specific incorporation of Vtg into the growing
oocytes (28,37). The concentration in CHOL did not
change much during this period, probably indicating that
plasma CHOL is actively stored as a precursor for ovarian
synthesis of the 17β-estradiol (1,38), estrogen known to be
responsible for the hepatic production of Vtg (35,36).
CHOL might also be directly taken up by the gonads for
tissue generation as cholesterol is a structural component
(23).
During RD (September–November), the lipidic flux
coming from the PAT directed towards the oocytes is gener-
ally interrupted, and most lipids mobilized from visceral fat
stores are used to supply the energy required rather than for
making the yolk (27). Consequently, the great increase in
GSI (up to 16%) observed at the end of this period cannot
be related to the incorporation of visceral lipids in the oo-
cyte, but rather to the very large amounts of Vtg which are
taken up by the growing oocytes during this period (28,34).
At this point of maturation, lipids bound to Vtg are the
major source of lipids for growing oocytes (33). Indeed, rain-
bow trout Vtg contains up to 20% of lipids of which over
two-thirds are phospholipids, while the remainder are tri-
glycerides and cholesterol (18,33). In rainbow trout, the last
stage of Type II-vitellogenesis is generally associated with
a considerable rise in the plasma levels of testosterone and
of 17α-20β progesterone which is the main sex steroid in-
volved in the final oocyte maturation process (15,25). The
Plasma Lipids and Vitellogenesis
transient decrease in CHOL levels we observed at the end of
RD may reflect an intensive production of these sex steroids.
CHOL may also serve in the composition of cell membranes
which are formed at higher rates during this late gonadal
development (10).
All of these observations suggest that late SD probably
corresponds to the beginning of Type II-vitellogenesis, dur-
ing which there is coincidence of ‘‘endogenous’’ and ‘‘exog-
enous’’ vitellogenesis (22), and that RD probably corre-
sponds to the intensive Type II-vitellogenesis, during which
‘‘exogenous’’ vitellogenesis is predominant.
Under the accelerated photoperiod regimes (the S9 and
S6 groups), neither the duration of Type II-vitellogenesis
nor the plasma and the morphometric parameter profiles
were affected by photoperiod changes. These results suggest
that during Type II-vitellogenesis, the biochemical pro-
cesses involved in reproduction are probably controlled by
an endogenous biological clock which is synchronized by
the photoperiod. Indeed, the RD phase never took place
before the decreasing photoperiod reached 10L:14D. Sev-
eral authors have also reported, in rainbow trout, the exis-
tence of endogenous biological clocks involved in the circ-
annual variations of lipids (29), lipoproteins (38), steroids
(12) and calcium, the latter being an indirect indicator of
Vtg synthesis (12). Our results show that during Type II-
vitellogenesis, this hypothetic endogenous biological clock
was able to compensate for some of the biochemical alter-
ations which occurred during previtellogenesis. For exam-
ple, the significant increase in TG levels (peak B) and in
HSI values that we observed in the S9 and S6 groups, is
probably related to the significant increase of Vtg biosyn-
thesis. Indeed, Vtg levels reached 83 mg/ml in the S9 group
and 116 mg/ml in the S6 group, as contrasted to only 60
mg/ml in controls. However, this corrective action was not
sufficient to compensate for the metabolic deficiencies in-
duced by photoperiod changes in the early stages of oocyte
development, leading to smaller egg sizes at spawning.
CONCLUSION
The compression of the reproductive cycle by accelerated
photoperiod regimes demonstrated that previtellogenesis
and Type I-vitellogenesis were photosensitive periods. Dur-
ing these, any modification of the photoperiod regime, i.e.,
an abrupt increase in the light from short days to long days,
could produce an alteration of the biochemical processes
involved in reproduction. This study also demonstrated that
Type II-vitellogenesis was probably under the control of en-
dogenous biological rhythms synchronized by the photope-
riod. However, the degree to which endogenous biological
rhythms exert an independent effect on variations in lipids
and vitellogenin concentrations are presently unknown, as
are the molecular mechanisms involved. Finally, this study
demonstrated that when the time of spawning is advanced,
the reduction of egg size is due neither to an alteration in
189
lipid metabolism nor to an alteration in vitellogenin synthe-
sis during Type II-vitellogenesis. Additional studies have
been conducted to verify the possible modification of the
kinetics of vitellogenin incorporation into the growing oo-
cytes (to be published).
This work was financially supported by a scientific research grant from
the French government (ANRT, Association Nationale de la Recherche
et de la Technique) and from a private fishfarm ‘‘Salmonidés d’Aqui-
taine.’’ We wish to thank Laurence Larroquet at the Institut National
de la Recherche Agronomique (INRA), St Pée sur Nivelle, France,
for valuable help during the lipid analyses, and Ullah Barbe, Daniel
Brajat and Augustin Bawala at the L.B.R.P., U.A. INRA, University
of Bordeaux I, France, for their excellent technical assistance. The au-
thors also wish to thank the staff from the private fishfarm ‘‘Pisciculture
des fontaines’’ for their assistance.
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