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Photoperiod-Dependent Changes in Melatonin

Synthesis in the Turkey Pineal Gland and Retina
Article in Poultry Science August 2007
DOI: 10.1093/ps/86.7.1397 Source: PubMed

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Photoperiod-Dependent Changes in Melatonin Synthesis

in the Turkey Pineal Gland and Retina
J. B. Zawilska,*1 A. Lorenc, M. Berezinska, B. Vivien-Roels, P. Pevet, and D. J. Skene
*Centre for Medical Biology, Polish Academy of Sciences, Lodz, 93-232 Poland; Department of Pharmacodynamics,
Medical University of Lodz, 90-151 Poland; Department of Pharmacology, Medical University of Lodz, 90-752 Poland;
Institut des Neurosciences Cellulaires et Integratives, Department de Neurobiologie des Rythmes,

Unite Mixte de Recherche 7168/LC2 Universite Louis PasteurCentre National de la Recherche Scientifique,
Strasbourg, 67000 France; and Centre for Chronobiology, School of Biomedical and Molecular Sciences,
University of Surrey, Guildford GU2 7XH, United Kingdom
ABSTRACT The effect of photoperiod on melatonin content and the activity of the melatonin-synthesizing enzymes, namely, serotonin N-acetyltransferase (AANAT)
and hydroxyindole-O-methyltransferase, were investigated in the pineal gland and retina of turkeys. The birds
were adapted to 3 different lighting conditions: 16L:8D
(long photoperiod), 12L:12D (regular photoperiod), and
8L:16D (short photoperiod). Pineal, retinal, and plasma
melatonin concentrations oscillated with a robust diurnal
rhythm, with high values during darkness. The duration
of elevated nocturnal melatonin levels in the turkey pineal
gland, retina, and plasma changed markedly in response
to the length of the dark phase, being longest during the
short photoperiod with 16 h of darkness. These photoperiodic variations in melatonin synthesis appear to be driven
by AANAT, because changes in the activity of this enzyme
were closely correlated with changes in melatonin. By con-

trast, pineal and retinal hydroxyindole-O-methyltransferase activities failed to exhibit any significant 24-h variation
in the different photoperiods. A marked effect of photoperiod on the level of melatonin production was also observed. Peak values of melatonin and AANAT activity in
the pineal gland (but not in the retina) were highest during
the long photoperiod. During the light phase, mean melatonin concentrations in the pineal gland and retina of turkeys kept under the long photoperiod were significantly
higher compared with those from birds maintained under
the regular and short photoperiods. In addition, mean circulating melatonin levels were lowest in the short photoperiod. Finally, the magnitude of the light-evoked suppression of nighttime pineal AANAT activity was also influenced by photoperiod, with suppression being smallest
under the long photoperiod. These findings show that in
the turkey, photoperiod plays an important role in regulating the melatonin signal.

Key words: melatonin, turkey, pineal gland, retina, photoperiod

2007 Poultry Science 86:13971405

INTRODUCTION
Life on earth is subjected to a strict regimen of cyclical
changes, the most prominent being the daily variations
between day and night, and the annual succession of seasons. In temperate latitudes, photoperiod appears to be
the most reliable parameter of all the seasonal cues, and
as such, is used by animals to indicate the time of year
in order to synchronize endogenous annual rhythms of
physiology and behavior to environmental conditions (reviewed by Goldman, 2001; Herzog and Schwartz, 2002;
and Hazlerigg and Wagner, 2006). In numerous vertebrates, environmental photoperiodic information is trans-

2007 Poultry Science Association Inc.

Received February 1, 2007.
Accepted March 7, 2007.
1
Corresponding author: jzawilska@pharm.am.lodz.pl

formed into a neuroendocrine signal, the hormone melatonin, which is synthesized in the pineal gland and, additionally, in the retina of several species (reviewed by Arendt,
1995; Goldman, 2001; Malpaux et al., 2001; and Iuvone et
al., 2005). It has been demonstrated that in mammals, the
melatonin rhythm (e.g., its amplitude, peak, and nadir; rate
of increase and decrease of synthesis; duration of elevated
levels; and sensitivity to light) is influenced by the photoperiod. Moreover, in several mammalian species a significant
correlation has been shown between photoperiod-dependent changes in circulating melatonin levels and seasonal
cycles of reproduction, prolactin levels, intensity of metabolic processes (fat metabolism), voluntary food intake,
body mass, immune system activity, and changes of pelage
(reviewed by Arendt, 1995; Goldman, 2001; and Malpaux
et al., 2001).
Although many avian species are photoperiodic, regulatory mechanisms responsible for adjusting their physiology and behavior to annual changes in day-length are

1397

1398

ZAWILSKA ET AL.

less well understood than in mammals. Initially, it was

postulated that birds, in contrast to mammals, do not use
seasonal changes in nocturnal melatonin production to
time their reproductive activity, and the role of melatonin
in avian physiology has remained enigmatic for many
years (Wilson, 1991; Juss et al., 1993; Pevet, 2001; Underwood et al., 2001). Despite this, evidence is accumulating for a relationship between melatonin and several seasonal processes, including gonadal size and activity (Ramachandran et al., 1996; Sudhakumari et al., 2001; Singh
and Haldar, 2007), sexual development (Guyomarch et
al., 2001), egg production (Siopes and Underwood, 1987),
cellular and humoral immune responses (Moore and Siopes, 2000, 2002; Moore et al., 2002; Singh and Haldar,
2007), cold resistance and thermogenesis (Saarela and
Heldmaier, 1987), and endocrine activity of the adrenal
glands (Ramachandran et al., 1996; Sudhakumari et al.,
2001). Moreover, it has recently been postulated that melatonin, by stimulating expression of a newly discovered
gonadotropin-inhibitory hormone, may provide an important photoperiodic signal to influence the reproductive
axis of birds (Ubuka et al., 2005; Tsutsui et al., 2006).
The turkey (Meleagris gallopavo), a representative of galliforms, is an avian species in which several physiological
processes (including growth, sexual development, egg production), and changes in plasma concentrations of luteinizing hormone and prolactin are potently influenced by photoperiod (Siopes et al., 1993; Siopes, 1994; Bedecarrats et
al., 1997; Yang et al., 1999; Noirault et al., 2006). Until
now, the regulatory mechanisms governing photoperioddependent responses in the turkey were unknown. One of
the likely candidates for transducing environmental photic
information might be melatonin. To begin to define a putative role for melatonin in photoperiodic time measurement
in the turkey, it is imperative to obtain data on the regulation of melatonin production. We previously demonstrated that the turkey pineal gland and retina synthesize
melatonin in a circadian manner (Zawilska et al., 2006).
In the present study, we investigated whether melatonin
production in the turkey is sensitive to changes in daylength. Melatonin content and the activity of the melatoninsynthesizing enzymes, serotonin N-acetyltransferase (AANAT; EC 2.3.1.87) and hydroxyindole-O-methyltransferase (HIOMT; EC 2.1.1.4), were thus measured in the pineal
gland, retina, and plasma at regular time intervals from
turkeys that were adapted to 3 different photoperiods,
namely, a long photoperiod of 16L:8D, a regular photoperiod of 12L:12D, and a short photoperiod of 8L:16D. In
addition, we analyzed whether the responsiveness of pineal and retinal AANAT activity to the suppressive action
of light at night is modified by the photoperiodic history
of the turkey.

MATERIALS AND METHODS

Birds and Experimental Design
A total of 264 White turkey poults (Meleagris gallopavo)
of both sexes were purchased on the day of hatching and

kept in temperature-controlled warmed brooders (34 1C

for the first 5 d, and 30 1C afterward). Birds were fed
a standard diet (commercial turkey prestarter) containing
28% CP and 2,800 kcal of ME/kg of feed. Feed and water
were supplied for ad libitum consumption. On the third
day, turkeys were divided into 3 experimental groups and
placed under the following photoperiodic regimes: 1) long
photoperiod (16L:8D), 2) regular photoperiod (12L:12D),
and 3) short photoperiod (8L:16D). The lighting cycle
(lights off at 2000 h) was produced by overhead cool fluorescent lamps providing light intensity at the level of the
birds heads of approximately 150 lx. The growth pattern
of the turkeys did not differ markedly among the 3 photoperiodic groups (data not shown). The experiments were
carried out in strict accordance with the Polish governmental regulations concerning experiments on animals
(Dz.U.05.33.289). All experimental protocols were approved by the Local Ethical Commission for Experimentation on Animals.
After adaptation for a month, the turkeys were decapitated at 3-h intervals over a 24-h period, starting from the
beginning of the dark phase of the imposed L:D cycle.
Additionally, groups of animals that were maintained under the long photoperiod and the short photoperiod were
killed at the end of the dark phase. Pineal glands and
retinas were quickly isolated, frozen on dry ice, and stored
at 70C (maximally for 3 d) before biochemical analysis.
Trunk blood was collected in heparinized tubes. After centrifugation (at 3,000 g for 30 min at 4C), the plasma was
stored at 70C until melatonin analysis. Decapitation of
the turkeys and tissue isolation during the dark phase of
an imposed L:D cycle were performed under dim (3-lx)
red light. In another set of experiments, in the middle of
the dark phase of the imposed L:D cycle (i.e., 4, 6, and 8
h after lights off in the long, regular, and short photoperiod,
respectively), groups of turkeys were exposed to white
light (150 lx) for 0.5, 2, 10, or 30 min, and killed immediately
afterward. Pineal glands and retinas were quickly isolated,
frozen on dry ice, and stored at 70C (maximally for 3
d) before measurement of AANAT activity.

Biochemical Assays
Pineal glands and retinas were sonicated in ice-cold 0.05
M sodium phosphate buffer (pH 6.8) in a proportion of 1
pineal/200 L, and 1 mg of wet retina/10 L. The homogenate was centrifuged at 10,000 g for 5 min at 4C, and
aliquots of the supernatant were assayed for AANAT activity, HIOMT activity, and melatonin.
Serotonin N-acetyltransferase activity was determined
according to our routine radioisotopic procedure (Nowak
et al., 1989), using acetyl coenzyme A (152 M) containing
16 nCi of [acetyl-1-14C]coenzyme A and tryptamine-HCl
as substrates. To measure HIOMT activity, aliquots of pineal and retinal supernatants were adjusted to pH 7.9 with
0.05 mM sodium phosphate buffer (pH 9.0). Hydroxyindole-O-methyltransferase activity was measured by a radiosotopic method (Nowak et al., 1993), using N-acetylserotonin and S-adenosyl-L-methionine (100 M) containing

PHOTOPERIOD AND MELATONIN SYNTHESIS IN TURKEY

20 nCi of S-[methyl-14C]adenosyl-L-methionine as substrates.

Melatonin was extracted from retinal supernatants and
plasma using dichloromethane as described previously
(Rudolf et al., 1992); melatonin standards were extracted
using the same procedure. To measure pineal melatonin
content, aliquots of tissue supernatant were diluted in icecold 0.1 M tricine buffer (pH 5.0, containing 0.9% NaCl
and 0.1% gelatin). Melatonin was measured by RIA, using
a rabbit antiserum (batch no. R19540; INRA, Nouzilly,
France) at a final dilution of 1:200,000 and [125I]iodomelatonin as a tracer (Rudolf et al., 1992). Polyethylene glycol
in combination with sheep antirabbit antiserum (INRA)
was used to separate the bound and free tracer. The limit
of sensitivity of the assay was 1.0 pg/tube. Measurements
of quality controls containing 6, 30, and 150 pg of melatonin/tube gave interassay coefficients of 13.6, 14.2, and
12.0%, respectively. The RIA was validated by assessing
the parallelism between serial dilutions of pineal supernatants or dichloromethane extracts of plasma and retinal
samples and the melatonin standard curve. Furthermore,
quantitative recovery experiments showed that the
amount of melatonin measured by RIA was closely correlated with the amount of standard melatonin (25 to 500
pg/mL) added to pools of the tissue samples analyzed
(Zawilska et al., 2006).
Acetyl coenzyme A, N-acetylserotonin, S-adenosyl-Lmethionine, and melatonin were purchased from Sigma
Chemical Co. (St. Louis, MO), tryptamine was obtained
from Serva (Heildelberg, Germany). [Acetyl-1-14C]Coenzyme A (specific activity, 60 mCi/mmol), and S-[methyl14
C]adenosyl-L-methionine (specific activity, 52.7 mCi/
mmol) were purchased from PerkinElmer Life Sciences
(Boston, MA). Other chemicals were of analytical purity
and were purchased mainly from Sigma Chemical Co.

Data Analysis
Data are expressed as the means SEM values and were
analyzed for statistical significance by one-way ANOVA
followed by a posthoc Student-Newman-Keuls test using
GraphPad InStat version 3.05 for Windows 95 (GraphPad,
San Diego, CA). To test for rhythmicity, we used the traditional F-test, which compares the (reparameterized) cosine
model [c = M + A cos(F t + ) + t, where c is the
observed mean concentration of the compound at a given
time t, F is the angular frequency, A is the amplitude,
is the acrophase expressed in radians, and t is the deviation of observed values from the fitted curve] with the
nonrhythmic model (c = M + t). The F-test was performed
using GraphPad InPlot version 4.00 for Windows
(GraphPad). The area under the curve (AUC) was calculated with the aid of SigmaPlot, version 8.02 (Systat Software Inc., Chicago, IL).

RESULTS
Effect of Photoperiod on Pineal
Melatonin Content and on AANAT
and HIOMT Activities
Turkeys kept under long (16L:8D), regular (12L:12D),
and short (8L:16D) photoperiods exhibited robust daily

1399

rhythms in pineal melatonin concentrations and AANAT

activity (Figure 1). Both melatonin content and AANAT
activity were high during the dark phase and low during
the light phase of the imposed L:D cycle. The nighttime
increase in pineal melatonin content and AANAT activity
in turkeys kept under a short photoperiod was slower
compared with changes observed in birds kept under the
long and regular photoperiods. During the long photoperiod, the maximum pineal AANAT activity was significantly greater (P < 0.05) than peak values found during
the regular and short photoperiods (Figure 1). During the
light phase of the L:D cycle, mean pineal melatonin concentrations in turkeys kept under the long photoperiod (172
19 pg/mg of pineal; n = 34) were significantly higher (P
< 0.001) than those found in pineals isolated from birds
maintained under the regular (75 7 pg/mg of pineal; n =
20) and short photoperiods (94 17 pg/mg of pineal;
n = 17). The dark-phase melatonin constituted 77% (long
photoperiod), 91% (regular photoperiod), and 94% (short
photoperiod) of the total amount of melatonin (AUC) produced during the 24-h period. Computer-assisted analysis
revealed that pineal HIOMT activity did not exhibit significant rhythmic variation throughout the 24-h period in
any of the photoperiods tested (Figure 1).

Effect of Photoperiod on Retinal

Melatonin Content and on AANAT
and HIOMT Activities
In retinas of turkeys kept under the different photoperiods tested, melatonin content and AANAT activity also
exhibited high-amplitude rhythmic oscillations, with high
values during the dark phase and low values during the
light phase of the imposed L:D cycle (Figure 2). During
the light phase of the L:D cycle, mean melatonin concentrations in the retinas of turkeys kept under the long photoperiod (17.3 1.9 pg/mg of retina; n = 34) were significantly
higher (P < 0.001) than those found in tissues isolated from
birds maintained under the regular (6.7 0.6 pg/mg of
retina; n = 19) and short photoperiods (7.1 0.9 pg/mg
of retina; n = 18). Melatonin produced during the dark
phase constituted 60% (long photoperiod), 88% (regular
photoperiod), and 92% (short photoperiod) of the total
amount of melatonin (AUC) produced during the 24-h
period.
Hydroxyindole-O-methyltransferase activity in retinas
of turkeys kept under the long photoperiod (8.7 0.3
pmol/h per mg of retina; n = 48) was significantly (P <
0.01) lower than that measured in retinas from turkeys
kept in the regular (11.3 0.3 pmol/h per mg of retina;
n = 34) and short photoperiods (10.4 0.4 pmol/h per mg
of retina; n = 51). Computer-assisted analysis revealed that
retinal HIOMT activity did not exhibit significant rhythmic
variation throughout the 24-h period in any of the photoperiods tested (Figure 2).

Effect of Photoperiod on Plasma

Melatonin Concentrations
In each photoperiod studied, plasma melatonin concentrations exhibited a clear diurnal change (Figure 3). During

1400

ZAWILSKA ET AL.

Figure 1. Diurnal variation in pineal melatonin concentrations and in serotonin N-acetyltransferase (AANAT) and hydroxyindole-O-methyltransferase (HIOMT) activities (act.) in turkeys kept under 3 different photoperiods: (A) long photoperiod (16L:8D), (B) regular photoperiod (12L:12D),
and (C) short photoperiod (8L:16D). The gray area indicates the dark phase of the imposed L:D cycle. Values shown are means SEM (n = 5 to
6 animals/time point).

the short photoperiod, the amplitude of the plasma melatonin rhythm (102 5 pg/mL) was significantly lower (P <
0.001) than the amplitude observed under the long (174
10 pg/mL) and regular (207 11 pg/mL) photoperiods.
In the long photoperiod, the total AUC of the melatonin
profile was markedly higher by 18 and 22% when compared with the regular and short photoperiods, respectively. In the long photoperiod, nighttime melatonin corresponded to only 48% of the total AUC; this proportion

was markedly lower compared with the 72 and 77% found

in the regular and short photoperiods, respectively.

Effect of Light at Night on Pineal and

Retinal AANAT Activity
Exposure of turkeys to light in the middle of the dark
phase of the imposed L:D cycle suppressed pineal and
retinal AANAT activity. The magnitude of the light-in-

PHOTOPERIOD AND MELATONIN SYNTHESIS IN TURKEY

1401

Figure 2. Diurnal variation in retinal melatonin concentrations and in serotonin N-acetyltransferase (AANAT) and hydroxyindole-O-methyltransferase (HIOMT) activities (act.) in turkeys kept under 3 different photoperiods: (A) long photoperiod (16L:8D), (B) regular photoperiod (12L:12D),
and (C) short photoperiod (8L:16D). The gray area indicates the dark phase of the imposed L:D cycle. Values shown are means SEM (n = 5 to
6 animals/time point).

duced changes was dependent on the analyzed tissue, the

duration of the light pulse, and the photoperiodic history
of the birds (Table 1). In turkeys maintained under the
regular and short photoperiods, a 2-, 10-, and 30-min pulse
of light produced a statistically significant decline in pineal
AANAT activity compared with the no-light condition. In
turkeys adapted to the long photoperiod, only a 30-min
exposure to light markedly decreased AANAT activity in
the pineal gland; shorter light pulses were ineffective. In

the retina, all tested light pulses produced statistically significant decreases of nighttime AANAT activity. The magnitude of the observed changes did not differ among the
3 different photoperiodic groups, with the exception of the
response to a 0.5-min pulse. The response to this light
pulse of very short duration was significantly weaker in
the retinas of turkeys kept under the long photoperiod
compared with those birds maintained in the regular and
short photoperiods. The present results also revealed that

1402

ZAWILSKA ET AL.

the pineal gland appeared to be less sensitive to the suppressive action of light than the retina (Table 1).

DISCUSSION

Figure 3. Diurnal variation in plasma melatonin concentrations in

turkeys kept under 3 different photoperiods: (A) long photoperiod
(16L:8D), (B) regular photoperiod (12L:12D), and (C) short photoperiod
(8L:16D). The gray area indicates the dark phase of the imposed L:D
cycle. Values shown are means SEM (n = 5 to 6 animals/time point).

As in mammals, the most important event for annual

changes in physiology and behavior in birds, including
plasma levels of several hormones (i.e., luteinizing hormone, prolactin, and melatonin), gonadal activity, fattening and body mass gain, moult and migration, is the
annual cycle of the photoperiod (reviewed by Dawson et
al., 2001). Increasing experimental evidence indicates that
in both mammals and birds, the circadian system plays
an important role in photoperiodic time measurement (reviewed by Dawson et al., 2001; Gwinner and Brandstatter,
2001; Pevet, 2001; Brandstatter, 2003; and Lincoln et al.,
2003, 2006). It has been postulated that the avian circadian
timing system might be more complex than in mammals,
and involves 3 autonomous and anatomically distinct multiple central oscillators that are localized in the eyes, the
hypothalamus, and the pineal gland. The functional significance of these oscillators for circadian rhythmicity varies markedly among avian species (reviewed by Gwinner
and Brandstatter, 2001; Underwood et al., 2001; and Brandstatter, 2003).
The indoleamine hormone melatonin is synthesized by
the vertebrate pineal gland and, additionally, in some species, by the retina in a light-dependent rhythmic fashion
controlled by an endogenous circadian clock (reviewed
by Arendt, 1995; and Iuvone et al., 2005). We previously
showed that the turkey pineal gland and retina produce
melatonin in a high-amplitude circadian rhythm (Zawilska
et al., 2006). The present study extends these findings and
demonstrates that the duration of elevated melatonin in
the turkey pineal gland and retina changes potently in
response to the length of the dark phase of the imposed
L:D cycle. Thus, in both tissues the duration of melatonin
production was significantly longer under the short photoperiod compared with the long and regular photoperiods.
Similarly, the duration of elevated plasma melatonin levels
(reflecting primarily pineal melatonin; Siopes and Underwood, 1987) gradually increased with the lengthening
of the dark phase. These photoperiodic-dependent variations in melatonin synthesis appear to be driven by the
enzyme AANAT, because changes in AANAT activity
were closely correlated with changes in melatonin. By contrast, the activity of HIOMT, the final enzyme in the melatonin-synthesizing pathway, did not exhibit any significant
variation throughout the 24-h period in the pineal glands
and retinas isolated from turkeys kept under the 3 different photoperiods.
The photoperiod-dependent changes observed in the
turkey are in agreement with the existing literature. Photoperiodic modifications of the melatonin (or AANAT activity) rhythm in the pineal gland, retina, and blood have
been reported in several avian species, namely, the Japanese quail (Underwood and Siopes, 1985; Zeman and Illnerova, 1988), chicken (Binkley et al., 1977), European starling (Dawson and Vant, 2002), house sparrow (Brandstat-

1403

PHOTOPERIOD AND MELATONIN SYNTHESIS IN TURKEY

Table 1. Effect of light at night on serotonin N-acetyltransferase activity (pmol/h per mg of tissue) in the pineal
gland and retina of turkeys adapted to different photoperiodic regimes1
Light-exposed

Dark
control

0.5 min

1,097 23

1,109 191

Regular (12L:12D)

1,117 77

1,182 36

Short (8L:16D)

1,183 109

1,097 130

Retina
Long (16L:8D)

210 9

Photoperiod
Pineal gland
Long (16L:8D)

Regular (12L:12D)

185 16

Short (8L:16D)

197 14

170 11a
(81%)
125 21ab
(68%)
117 14ab
(59%)

2 min
1,180 170
780 33ab
(70%)
792 128ab
(67%)

10 min

30 min

1,026 196

672 75a
(61%)
516 63a
(46%)
357 48abc
(30%)

649 45ab
(58%)
375 32abc
(32%)

113 6a
(54%)

94 9a
(45%)

75 7a
(35%)

91 9a
(49%)
113 8a
(57%)

87 4a
(47%)
71 5a
(36%)

82 12a
(44%)
75 4a
(38%)

P < 0.01 vs. dark control; bP < 0.01 vs. long photoperiod; cP < 0.01 vs. regular photoperiod.
In the middle of the dark phase, that is, 4 (long photoperiod), 6 (regular photoperiod), or 8 (short photoperiod)
h after lights off, turkeys were exposed to white light (150 lx) for 0.5, 2, 10, or 30 min and killed immediately
afterward. Values are means SEM (n = 5 to 6 animals/group). Values shown in parentheses represent the
percentage of the respective dark control value.
a
1

ter et al., 2000), emperor penguin (Miche et al., 1991), and

Svalbard ptarmigan (Reierth et al., 1999), with the nighttime increase reflecting the duration of the dark phase.
Additionally, lengthening of the dark phase has been
shown to result in reducing the amplitude of the melatonin
rhythm (Underwood and Siopes, 1985; Miche et al., 1991;
Reierth et al., 1999; Brandstatter et al., 2000).
Despite this agreement across studies, the physiological
relevance of these photoperiod-induced changes in the
melatonin signal is less clear. Changes in both the intensity
and duration of the nocturnal melatonin profile might provide an important seasonal signal to the turkey. In addition, it may be that by manipulating the day-length, and
thus the melatonin level, beneficial modulation of the immune system can be achieved. There is increasing experimental evidence demonstrating a functional link between
melatonin and immune responses, not only in mammals
(reviewed by Carrillo-Vico et al., 2006), but also in birds.
Pinealectomy has potently reduced immune responses in
the ring-dove and Japanese quail (Rodriguez and Lea,
1994; Moore et al., 2002), suggesting that immune function
may be regulated by melatonin. In the Japanese quail,
exogenous melatonin counteracted the immunosuppressive effect of pinealectomy or constant-light treatment
(Moore and Siopes, 2000; Moore et al., 2002). Exogenous
melatonin has also been shown to enhance white blood
cell counts and activate B and T lymphocytes in immature
chickens (Brennan et al., 2002). In addition, both embronic
and posthatched exposure to melatonin accelerated the
development of cell-mediated and humoral immune activities in the turkey (Moore and Siopes, 2002, 2005). Noticeably, in the turkey and chicken, mortality caused by infectious diseases is prevalent during the first few weeks posthatch (Alexander, 1990; Law and Payne, 1990; Cook, 2000),
and a strong correlation between immune dysfunction and
the pathogenesis of certain diseases in neonatal birds has

been demonstrated (Sharma et al., 1994; Bounous et al.,

1995; Qureshi et al., 1997). Thus, as already emphasized
by Moore and Siopes (2005), the immune-enhancing effect
of melatonin appears to be especially important in the early
stage of a turkeys life. Whether endogenous melatonin has
a similar immune-enhancing function remains to be determined.
Photoperiod-dependent changes in melatonin level
might also play a regulatory role in the turkey reproductive
system. Involvement of melatonin in the regulation of seasonally changing gonadal activity and gonadotropin secretion has been demonstrated for some avian species (e.g.,
Ohta et al., 1989; Ramachandran et al., 1996; Guyomarch
et al., 2001; Sudhakumari et al., 2001; Trivedi et al., 2004,
Singh and Haldar, 2007). In the turkey, pinealectomy of
breeder hens significantly delayed the onset of egg laying
and depressed egg production (Siopes and Underwood,
1987). It has recently been proposed that in birds (and
likely also in mammals), melatonin regulates gonadotropin
release by stimulating expression of gonadotropin-inhibitory hormone (Ubuka et al., 2005; Tsutsui et al., 2006).
Gonadotropin-inhibitory hormone is the first hypothalamic neuropeptide shown to directly inhibit gonadotropin
release at the pituitary level (Yin et al., 2005). Gonadotropin-inhibitory hormone may also act on the hypothalamus
to inhibit release of gonadotropin-releasing hormone (Yin
et al., 2005; Bentley et al., 2006). However, whether melatonin can also act directly on the pituitary to regulate changes
in prolactin secretion, as has already been demonstrated
in mammals (reviewed by Lincoln et al., 2003; and Hazlerigg and Wagner, 2006), remains to be elucidated.
Light is the dominant environmental factor controlling
melatonin biosynthesis in both the pineal gland and retina.
Light at night acutely suppresses melatonin synthesis. In
addition, pulses of light properly timed adjust and reset
the circadian oscillator generating the melatonin rhythm

1404

ZAWILSKA ET AL.

(Arendt, 1995; Zawilska et al., 2000; Iuvone et al., 2005).

The responsiveness of the mammalian circadian clock to
these phase-shifting effects of light is known to be altered
by the photoperiodic history of the animal, with phase
shifts being larger after entrainment to short days (e.g.,
Vuillez et al., 1996; Evans et al., 2004). Studies in humans
have also shown that light responses (e.g., light-induced
melatonin suppression) are affected by an individuals previous light history (Owen and Arendt, 1992; Hebert et
al., 2002; Smith et al., 2004). Results of the present study,
indicating that the suppressive action of light on nighttime
melatonin synthesis is modified by the birds photoperiodic history, support the previous studies in mammals. In
addition to its acute effect on melatonin synthesis, the
photoperiod may modulate other light responses, including induction of early response genes and clock genes (e.g.,
Meddle and Follet, 1997; Sumova et al., 2004).
In conclusion, photoperiod-induced changes in melatonin synthesis were observed in the turkey pineal gland
and retina. In addition, the light-responsiveness of the melatonin-generating system was shown to be affected by the
photoperiodic history. It is suggested that these photoperiod-dependent changes in the melatonin signal may play
an important role in modulating the immune function and
reproductive status of the turkey.

ACKNOWLEDGMENTS
This work was supported by grant no. 2 PO6D 025
29 from the Ministry of Science and Higher Education,
Warsaw, Poland. The authors thank Jean-Paul Ravault
(INRA, Nouzilly, France) for kindly providing the melatonin antibody. The technical assistance of Teresa Kwapisz
(Centre for Medical Biology, Lodz, Poland) and Karolina
Czarnecka (Medical University of Lodz, Lodz, Poland) is
highly appreciated.

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