MARGEN-00181; No of Pages 8

Marine Genomics xxx (2014) xxx–xxx

Contents lists available at ScienceDirect

Marine Genomics
journal homepage: www.elsevier.com/locate/margen

Impacts of moonlight on fish reproduction

Taro Ikegami, Yuki Takeuchi, Sung-Pyo Hur, Akihiro Takemura ⁎
Department of Chemistry, Biology and Marine Science, University of the Ryukyus, Senbaru 1, Nishihara, Okinawa 903-0213, Japan

a r t i c l e i n f o a b s t r a c t

Article history: The waxing and waning cycle of the moon is repeated at approximately 1-month intervals, and concomitant
Received 24 June 2013 changes occur in the levels of moonlight and cueing signals detected by organisms on the earth. In the goldlined
Received in revised form 27 November 2013 spinefoot Siganus guttatus, a spawner lunar-synchronized around the first quarter moon, periodic changes in
Accepted 29 November 2013
moonlight are used to cue gonadal development and gamete release. Rearing of mature fish under artificial
Available online xxxx
constant full moon and new moon conditions during the spawning season leads to disruption or delay of
Keywords:
synchronous spawning around the predicted moon phase. Melatonin, an endogenous transducer of the environ-
Clock gene mental light/dark cycle, increases in the blood and in the pineal gland around the new moon period and de-
Cryptochrome creases around the full moon period. In synchrony with melatonin fluctuation, melatonin receptor(s) mRNA
Melatonin abundance is higher during the new moon period than during the full moon. The melatonin/melatonin receptor
Moonlight system is likely affected by moonlight. Measurements of the expression patterns of clock genes in neural tissues
Period demonstrate that Cryptochrome (Cry1 and Cry3) and Period (Per2) fluctuate with lunar periodicity, the former
Spinefoot peaking in the medial part of the brain around the first quarter moon period, and the latter peaking in the pineal
gland around the full moon. Some clock genes may respond to periodic changes in moon phase and appear to be
involved in the generation of lunar-related rhythmicity in lunar spawners. Thus, some fish use moonlight-related
periodicities as reliable information for synchronizing the timing of reproductive events.
© 2013 Elsevier B.V. All rights reserved.

1. Introduction of biological activities at intervals of 12.4 h are in synchrony with the

The moon, the sole natural satellite of the earth, orbits the planet at
intervals of 27.3 days. The cycles of the moon drive diverse periodicities
in environments on the earth, and these influence biological activities of
organisms. The phases of the moon drive periodic changes in nighttime
illumination (moonlight intensity), geomagnetic fields, gravitational
pull, tidal amplitude, and many other factors of major biological influ-
ence. Among these factors, moonlight intensity and geomagnetic fields
fluctuate with peaks around the full moon and last quarter moon period,
respectively. In contrast, the peaks of gravitational pull and tidal ampli-
tude occur at times of new moon and full moon. The former and the
latter factors are periodicities repeated at intervals of once and twice a
month, respectively. In addition, semidiurnal changes in tidal ampli-
tude, which repeat at intervals of 12.4 h, also occur. Most marine organ-
isms inhabiting shallow water entrain biological activities to these
moon-related periodicities as components of their adaptive strategies
for persistence in aquatic habitats. In some organisms, endogenous
clocks tracking moon-related periodicities have evolved to anticipate
environmental changes (Takemura et al., 2004a, 2010).
At least three categories (lunar cycle, semilunar cycle, and tidal
cycle) of moon-related periodicities exist in organisms (Takemura
et al., 2010). Lunar and semilunar biological activities are repeated at in-
tervals of approximately 1 month and 2 weeks, respectively. Repetitions
⁎ Corresponding author. Tel.: +81 98 895 8993; fax: +81 98 895 8576.
E-mail address: takemura@sci.u-ryukyu.ac.jp (A. Takemura).
tidal cycle (Leatherland et al., 1992). Driving factors in the lunar cycle
are likely to be associated primarily with changes in nighttime moon-
light intensity and geomagnetic fields, while those of the semilunar
cycle are associated with gravitational pull and tidal amplitude. However,
since changes in these factors are coincident in marine environments,
organisms may have integrated responses to the different cycles
expressed in their periodic activities.
Recent reports demonstrate that change in moonlight intensity is a
possible entrainer of gonadal development and gamete release under-
going around selected periods of the lunar phase (Takemura et al.,
2010). Accumulating experimental evidence indicates that moonlight
intensity influences physiological and behavioral activities of fish
inhabiting tropical and subtropical waters, in which cueing from the
moon is relatively strong and functions as a reliable zeitgeber in aquatic
environments having less variability of photoperiod and water temper-
ature (Takemura et al., 2010). The mechanism whereby lunar-related
activity is regulated endogenously and the ways in which biological
clocks are involved in its expression remain unclear (Takemura et al.,
2010). A recent contribution from physiological and molecular ap-
proaches may help resolve these open questions. This review focuses
on moonlight effects on the reproductive activity of teleost fish. The
influence of tidal stimuli in the expression of reproductive activity in
fish will be a component of our discussion. We first present diverse
lunar-related examples of reproductive and behavioral activities in
fish, mostly in tropical and subtropical waters, and discuss direct and in-
direct effects of moonlight intensity on these activities. Subsequently,

1874-7787/$ – see front matter © 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.margen.2013.11.007

Please cite this article as: Ikegami, T., et al., Impacts of moonlight on fish reproduction, Mar. Genomics (2014), http://dx.doi.org/10.1016/
j.margen.2013.11.007
2 T. Ikegami et al. / Marine Genomics xxx (2014) xxx–xxx
we review physiological and molecular mechanisms on lunar-related
reproduction in fish. Our aim is to promote better overall understanding
of this fascinating biological activity.
2. Moonlight
Since the moon is not a celestial star, it does not generate light but
instead reflects sunlight. The portion of sunlight intercepted by the
moon's surface and the amount reflected change according to relative
positions of the sun, earth, and moon. The intercepted portion of sun-
light on the moon's surface is expressed by the moon's phase (Fig. 1):
new moon (lunar age 0), waxing crescent moon (lunar age 1–6), first
quarter moon (lunar age 7), waxing gibbous moon (lunar age 8–14),
full moon (lunar age 15), waning gibbous moon (lunar age 16–21),
last quarter moon (lunar age 22), and waning crescent moon (lunar
age 23–29). These phase stages are seen as periodic changes in moon
form by an observer on the earth. One cycle of moon-form change
between phases is repeated at an interval of 29.53 days. The amount
of sunlight reflected from the moon depends on the form of the
moon; when moonlight intensity on the full moon day is expressed as
100%, the intensity during the waxing and waning gibbous moon days
is between 51% and 99%. It is 50% during the first and last quarter
moon days, 1–49% during the waxing and waning crescent moon
days, and 0% on the new moon day.
Depending on the lunar phase, the times of moonrise and moonset
and the durations of moonlight vary on a daily basis (Fig. 1). For exam-
ple, in Okinawa, Japan (26°38′N, 127°51′E) during June, darkness occurs
in the nights of the new moon period. Moonlight lasts for 1–5 h after
sunset during the waxing crescent moon period, about 6 h from sunset
to midnight during the first quarter moon period, 7–11 h after sunset
during the waxing gibbous moon period, about 12 h from sunset to
dawn during the full moon, 11–7 h before sunrise during the waning
gibbous moon period, about 6 h from midnight to dawn during the
last quarter moon period, and 5–1 h before sunrise during the waning
crescent moon period (Fig. 1). The intensity and duration of moonlight
may synergistically influence the lunar-related activities of earth's
organisms.
3. Moonlight-related reproduction in fish
Lunar-related rhythmicity may be categorized roughly into two
classes: ‘innate’ and ‘apparent’ activities. Fish in the innate category
have active lunar-related rhythmicity in response to periodic changes
in moonlight reaching their habitats. In such cases, individuals are able
to perceive cues from the moon and transduce them as internal signals
to activate their endocrine systems. On the other hand, fish in the appar-
ent category have an active response to innately driven behavioral
changes in other species. Thus, lunar-related rhythmicity in fish with
apparent activity may disappear, if other influential species with lunar
periodicity are removed from habitats. Alternatively, fish with apparent
Fig. 1. Schematic pattern of solar light phase, moonlight phase, and moonlight intensity in
Okinawa, Japan (26°38′N, 127°51′E) during June.
Modified from Fukushiro et al. (2011).
Please cite this article as: Ikegami, T., et al., Impacts of moonlight on fish reproduction, Mar. Genomics (2014), http://dx.doi.org/10.1016/
j.margen.2013.11.007
activity may only respond to periodical changes in habitat environ-
ments with lunar cycle. Overall understanding of lunar-related rhyth-
micity in fish is incomplete and we are therefore unable to rule out
the possibility that fish with apparent activities also have their own
rhythmicity in relation to moonlight.
Field studies have demonstrated that periodic change in moonlight
influences the processes of spawning migration in certain teleost fish.
Downstream migration of the European eel Anguilla anguilla is triggered
by moonless nights from the last quarter moon to the new moon period
(Miyai et al., 2004). During the migration toward the spawning ground
around the Sargasso Sea, this eel species swims below the cold water
layer and above the warm water layer of the ocean thermocline at
night in the presence and absence of moonlight, respectively, suggest-
ing that the eels during spawning migration are more sensitive to
moonlight than temperature (Tesch, 1978). Research cruises show
that just-hatched larvae of the Japanese eel Anguilla japonica accumu-
late in the North Equatorial Current west of the Mariana Islands around
the time of the new moon, suggesting that this species spawns accord-
ing to the lunar cycle (Tsukamoto, 2006). During homeward migration
of the chum salmon Oncorhynchus keta, swimming depth and speed
during the full moon period are shallower and faster, respectively,
than during other moon phases (Hasegawa, 2012). Thus, moonlight
may be required for correct navigation during homeward migration
(Hasegawa, 2012). Many Serranidae species inhabiting coral reefs
utilize periodic changes in moonlight intensity for their spawning mi-
gration (Colin et al., 1987). A typical case is migration of the leopard
coral grouper Plectropomus leopardus, whose movement toward and
aggregation at the species-selective spawning ground occur around
the new moon period (Johannes, 1978). Courtship by many mature
fish and simultaneous release of gametes occur at this site. Lunar syn-
chrony during this spawning migration and aggregation has advantages
realized in enhanced spawning success and decreased predation risk
(Johannes, 1978). Therefore, this fish is likely able to perceive differ-
ences in moon phase as changes in moonlight intensity and then choose
the appropriate moment for spawning migration. This lunar-related
migration may be an innate activity in the species.
Synchronization of gonadal development through the approach to
the species-selective lunar phase and spawning thereafter occurs in
many species (DeVries et al., 2004; Johannes, 1978; Takemura et al.,
2004b). Histological observations demonstrate that yolk-laden oocytes
develop toward the new moon and post-ovulatory follicles (a useful in-
dicator of spawning) appear afterward in ovaries of the little spinefoot
(formerly spiny rabbitfish) Siganus spinus (Harahap et al., 2001) and
the white-spotted spinefoot (seagrass rabbitfish) Siganus canaliculatus
(Hoque et al., 1999). Similar relationships exist between oocyte devel-
opment and lunar phase in the goldlined spinefoot (golden rabbitfish)
Siganus guttatus and the streamlined spinefoot (forktail rabbitfish)
Siganus argenteus; the goldlined spinefoot synchronously spawns
around the first quarter moon (Rahman et al., 2000), while the stream-
lined spinefoot spawns around the last quarter moon (Park et al.,
2006a). The roles of moonlight intensity in lunar-related gonadal devel-
opment and spawning have been examined in the goldlined spinefoot
(Takemura et al., 2004a). When mature goldlined spinefoot is reared
in tanks under artificial constant full moon and new moon conditions
over 1 month preceding the predicted spawning lunar phase, expected
spawning is disrupted or delayed. This response may be compared to
the control, in which the fish in tanks under natural night conditions
spawned around the expected lunar phase (from 6th to 8th lunar calen-
dar date; Table 1). Thus, changes in moonlight influence synchroniza-
tion of gonadal development and spawning in certain fish (Takemura
et al., 2010). Since fish likely perceive changes in moonlight, they may
have innate activity in synchronous gonadal development and gamete
release.
Lunar-related reproductive activity also occurs in the spotted tilapia
Tilapia mariae in the Ethiop River, Nigeria (Schwanck, 1987) and in the
cichlid fish Neolamprologus moorii in Lake Tanganyika (Rossiter, 1991).
 T. Ikegami et al. / Marine Genomics xxx (2014) xxx–xxx
Table 1
Spawning evidence in the three different experimental conditions during the reproductive
season.
Modified from Takemura et al. (2004a).
Lunar calendar datea
5 6 7 8
First spawning period
Natural night − + ++ + −
Artificial constant new moon − − − −
Artificial constant full moon − − − −
Second spawning period
Natural night − − − ++
Artificial constant new moon − − + ++
Artificial constant full moon − − − −
−, no spawning; +, minor spawning; ++, major spawning.
a
A lunar calendar is a calendar that is based on cycles of the lunar phase. Lunar calendar
dates of new moon, first quarter moon, full moon and last quarter moon are 0, 7, 15, and
22, respectively.
Both species have efficient parental care in their nests after spawning,
and a sequence of parental care processes is completed around the full
moon period. An ecological advantage of this lunar-related activity is
that ‘brightness at night’ during the full moon period allows a parent
to visually detect predators before they reach the nest, and then to
repel them on their approach (Moyer, 1975; Rossiter, 1991). The lunar-
related cycle in these species is likely an apparent activity. However, if
the lunar-related cycle is demonstrable even under conditions without
predators, it may be a form of innate activity.
4. Endocrine processes in lunar-related reproductive activity
When environmental parameters are in a range suitable for the ini-
tiation of reproduction, the hypothalamus–pituitary–gonadal endocrine
axis (HPG axis) is activated, triggering gonadal development (Sower
et al., 2009). The hypothalamus, pituitary, and gonads synthesize
gonadotropin-releasing hormone (GnRH), gonadotropins (GtH), and
sex steroids, respectively (Sower et al., 2009). In teleost fish, GtH I
(known as follicle stimulating hormone; FSH) and GtH II (known as
luteinizing hormone; LH) are involved in vitellogenesis and final oocyte
maturation through productions of estradiol-17β (E2) and 17α,20β-
dihydroxy-4-pregnen-3-one (DHP), respectively, both of which are
synthesized in the ovarian follicles (Nagahama, 1994; Patiño and
Sullivan, 2002).
Similar to processes in the little spinefoot (described in Section 3), a
close relationship exists between gonadal development and the lunar
cycle in the goldlined spinefoot; in this species, oocytes at vitellogenic
stages in an ovary increase toward the new moon period. Oocytes ma-
ture around the first quarter moon, and spawning follows (Rahman
et al., 2000). Measurements of weekly changes in plasma steroid hor-
mone levels using enzyme-linked immunosorbent assays demonstrate
that E2 and DHP increase toward the first quarter moon period
(Rahman et al., 2000). In vitro studies indicate that incubation of intact
follicles of oocytes with human chorionic gonadotropin (hCG) stimu-
lates production of E2 and DHP around the new moon and the first
lunar quarter moon period, respectively (Fig. 2a, b). A dose-dependent
effect of DHP and hCG on induction of germinal vesicle breakdown
(GVBD) in oocytes occurs around the first quarter moon; conversions
of testosterone to E2 and 17α-hydroxyprogesterone to DHP occur
around the new moon and the first quarter moon, respectively
(Rahman et al., 2002). These data suggest that oocyte development
depends on aromatase and 20β-hydroxysteroid dehydrogenase (key
enzymes that biosynthesize estrogen and oocyte maturation inducing
hormone (MIH), respectively), which are activated in the follicles in
response to GtH. This endocrine regulation of oocyte development in
the goldlined spinefoot is closely similar to that in other teleosts studied
(Nagahama, 1994). A sequence of endocrine changes in the HPG axis
Please cite this article as: Ikegami, T., et al., Impacts of moonlight on fish reproduction, Mar. Genomics (2014), http://dx.doi.org/10.1016/
j.margen.2013.11.007
3
9 10
−
− −
− −
++ −
+ −
− −
Fig. 2. In vitro production of steroid hormones by intact follicles of the goldlined spinefoot
in response to different concentrations of human chorionic gonadotropin (hCG).
(a) Estradiol-17β (E2) production 1 week before spawning (new moon, NM) and imme-
diately before spawning (first quarter moon, FQM). (b) 17α,20β-Dihydroxy-4-pregnen-3-
one (DHP) production 1 week before spawning (NM) and immediately before spawning
(FQM). Asterisks indicate significant differences (p b 0.05) in E2 and DHP productions
between the new moon and the first quarter moon.
Modified from Rahman et al. (2002).
occurring in concert with changes in the lunar phase is a characteristic
of lunar-related spawners. Thus, perception and transduction of cues
from the moon are processed in higher parts of the brain than the
HPG axis.
5. Melatonin as a possible transducer in lunar rhythmicity
Melatonin is a neurohormone synthesized primarily by the pineal
gland and the retina (Bromage et al., 2001); it is termed the ‘hormone
of darkness’ because it increases in concentration during scotophase
and decreases during photophase (Hardeland, 2008). Melatonin syn-
thesized in the pineal gland is released into the blood circulation and
plays a role in transduction of photoperiodic information not only to
the central nervous system (CNS) but also to the peripheral tissues.
Melatonin in the retina is involved in regulating retinal physiological
processes, including outer segment disk shedding, retinomotor move-
ments, dopamine release, and horizontal cell sensitivity, as demonstrated
by electroretinography (Boatright et al., 1994; Cahill and Besharse,
1989; Iuvone and Gan, 1995). Melatonin synthesis in the pineal gland
of mammals is regulated via the sympathetic nervous system by the
master circadian clock located in the suprachiasmatic nuclei (SCN). Ex-
cept in the case of the bastard halibut Paralichthys olivaceus (Watanabe
et al., 2012), master circadian clocks in the SCN have not been identified
in fish. Since the pineal gland of most fish shares abilities of melatonin
synthesis, the biological clock, and photoperception (Ekström and
Meissl, 1997, 2003), the gland functions as a ‘stand-alone’ master in
transducing diurnal and circadian information into the whole body
(Ekström and Meissl, 1997).
The threshold of light intensity for suppressing melatonin produc-
tion likely depends on diverse factors, including experimental condi-
tions, light quality, and photophase duration (Bromage et al., 2001;
Falcón et al., 2010). When the European sea bass Dicentrarchus labrax
is subjected to a 1-h light pulse at an irradiance of 6 μW/cm2 (about
10 lx) at midnight, plasma melatonin levels are suppressed (Bayarri
et al., 2002). The tench Tinca tinca is very sensitive to low light intensity;
a 1-h light pulse at an irradiance of 3.3 μW/cm2 (about 0.3 lx) is suffi-
cient to suppress plasma melatonin concentration to midday levels
4 T. Ikegami et al. / Marine Genomics xxx (2014) xxx–xxx
(Vera et al., 2005). Thus, sensitivity to light intensity differs among
species (Falcón et al., 2010).
Irradiant intensities during the full moon and new moon periods
are approximately 2 lx and 0.05 lx, respectively. The impact of light
at moonlight intensities on plasma melatonin levels has been demon-
strated in spinefoot species. Maximum plasma melatonin levels are
higher during the new moon period than during the full moon period
(Rahman et al., 2004; Takemura et al., 2004a) (Fig. 3a). When the
goldlined spinefoot is adapted to darkness and then exposed to moon-
light during the full moon and new moon periods, plasma melatonin
levels decrease to basal levels of naturally reared fish within 30 min
(Takemura et al., 2004a) (Fig. 4a, b). Exposing the isolated pineal
gland to moonlight during the full moon period suppresses melatonin
synthesis (Takemura et al., 2006). Notably, mRNA expression of
arylalkylamine N-acetyltransferase (AANAT1) (a rate-limiting enzyme
of melatonin) in the retina of the goldlined spinefoot is suppressed
immediately following moonlight exposure (Kashiwagi et al., 2013).
A similar influence of moonlight occurs in the Mozambique tilapia
Fig. 3. Comparisons of melatonin concentration in the plasma (a), melatonin receptor,
(MT1) gene expression in the pineal gland (b), and melatonin receptor (Mel1c) gene
expression in the pineal gland (c) of the goldlined spinefoot between the new moon
(NM) and full moon (FM). In the case of melatonin receptor in teleost fish, there are
three different types of melatonin receptors, MT1 (formerly Mel1a), MT2 (formerly
Mel1b), and Mel1c. The plasma sample and the pineal gland were taken from fish at mid-
night during the new moon (■) and full moon (□). Melatonin levels in the plasma were
measured by time-resolved immunofluoroassay. Expression of MT1 and Mel1c mRNA in
the pineal gland was determined by quantitative real-time PCR and normalized by the
amount of β-actin mRNA. Values are means ± SEM. Asterisk indicates a significant differ-
ence (p b 0.05) between the new moon and full moon.
Modified from Park et al. (2014) and Takemura et al. (2004a).
Please cite this article as: Ikegami, T., et al., Impacts of moonlight on fish reproduction, Mar. Genomics (2014), http://dx.doi.org/10.1016/
j.margen.2013.11.007
Fig. 4. Changes in plasma melatonin concentration after moonlight exposure at full moon
(a) and new moon (b). Fish were reared under normal moonlight conditions during the
full moon and new moon (naturally reared fish, NR). Fish tanks were covered with a
black sheet at 12:00 to block moonlight. At 24:00, the fish in one tank were exposed to
moonlight (moonlight-exposed fish, ME), while the fish in the other tank were kept
under constant darkness (moonlight-shaded fish, MI). The bloods from each group
(n = 6) were sampled at 24:00, 0:30, and 1:00. Values are means ± SEM. Asterisks *
and ** indicate significant differences at p b 0.05 and p b 0.001, respectively.
Modified from Takemura et al. (2004a).
Oreochromis mossambicus (Nikaido et al., 2009) and the tench (Vera
et al., 2005). Certain fish appear to be very sensitive to changes in noc-
turnal light signals. Periodic changes in moonlight are expressed as
monthly changes in plasma melatonin levels, which signal differences
in moon phase. Since night-high and day-low fluctuations of plasma
melatonin levels occur in spinefoots (Takemura et al., 2004a), a noctur-
nal difference in melatonin concentration amplitude may be important
in the expression of lunar-related rhythmicity: e.g., a small difference
during the full moon period, a large difference during the new moon
period, small and large differences in the first and last period of the
night during the first quarter moon, and large and small difference in
the first and last period of the night during the last quarter moon.
Cellular actions of melatonin are mediated via melatonin receptors,
which are members of the G-protein-coupled receptor superfamily
(Reppart et al., 1996). Vertebrates have three kinds of melatonin recep-
tor types: MT1 (Mel1a), MT2 (Mel1b), and Mel1c (Ikegami et al., 2009a,
b). MT1 and MT2 have been identified in various tissues of all verte-
brates investigated, whereas Mel1c is found only in the CNS of nonmam-
malian species (Ebisawa et al., 1994; Reppert et al., 1995). Real-time
quantitative polymerase chain reaction (PCR) analyses demonstrate
daily (or day/night) and seasonal variations in melatonin receptor
mRNA abundance in certain fish species (Confente et al., 2010;
Ikegami et al., 2009a,b; Park et al., 2006b, 2007a,b). Bimodal (midday
and midnight) and single (midnight) peaks in MT1 and MT2 mRNA
abundances, respectively, occur in the brains of 180-day-old chum
salmon (Shi et al., 2004). In addition to daily variations, circadian varia-
tions in MT1, MT2, and Mel1c mRNA abundance occur in the pineal
gland of the goldlined spinefoot (Park et al., 2006b, 2007a,b) and in
the diencephalon of the grass puffer Takifugu niphobles (Ikegami et al.,
2009a). The mRNA abundances of MT1 and MT2, but not of Mel1c,
vary by day and night in the optic tectum of the Senegalese sole Solea
senegalensis (Confente et al., 2010). In this species, seasonal variations
occur in MT1 and MT2, and these increase during the winter and sum-
mer solstices (Confente et al., 2010). Increases at night are associated
with involvement of melatonin in the regulation of melatonin receptor
expression of the CNS. In vitro experiments demonstrate that exposing
the pineal gland of the goldlined spinefoot to light during the dark
 T. Ikegami et al. / Marine Genomics xxx (2014) xxx–xxx 5

period results in suppressions of melatonin synthesis and MT1 mRNA

expression and vice versa (Park et al., 2007a).
Moonlight is likely to have impacts on expression pattern of melato-
nin receptor subtypes in the pineal gland of the goldlined spinefoot;
mRNA abundances of MT1 and Mel1c during the new moon period
are higher than during the full moon period (Fig. 3b, c). In addition,
moonlight suppresses mRNA abundances of MT1 and Mel1c (Park
et al., 2014). These expression patterns of variations in melatonin recep-
tors are quite similar to those of melatonin in the blood circulation
(Takemura et al., 2004a, 2006). Melatonin and its receptors may fluctu-
ate monthly with the lunar cycle and their interplay might contribute to
the expression of lunar-related reproductive rhythmicity in fish.

6. Are clock genes involved in lunar rhythmicity?
The core molecular mechanism of the circadian rhythm is an
autoregulatory feedback loop. Two transcriptional activators in this
loop, circadian locomotor output cycles kaput (CLOCK) and brain and
muscle ARNT-like 1 (BMAL1) proteins, form heterodimers in the cyto-
plasm and enter the nucleus where they bind the E-box sequence in
the promoters of Period (Per) and Cryptochrome (Cry) genes; transcrip-
tions of Per and Cry are activated subsequently. Synthesized PER and
CRY proteins form complexes, enter the nucleus, bind to the CLOCK/
BMAL1 complex, and inhibit transcription of their own genes (Allada
et al., 2001; Okamura et al., 2002; Young and Kay, 2001). One cycle of
this autoregulatory feedback loop takes about 24 h and generates circa-
dian rhythmicity in organisms. A spectrum of knowledge on circadian
rhythm molecular mechanism has accumulated for model species
(e.g., fruit fry Drosophila melanogaster, mouse Mus musculus and
zebrafish Danio rerio).
The molecular mechanism of the circalunar rhythm is poorly under-
stood because few model species have lunar periodicity under laborato-
ry conditions. Limited studies have been performed on stony corals
(Hoadley et al., 2011; Levy et al., 2007) and one fish (Fukushiro et al.,
2011; Sugama et al., 2008), which are based on knowledge of the regu-
latory mechanism of the circadian rhythm. In Acropora millepora, a full
moon spawner of a stony coral, mRNA abundance of Cry2, but not
Cry1, is higher at midnight in the full moon period than in the new
moon period, suggesting that Cry2 entrains the intrinsic clock of corals
to the lunar phase (Levy et al., 2007). This outcome contrasts with the
case of another coral, Favia fragum, in which no expression patterns of
Cry1, Cry2, Clock, and Cycle related to the lunar cycle occur. Thus, a diver-
sity of molecular mechanisms may exist among scleractinian corals.
Alternatively, photopigments such as opsins may play a common role
in perception of moonlight, which can be a trigger of lunar-related
mass spawning (Levy et al., 2007).
Several clock genes, including Cry and Period2 (Per2), are light-
responsive and may play a crucial role in the entrainment of fish rhyth-
micity (Tamai et al., 2007; Vatine et al., 2009). Since moonlight is likely
an informative nighttime light source for certain species, recent empha-
sis has been placed on the elucidation of these clock genes as a possible
state variable against the moonlight in the goldlined spinefoot
(Fukushiro et al., 2011; Sugama et al., 2008). These studies revealed
that some clock genes fluctuate with changes in the lunar phase: In
the pineal gland, Per2 mRNA abundance is higher at midnight during
the full moon period than during the new moon period (Fig. 5); expo-
sure of fish to moonlight during the full moon period stimulates Per2
mRNA abundance (Sugama et al., 2008) (Fig. 6). Fukushiro et al.
(2011) examined weekly changes in mRNA abundance of Cry1 and
Cry3 in the medial part of the brain and demonstrated a lunar phase-
dependency: a decrease in abundance occurs around the full moon
period, whereas its increase occurs around the first quarter moon
period, which is the spawning lunar phase of this fish (Fig. 7a, b). To ex-
plain this mRNA profile, it is proposed that putative photorepressive
phase (ZT18–ZT21) exists in the late nights from the waxing gibbous
moon to the last quarter moon period: that is, moonlight exposure
Fig. 5. Comparisons of Period2 (Per2) gene expression in the pineal gland of the goldlined
spinefoot between the new moon (NM) and full moon (FM). Expression of Per2 mRNA in
the pineal gland was determined by quantitative real-time PCR and normalized by the
amount of β-actin mRNA. Values are means ± SEM. Asterisk indicates a significant differ-
ence (p b 0.05) between the new moon and full moon.
Modified from Sugama et al. (2008).
during the photo-repressive phase leads to a decrease of Cry mRNA
abundance in the medial part of the brain (Fukushiro et al., 2011). In a
seasonally breeding bird, Japanese quail Coturnix coturnix japonica,
long-day is recognized according to the existence of light during the
photo-inducible phase in early night and activates their reproduction
(Follett and Sharp, 1969). Moreover, Yasuo et al. (2004) demonstrated
that light-inducible Cry1 mRNA expression in the pars tuberalis (PT) of
the bird occurs upon light exposure only during the photo-inducible
phase. Such molecular event in the photoperiodism seems similar to
lunar phase dependent Cry expression, although the response of Cry
mRNA expression against the light cue is reversible.
Lunar-specific genes have not been identified in organisms to date. A
novel type (circalunar clocks) of clock gene or clock system cannot be
ruled out at this stage because molecular studies on lunar cycle have
barely begun.
7. Closing remarks
The purpose of this review is to demonstrate involvement of moon-
light in the expression of the lunar-related reproductive cycle and
to outline underlying physiological and molecular mechanisms. In
our summary of possible associations of melatonin/melatonin receptors
and clock genes with lunar-related reproductive cycles in fish (e.g., the
goldlined spinefoot), we suggest that rhythmic fluctuations of
these players are involved in the repetition of gonadal development
at 1-month intervals (Fig. 8).
Fig. 6. Effect of moonlight on the expression of Per2 mRNA in the pineal gland of goldlined
spinefoot. Fish were reared under normal moonlight conditions during the full moon
(naturally reared fish, NR). Fish tanks were covered with a black sheet at zeitgeber time
(ZT) 12 to block moonlight. At ZT18, the fish in one tank were exposed to moonlight
(moonlight-exposed fish, ME), while the fish in the other tank were kept under constant
darkness (moonlight-shaded fish, MI). The pineal glands from each group (n = 7–8) were
sampled at ZT18, ZT19, and ZT20. Values are means ± SEM. Data were normalized by
determining the amount of β-actin mRNA. Asterisk indicates a significant difference at
p b 0.05.
Modified from Sugama et al. (2008).

Please cite this article as: Ikegami, T., et al., Impacts of moonlight on fish reproduction, Mar. Genomics (2014), http://dx.doi.org/10.1016/
j.margen.2013.11.007
6 T. Ikegami et al. / Marine Genomics xxx (2014) xxx–xxx

Endocrine roles of melatonin in reproduction are still moot (Migaud

Fig. 7. Lunar-dependent variations in the expression of genes encoding Cryptochrome,
(a) Cry1 and (b) Cry3 in the brain of the goldlined spinefoot. Each Cry mRNA value was
calculated as a value relative to β-actin mRNA. Values are means ± SD. Lunar phases are
indicated above the graph by schematic moon images. Probability values (p) for statisti-
cally significant differences among dates are indicated (ANOVA). Values with different
characters are significantly different among lunar phases (p b 0.05, Tukey's test).
Modified from Fukushiro et al. (2011).
et al., 2010). Melatonin administration attenuates ovarian and testicular
development in the goldfish Carassius auratus (Fenwick, 1970), and its
injection inhibits expression of gnrh1, gnrh3, and gnrh receptor genes
in their brain (Servili et al., 2013). In contrast, immersing the zebrafish
in melatonin-containing water stimulates ovarian development
(Carnevali et al., 2011). Concomitant increases in kiss1, kiss2, and
gnrh3 gene expression occur in zebrafish brains. Therefore, melatonin
likely acts on the HPG axis and different effects are probably related to
diverse reproductive strategies among fish (Carnevali et al., 2011). In
the case of the three-spined stickleback, Gasterosteus aculeatus, matura-
tion pace was not inhibited by melatonin treatment under long photo-
periodic stimulation in winter, suggesting that melatonin plays a
relatively minor role in the control of the reproductive activity and
that photoperiodic effects are largely mediated via mechanisms other
than circulating melatonin levels (Bornestaf et al., 2001). We still do
not have a clear understanding of the link between the light induced
melatonin signal and the reproductive endocrine system in fish. Never-
theless, this train of reasoning may be applicable to lunar spawning
cycles. Monthly changes in moonlight from the full moon to the new
moon promote gradual increases in melatonin levels at night and stim-
ulate gonadal development of lunar synchronizers around the new
moon period. Moonlight changes from the new moon to the full moon
promote reverse effects in lunar synchronizers around the full moon
period (e.g., Rahman et al., 2003 for forktail rabbitfish; Lee et al., 2002
for the honeycomb grouper Epinephelus merra). Moreover, mRNA abun-
dances of melatonin receptors and the neuropeptide LPXRF amide track
diurnal and circadian variations in the diencephalon, including the
hypothalamus, of the grass puffer, which is a semilunar spawner
(Ikegami et al., 2009a; Shahjahan et al., 2011); LPXRF amide acts as a

Fig. 8. Schematic patterns (for goldlined spinefoot reproduction) of changes in environmental factors (a), possible players in the lunar-related rhythm (b), and reproductive activity (c).
Photoperiod increases from spring to summer and peaks at the summer solstice (June). Moonlight illumination fluctuates in a 1-month cycle and peaks around the full moon. Plasma levels
of melatonin and expression levels of melatonin receptor genes fluctuate in 1-month cycles and peak around the new moon. The mRNA abundance of Cryptochrome (Cry1, Cry3) and
Period2 (Per2) fluctuate in a 1-month cycle and peak around the first quarter moon and full moon, respectively. Gonadal activity increases toward and decreases after the spawning
day (first quarter moon). Oocytes at vitellogenic stages appear among immature oocytes around the full moon and develop fully around the new moon period. Following final oocyte
maturation, synchronous spawning occurs around the first quarter moon. This cycle is repeated through the reproductive season.

Please cite this article as: Ikegami, T., et al., Impacts of moonlight on fish reproduction, Mar. Genomics (2014), http://dx.doi.org/10.1016/
j.margen.2013.11.007
 T. Ikegami et al. / Marine Genomics xxx (2014) xxx–xxx
gonadotropin-inhibitory hormone (GnIH) in birds and is regulated by
melatonin via Mel1c (Tsutsui et al., 2006).
Few studies have examined the involvement of clock genes in fish
reproduction. Data are available only for Salmo salar, in which Clock is
upregulated in sneaker strategy males, but not in immature males
(Aubin-Horth et al., 2005). Periodic changes in Cry and Per2 abundance
may act as timekeepers for identifying lunar phases and help synchro-
nize endocrine drivers for reproductive events.
We conclude that information on the brightness at night generated
by the moon may be used in synchronization of various reproductive
events in fish. Information received is acted upon by the endocrine
network operating in concert with melatonin/melatonin receptors and
the endogenous clock system. Alternatively, there may be the endoge-
nous clock system attributing to operation through an independent
(non-melatonin) pathway as suggested by some researchers (Amano
et al., 2000; Bornestaf et al., 2001). Further physiological and molecular
studies are required for a more comprehensive understanding of lunar
synchronization in fish.
Acknowledgments
This study was supported in part by a Grant-in-Aid for Scientific
Research (JSPS KAKENHI Grant Numbers 23248033 and 25304032)
from the Japan Society for the Promotion of Science (JSPS) to AT and a
Joint Research Project under the Japan–Korea Basic Scientific Coopera-
tion Program from Japan Society for the Promotion of Science to AT.
References
Allada, R., Emery, P., Takahashi, J.S., Rosbash, M., 2001. Stopping time: the genetics of fly
and mouse circadian clocks. Annu. Rev. Neurosci. 24, 1091–1119.
Amano, M., Iigo, M., Ikuta, K., Kitamura, S., Yamada, H., Yamamori, K., 2000. Roles of
melatonin in gonadal maturation of underyearling precocious male masu salmon.
Gen. Comp. Endocrinol. 120, 190–197.
Aubin-Horth, N., Landry, C.R., Letcher, B.H., Hofmann, H.A., 2005. Alternative life histories
shape brain gene expression profiles in males of the same population. Proc. R. Soc.
Lond. B 272, 1655–1662.
Bayarri, M.J., Madrid, J.A., Sánchez-Vázquez, F.J., 2002. Influence of light intensity, spec-
trum and orientation on sea bass plasma and ocular melatonin. J. Pineal Res. 32,
34–40.
Boatright, J.H., Rubim, N.M., Iuvone, P.M., 1994. Regulation of endogenous dopamine
release in amphibian retina by melatonin: the role of GABA. Vis. Neurosci. 11,
1013–1018.
Bornestaf, C., Mayer, I., Borg, B., 2001. Melatonin and maturation pace in female three-
spined stickleback, Gasterosteus aculeatus. Gen. Comp. Endocrinol. 122, 341–348.
Bromage, N., Porter, M., Randall, C., 2001. The environmental regulation of maturation
in farmed finfish with special reference to the role of photoperiod and melatonin.
Aquaculture 197, 63–98.
Cahill, G.M., Besharse, J.C., 1989. Retinal melatonin is metabolized within the eye of
Xenopus laevis. Proc. Natl. Acad. Sci. U. S. A. 86, 1098–1102.
Carnevali, O., Gioacchini, G., Maradonna, F., Olivotto, I., Migliarini, B., 2011. Melatonin
induces follicle maturation in Danio rerio. PLoS ONE 6, e19978.
Colin, P.L., Shapiro, D.Y., Weiler, D., 1987. Aspects of the reproduction of two groupers,
Epinephelus guttatus and E. striatus in the West Indies. Bull. Mar. Sci. 40, 220–230.
Confente, F., Rendón, M.C., Besseau, L., Falcón, J., Muñoz-Cueto, J.A., 2010. Melatonin re-
ceptors in a pleuronectiform species, Solea senegalensis: cloning, tissue expression,
day–night and seasonal variations. Gen. Comp. Endocrinol. 167, 202–214.
DeVries, P., Goetz, F., Fresh, K., Seiler, D., 2004. Evidence of a lunar gravitation cue on
timing of estuarine entry by Pacific salmon smolts. Trans. Am. Fish. Soc. 133,
1379–1395.
Ebisawa, T., Karne, S., Lerner, M.R., Reppert, S.M., 1994. Expression cloning of a high-
affinity melatonin receptor from Xenopus dermal melanophores. Proc. Natl. Acad.
Sci. U. S. A. 91, 6133–6137.
Ekström, P., Meissl, H., 1997. The pineal organ of teleost fishes. Rev. Fish Biol. Fish. 7,
199–284.
Ekström, P., Meissl, H., 2003. Evolution of photosensory pineal organs in new light: the
fate of neuroendocrine photoreceptors. Philos. Trans. R. Soc. B 358, 1679–1700.
Falcón, J., Migaud, H., Muñoz-Cueto, J.A., Carrillo, M., 2010. Current knowledge on the
melatonin system in teleost fish. Gen. Comp. Endocrinol. 165, 469–482.
Fenwick, J.C., 1970. Demonstration and effect of melatonin in fish. Gen. Comp. Endocrinol.
14, 86–97.
Follett, B.K., Sharp, P.J., 1969. Circadian rhythmicity in photoperiodically induced gonado-
trophin release and gonadal growth in the quail. Nature 223, 968–971.
Fukushiro, M., Takeuchi, T., Takeuchi, Y., Hur, S.P., Sugama, N., Takemura, A., Kubo, Y.,
Okano, K., Okano, T., 2011. Lunar phase-dependent expression of cryptochrome and
a photoperiodic mechanism for lunar phase-recognition in a reef fish, goldlined
spinefoot. PLoS ONE 6, e28643.
Please cite this article as: Ikegami, T., et al., Impacts of moonlight on fish reproduction, Mar. Genomics (2014), http://dx.doi.org/10.1016/
j.margen.2013.11.007
7
Harahap, A.P., Takemura, A., Nakamura, S., Rahman, M.S., Takano, K., 2001. Histolog-
ical evidence of lunar-synchronized ovarian development and spawning in the
spiny rabbitfish Siganus spinus (Linnaeus) around the Ryukyus. Fish. Sci. 67,
888–893.
Hardeland, R., 2008. Melatonin, hormone of darkness and more — occurrence, control
mechanisms, actions and bioactive metabolites. Cell. Mol. Life Sci. 65, 2001–2018.
Hasegawa, E.I., 2012. Chum salmon Oncorhynchus keta respond to moonlight during
homeward migrations. J. Fish Biol. 81, 632–641.
Hoadley, K.D., Szmant, A.M., Pyott, S.J., 2011. Circadian clock gene expression in the coral
Favia fragum over diel and lunar reproductive cycles. PLoS ONE 6, e19755.
Hoque, M.M., Takemura, A., Matsuyama, M., Matsuura, S., Takano, K., 1999. Lunar
spawning in Siganus canaliculatus. J. Fish Biol. 55, 1213–1222.
Ikegami, T., Motohashi, E., Doi, H., Hattori, A., Ando, H., 2009a. Synchronized diurnal
and circadian expressions of four subtypes of melatonin receptor genes in the
diencephalon of a puffer fish with lunar-related spawning cycles. Neurosci. Lett.
462, 58–63.
Ikegami, T., Azuma, K., Nakamura, M., Suzuki, N., Hattori, A., Ando, H., 2009b. Diurnal
expressions of four subtypes of melatonin receptor genes in the optic tectum and
retina of goldfish. Comp. Biochem. Physiol. A 152, 219–224.
Iuvone, P.M., Gan, J., 1995. Functional interaction of melatonin receptors and D1 dopa-
mine receptors in cultured chick retinal neurons. J. Neurosci. 15, 2179–2185.
Johannes, R.E., 1978. Reproductive strategies of coastal marine fishes in the tropics.
Environ. Biol. Fish. 3, 65–84.
Kashiwagi, T., Park, Y.J., Park, J.G., Imamura, S., Takeuchi, Y., Hur, S.P., Takemura, A., 2013.
Moonlight affects mRNA abundance of arylalkylamine N-acetyltransferase in the ret-
ina of a lunar-synchronized spawner, the goldlined spinefoot. J. Exp. Zool. A 319,
505–516.
Leatherland, J.F., Farbridge, K.J., Boujard, T., 1992. Lunar and semi-lunar rhythms in fishes.
In: Ali, M.A. (Ed.), Rhythms in Fishes. Plenum Press, New York, pp. 83–107.
Lee, Y.D., Park, S.H., Takemura, A., Takano, K., 2002. Histological observations of seasonal
reproductive and lunar-related spawning cycles in the female honeycomb grouper
Epinephelus merra in Okinawan waters. Fish. Sci. 68, 872–877.
Levy, O., Appelbaum, L., Leggat, W., Gothlif, Y., Hayward, D.C., Miller, D.J., Hoegh-Guldberg,
O., 2007. Light-responsive cryptochromes from a simple multicellular animal, the
coral Acropora millepora. Science 318, 467–470.
Migaud, H., Davie, A., Taylor, J.F., 2010. Current knowledge on the photoneuroendocrine
regulation of reproduction in temperate fish species. J. Fish Biol. 76, 27–68.
Miyai, T., Aoyama, J., Sasai, S., Inoue, J.G., Miller, M.J., Tsukamoto, K., 2004. Ecological
aspects of the downstream migration of introduced European eels in the Uono
river, Japan. Environ. Biol. Fish. 71, 105–114.
Moyer, J.T., 1975. Reproductive behavior of the damselfish Pomacentrus nagasakiensis at
Miyake-jima, Japan. Jpn. J. Ichthyol. 22, 151–163.
Nagahama, Y., 1994. Endocrine regulation of gametogenesis in fish. Int. J. Dev. Biol. 38,
217–229.
Nikaido, Y., Ueda, S., Takemura, A., 2009. Photic and circadian regulation of melatonin
production in the Mozambique tilapia Oreochromis mossambicus. Comp. Biochem.
Physiol. A 152, 77–82.
Okamura, H., Yamaguchi, S., Yagita, K., 2002. Molecular machinery of the circadian clock
in mammals. Cell Tissue Res. 309, 47–56.
Park, Y.J., Takemura, A., Lee, Y.D., 2006a. Annual and lunar-synchronized ovarian activity
in two rabbitfish species in the Chuuk lagoon, Micronesia. Fish. Sci. 72, 166–172.
Park, Y.J., Park, J.G., Kim, S.J., Lee, Y.D., Rahman, M.S., Takemura, A., 2006b. Melatonin
receptor of a reef fish with lunar-related rhythmicity: cloning and daily variations.
J. Pineal Res. 41, 166–174.
Park, Y.J., Park, J.G., Hiyakawa, N., Lee, Y.D., Kim, S.J., Takemura, A., 2007a. Diurnal and
circadian regulation of a melatonin receptor, MT1, in the golden rabbitfish, Siganus
guttatus. Gen. Comp. Endocrinol. 150, 253–262.
Park, Y.J., Park, J.G., Jeong, H.B., Takeuchi, Y., Kim, S.J., Lee, Y.D., Takemura, A., 2007b.
Expression of the melatonin receptor Mel1c in neural tissues of the reef fish Siganus
guttatus. Comp. Biochem. Physiol. A 147, 103–111.
Park, Y.J., Park, J.G., Takeuchi, Y., Hur, S.P., Lee, Y.D., Takemura, A., 2014. Influence of moon-
light on mRNA expression patterns of melatonin receptor subtypes in the pineal
organ of a tropical fish. Mar. Genomics.
Patiño, R., Sullivan, C.V., 2002. Ovarian follicle growth, maturation, and ovulation in
teleost fish. Fish Physiol. Biochem. 26, 57–70.
Rahman, M.S., Takemura, A., Takano, K., 2000. Correlation between plasma steroid
hormones and vitellogenin profiles and lunar periodicity in the female golden
rabbitfish, Siganus guttatus (Bloch). Comp. Biochem. Physiol. B 127, 113–122.
Rahman, M.S., Takemura, A., Takano, K., 2002. Lunar synchronization of in vitro steroido-
genesis in ovaries of the golden rabbitfish, Siganus guttatus (Bloch). Gen. Comp.
Endocrinol. 125, 1–8.
Rahman, M.S., Morita, M., Takemura, A., Takano, K., 2003. Hormonal changes in relation to
lunar periodicity in the testis of the forktail rabbitfish, Siganus argenteus. Gen. Comp.
Endocrinol. 131, 302–309.
Rahman, M.S., Kim, B.H., Takemura, A., Park, C.B., Lee, Y.D., 2004. Effects of moonlight
exposure on plasma melatonin rhythms in the seagrass rabbitfish, Siganus
canaliculatus. J. Biol. Rhythms 19, 325–334.
Reppart, S.M., Weaver, D.R., Godson, C., 1996. Melatonin receptors step into the light:
cloning and classification of subtypes. Trends Pharmacol. Sci. 17, 100–102.
Reppert, S.M., Weaver, D.R., Cassone, V.M., Godson, C., Kolakowski Jr., L.F., 1995. Melatonin
receptors are for the birds: molecular analysis of two receptor subtypes differentially
expressed in chick brain. Neuron 15, 1003–1015.
Rossiter, A., 1991. Lunar spawning synchroneity in a freshwater fish. Naturwissenschaften
78, 182–184.
Schwanck, E., 1987. Lunar periodicity in the spawning of Tilapia mariae in the Ethiop river,
Nigeria. J. Fish Biol. 30, 533–537.
8 T. Ikegami et al. / Marine Genomics xxx (2014) xxx–xxx
Servili, A., Herrera-Pérez, P., del Carmen Rendón, M., Muñoz-Cueto, J., 2013. Melatonin in-
hibits GnRH-1, GnRH-3 and GnRH receptor expression in the brain of the European
sea bass, Dicentrarchus labrax. Int. J. Mol. Sci. 14, 7603–7616.
Shahjahan, M., Ikegami, T., Osugi, T., Ukena, K., Doi, H., Hattori, A., Tsutsui, K., Ando, H.,
2011. Synchronised expressions of LPXRFamide peptide and its receptor genes:
seasonal, diurnal and circadian changes during spawning period in grass puffer.
J. Neuroendocrinol. 23, 39–51.
Shi, Q., Ando, H., Coon, S.L., Sato, S., Ban, M., Urano, A., 2004. Embryonic and post-
embryonic expression of arylalkylamine N-acetyltransferase and melatonin receptor
genes in the eye and brain of chum salmon (Oncorhynchus keta). Gen. Comp.
Endocrinol. 136, 311–321.
Sower, S.A., Freamat, M., Kavanaugh, S.I., 2009. The origins of the vertebrate hypothalamic–
pituitary–gonadal (HPG) and hypothalamic–pituitary–thyroid (HPT) endocrine
systems: new insights from lampreys. Gen. Comp. Endocrinol. 161, 20–29.
Sugama, N., Park, J.G., Park, Y.J., Takeuchi, Y., Kim, S.J., Takemura, A., 2008. Moonlight
affects nocturnal Period2 transcript levels in the pineal gland of the reef fish Siganus
guttatus. J. Pineal Res. 45, 133–141.
Takemura, A., Susilo, E.S., Rahman, M.S., Morita, M., 2004a. Perception and possible utili-
zation of moonlight intensity for reproductive activities in a lunar-synchronized
spawner, the golden rabbitfish. J. Exp. Zool. A 301, 844–851.
Takemura, A., Rahman, M.S., Nakamura, S., Park, Y.J., Takano, K., 2004b. Lunar cycles and
reproductive activity in reef fishes with particular attention to rabbitfishes. Fish
Fish. 5, 317–328.
Takemura, A., Ueda, S., Hiyakawa, N., Nikaido, Y., 2006. A direct influence of moonlight in-
tensity on changes in melatonin production by cultured pineal glands of the golden
rabbitfish, Siganus guttatus. J. Pineal Res. 40, 236–241.
Please cite this article as: Ikegami, T., et al., Impacts of moonlight on fish reproduction, Mar. Genomics (2014), http://dx.doi.org/10.1016/
j.margen.2013.11.007
Takemura, A., Rahman, M.S., Park, Y.J., 2010. External and internal controls of lunar-
related reproductive rhythms in fishes. J. Fish Biol. 76, 7–26.
Tamai, T.K., Young, L.C., Whitmore, D., 2007. Light signaling to the zebrafish circadian
clock by Cryptochrome 1a. Proc. Natl. Acad. Sci. U. S. A. 104, 14712–14717.
Tesch, F.W., 1978. Telemetric observations on the spawning migration of the eel
(Anguilla anguilla) west of the European continental shelf. Environ. Biol. Fish. 3,
203–209.
Tsukamoto, K., 2006. Oceanic biology: spawning of eels near a seamount. Nature 439, 929.
Tsutsui, K., Ubuka, T., Yin, H., Osugi, T., Ukena, K., Bentley, G.E., Ciccone, N., Inoue, K.,
Chowdhury, V.S., Sharp, P.J., Wingfield, J.C., 2006. Mode of action and functional
significance of avian gonadotropin-inhibitory hormone (GnIH): a review. J. Exp.
Zool. A 305, 801–806.
Vatine, G., Vallone, D., Appelbaum, L., Mracek, P., Ben-Moshe, Z., Lahiri, K., Gothilf, Y.,
Foulkes, N.S., 2009. Light directs zebrafish period2 expression via conserved D and E
boxes. PLoS Biol. 7, e1000223.
Vera, L.M., López-Olmeda, J.F., Bayarri, M.J., Madrid, J.A., Sánchez-Vázquez, F.J., 2005. Influ-
ence of light intensity on plasma melatonin and locomotor activity rhythms in tench.
Chronobiol. Int. 22, 67–78.
Watanabe, N., Itoh, K., Mogi, M., Fujinami, Y., Shimizu, D., Hashimoto, H., Uji, S., Yokoi, H.,
Suzuki, T., 2012. Circadian pacemaker in the suprachiasmatic nuclei of teleost fish
revealed by rhythmic period2 expression. Gen. Comp. Endocrinol. 178, 400–407.
Yasuo, S., Watanabe, M., Tsukada, A., Takagi, T., Iigo, M., Shimada, K., Ebihara, S.,
Yoshimura, T., 2004. Photoinducible phase-specific light induction of Cry1 gene in
the pars tuberalis of Japanese quail. Endocrinology 145, 1612–1616.
Young, M.W., Kay, S.A., 2001. Time zones: a comparative genetics of circadian clocks. Nat.
Rev. Genet. 2, 702–715.