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Behavioural Brain Research 189 (2008) 387391

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Maternal pinealectomy increases depressive-like responses in Siberian hamster offspring

Joanna L. Workman a, , Zachary M. Weil b , Christiana R. Tuthill d , Randy J. Nelson a,b,c
a

Department of Psychology, The Ohio State University, 1835 Neil Avenue, Columbus, OH 43210, USA b Department of Neuroscience, The Ohio State University, Columbus, OH 43210, USA c Institute for Behavioral Medicine Research, The Ohio State University, Columbus, OH 43210, USA d Department of Psychology, University of California, Berkeley, CA 94720, USA Received 13 December 2007; received in revised form 23 January 2008; accepted 28 January 2008 Available online 5 February 2008

Abstract This study investigated the effect of maternal pinealectomy and postnatal pinealectomy on affective responses. Siberian hamsters were born to either pinealectomized or sham-operated dams and then underwent pinealectomy or a sham operation. Maternal pinealectomy increased depressivelike responses of offspring in the forced swim test. Maternal pinealectomy increased rearing behaviour and postnatal pinealectomy increased locomotor behaviour in the open eld test. These results suggest that prenatal melatonin organizes adult affective responses. 2008 Elsevier B.V. All rights reserved.
Keywords: Pineal gland; Melatonin; Forced swim; Open eld; Photoperiod; Seasonal affective disorder

1. Introduction Siberian hamsters are seasonally breeding rodents that adjust physiology and behaviour throughout the year to take advantage of increased food availability during the spring and summer and to survive harsh conditions during the winter [7]. Hamsters use two pieces of information to synchronize internal processes with environmental conditions: (1) day length, and (2) the direction of change in day lengths [20]. Melatonin, a neurohormone that transduces night length information, is secreted primarily from the pineal gland at night and duration of secretion is inversely related to day length [7]. For Siberian hamsters, exposure to short days results in regression of the reproductive tract, altered metabolism [24,30], enhancement of some components of the immune system (for review, see [15]), elevated aggressive behaviour [3,34], and altered affective responses [21,23]. In addition, duration of maternal melatonin can organize physiology and behaviour of offspring in adulthood. Although Siberian hamsters regress their reproductive tracts and cease

Corresponding author. Tel.: +1 614 688 4674; fax: +1 614 688 4733. E-mail address: workman.1113@osu.edu (J.L. Workman).

reproductive behaviour in the winter, social cues in the laboratory can override the effect of short day lengths on reproduction [10]. Heterosexual pairs will continue to breed when housed in short photoperiods [10]. Further, because maternal melatonin is communicated to the fetus [25,26], melatonin duration can be manipulated in utero. Importantly, maternal melatonin inuences reproductive development prior to puberty [9,25] which presumably allows fetuses to anticipate environmental conditions before birth to develop traits that favor either winter survival or spring and summer breeding. In addition to the reproductive system, duration of maternal melatonin may organize components of the immune system [33] and affective responses [23] of offspring. Specically, Siberian hamsters increase depressive- and anxiety-like responses when housed in short day lengths [21]. These responses may be adaptive for surviving winter conditions. It may conserve considerable energy to disengage early from a task in which success is unlikely [16]. For example, adult hamsters housed in short day lengths oat more in the forced swim test than their long-day counter parts [21,23]. Such a response might save energy that can be shunted towards biological processes, such as immune function, that promote winter survival. Affective responses in Siberian hamsters may

0166-4328/$ see front matter 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.bbr.2008.01.016

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approximate symptoms of seasonal affective disorder (SAD), which is characterized by symptoms of depression in the winter that are alleviated in the summer, by morning light, or by treatment with selective serotonin reuptake inhibitors. One notable hypothesis regarding SAD, the Phase Shift Hypothesis (PSH), implicates melatonin in the etiology of the disorder [14]. The PSH stipulates that SAD may be a consequence of lengthened melatonin secretion in the winter when dim morning light is insufcient to inhibit melatonin secretion. Others have suggested that symptoms of SAD are vestigial adaptations that conferred energetic savings during winter conditions [6,12,16]. In addition, some psychological disorders are more common among individuals born at specic times across the year. For example, schizophrenia, bipolar disorder, and major depressive disorder are more prevalent among people born in winter and spring [27]. SAD is more prevalent among those born in spring and summer [1,18]. This information suggests that in humans, melatonin duration may contribute to the development of psychological disorders. In hamsters, perinatal photoperiods can organize adult affective responses [23] and this phenomenon may resemble season-of-birth variations in human psychological disorders. Taken together, this information suggests that Siberian hamsters may provide a means of studying the neuroendocrine substrates of SAD and other disorders that uctuate seasonally in incidence or severity. Further, manipulation of perinatal photoperiod or perinatal melatonin may provide an approximation for studying mechanisms by which season of birth inuences the incidence of disorders. We hypothesized that hamster offspring would respond to the relative change in melatonin duration rather than the absolute length of duration. Therefore, hamsters that gestate in dams that lack pineal melatonin, but have an intact pineal gland throughout development would interpret this transition as a transfer to short day lengths and display more depressive- and anxious-like responses (i.e., more oating in the forced swim test and less central tendency and rearing in the open eld). We hypothesized that all other hamster groups would behave similarly to long-day hamsters and not elevate depressive-like responses because they would undergo either a reduction of melatonin duration or no change in melatonin duration. Twenty-two male Siberian hamsters (Phodopus sungorus) were used in this study. Hamsters were born to dams that underwent either pinealectomy (PNX; n = 8) or sham operation (n = 8) as previously described [29]. Dams and offspring were housed in 14L:10D with lights off at 1800 h Pacic Standard Time (PST). On postnatal day 3, offspring underwent either PNX or sham operation such that there were 4 groups: maternal PNX, postnatal PNX (PNXPNX; n = 6), maternal PNX, postnatal sham (PNXsham; n = 6), maternal sham, postnatal PNX (shamPNX; n = 5), and maternal sham, postnatal sham (shamsham; n = 9). Surgeries were performed in Dr. Irving Zuckers lab at the University of California at Berkeley and hamsters were transferred to animal facilities at The Ohio State University for testing when 6090 days of age. Hamsters were housed with siblings in polypropylene cages (28 cm 17 cm 12 cm) in rooms with constant temperature and humidity (21 4 C and 50 10%, respectively). The light

cycle was 14L:10D with lights-off at 1500 Eastern Standard Time (EST) and behavioural testing began at 1500 EST after 9 weeks of habituation. Hamsters were allowed to habituate to testing rooms for 15 min before behavioural testing. Food (Harlan Teklad 8640 Rodent Diet, Indianapolis, IN) and tap water were available ad libitum. All procedures were conducted in accordance with standards of the University of California at Berkeley and The Ohio State University Institutional Lab Animal Care and Use Committees. To assess total locomotor behaviour and anxiety-like responses, hamsters were placed in a 40 cm 40 cm clear acrylic chamber lined with corncob bedding, inside a ventilated cabinet. The center of the open eld was dened as the central 30 cm 30 cm. A frame at the base of the chamber consisting of 32 photobeams in a 16 16 arrangement, in addition to a row of beams above, detected the location of horizontal movements and rearing, respectively (Open Field Photobeam Activity System, San Diego Instruments Inc., San Diego, CA). Total movement was tracked for 60 min and analyzed for (1) the percentage of time spent in the center of the open eld, (2) number of rears, and (3) total locomotor behaviour. An increase in central tendency and rearing are generally interpreted as a decreased anxiety-like response [22,23]. To assess depressive-like responses [19], hamsters were placed in room-temperature water 17 cm deep within an opaque, cylindrical tank (24 cm diameter, 53 cm height). Swimming behaviour was recorded on video for 7 min and scored by a blind observer with The Observer software (Version 5, Exeter Software, Setauket, NY) to quantify (1) latency to oat, (2) total number of oating bouts, and (3) total time spent oating. More oating is interpreted as an increased depressive-like response. Data were analyzed using 2 2 ANOVAs with maternal condition and neonatal condition as the independent variables and behavioural measures as the dependent variables. To determine simple main effects, t-tests were conducted based on a priori hypotheses and were limited to (1) within maternal condition, between postnatal conditions and (2) within postnatal condition, between maternal conditions. All data were analyzed using StatView software, version 5.0.1 (Cary, NC, USA). Mean differences were considered statistically signicant when p 0.05. The main effect of maternal condition was signicant for all three measures in the forced swim test. Maternal pinealectomy reduced the latency to oat (F1,22 = 19.208; p < 0.0005; Fig. 1A), increased oat bouts (F1,22 = 14.602; p < 0.001; Fig. 1B), and increased the time spent oating (F1,22 = 4.481; p < 0.05; Fig. 1C). The main effect of postnatal condition on number of oat bouts was also signicant; postnatal pinealectomy reduced the number of oat bouts (F1,22 = 4.351; p < 0.05; Fig. 1B). Within the maternal pinealectomy group, postnatal pinealectomy signicantly reduced the number of oat bouts (Fig. 1B), but increased time spent oating (Fig. 1C). There was a signicant main effect of postnatal condition on total locomotor activity and percent central tendency. Postnatal pinealectomy increased basal locomotor activity (F1,22 = 5.207; p < 0.05; Fig. 2B) and the percent of beams broken in the center of the open eld (F1,22 = 13.311; p < 0.005; Fig. 2C). Within the maternal sham condition, postnatal pinealectomy signi-

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the PNXPNX group. Pair-wise comparisons revealed that the PNXsham group oated more often than the PNXPNX group, but the PNXPNX group spent a greater amount of time oating. Although the PNXPNX group had a lower number of oat bouts, they spent a greater amount of time oating, suggesting that they failed to resume swimming. Taken together, these three measures suggest that a lack of melatonin in utero leads to an increase in depressivelike responses in adulthood. These data are consistent with previous work conducted in our lab with melatonin receptor knockout (MT1/) mice [32]. Mice lacking this gene throughout life display elevated depressive-like responses in the forced swim test. The current study extends the results

Fig. 1. Maternal pinealectomy (A) reduced the latency to oat (mean S.E.M.), (B) increased the number of oat bouts (mean S.E.M.), and (C) increased the time spent oating (mean S.E.M.). Hamsters born to pinealectomized dams (B) had a greater number of oat bouts if they had an intact pineal gland and (C) spent more time oating if they underwent a postnatal pinealectomy. ( indicates main effect; * indicates simple main effect within maternal condition; p < 0.05).

cantly increased basal locomotor activity (Fig. 2B) and percent central tendency (Fig. 2C). There was a signicant effect of maternal condition on rearing behaviour in the open eld. Maternal pinealectomy increased number of rears in the open eld (F1,22 = 37.4; p < 0.0001; Fig. 2A). Overall, maternal pinealectomy increased depressive-like responses of adult offspring. Specically, maternal pinealectomy decreased the latency to oat, increased the number of oat bouts, and increased the time spent oating (Fig. 1). The effect of maternal pinealectomy on oat bouts was largely due to the PNXsham group. In addition, the effect of maternal pinealectomy on time spent oating was largely due to

Fig. 2. Maternal pinealectomy (A) increased the number of rears in the open eld (mean S.E.M.). Postnatal pinealectomy increased (B) total locomotor activity (mean S.E.M.) and (C) percent of time in the center of the open eld (mean S.E.M.). Of hamsters born to dams with intact pineal glands, postnatal pinealectomy increased (B) total locomotor activity and (C) percent of time spent in the center of the open eld. ( indicates main effect; * indicates simple main effect within maternal condition; p < 0.05).

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of the previous study. Gene knockout studies cannot elucidate whether differences are due to lacking the gene in utero or in adulthood, but when considered in conjunction with this study, it suggests that melatonin signaling in utero is responsible for organizing adult behaviour, and probably normal brain development. Indeed, early pinealectomy can alter morphological development of cerebellar granule cells in chicks [28]. Because they would be exposed to a relative lengthening of nightly melatonin secretion, we expected the PNXsham group to display more depressive-like responses than either of the maternal sham groups. We did not expect, however, the PNXPNX group also to display increases in depressivelike responses. Hamsters that gestate in pinealectomized dams receive no photoperiod information as assessed by reproductive development [11] and pineal maturation does not occur until the 3rd week of life [35]. Therefore, these data suggest that, rather than a relative change in melatonin duration, a complete lack of pineal melatonin in utero organizes adult behaviour independently of the postnatal condition. The organizational versus activational effects of steroid hormones on adult reproductive behaviour are well established [8,17]. Perinatal exposure to androgens organizes adult masculine reproductive behaviours, but these behaviours must be activated by the presence of androgens in adulthood. Likewise, lack of androgens permits female reproductive behaviours in adulthood, but female reproductive behaviours must also be activated by the presence of estrogens and progestins. In common with steroid hormones, melatonin signaling early in life organizes reproductive development [29] and organizes adult behavior. The effect of maternal pinealectomy in hamsters may be relevant for humans because individuals born in spring and summer (a time when pineal melatonin duration is reduced) are at a greater risk for developing SAD. Because developmental events in humans are distributed across 9 months during which melatonin duration would uctuate, it seems likely that there is a developmental window during which low melatonin may have adverse effects on adult affect. Although reproduction appears unaffected by photoperiod in humans, photoperiodic responsiveness in nonreproductive traits persist in primates, including humans (for review, see [31]). In the open eld test (Fig. 2), maternal pinealectomy increased the number of rearing events in the chamber. These data are consistent with another study reporting that maternal melatonin treatment decreases rearing behaviour in offspring [13]. Postnatal pinealectomy increased both basal locomotor activity and percent time in the center of the open eld. These data must be interpreted cautiously. Percent of time spent in the center of the open eld is generally interpreted as a measure of anxiety-like behaviour. In this measure, infrared beams broken in the center are normalized to total of infrared beams broken. Although these data suggest an altered anxiety-like response, we cannot solidly conclude that postnatal pinealectomy decreases anxiety-like responses because we cannot tease apart locomotor behaviour from central tendency. In addition, maternal pinealectomy increased rearing; a behaviour that is often associated with increased central tendency [22,23]. Our data, however, do

not reect that relationship. Further research will be useful in differentiating the effects of melatonin duration, in utero and postnatally, on anxiety-like responses and locomotor behaviour. Maternal melatonin duration may alter behaviour by altering the development of neurotransmitter systems. Ambient photoperiod can alter serotonin turnover [5] and manipulation of the serotonergic system in early life may alter its development [4]. Season of birth in humans may also alter development of the serotonin system [2]. Although we do not rule out litter effects on behaviour, we believe that these results are due to maternal and postnatal melatonin. The data presented in this study provide a starting point for investigating the effect of melatonin on affective behaviours that might model symptoms of psychological disorders with season-of-birth variations. Acknowledgements The authors thank Emily Webber for technical assistance, Lynn Martin for helpful discussions and comments on an earlier version of this manuscript, and Irving Zucker for conceptual discussions regarding this study and for generously providing the animals. This research was supported by NSF grant OIS 04-16897. References
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