Co-localization of serine/threonine kinase 33 (Stk33) and vimentin in the hypothalamus

We investigate the immunoreactivity of serine/threonine kinase 33 (Stk33) and of vimentin in the brain of mouse, rat and hamster. Using a Stk33-specific polyclonal antibody, we show by immunofluorescence staining that Stk33 is present in a variety of brain regions. We found a strong staining in the ependymal lining of all cerebral ventricles and the central canal of the spinal cord as well as in hypothalamic tanycytes. Stk33 immunoreactivity was also found in circumventricular organs such as the area postrema, subfornical organ and pituitary and pineal glands. Double-immunostaining experiments with antibodies against Stk33 and vimentin showed a striking colocalization of Stk33 and vimentin. As shown previously, Stk33 phosphorylates recombinant vimentin in vitro. Co-immunoprecipitation experiments and co-sedimentation assays indicate that Stk33 and vimentin are associated in vivo and that this association does not depend on further interacting partners (Brauksiepe et al. in BMC Biochem 9:25, 2008). This indicates that Stk33 is involved in the dynamics of vimentin polymerization/depolymerization. Since in tanycytes the vimentin expression is regulated by the photoperiod (Kameda et al. in Cell Tissue Res 314:251–262, 2003), we determine whether this also holds true for Stk33. We study hypothalamic sections from adult Djungarian hamsters (Phodopus sungorus) held under either long photoperiods (L:D 16:8 h) or short photoperiods (L:D 8:16 h) for 2 months. In addition, we examine whether age-dependent changes in Stk33 protein content exist. Our results show that Stk33 in tanycytes is regulated by the photoperiod as is the case for vimentin. Stk33 may participate in photoperiodic regulation of the endocrine system.     

Effect of photoperiod on vasopressin-induced aggression in Syrian hamsters

Syrian hamsters are photoperiodic and become sexually quiescent when exposed to short "winter-like" photoperiods. In short photoperiods, male hamsters display significantly higher levels of aggression than males housed in long photoperiods. Arginine-vasopressin (AVP) within the anterior hypothalamus (AH) has been reported to modulate aggression in hamsters housed in long photoperiods. Previous studies have shown that AVP can facilitate aggression and its effects appear to be mediated by AVP V(1a) receptors (V(1a)R). In the present study, we investigated whether the increased levels of aggression observed after exposure to short photoperiod were the result of an increased responsiveness to AVP within the AH. Injections of AVP into the AH significantly increased aggression in hamsters housed in a long photoperiod, but had no effect in hamsters housed in a short photoperiod. In addition, injection of a V(1a)R antagonist into the AH significantly inhibited aggression in hamsters housed in long photoperiod, but had no effect in hamsters housed in a short photoperiod. These findings indicate that AVP within the AH increases aggression in hamsters housed in long photoperiods, but not in hamsters housed in short photoperiods.     

European hamsters (Cricetus cricetus) show a transient phase of insensitivity to long photoperiods after gonadal regression

Annual rhythms of body weight and reproduction in the European hamster (Cricetus cricetus) are the result of an interaction between seasonal changes in day length (photoperiod) and seasonal changes in the responsiveness of animals to these photoperiods. The present study demonstrates that under natural conditions European hamsters are not able to perceive long photoperiods (i.e., a 16L:8D cycle) before mid-November. This is an important difference to other hamster species, in which regrowth of the gonads can be stimulated by exposure to long photoperiods at any stage of gonadal regression. The experiments also demonstrate the existence of an annual phase of sensitivity to long photoperiods that starts around mid-November and extends until March/April. During this phase of sensitivity, exposure to a long photoperiod (16L:8D) induced gonadal regrowth within 3 wk. Additional experiments with an accelerated photoperiodic lighting regimen indicated that a photoperiod of approximately 13 h is necessary to stimulate gonadal regrowth. Under natural light conditions in Stuttgart (48.46 degrees N), a photoperiod of 13 h is reached by the beginning of April, which fits well with the finding that the majority of animals kept under a natural light:dark cycle had well-developed gonads by the end of April. Nevertheless, these animals showed a rather variable timing of gonadal regrowth, ranging from early January to late April. This is most likely the result of two processes: first, an endogenous mechanism (photorefractoriness) that induces gonadal recrudescence without any photoperiodic information while the animals are still in their hibernation burrows, and second, a direct stimulatory effect of long photoperiods.     

Twenty-four-hour profiles of serum leptin in siberian and golden hamsters: photoperiodic and diurnal variations

Serum leptin concentrations were obtained from male Siberian hamsters (Phodopus sungorus) and golden hamsters (a.k.a. Syrian, Mesocricetus auratus) housed on long [light:dark (LD) 16:8] and short (LD 6:18) photoperiods for 10-11 weeks. Blood samples were collected at 45-min intervals for 24 h from individual animals using an in-dwelling atrial catheter. In Siberian hamsters, exposure to short photoperiods as compared to long photoperiods reduced body weight (32.5 +/- 1.5 vs 47.7 +/- 1.1 g) and leptin (24-h mean: 5.3 +/- 0.4 ng/ml vs 18.6 +/- 2.1 ng/ml). Although photoperiod influenced the temporal distribution of leptin in golden hamsters, the main effect of photoperiod on leptin levels in golden hamsters did not reach significance (24-h mean: 7.1 +/- 1.0 ng/ml vs 5.1 +/- 0.8 ng/ml.). Body weights of golden hamsters did not vary significantly following exposure to short photoperiod for 11 weeks (178.3 +/- 3.6 g in LD 6:18 vs 177.8 +/- 7.3 g in LD 16:8). There was no nocturnal increase in serum leptin in either species. Marked interindividual differences were apparent in individual leptin profiles. Periodogram analysis revealed that only a few animals exhibited 24-h periodicities; the presence of a significant 24-h periodicity was more common in hamsters exposed to short days. Photoperiod-associated differences in the 24-hour profile of leptin secretion may be the result of photoperiod-associated changes in feeding behavior or metabolism. A full understanding of the regulation of leptin secretion in multiple time domains may enhance our understanding of the function of leptin.     

Hypothalamic expression of thyroid hormone-activating and -inactivating enzyme genes in relation to photorefractoriness in birds and mammals

Photorefractoriness is the insensitivity of gonadal development to the stimulatory effects of long photoperiods in birds and to the inhibitory effects of short photoperiods in small mammals. Its molecular mechanism remains unknown. Recently, it has been shown that reciprocal expression of thyroid hormone-activating enzyme [type 2 deiodinase (Dio2)] and -inactivating enzyme [type 3 deiodinase (Dio3)] genes in the mediobasal hypothalamus is critical for photoperiodically induced gonadal growth. Since thyroid hormones are required not only for photoinduction, but also for the induction of photorefractoriness, we examined the expression of these genes in relation to photorefractoriness in birds and mammals. Transfer of birds to long photoperiods induced strong expression of Dio2. This was maintained in tree sparrow when they later became photorefractory, but decreased somewhat in quail. In hamsters, transfer to long photoperiods also induced strong expression of Dio2. High values were not maintained under long photoperiods, and, indeed, expression decreased at the same rate as in animals transferred to short photoperiods. There was no renewed expression of Dio2 associated with testicular growth as animals became refractory to short photoperiods. Expression of Dio3 was high under short photoperiods and low under long photoperiods in all the animals examined, except for the short photoperiod-refractory hamsters. Our present study revealed complex regulation of deiodinase genes in the photoinduction and photorefractory processes in birds and mammals. These gene changes may be involved in the regulation of photorefractoriness, as well as photoinduction.     

Photoperiodic regulation of pulsatile luteinizing hormone secretion and adenohypophyseal gene expression in female golden hamsters.

LH concentrations were measured in serum collected at 10-min intervals from chronically ovariectomized female Syrian hamsters that had been maintained for 9 wk in stimulatory (long) or inhibitory (short) photoperiods. Short days reduced the number of detectable LH pulses during both the morning and the afternoon. Most short-day hamsters experienced a gradual afternoon rise in serum LH concentrations; this rise was not composed of multiple pulses. In separate groups of similarly treated hamsters, pituitary LH-beta mRNA abundance was significantly reduced by short-day exposure at both times of day even though serum LH concentrations rose in the afternoon. Estradiol treatment induced an afternoon surge of serum LH in both photoperiods, and eliminated the effect of photoperiod on LH-beta mRNA abundance in the afternoon. Serum prolactin (PRL) concentrations were not consistently influenced by day length in castrated hamsters with or without estrogen treatment, but PRL mRNA abundance was significantly suppressed by short-day exposure in all groups. The results indicate that day length exerts profound steroid-independent effects upon hypophyseal gene expression, and that the regulation of LH-beta mRNA abundance may be due to photoperiodic control of the neural GnRH pulse generator.     

Clock gene daily profiles and their phase relationship in the rat suprachiasmatic nucleus are affected by photoperiod

Rhythmicity of the rat suprachiasmatic nucleus (SCN), a site of the circadian pacemaker, is affected by daylength; that is, by the photoperiod. Whereas various markers of rhythmicity have been followed, so far there have been no studies on the effect of the photoperiod on the expression of the clock genes in the rat SCN. To fill the gap and to better understand the photoperiodic modulation of the SCN state, rats were maintained either under a long photoperiod with 16 h of light and 8 h of darkness per day (LD16:8) or under a short LD8:16 photoperiod, and daily profiles of Per1, Cry1, Bmal1 and Clock mRNA in darkness were assessed by in situ hybridization method. The photoperiod affected phase, waveform, and amplitude of the rhythmic gene expression as well as phase relationship between their profiles. Under the long period, the interval of elevated Per1 mRNA lasted for a longer and that of elevated Bmal1 mRNA for a shorter time than under the short photoperiod. Under both photoperiods, the morning and the daytime Per1 and Cry1 mRNA rise as well as the morning Bmal1 mRNA decline were closely linked to the morning light onset. Amplitude of Per1, Cry1, and Bmal1 mRNA rhythms was larger under the short than under the long photoperiod. Also, under the short photoperiod, the daily Clock mRNA profile exhibited a significant rhythm. Altogether, the data indicate that the whole complex molecular clockwork in the rat SCN is photoperiod dependent and hence may differ according to the season of the year.     

Photoperiodic regulation of adrenal hormone secretion and aggression in female Syrian hamsters

Seasonal changes in the length of the daily photoperiod induce significant changes in social behavior. Hamsters housed in winter-like short photoperiods (SP) can express significantly higher levels of aggression than hamsters housed in long photoperiods (LP) that mimic summer. The mechanisms responsible for increasing aggressiveness in SP-exposed female hamsters are not well understood but may involve seasonal changes in the endocrine system. In experiment 1, the effects of SP exposure on the circulating levels of three adrenal hormones were determined. Short photoperiod exposure was found to significantly depress the circulating levels of cortisol and the adrenal androgen dehydropiandrosterone (DHEA) but significantly increased the circulating levels of the sulfated form of DHEA, DHEAS. Experiment 2 examined the effects of gonadal hormones on several different measures of aggression including its intensity in females housed in both long and short photoperiod. Exposure to SP resulted in high levels of aggression regardless of the endocrine state of the animal or the measure used to quantify aggression. In contrast, administration of estradiol to hamsters housed in LP significantly reduced aggression. The data of the present study support the hypothesis that SP-housed females are more aggressive than LP-housed females because SP exposure renders females insensitive to the aggression-reducing effects of ovarian hormones.     

Long day photoperiods and temperature of 20 degrees C induce spermatogenesis in blinded and non-blinded marbled newts during the period of testicular quiescence

Adult male marbled newts (Triturus marmoratus) were collected at the end of the spermatogenesis period and exposed to different photoperiods (natural-daylength-simulated photoperiod, total darkness, 8L:16D, 12L:12D, 16L:8D, and continuous light) for 3 mo. Temperature was maintained at 20 degrees C. Two additional groups of newts were blinded and exposed to either the natural-simulated photoperiod and to 16 h of light per day respectively. Quantitative histologic studies on testicular development and germ cell volume per testis were performed. The newts captured in the field at the beginning (initial controls) or at the end of the experiments (final controls) were in the period of testicular quiescence. Newts kept in total darkness or exposed to a short photoperiod (8L:16D) showed germ cell development up to primary spermatocytes, whereas germ cell development in the newts exposed to long photoperiods (12L:12D or 16L:8D) progressed to elongated spermatids. The newts exposed either to intermediate photoperiods (natural-simulated photoperiod) or to constant light showed an intermediate degree of germ cell development (up to round spermatids). No significant differences between non-blinded and blinded animals were found. These results suggest that (1) mild temperature initiates testicular development in the period of testicular quiescence, (2) long photoperiods associated with mild temperatures produce spermatogenesis in this period, (3) complete darkness or constant light are less effective than some intermediate photoperiod, and (4) the effect of photoperiod on testicular function in newts is not related to ocular photoreception.     

Photoperiod-peptide interactions in the energy intake of Siberian hamsters

Siberian hamsters (Phodopus sungorous sungorous) decrease their food intake when exposed to short (“winter-like”) photoperiods. The cause of this naturally-occurring hypophagia is unknown, but it may be due to a heightened sensitivity to the factors that normally terminate food intake in long photoperiods, such as the putative satiety peptides. The purpose of the present investigation was to test whether there would be an enhanced sensitivity to the inhibitory effects of some of these peptides on food intake in short relative to long days. Ad lib-fed, adult female Siberian hamsters were housed in a long photoperiod (LD 14:10) and injected with bombesin, glucagon, cholecystokinin octapeptide (CCK-8) and calcitonin (CT). Food intake was monitored 1, 2, 4, 6, and 24 hr post-injection. Bombesin and glucagon had no effect on food intake in long day-housed hamsters. CCK-8 and CT inhibited food intake; however, CCK-8 did so without any apparent behavioral disruption, while CT produced a marked and prolonged depression of behavior. After 10 weeks of exposure to a short photoperiod (LD 8:16) the hamsters were tested again. The previously ineffective dose of bombesin greatly inhibited food intake following short photoperiod exposure. In addition, an increased inhibition of food intake by CCK-8 was also found. In contrast, glucagon did not decrease food intake and CT still produced its non-specific, behaviorally disruptive effects. To our knowledge, this is the first demonstration that the effectiveness of a putative satiety peptide can be dependent upon a change in the photoperiod. This heightened responsiveness of short photoperiod-exposed Siberian hamsters to the inhibitory effects of bombesin and cholecystokinin may account for the reduction in food intake that accompanies short day exposure in this species.     

Testicular prolactin receptors and serum growth hormone in golden hamsters: effects of photoperiod and time of day

Investigations were conducted to determine effects of exposure to short photoperiod--with its accompanying reductions in serum prolactin (Prl) concentrations--for various durations on testicular Prl receptors. An additional study investigated the possibility of nyctohemeral fluctuations in testicular Prl receptors and serum growth hormone (GH) concentrations and their alteration by photoperiod. After 10 and 28 days of exposure to a short photoperiod consisting of 5 h of light and 19 h darkness (5L:19D) (and prior to changes in testicular weight), there were progressive and significant reductions in the concentration of testicular Prl receptors (fmol/mg protein) when compared with long-photoperiod controls (14L:10D). After 12 weeks of 5L:19D, when testicular weights were dramatically decreased, Prl receptor concentration was reduced to 39% of long-photoperiod controls in one study, without alteration of affinity of Prl receptors for their labeled ligand. When measured at 6-h intervals in hamsters on 14L:10D, and on 5L:19D for 12 weeks, there were no significant changes in concentration or total content (fmol/testes) of testicular Prl receptors throughout the day. Although serum GH concentrations fluctuated markedly in hamsters on both photoperiods, no definitive nyctohemeral patterns were detected. These data provide indirect evidence for the ability of Prl to regulate its own testicular receptors, and demonstrate that diurnal fluctuations in testicular sensitivity to injected Prl are not a consequence of changes in Prl receptors. The data also suggest the absence of effects of photoperiod on serum GH concentrations in male golden hamsters.     

Effects of a circadian mutation on seasonality in Syrian hamsters (Mesocricetus auratus).

In Syrian hamsters, exposure to short photoperiods or constant darkness induces a decrease in gonadotrophin secretion and gonadal regression. After 10-12 weeks, animals undergo spontaneous gonadal reactivation, gonadotrophin concentrations rise, and in males, testes size increases and spermatogenesis resumes. The tau mutation shortens the period of circadian wheel-running activity by 4 h in the homozygote. Here, we examine the impact of this mutation on the reproductive response to photoperiod change. Seventeen adult tau mutant and nine adult wild-type males were housed in complete darkness for 25 weeks and testes size determined at weekly intervals. Gonadal regression and subsequent recrudescence occurred in both groups of animals. Regression occurred more rapidly in tau mutants, with a nadir significantly earlier than wild-types but after a similar number of circadian cycles. Rates of testicular recrudescence were similar in both groups. Our data suggest that an acceleration of the circadian period increases the rate of reproductive inhibition in animals exposed to inhibitory photoperiods. Once initiated, the rate of spontaneous reactivation may be independent of the circadian axis.     

Development and seed number in indeterminate soybean as affected by timing and duration of exposure to long photoperiods after flowering

BACKGROUND AND AIMS: Long photoperiods from flowering to maturity have been found to delay reproductive development in soybean (Glycine max) and to increase the number of seeds per unit land area. This study was aimed to evaluate whether sensitivity to photoperiod after flowering (a) is quantitatively related to the length of exposure to long days and (b) persists throughout the whole pod-setting period. It was also evaluated whether seed number was related to changes in the duration of post-flowering phenophases. METHODS: Two field experiments were conducted with an indeterminate cultivar of soybean of maturity group V. In expt 1, photoperiods 2 h longer than natural daylength were applied during different numbers of days from the beginning pod stage (R3) onwards, while in expt 2 these photoperiod extensions were imposed during 9 consecutive days starting at different times between R3 and R6 (full seed) stages. KEY RESULTS: There was a quantitative response of development to the number of cycles with a long photoperiod. The exposure to long photoperiods from R3 to R5 (beginning of seed growth) increased the duration of R3-R6 regardless of the timing of exposure. The stages of development comprised in the R3-R6 phase were delayed by current as well as by previous exposure to long days. A positive relationship was found between seed number and the duration of R3-R6, irrespective of the timing and length of exposure to the long photoperiod. CONCLUSIONS: Sensitivity to photoperiod remained high during the reproductive period and was highly and positively coupled with the processes of generation of yield.     

Social cues attenuate photoresponsiveness of the male reproductive system in Siberian hamsters (Phodopus sungorus)

Transfer of adult Siberian hamsters (Phodopus sungorus) from long (16 h light and 8 h dark, 16L:8D) to short (8L:16D) daily photoperiods induces an involution of the gonads and a cessation of reproductive behavior 8 to 10 weeks later. However, when male and female long-day hamsters were paired on transfer to short photoperiods, the males' gonads did not undergo the typical short-day response. Similarly, when male long-day hamsters were paired with refractory females (i.e., females housed in short photoperiods for at least 28 weeks so that they became unresponsive to short photoperiods), the response of the males' reproductive system to short photoperiods also was attenuated. Thus, social cues can override or delay the effects of photoperiod on the testes of this species. These results suggest that the inhibitory effects of long durations of melatonin secretion (in response to short photoperiods) on the male hypothalamic-pituitary-gonadal axis may be attenuated by social cues such as contact with the opposite sex.     

Circadian Changes in Anxiolysis-Related Behavior of Syrian Hamsters. Correlation with Hypothalamic GABA Release

The diurnal variations in anxiolysis and exploratory behavior were examined in a plus-maze paradigm in Syrian hamsters exposed to 14 h light: 10 h dark photoperiods or to constant darkness for 3 days. The percent of time spent in open arms and the percent of entries to open arms (both indexes of anxiety-related behavior) as well as the number of crosses to both arms (an index of locomotor behavior) showed significant daily variations under the two lighting conditions, maxima being found at night (2400-0400 h). Flumazenil (5 mg/kg) injected at the middle of the light (at 1600 h) or dark period (at 0400 h) decreased by 39-54 % anxiolysis-related behavior without affecting locomotor activity significantly. 3 H-?-aminobutyric acid (GABA) release from preoptic-medial basal hypothalamic explants obtained from hamsters exposed to 14 h light: 10 h dark photoperiods attained maximal values at 2400-0400 h. The results further supported the existence of circadian changes in anxiolysis-related behavior in Syrian hamsters that correlated with an increased hypothalamic GABA release.     

Role of photoperiod and melatonin in seasonal acclimatization of the djungarian hamster,Phodopus sungorus

The Djungarian hamster,Phodopus sungorus, shows a clear annual cycle in some thermogenic parameters such as nonshivering thermogenesis (NST) and cold resistance. These seasonal changes were found to be basically controlled by natural changes in photoperiod. Further support for this view was obtained by exposing the hamsters to artificial long and short photoperiods.Implantation of melatonin during fall and winter results in an increased thermogenic capacity in both short and long day hamsters comparable to that shown by values of control hamsters exposed to short photoperiods during winter. This thermotropic action of melatonin and of short photoperiod could be found only in fall and winter whereas during spring and summer, melatonin, like photoperiod, had no influence on thermogenic capacities. These results show that the actions of melatonin and photoperiod vary with the season and that they depend upon the photoperiodic history of the hamsters. Our results further indicate that the pineal gland with its hormone melatonin is involved in mediation of photoperiodic control of seasonal acclimatization.     

Photoperiod controls the induction,retention, and retrieval of antigen-specific immunological memory

Changes in day length affect several measures of immunity in seasonally breeding mammals. In Siberian hamsters (Phodopus sungorus), short day lengths suppress specific secondary antibody responses to the keyhole limpet hemocyanin (KLH) antigen and enhance cutaneous delayed-type hypersensitivity (DTH) responses to dinitrofluorobenzene (DNFB). These experiments tested whether day length affects secondary antibody and DTH responses by altering immune function solely during the interval after the initial exposure to each antigen, solely during the interval after the second exposure, or during both stages of the respective immune responses. Adult male Siberian hamsters were exposed to either a long (16 h light/day; LD) or a short (8 h light/day; SD) photoperiod for 7.5 wk before receiving an initial exposure to each antigen (KLH injection, cutaneous DNFB treatment; separate groups of animals for each antigen). A subset of LD hamsters was transferred to the SD photo-period, and a subset of SD hamsters was transferred to the LD photoperiod. Other hamsters remained in LD or SD. Eight weeks later, all hamsters were challenged with a second subcutaneous injection of KLH or a second application of DNFB to the ear, and immune responses were measured. Exposure to SD during the primary antibody response did not affect secondary IgG responses, but SD exposure during the secondary response significantly suppressed IgG production independent of day length during the initial KLH treatment. In contrast, exposure to SD during the DNFB challenge enhanced the ensuing DTH response, but this enhancement depended on the photoperiod prevailing during the initial exposure. Exposure to SD during the sensitization stage did not enhance DTH in hamsters subsequently exposed to LD. The data suggest that short photoperiods have enduring effects on immune responsiveness and on the establishment and retention of immunological memory.