Protein Phosphorylation in Rat Pineal Gland and Its Regulation in Supersensitive and Subsensitive States

The phosphorylation of specific proteins in pineal homogenate was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Cyclic AMP had the capacity to stimulate in a dose-dependent manner the incorporation of 32P in protein bands of apparent molecular weights 59K, 56K, and 35K with a maximal effect at 1 microM. On the other hand, calcium alone did not induce a marked increase in 32P incorporation with the exception of a dose-dependent phosphorylation of a 46K protein with a peak effect at 0.2 mM calcium concentration. The addition of exogenous calmodulin enhanced 32P incorporation in proteins migrating in the 62K and 52K regions, an effect that was antagonized by the calmodulin inhibitor trifluoperazine. However, also under these conditions, the stimulation of pineal protein phosphorylation was rather weak compared to that observed in other brain areas. In an attempt to investigate the functional changes of these biochemical processes during environmental lighting and adrenergic stimulation, it was found that the administration of (-)-isoproterenol (5 mg/kg, s.c.), a beta-receptor agonist, induced a clear-cut enhancement of 32P incorporation into the cyclic AMP-sensitive 59K and 56K proteins only in animals exposed for 18 h to the light, whereas it was almost ineffective in those kept in the dark for the same period. This effect was antagonized by (-)-propranolol pretreatment (20 mg/kg), suggesting that the changes in cyclic AMP-dependent protein phosphorylation observed in supersensitive pineals may represent a beta-receptor mediated process.(ABSTRACT TRUNCATED AT 250 WORDS)     

Infanticide in chimpanzees: Review of cases and a new within-group observation from the Kanyawara study group in Kibale National Park

A prolonged attack on a mother and 2-year-old infant that resulted in the death of the infant was observed in the Kanyawara study group in Kibale National Park. The mother was a border-area resident who was first observed associating with unit-group males six years previously. The attackers were an adult male and an adult female with a 6-week-old infant clinging ventrally to her. The attack was unusual in several respects: it is the first time a male and a female chimpanzee have been observed cooperating closely in an infanticidal attack; the adult female initially attempted to intervene in the victim's behalf, but later joined in the attack after receiving aggression from the male; and the episode was longer in duration than other reported cases. In the year following the incident, the mother did not increase her association with community males, but was seen with the male who killed her infant. The relevance of these observations to sexual selection-based explanations for infanticide in chimpanzees is discussed.     

Biphasic 24-Hour Variations in Cyclic GMP Accumulation in the Rat Pineal Gland Are Due to Corresponding Changes in the Activity of Cytosolic and Particulate Guanylate Cyclase

Various parameters of the rat pineal gland display a 24-h rhythm. However, nothing is known about possible 24-h variations in cyclic GMP (cGMP) metabolism. In the present study, 24-h variations in pineal gland cGMP accumulation were investigated by determining the increase in cGMP level with and without inhibitors of phosphodiesterase at different time points over a light/dark cycle (12/12 h). Furthermore, the activity of guanylate cyclase (GC) was determined under substrate-saturated conditions regarding the cytosolic and particulate forms of the enzyme. It has been found that cGMP accumulation and GC activity display biphasic 24-h variations with two peaks--one approximately 7 h after lights "on" and the other approximately 7 h after lights "off." The activity of cytosolic GC remains unchanged in the presence of the nitric oxide (NO) synthesis inhibitor N-monomethyl-L-arginine, indicating that 24-h variations in the activity do not reflect changes in the synthesis of the GC stimulator NO.     

Parental care and EGG size in salamanders: An examination of the safe harbor hypothesis

The safe harbor hypothesis includes the suggestion that parental care causes the embryonic stage to be the safest harbor, and, therefore, egg size will increase in populations with parental care to decrease the duration of subsequent, higher risk stages. Neither the safe habor hypothesis nor r and K theory seem adequate to explain the correlation between egg size and the presence/absence of parental care among salamanders, a group in which there is a further correlation between the larval (hatchling) habitat and egg size/parental care. Pond-breeding salamanders generally have small eggs and lack parental care, and stream-breeding salamanders generally have large eggs and parental care. I argue that the fundamental difference in the food available to hatchling salamanders between lentic (plankton-rich) and lotic (plankton-poor) environments selects for relatively lower parental investment in the lentic environment. From the standpoint of parental fitness, small (more numerous) hatchlings have a greater payoff where the available food is mall and dense (zooplankton in lentic environments), and large hatchlings are selectively advantageous where the food is of large size and less dense (benthic invertebrates in lotic environments). Selection for larger hatchlings in lotic environments results in longer embryonic periods and, ceteris paribus, greater total embryonic mortality. Embryo hiding and guarding have evolved among lotic-breeding salamanders as compensatory mechanisms to reduce the rate of embryonic mortality. In this view, parental care is a consequence of selection for larger egg size and not an umbrella that allows egg size to increase, contrary to the safe harbor hypothesis. The relationship between variance in parental investment and food available to offspring, developed here for salamanders, may be of general significance. YosiakiItô , a critic of r and K theory, independently arrived at a similar conclusion from a broader data base.     

Differential Expression of mRNA and Protein Encoding Retinal and Pineal S-Antigen During the Light/Dark Cycle

S-Antigen is a soluble cell protein unique to the retina and pineal gland. In the former, it is a well-characterized molecule that participates in light-induced signal transduction in photoreceptor cells. In the latter, the functional role is presently not known. The expression of S-antigen and its mRNA was examined in the rat retina and pineal gland throughout the diurnal cycle and with light interruption of the dark cycle. A cDNA for rat S-antigen was isolated from a pineal gland library to examine the mRNAs. A 1.7-kb mRNA for S-antigen was observed in both the pineal gland and the retina. Retinal S-antigen mRNA was expressed throughout the diurnal cycle and increased with light interruption of the dark cycle. In contrast, pineal gland S-antigen mRNA levels were detectable only during the dark and were absent preceding and during light. The phenotypic expression of immunoreactive S-antigen, identified with two S-antigen monoclonal antibodies (MAbs), MAb A9C6 and MAb C10C10, was analyzed by sodium dodecyl sulfate (SDS)-polyacrylamide gel (PAGE) and isoelectric focusing (IEF) electrophoresis. Immunoblot analysis of gels after SDS-PAGE revealed a single 46-kDa protein in retina. In contrast, two bands of approximately 43 and 46 kDa were identified in the pineal gland. Immunoblots of the retinal extracts separated by IEF electrophoresis revealed five S-antigen isomers, which vary quantitatively throughout the diurnal cycle and when light interrupted the dark cycle. Immunoblots of the pineal gland samples separated by IEF electrophoresis indicated that the pineal gland possesses four pineal gland-specific forms of S-antigen in addition to the five forms present in the retina. The differences observed in the mRNA and protein analyses suggest tissue-specific structural components for S-antigen in the retina and pineal gland that are not regulated in the same manner.     

Resynchronization of the Circadian Corticosterone Rhythm After a Light/Dark Shift in Juvenile and Adult Mice

Most of the extensive literature concerning the resynchronization of circadian rhythms after a Zeitgeber shift is devoted to the dependence of resynchronization on the mode of the shift and the strength of the Zeitgeber, as well as on the circadian function investigated. Ontogenetic influences have rarely been investigated. Therefore, we studied the resynchronization of several circadian rhythms in juvenile and adult female laboratory mice. We present here the results concerning the corticosterone rhythm. The daily rhythms were determined as transverse profiles (2-h intervals) before as well as 3, 7, and 14 days after an 8-h phase delay of the light/dark cycle produced by a single prolongation of dark time. The corticosterone concentration in serum was determined radioimmunologically. In the control animals the daily patterns were bimodal, with main maxima at the end of the light time and secondary ones just after lights on. Ontogenetic differences were small. In adult mice the amplitude was slightly increased due to an increase in the maximum values, and the time of highest hormone concentrations was slightly phase advanced. In juvenile mice, a distinct daily pattern with a phase position in relation to the light/dark cycle corresponding to that of control animals was present on the 3rd day after the Zeitgeber shift. The daily mean as well as the minimum and maximum values increased initially and reached the values of control animals during the second week. In adult animals, a pronounced daily rhythm with the normal phase position was present only at the 7th postshift day. The amplitude, daily mean, and maximum values were decreased, and the minimum values were increased. The initial values were not reached even after 2 weeks. The results show that resynchronization was faster in juvenile mice compared with adult mice. As a possible cause for the observed age-related differences, a not yet stabilized phase-coupling between various circadian rhythms is supposed.     

Epigenetic Basis of Regeneration: Analysis of Genomic DNA Methylation Profiles in the MRL/MpJ Mouse

Epigenetic regulation plays essential role in cell differentiation and dedifferentiation, which are the intrinsic processes involved in regeneration. To investigate the epigenetic basis of regeneration capacity, we choose DNA methylation as one of the most important epigenetic mechanisms and the MRL/MpJ mouse as a model of mammalian regeneration known to exhibit enhanced regeneration response in different organs. We report the comparative analysis of genomic DNA methylation profiles of the MRL/MpJ and the control C57BL/6J mouse. Methylated DNA immunoprecipitation followed by microarray analysis using the Nimblegen ‘3 × 720 K CpG Island Plus RefSeq Promoter’ platform was applied in order to carry out genome-wide DNA methylation profiling covering 20 404 promoter regions. We identified hundreds of hypo- and hypermethylated genes and CpG islands in the heart, liver, and spleen, and 37 of them in the three tissues. Decreased inter-tissue diversification and the shift of DNA methylation balance upstream the genes distinguish the genomic methylation patterns of the MRL/MpJ mouse from the C57BL/6J. Homeobox genes and a number of other genes involved in embryonic morphogenesis are significantly overrepresented among the genes hypomethylated in the MRL/MpJ mouse. These findings indicate that epigenetic patterning might be a likely molecular basis of regeneration capability in the MRL/MpJ mouse.     

Parental effects: physiological age, mating pattern, and diapause duration on diapause incidence of progeny in the cabbage beetle, Colaphellus bowringi Baly (Coleoptera: Chrysomelidae)

The cabbage beetle, Colaphellus bowringi Baly, is a short-day species that has both a facultative summer and winter adult diapause. In this study, we systematically investigated the effects of parental physiological age, mating pattern (including mating frequency and delayed mating), and diapause duration on the diapause incidence of progeny in the beetle. As parents aged, progeny diapause incidences showed considerable inter-parental variability and an abnormally varying pattern, but their predicting line was approximate to horizontal when reared under constant conditions. There was significant difference in progeny diapause incidence between multiple-mating and once-mating treatments. Higher diapause incidences in the progeny were observed in delayed-mating treatments. Progeny diapause incidences increased as diapause duration in the parental generation increased, and hibernating individuals produced more diapausing progeny than aestivating ones. Reciprocal cross tests between the post-diapause adults and non-diapause adults with no diapause history showed that the effect of parental diapause duration on the progeny diapause incidence was determined by both mothers and fathers but the mothers appeared to have a stronger effect than the fathers. Our results suggest that the parental age effect in Colaphellus bowringi might not be simply age-dependent, but may be controlled by the interaction of multiple factors, including physiological, environmental, behavioral and genetic factors. We discuss the ecological and physiological significance of these findings.     

Parental care behavior in the monogamous,sexually dimorphic Madagascar paradise flycatcher: sex differences and the effect of brood size

I studied the parental care behavior of the Madagascar paradise flycatcher Terpsiphone mutata in northwestern Madagascar. I especially focused on feeding, brooding and vigilance behaviors. Feeding rate did not differ between males and females, but females spent more time at the nest than males. Females dedicated their time to brooding, while males perched on the nest and were vigilant. Both parents changed the feeding rate in relation to brood size, so the feeding rate per nestling was not different among nests of different brood size. Duration of brooding by females increased with decreasing brood size, suggesting that the Royama effect, the pattern of lower feeding rate per nestling in larger broods, did not apply in this study. Males spent more time on vigilance than females. Anti-predator vigilance by males should be important for nestling survival given the high predation pressure typical of this population. In conclusion, males provide considerable parental care probably to minimize nestling starvation and to avoid nest predation. My results are not consistent with the general pattern of less parental effort by males in monogamous, sexually dimorphic species.     

Scrapie infection activates the replication of ecotropic, xenotropic, and polytropic murine leukemia virus (MuLV) in brains and spinal cords of senescence-accelerated mice: implication of MuLV in progression of scrapie pathogenesis

Senescence-accelerated mice (SAMP8) have a short life span, whereas SAMR1 mice are resistant to accelerated senescence. Previously it has been reported that the Akv strain of ecotropic murine leukemia virus (E-MuLV) was detected in brains of SAMP8 mice but not in brains of SAMR1 mice. In order to determine the change of MuLV levels following scrapie infection, we analyzed the E-MuLV titer and the RNA expression levels of E-MuLV, xenotropic MuLV, and polytropic MuLV in brains and spinal cords of scrapie-infected SAM mice. The expression levels of the 3 types of MuLV were increased in scrapie-infected mice compared to control mice; E-MuLV expression was detected in infected SAMR1 mice, but only in the terminal stage of scrapie disease. We also examined incubation periods and the levels of PrPSc in scrapie-infected SAMR1 (sR1) and SAMP8 (sP8) mice. We confirmed that the incubation period was shorter in sP8 (210+/-5 days) compared to sR1 (235+/-10 days) after intraperitoneal injection. The levels of PrPSc in sP8 were significantly greater than sR1 at 210+/-5 days, but levels of PrPSc at the terminal stage of scrapie in both SAM strains were virtually identical. These results show the activation of MuLV expression by scrapie infection and suggest acceleration of the progression of scrapie pathogenesis by MuLV.     

Taurine Release in the Developing and Adult Mouse Hippocampus: Involvement of Cyclic Guanosine Monophosphate

The inhibitory neuromodulator taurine is involved in osmoregulation and cell volume adjustments in the central nervous system. In addition, taurine protects neural cells from excitotoxicity and prevents harmful metabolic events evoked by cell-damaging conditions. The release of taurine in nervous cell preparations is greatly enhanced by glutamate receptor agonists and various cell-damaging conditions. NO-generating compounds also increase taurine release in the mouse hippocampus. The further involvement of the NO/cGMP pathway and protein kinases in preloaded [³H]taurine release from hippocampal slices from adult (3-month-old) and developing (7-day-old) mice in normoxia and in ischemia was now studied using a superfusion system. The release was enhanced by 8-Br-cGMP and the phosphodiesterase inhibitor 2-(2-propyloxyphenyl)-8-azapurin-6-one (zaprinast), particularly in the immature hippocampus, indicating that increased cGMP levels induce taurine release. The release was also increased by the inhibitor of soluble guanylyl cyclase, 1H-(1,2,4)oxadiazolo-(4,3a)quinoxalin-1-one (ODQ) and the protein kinase C activator 4-phorbol 12-myristate 13-acetate (PMA), but only in the adult hippocampus. The ischemia-induced release was also enhanced by increased cGMP levels in both adult and developing mice, whereas protein kinase inhibitors had no effects in any conditions. The results demonstrate that cGMP is able to modulate hippocampal taurine release in both adult and developing mice, the rise in cGMP levels evoking taurine release in normoxia and in ischemia. This could be part of the neuroprotective properties of taurine, being thus important particularly in cell-damaging conditions and in preventing excitotoxicity.     

Regulation by Photoperiod of Seasonal Changes in Body Mass and Reproductive Function in Gray Mouse Lemurs (Microcebus murinus): Differential Responses by Sex

Microcebus murinus exhibits highly seasonal biological rhythms to cope with extreme seasonality in availability of resources. To study the role of daylength on seasonal changes in body mass and reproductive function, we exposed male and female gray mouse lemurs to natural, constant, or alternating light cycles for 2 years under constant environmental conditions. When exposed to either constant short (SD: 10 h light/day), long (LD: 14 h light/day), or intermediate (ID: 12 h light/day) daylength, males and females maintained a constant body mass with no spontaneous cyclic variation. We only observed typical seasonal body mass changes in subjects exposed to alternating periods of SD and LD, the weight gain being triggered by SD, whereas weight loss occurred under LD. Reproductive activity in females proceeded from an endogenous rhythm that was expressed under constant daylengths. In contrast, changes in reproductive activity in males depended on daylength variation. In both sexes, SD and LD have direct inhibitory or stimulatory effects on reproductive activity. In females, daylength regulates breeding season by synchronizing an endogenous sexual rhythm with the season, whereas in males, the perception of a critical photoperiod is used to determine the subsequent onset or arrest of their breeding season. These sexual differences in the effect of daylength could be related to sex-specific differences in reproductive constraints.     

Stimulatory Effects of Exogenous Thyroid Hormone and Growth Hormone on sn-Glycerol-3-Phosphate Dehydrogenase Activity in the Genetic-Hypothyroid Mutant Mouse Cerebellum: Restricted to the Second 20 Days of Postnatal Life

We recently reported that by postnatal day 40 the activity of sn-glycerol-3-phosphate dehydrogenase (GPDH) was significantly depressed in the cerebellum of genetic-hypothyroid mutant mice. This mutant mouse-GPDH combination was used in the present study to define the critical time period during which thyroid hormone (T4) and growth hormone (GH) are essential for maturation of Bergmann glial cells. Our findings are that (a) induction of GPDH activity in the Bergmann glial cell is dependent on T4, (b) T4 is most effective when administered during the second 20 days of postnatal life, (c) the effect of GH on GPDH activity is complementary to or synergistic with that of T4, and (d) Bergmann glial cells and radial glial fibers of the mutant mice contain immunoreactive GPDH following various hormonal treatments. These results suggest that T4 is indispensable for the maturation of Bergmann glial cells.     

The Behavioral Ecology of Libellula luctuosa (Burmeister) (Anisoptera: Libellulidae): I. Temporal Changes in the Population Density and the Effects on Male Territorial Behavior

A single population of a common pond dragonfly, Libellula luctuosa, was studied at a site where the density of males increased dramatically during the breeding season. Early in the summer one active male was found on each territory on the pond. Satellite males were only occasionally found on the territories. Later in the season the number of males per territory increased so that two or more males simultaneously defended on many of the territories, and several satellite males occupied each of the territories. The number and rate of female visitations per day did not change over the summer. These factors resulted in a change in the operational sex ratio with variations in male density. Male behavior was also altered with increasing population density. As male density increased, males were less likely to be seen perching on their territories and more likely to be seen performing aggressive acts such as chasing nearby territorial males and chasing intruders. At high male density, the duration of territorial behaviors was shorter than at low male density. Thus, the percent of a time budget spent in any one activity did not change despite the change in number of males present. Male activity in L. luctuosa is not strictly determined by the opportunity for aggression. Costs of aggression associated with territoriality are minimized by maintaining flexible territorial behaviors.     

Male Behavior and Regulation of Worker Mating in a Ponerine Ant, Pachycondyla (Bothroponera) sp. (Hymenoptera: Formicidae)

Male behavior and regulation of worker mating were observed in Indonesian Pachycondyla sp. The colonies had some mated workers but only one gamergate (mated and egg-laying workers). Males frequently tried to mate with workers in their natal nests irrespective of worker age and dominance rank, however, workers never accepted mounting of nestmate males. Worker mating occurred only under the gamergateless condition with alien males.     

Biochemical Correlates of Epilepsy in the El Mouse: Analysis of Glial Fibrillary Acidic Protein and Gangliosides

The E1 (epileptic) mouse is considered a model for complex partial seizures in humans. Seizures in E1 mice begin around 7-8 weeks of age and persist throughout life. To determine if astrocytic gliosis was present in adult seizing E1 mice, the distribution of glial fibrillary acidic protein (GFAP) was studied in the hippocampus using an antibody to GFAP. The mean number of GFAP-positive cells per square millimeter of hippocampus was approximately 15- to 40-fold higher in adult E1 mice than in nonseizing control C57BL/6J (B6) mice or in young nonseizing E1 mice. Relative GFAP concentration (expressed per milligram of total tissue protein) in hippocampus and cerebellum was estimated by densitometric scanning of peroxidase-stained western blots. GFAP concentration was 2.7-fold greater in hippocampus of adult seizing E1 mice than in the control B6 mice. No differences in GFAP content were detected between the strains in the cerebellum. Because gangliosides can serve as cell surface markers for changes in neuronal cytoarchitecture, they were analyzed to determine if the gliotic response in E1 mice was associated with changes in neural composition. Although the total ganglioside concentration of hippocampus, cerebral cortex, and cerebellum was similar in adult E1 and control B6 mice, a synaptic membrane enriched ganglioside, GD1a, was elevated in the adult E1 cerebral cortex and hippocampus. The findings indicate that E1 mice express a type of gliosis that is not accompanied by obvious neuronal loss.     

Circamensal Rhythmicity and Muscle Function: The Role of Reproductive Hormones in the Regulation of Strength

The human female menstrual cycle is characterised by fluctuations in reproductive and pituitary hormones which regulate the monthly release of an ovum for fertilisation. The circamensal changes in the hormonal milieu may influence exercise performance. The generation of maximal voluntary muscle strength, mediated by changes in the steroid hormones, has been the focus of recent research, particularly ergogenic effects of oestrogen. Peak force of the adductor pollicis, normalised for muscle size, has been reported mid-cycle in concert with the rise in oestrogen. The removal of an oestrogen and progesterone environment results in muscle weakness in humans and animals. These hormones decline concomitantly following natural ovarian failure (i.e., the menopause) and surgical extraction of the ovaries (i.e., bilateral oophorectomy). To overcome the problem of controlling for rapid cyclical changes in reproductive hormones across the menstrual cycle, and to isolate the effects of oestrogen when measuring strength, other models have been adopted. Treatment of in vitro fertilisation patients involves, in part, the down-regulation of gonadotropin releasing hormone receptors followed by up-regulation with exogenous pituitary hormones. This presents a very low and very high oestrogenic milieu while progesterone remains rel atively stable. Under these conditions, maximal strength of the first dorsal interosseus muscle has been unchanged. The rapid loss in strength observed post-menopause is preserved, and possibly restored, in women taking hormone replacement therapy. These findings are significant for the preservation of skeletal and muscular health of post-menopausal women. There is increasing evidence to support the role of reproductive hormones in the regulation of muscle strength. Since the link between the hormone milieu during the menstrual cycle and strength changes is weak, other models must be employed. Strength declines rapidly at the menopause, and offsetting muscle weakness in post-menopausal women has implications in reducing the risk of muscloskeletal injury. Female athletes who are prone to amenorrhoea constitute another target group. A hormone-deficient state may not be beneficial to performance, but may compromise muscle function.     

HMGB1对小鼠系膜细胞细胞周期及细胞周期相关蛋白表达的影响

该实验以小鼠系膜细胞MMC为研究对象,以重组HMGB1为刺激物,通过检测细胞周期的变化及细胞PCNA、CyclinD1、CDK4和p16的表达水平,初步探讨HMGB1对系膜细胞的细胞周期及其相关调控因子的影响。选取小鼠系膜细胞MMC为研究对象,随机分为对照组及0.05mg/LHMGB1刺激组,经流式细胞术检测发现HMGB1能够上调小鼠系膜细胞中S期细胞所占比例;免疫细胞化学检测显示,PCNA蛋白在小鼠系膜细胞中的表达上调;通过RT-PCR技术及Western blot技术检测到小鼠系膜细胞中CyclinD1 mRNA和蛋白以及CDK4蛋白的高表达情况,而p16蛋白的表达呈时间依赖性降低。由此可见,HMGB1可能是通过上调CyclinD1/CDK4的表达,并下调p16的表达,促进细胞从G_0/G_1期进入S期,介导了小鼠系膜细胞的异常增殖,可能是HMGB1参与狼疮性肾炎发病的可能机制之一。