본 연구는 돼지의 난포란을 체외에서 성숙, 수정시킨 체외수정란의 체외배양 체계를 확립하고 그 기작을 규명하기 위하여 체외배양액에 항산화제인 melatonin의 첨가 및 melatonin과 sodium nitroprusside(SNP)의 첨가배양이 체외수정란의 체외발육에 미치는 영향을 검토하고자 실시하였다. NCSU 23 배양액에 melatonin을 0, 1, 5 및 10nM을 첨가하여 체외배양을 실시한 결과, 배반포기까지 발육율은 17.8%, 26.1%, 20.0% 및 16.3%로서 melatonin 1nM 첨가구가 여타구에 비해 통계적으로 유의하게 높은 성적을 나타냈으며(P<0.05), 상실배기 이상 발육 성적에서도 melatonin 1 nM 첨가구가 39.1%로서 대조구 33.3%, 5 nM 첨가구의 33.3% 및 10 nM 첨가구의 27.9%보다 높은 발육율을 나타냈다(P<0.05). NCSU 23 배양액에 SNP를 0, 50 및 100 μM을 첨가하여 체외 배양한 결과, 상실배 이상 발육성적은 각각 41.9%, 25.6% 및 28.4%로서 SNP 첨가구가 대조구보다 유의적으로 낮은 성적을 나타내었다(P<0.05). NCSU 23 배양액에 대조구, SNP 50 μM, SNP 50 μM에 melatonin 1, 5 및 10nM을 혼합첨가하여 체외 발육율을 조사한 결과, 배반포기 발육율은 각각 2.5%, 1.2%, 9.9%, 5.1% 및 3.7%로서 SNP 50μM + Mel. 1nM 첨가구가 여타구 보다 높은 성적을 나타냈으며, 상실배기 이상 체외 발육율은 31.3%, 34.1%, 39.5%, 29.4% 및 39.5%로서 SNP 50μM + Mel. 1 nM 첨가구와 SNP 50 μM + Mel. 10 nM 첨가구가 여타구보다 높은 발육율을 나타냈다. 모든 처리구에서 배반포까지 발육된 체외수정란의 세포수는 커다란 차이가 인정되지 않았다.
광주기(하루 중 빛의 길이)는 골든 햄스터의 생식을 조절하는 주된 요인이다. 광주기 정보는 멜라토닌을 통하여 생식 내분비계로 전달된다. 따라서 멜라토닌이 생식에 미치는 효과를 여러 광주기에 노출시킨 햄스터에서 조사하였다. 단주기(하루 중 12시간 이하의 조명)에 노출시킨 동물들과 저녁에 멜라토닌을 주사한 동물들의 정소 무게는 현저하게 줄어들었으나, 장주기 (하루 중 12.5시간 이상의 조명)에 유지된 동물과 오전에 멜라토닌을 투여한 동물들의 정소 무게는
The purpose of this study was to examine the effectiveness of the forest healing program for improving the mental health of urban elderly people. Most participants were over 70 years old (87%) among total participants (46 people). The period was from May 5, 2015 to September, 4, 2015. During the experiment, research was stopped for one month due to the spread of Mers. The 46 subjects who were over 60 years old were divided into 4 groups by the number of participation (0, 4, 8, 12 times). Psychiatric tests and blood melatonin concentration test were performed before and after the experiment and the mean difference was determined by the corresponding sample t-test. Differences between the groups were analysed by ANOVA analysis. The change of melatonin concentration was examined by enzyme immunoassay. According to the result of the psychiatric test, mental health of the aged people who did not participate in the forest healing program was worse than 16 weeks ago. People who participated 4 times also had worse mental health. 8 and 12 times participated people showed improved mental health which indicate the need of long-term forest healing for mental health. The result of melatonin test showed that blood melatonin did not increase in the elderly people who did not participate, whereas blood melatonin of the elderly who participated for 12 times showed an increase tendency. This result is similar with the previous study, which suggested that melatonin can be increased by activity of body stimulation or mood change. In conclusion, we can say that forest healing programs have a positive effect on the mental health of elderly people. However, this study had several limitations. There is a need for follow-up studies to study age and individual differences, and to identify differences between periods
The changes of approximate composition, antioxidant activity and melatonin content in rapeseed were monitored during 10 days germination to choose the optimum sprouts with beneficial functionalities. The moisture content of seeds increased from 3.23% to 17.50% during 10 days germination period, while ash content was similar (4.03~4.91%). The fat content was reduced from 38.18% to 22.65%, however, protein content increased from 21.43% to 26.72%. The carbohydrate content increased from 32.71% to 35.84% at 8th day. The reducing sugar and fiber contents reached 10.47% and 15.12% at 6th day from 4.04% and 12.82%, respectively. Eighth day sprouts contained highest level of polyphenol (gallic acid equivalent, 19.2 g/kg). Germination increased antioxidant activities (trolox equivalent, mmol/kg) of seeds with slight difference depending on assay. That is, antioxidant activity was highest at 6th day in DPPH radical scavenging (55.9), 10th day both in ABTS radical scavenging (71.6) and Fe(III) reducing (125.3), and 4th day in peroxy radical scavenging (116.7) assay. Melatonin content of seeds (0.85 μg/kg) also increased and reached up to 14.93 μg/kg at 6th day. In conclusion, germination more than 4 days raised antioxidant activity of seeds highly, and 6th day sprouts had additional benefit in terms of melatonin, reducing sugar and crude fiber contents.
Caffeic acid O-methyltransferase (COMT) methylates N-acetylserotonin into melatonin; that is, it has N-acetylserotonin O-methyltransferase (ASMT) activity. The ASMT activity of COMT was first detected in Arabidopsis thaliana COMT (AtCOMT). To confirm the ASMT activity of COMT in other plant species, we evaluated the ASMT activity of a COMT from rice (Oryza sativa) (OsCOMT). Purified recombinant OsCOMT protein from Escherichia coli was used to validate the high ASMT activity of OsCOMT, similar to that of AtCOMT. The Km and Vmax values for the ASMT activity of OsCOMT were 243 μm and 2,400 pmol/min/mg protein, which were similar to those of AtCOMT. Similar to AtCOMT, OsCOMT was localized in the cytoplasm. In vitro ASMT activity was significantly inhibited by either caffeic acid or quercetin in a dose-dependent manner. Analogously, in vivo production of melatonin was significantly inhibited by quercetin in 4-week-old detached rice leaves, suggestive of a positive role of COMT in melatonin biosynthesis in plants.
Although melatonin biosynthetic genes from plants have been cloned, the melatonin catabolism mechanisms remain unclear. To clone the genes responsible for melatonin metabolism, we ectopically expressed 35 fulllength cDNAs of rice 2-oxoglutarate-dependent dioxygenase (2-ODD) in Escherichia coli and purified the corresponding recombinant proteins. In vitro 2-ODD assays showed four independent 2-ODD proteins that were able to catalyze melatonin into 2-hydroxymelatonin, exhibiting melatonin 2-hydroxylase (M2H). These M2H proteins had peak activities at pH 8.0 and 30°C. The Km ranged from 121 μM to 371 μM with the Vmax ranging from 1.7 to 18.5 pkat/mg protein, respectively. The M2H enzyme activities were dependent on cofactors such as α-ketoglutarate, ascorbate, and Fe2+, similar to the 2-ODD enzymes. M2H activity was inhibited by prohexadione-Ca, an inhibitor of 2-ODD, in a dose-dependent manner. M2H activity was high in the roots of rice seedlings, concurrent with high transcription levels of 2-ODD 21, suggesting that 2-ODD 21 was a major gene for M2H activity. Analogous to the high M2H activity in the roots, 2-hydroxymelatonin was found in large quantities in roots treated with melatonin. These results suggest that melatonin was metabolized into 2-hydroxymelatonin by the M2H genes in plants, but the physiological significance of 2-hydroxymelatonin remains to be examined in the future.
Melatonin has several known physiological functions, the main one being synchronization of daily and seasonal rhythms. In addition, melatonin has been reported to influence reproduction and behavioral rhythms with varying results depending on the species. To date, it remains unknown how this rhythm in locomotor activity is controlled endogenously, although there must be coordination of chemical and molecular drivers. However, the species is poorly characterized at molecular level with little sequence information available in public databases. The aim of study was to clarify involvement of endogenous melatonin rhythms and locomotor activity in day-night activity of the eel, Anguilla japonica which is an economically important but endangered species. The levels during daytime (zeitgeber time; ZT 6) were significantly (P<0.05) lower than those during nighttime (ZT 18). A similar pattern was persisted under DD conditions, whereas it disappeared under LL conditions and ocular melatonin levels remained low. Therefore, it is likely that ocular melatonin levels of the nocturnal eel reared under LD and DD conditions fluctuate in a daily/circadian manner and night-related physiological processes are dependent on eel locomotor activities which is a nocturnal species. We found that similar number of genes were differentially expressed between day (ZT6) and nighttime (ZT18), suggesting that during the nighttime also important in differential gene expression with daytime. This work also provides essential information for further studies investigating the molecular basis of daily/circadian system in this species.
Melatonin plays pleiotropic roles in both animals and plants. Among them, the possible role of melatonin in the innate immune response in plants was emerging recently. As an initial study, we employed Arabidopsis to see whether melatonin is involved in the defense system against a virulent bacterial pathogen Psudomonas syringae DC3000. It was obviously observed that melatonin application of 10 μm concentration onto Arabidopsis and tobacco leaves induced various pathogenesis-related (PR) genes as well as a series of defense genes activated by salicylic acid (SA) and ethylene (ET), two key factors involved in the plant defense response compared to the mock-treated Arabidopsis and tobacco leaves, respectively. The induction of these defense-related genes in the melatonin treated Arabidopsis was well matched with an increase in resistance against pathogenic bacterium by suppressing its multiplication with about 10 fold relative over the mock-treated Arabidopsis. Furthermore, melatonin induced PR genes were almost completely or partially suppressed in npr1, ein2, and mpk6 Arabidopsis mutants indicative of SA and ET dependency of melatonin in plant defense signaling. These results suggest that melatonin may play a novel defense signaling molecule in plant-pathogen interaction
Living beings are surrounded by various changes exhibiting periodical rhythms in environment. The environmental changes are imprinted in organisms in various pattern. The phenomena are believed to match the external signal with organisms in order to increase their survival rate. The signals are categorized into circadian, seasonal, and annual cycles. Among the cycles, the circadian rhythm is regarded as the most important factor because its periodicity is in harmony with the levels of melatonin secreted from pineal gland. Melatonin is produced by the absence of light and its presence displays darkness. Melatonin plays various roles in creatures. Therefore, this review is to introduce the diverse potential ability of melatonin in manifold aspects in living organism.
Golden hamsters reproduce in a limited time of a year. Their sexual activities are active in summer but inactive in winter during which day length does not exceed night time and environmental conditions are severe to them. The reproductive activities are determined by the length of light in a day (photoperiod). Melatonin is synthesized and secreted only at night time from the pineal gland. Duration of elevated melatonin is longer in winter than summer, resulting in gonadal regression. The present study aimed at the influences of continuous melatonin treatments impinging on the gonadal function in male golden hamsters. Animals received empty or melatonin-filled capsules for 10 weeks. They were divided into long photoperiod (LP) and short photoperiod (SP). All the animals maintained in LP (either empty or melatonin-filled capsules) showed large testes, implying that melatonin had no effects on testicular functions. Animals housed in SP displayed completely regressed testes. But animals kept in SP and implanted with melatonin capsules exhibited blockage of full regression by SP. These results suggest that constant release of melatonin prohibits the regressing influence of SP.
Fish reproduction is influenced by various external environmental factors. Among these factors, changes of photic conditions are thought be most closely related to the reproduction of many fish species. In recent studies, melatonin that mediates the changes of photic condition outside has been used in an attempt to get a similar effect to photoperiod manipulation without changing photoperiod. However, the effects of exogenous melatonin vary depending on life stages. We have previously investigated the effect of melatonin on reproductive activity in mature Nile tilapia (Oreochromis niloticus). In this study, effects of exogenous melatonin on the spawning activity, the rate of mouth brooding success rate and early survival rate of lavae produced from melatonin treated Nile tilapia were investigated. Nile tilapia at the time of puberty were randomly divided into three groups and fed on control (0 ㎍/g), low-dose melatonin (2.5 ㎍/g BW) or high-dose melatonin (25 ㎍/g BW) diet. Fish were reared indoor tanks at 27~28℃ under natural-photoperiod and fed twice a day at 10:00 and16:00 during the experiment (120 days). Fish were sampled to examine growth and gonadal development on Day 30, 60, 90 and 120. There were no significant differences of somatic growth between control (no melatonin) and melatonin-treated groups, indicating that melatonin administration did not cause any negative effects on the growth of experimental fish. The patterns of gonadal development were almost the same between control and melatonin-treated groups. In juvenile tilapia, treatment with low-dose melatonin resulted in the increase of spawning rate, the rate of mouth brooding success and early survival rate of laval tilapia. However, high-dose melatonin resulted in the decrease of spawning rate, the rate of mouth brooding success and early survival rate of laval tilapia, although the earliest spawning occurred in one of the high-dose group fish. Fish in high-dose melatonin group failed to recover from reproductively suppressed state even after the cease of melatonin treatment, indicating that too much melatonin might act negatively on spawning activities and gonadal maturation in Nile tilapia. In conclusion, the results from this study suggest the possibility that a pertinent dose of melatonin treatment can effectively accelerate the spawning rate and the rate of mouth brooding success, and bring positive effects on early survival rate of tilapia lavae.
This study was to investigate the effect of fermentation extracts on the concentration of serotonin and melatonin in the serum of the ICR mice. The ICR mice were divided into water control group, lactobacillus fermentation solution including (Lactobacillus paracasei and Bifidobacterium longum B6) control group, positive control group (milk and doxylamine succinate), negative control group (caffein) and the groups treated with the extracts of Berberis koreana bark (WE: water extracts, FE-L.P: fermentation extracts of Lactobacillus paracasei, FE-B.L: fermentation extracts of Bifidobacterium longum B6). After ten-day feeding treatment, the mean concentration of serotonin for water control, WE, FE-L.P and FEB. L group was 134.72, 183.01, 232.09 and 223.78 ng/ml, respectively. The mean concentration for FE-L.P and FE-B.L group were approximately 66% larger than that for water control group. The mean concentration of melatonin for water control, WE, FE-L.P and FE-B.L group was 76.92, 106.66, 157.56 and 141.81pg/ml, respectively. The mean concentration of melatonin for FE-L.P and FE-B.L group were also larger than that for water control group. Our results indicated that the fermentation extracts of Berberis koreana bark have relatively greater potential to induce secretion of serotonin and melatonin. Therefore, the fermentation extracts have antidepressant effect.
Photoperiod is known to be the most potent environmental stimulus for reproduction in many lower vertebrates, and it exerts an endogenous effect by causing a rhythmic change in melatonin level, which increases during the dark phase and decreases during the light phase. In recent studies, melatonin administration was capable of changing seasonal reproductive activities in several fish species, and also resulted in modulating the timing of smoltification in a salmonid fish (Amano et al., 2004; Ligo et al., 2005). In this study, we have investigated the changes of spawning activities (number of eggs, frequency of spawning) by oral administration of melatonin in the Nile tilapia, Oreochromis niloticus. Fish were obtained from a local fish farm (Chungju, Chung Buk) and acclimatized in fish rearing facilities in Sunmoon University (Asan, Chung Nam). Randomly selected 24 fishes (12 males, 12 females; 200~400 g in body weight, BW) were divided into three groups and each fish was accommodated into glass aquarium individually. Fish were fed either with control diet (0 ㎎ melatonin) or low-dose melatonin diet (0.3 ㎎ melatonin/㎏, BW) or high-dose melatonin diet (3 ㎎/㎏, BW). Fish were individually checked everyday for the presence of eggs in their mouths, and the eggs were removed and counted when found. Fish were fed twice a day, but melatonin treated diet was given only once a day at 10:00a.m.. The daily feeding ration was set at 2% of the body weight so that the fish ate all the pellets provided. Fish were sampled for blood parameters (Hb, Glu, GOT, GPT) on March 2 (Day 30), April 2 (Day 60), May 3 (Day 90), June 3 (Day 120) and July 14 (Day 160). The sampling was done between 14:00 and 16:00 p.m.. For the sampling, fish were anesthetized in 50 ppm benzocaine, and body length (BL) and BW were measured. Blood sample was collected using a heparinized syringe from the caudal blood vessel. Melatonin affects several photoperiodic signals including reproductive activities in fish. Low-dose melatonin administration appeared to disturb spawning and sexual maturation in Nile tilapia in this study. Fish treated with low-dose melatonin spawned less numbers of egg than the fish treated with high-dose. In contrast, high-dose melatonin administration caused more frequent spawning than low-dose melatonin. On the other hand, somatic growth and blood parameter were not affected by melatonin treatments. These results suggest that pertinent melatonin could be involved in providing photoperiodic information as short day photoperiod. Fish treated with over dose melatonin seemed to have no melatonin-free interval. In that circumstance the melatonin message might have not been conveyed as a proper signal.
멜라토닌(N-acetyl-5-methoxytryptamine)은 포유동물의 뇌의 송과선에서 분비되는 호르몬으로 수면과 생체 리듬 등을 조절하고 난소 기능과 번식에도 영향을 미친다. 또한, 강력한 scavenger로서 항산화제의 역할을 한다. 이 연구의 목적은 멜라토닌이 생쥐 난구세포-핵낭(germinal vesicle, GV) 시기 난자 복합체의 체외성숙에 미치는 영향을 알아보는 것이다. 3주령의 ICR 암컷 생쥐의 난소에서 회수된 난자-난구세포 복합체
골든 햄스터의 생식활동은 광주기에 의해 조절된다. 그들의 생식능력은 여름에 왕성하고 겨울에는 퇴화한다. 송과선에서 분비되는 멜라토닌은 계절적 번식동물에서 생식활동을 중재한다. 멜라토닌 수용체가 최근에 사람을 포함하는 몇몇 동물에서 확인되었지만 골든 햄스터의 생식능력과 관련하여 알려진 바가 많지 않다. 역전사 PCR 방법을 사용하여 멜라토닌 수용체의 일부 유전자를 동정하였다(309 염기). 멜라토닌 수용체의 핵산 서열과 추론된 아미노산 서열을 보고된 다른