褪黑素对男性下丘脑一垂体一性腺轴的调节作用

褪黑素（Melatonin,Mel）对生殖调控有重要意义,本文主要介绍了褪黑素对男性下丘脑-垂体-性腺轴（HPG）三个水平的调节作用,同时探讨了褪黑素调节生殖活动的可能作用机制,并展望了其在男性生殖中的研究发展趋势及应用。     

褪黑素和褪黑素受体对下丘脑-垂体-肾上腺轴作用的研究进展

松果体于儿童中期可发育至最高峰,普遍在7岁之后开始呈逐渐萎缩,并在成年后逐渐有钙盐沉着。褪黑素主要是由松果体进行合成和分泌所形成,存在较好的昼夜节律性,且通常是通过下丘脑的视交叉上核进行控制,并与环境中的光-暗呈现的周期改变存在密切关联。此外,褪黑素具有极其广泛的生物学作用,且其发挥作用的首站便是与特异性褪黑素受体相关结合,随后经由信号转导系统发挥相应的生物效应。褪黑素受体属于G蛋白耦联受体超家族重要成员之一,其主要是通过百日咳毒素敏感G蛋白的一致性G蛋白通路,减少环腺苷酸的急剧或(和)抑制毛喉菇素刺激的环腺苷酸升高,从而间接影响黑色素活动。下丘脑-垂体-肾上腺轴(HPA轴)是机体在发生应激反应过程中具有一定影响的系统,其所分泌的激素也会表现出昼夜节律性的改变,且此种改变与褪黑素的有关变化呈现出明显的相反性。提示了两者可能存在一定的相关,在机体免疫功能的调控中扮演着不同的角色。本文通过阐述褪黑素和褪黑素受体对HPA轴作用的最新研究进展,旨在明确三者存在的错综复杂的相互作用关系,继而为机体免疫功能调控的一系列疾病研究提供参考依据。     

光周期—褪黑素信号调控高原鼠兔精原细胞分化和季节性精子发生

高原鼠兔是青藏高原特有的小型哺乳动物,其繁殖活动呈现明显的季节性。成年雄性高原鼠兔在繁殖期睾丸重量显著增加,精子发生正常进行,而在非繁殖期睾丸退化,精子发生阻断在未分化精原细胞阶段。光周期控制实验显示,长光照（16h∶8h）诱导非繁殖期高原鼠兔重新启动精原细胞分化和精子发生;而短光照（8h∶16h）显著抑制繁殖期高原鼠兔精子发生。酶联免疫分析发现,褪黑素分泌水平在长日照条件下降低而在短日照条件下升高。非繁殖期高原鼠兔连续注射褪黑素拮抗剂能诱导生殖细胞发育和精子发生恢复。促性腺激素释放激素(GnRH)与黄体生成素(LH)在繁殖期鼠兔下丘脑垂体显著升高,促卵泡素（FSH）水平无显著差异。注射GnRH可以促进非繁殖期高原鼠兔精原细胞分化和精子发生,而褪黑素注射后抑制GnRH的分泌进而负调控性腺轴。综上,高原鼠兔季节性精子发生受光周期-褪黑素信号控制,后者主要通过控制GnRH、LH水平影响精原细胞分化。本研究对理解季节性动物精子发生的调控机制有重要借鉴意义。     

哺乳动物松果腺:其生物合成能力和生理意义

哺乳动物松果腺是视觉系统的一个退化器官。主要的明-暗周期与松果腺褪黑素生成的昼夜波动同步。明-暗周期的信息通过中枢和外周交感神经的一系列复杂神经元从眼传递到松果腺。在松果腺内,神经信号以褪黑素合成和释放在夜间升高为标志的内分泌信号表现出来。褪黑素是一种高活力的激素,它在调制光照周期对各种哺乳动物的季节性生殖中起着重要作用。褪黑素除影响生殖外,在机体内作用广泛。     

褪黑素激活大鼠大脑中动脉BKCa通道的受体依赖和非受体依赖途径

目的：研究褪黑素通过大电导Ca²⁺激活K⁺（BK_Ca）通道介导大脑中动脉张力变化的作用机制。方法：8周龄雄性Wistar大鼠,麻醉后取大脑中动脉,酶消化法急性分离脑中动脉平滑肌细胞,采用膜片钳技术全细胞记录模式检测细胞外液加入褪黑素前后BK_Ca通道和电压门控钾（K_V）通道的电流密度,褪黑素受体抑制剂2-苯基-N-乙酰色胺（luzindole）孵育后,全细胞记录模式记录加入褪黑素后BK_Ca通道电流幅值和贴附式单通道记录模式记录加入褪黑素后BK_Ca通道Po值,内面向外记录模式检测加入褪黑素后BK_Ca通道电导（G）,开放概率（Po）,平均开放时间（To）和关闭时间（Tc）。结果：①褪黑素（100 μmol/L）显著增加全细胞BK_Ca通道电流密度,但对K_V通道电流密度无显著影响;②luzindole （1 μmol/L）显著抑制褪黑素引起的BK_Ca通道电流密度增加;③贴附式单通道记录模式下,褪黑素（100 μmol/L）增加BK_Ca单通道Po值,luzindole （1 μmol/L）显著抑制褪黑素引起的Po增加;④内面向外单通道记录模式下,褪黑素（1 μmol/L,100 μmol/L）缩短BK_Ca单通道的To和Tc值,且Tc较To显著缩短;结论：褪黑素通过受体依赖和非受体依赖途径激活BK_Ca通道,介导大脑中动脉血管舒张。     

肾上腺素对幼龄小鼠胸腺褪黑素受体的调节

应用放射配体结合法检测幼龄小鼠胸腺褪黑素受体（ＭＲ）,并以此为实验模型研究肾上腺素（Ｅ）对胸ＭＲ的影响及作用机制。结果表明,生理浓度的Ｅ即对胸ＭＲ有明显抑制作用,其抑制效应具时间依赖性及剂量依赖性。β－肾上腺素能受体拮抗剂普萘洛尔可逆转引抑制效应,ＣＡＭＰ对ＭＲ也有明显抑制作用,表现Ｅ对ＭＲ的抑制是通过β受体而实现的。这些结果提示,Ｅ在生理情况下即对胸腺ＭＲ有调节作用。     

褪黑素调控动物繁殖功能的作用途径

褪黑素是由松果体分泌的一种多功能吲哚类激素,在动物繁殖过程中起了至关重要的作用.褪黑素可通过多条途径调控动物的繁殖功能,主要包括：G蛋白偶联受体途径;作为神经内分泌激素对动物繁殖进行调控;与其核受体结合在转录水平上调控动物繁殖;通过抗氧化作用调控卵泡发育.本文就褪黑素对动物繁殖功能的调控途径进行综述,旨在为褪黑素调控动物繁殖的相关研究提供帮助.     

Kisspeptins/GPR54神经内分泌系统对哺乳动物生殖活动的调控

哺乳动物的生殖活动受到神经内分泌系统的精确调控。近年来的研究表明,KISS-1基因的表达产物Kisspeptins及其G蛋白偶联受体（G protein coupled receptor 54,GPR54）在这一调控网络中发挥着重要作用,与哺乳动物青春期启动、发情排卵及季节性繁殖等过程密切相关。该文就近年来Kisspeptins/GPR54系统对哺乳动物生殖活动调控的研究进展进行简述。     

褪黑素对大鼠海马 CA3区长时程增强诱导的影响

目的：研究褪黑素对海马CA3区长时程增强（LTP）诱导阶段的影响及可能的机制。方法：用细胞外电生理记录方法。通过向大鼠海马CA3区分别微量注射褪黑素、Tacrine（胆碱酯酶抑制剂）和DNQX（非NMDA受体的竞争性拮抗剂）,以海马CA3区群体兴奋性突触后电位（fEPSP）斜率的改变为指标,观察PP-CA3通路的LTP变化。结果：①0．2μg/μl、1μg/μl和5μg/μl的褪黑素对CA3区的诱发电位和LTP均有抑制作用,随着剂量加大,抑制作用加强。②褪黑素能抑制Tacrine对LTP诱导的易化作用。③DNQx能部分阻断褪黑素对LTP诱导的抑制作用。结论：褪黑素对LTP诱导的抑制作用可能与胆碱能系统和非NMDA受体有关。     

雄性大熊猫褪黑素与睾酮季节性变化

褪黑素通过调控下丘脑-垂体-性腺内分泌轴使季节性繁殖动物在适宜的季节进行繁殖活动.大熊猫(Ailuropoda melanoleuca)在春季集中繁殖.为探究雄性大熊猫褪黑素和睾酮的季节性变化规律,本研究选取成都大熊猫繁育研究基地3只成年雄性大熊猫作为实验对象,在自然光照下对这3只大熊猫进行每周1次为期1年(2018年...     

Melatonin metabolism,signaling and possible roles in plants

Melatonin is a multifunctional biomolecule found in both animals and plants. In this review, the biosynthesis, levels, signaling, and possible roles of melatonin and its metabolites in plants is summarized. Tryptamine 5-hydroxylase (T5H), which catalyzes the conversion of tryptamine into serotonin, has been proposed as a target to create a melatonin knockout mutant presenting a lesion-mimic phenotype in rice. With a reduced anabolic capacity for melatonin biosynthesis and an increased catabolic capacity for melatonin metabolism, all plants generally maintain low melatonin levels. Some plants, including Arabidopsis and Nicotiana tabacum (tobacco), do not possess tryptophan decarboxylase (TDC), the first committed step enzyme required for melatonin biosynthesis. Major melatonin metabolites include cyclic 3-hydroxymelatonin (3-OHM) and 2-hydroxymelatonin (2-OHM). Other melatonin metabolites such as N¹-acetyl-N²-formyl-5-methoxykynuramine (AFMK), N-acetyl-5-methoxykynuramine (AMK) and 5-methoxytryptamine (5-MT) are also produced when melatonin is applied to Oryza sativa (rice). The signaling pathways of melatonin and its metabolites act via the mitogen-activated protein kinase (MAPK) cascade, possibly with Cand2 acting as a melatonin receptor, although the integrity of Cand2 remains controversial. Melatonin mediates many important functions in growth stimulation and stress tolerance through its potent antioxidant activity and function in activating the MAPK cascade. The concentration distribution of melatonin metabolites appears to be species specific because corresponding enzymes such as M2H, M3H, catalases, indoleamine 2,3-dioxygenase (IDO) and N-acetylserotonin deacetylase (ASDAC) are differentially expressed among plant species and even among different tissues within species. Differential levels of melatonin and its metabolites can lead to differential physiological effects among plants when melatonin is either applied exogenously or overproduced through ectopic overexpression.     

Desensitization of pigment granule aggregation in Xenopus leavis melanophores: melatonin degradation rather than receptor down-regulation is responsible

Xenopus laevis melanophores express a high density (B(max) 1224 fmol/mg protein) of high-affinity (K(d) 37 pm) cell membrane melatonin receptors. Treatment of melanophores with melatonin resulted in a loss of membrane melatonin receptors reaching a maximum (approximately 60%) by 6 h. In addition to receptor loss, a decline in the potency of melatonin to produce pigment aggregation was observed on prolonged treatment. However, the loss of potency (3.8-fold in 24 h and 162-fold in 96 h) was much slower than loss of receptors, and was completely prevented by inclusion of eserine (100 microm), an inhibitor of melatonin deacetylation in the culture medium. Incubation of melanophores with [(3)H]-melatonin showed that eserine prevented metabolism of melatonin to 5-methoxytryptamine. These results indicate that although receptor density does decline on prolonged treatment, this is not responsible for the diminishing melatonin potency, which is entirely due to degradation of melatonin by deacetylation and subsequent deamination in melanophores.     

Effects of melatonin and antagonists of MT1 and MT2 receptors on somatic pain induced within the fixed circadian rhythm

We studied behavioral pain-related reactions (PRRs) induced in mice by subcutaneous injections of 5% formalin within different phases of the fixed circadian illumination rhythm under conditions of administration of exogenous melatonin and of blocking of MT1 and MT2 melatonin receptors. It was demonstrated that modulation of experimentally induced somatic pain depends considerably on the phase of the preset circadian rhythm. In the norm, the duration of PRRs in the middle of the dark phase was 30% smaller than that in the middle of the light phase. Administration of exogenous melatonin in the middle of the light phase decreased the duration of episodes of noxious behavior by 43%, on average. Injections of melatonin within the dark phase resulted in no significant changes in the duration of PRRs. In the dark phase, the blockade of MT1 receptors by luzindole led to an increase in the duration of PRRs by 45%, as compared with the norm, while in the light phase we observed no significant alterations of this duration under conditions of blocking of the above-mentioned receptors. The blockade of MT2 receptors by prazocine in the middle of dark and light phases increased the durations of PRRs by 92 and 28%, respectively. Our data indicate that the analgesic effect of melatonin depends significantly on the level of this hormone in the organism; in turn, such a level is determined by the illumination conditions. The antinoxious effect of melatonin is mediated by MT receptors, in particular by MT2 receptors. Neirofiziologiya/Neurophysiology, Vol. 39, No. 3, pp. 255–259, May–June, 2007.     

The hockey-stick method to estimate evening dim light melatonin onset (DLMO) in humans

The onset of melatonin secretion in the evening is the most reliable and most widely used index of circadian timing in humans. Saliva (or plasma) is usually sampled every 0.5–1 hours under dim-light conditions in the evening 5–6 hours before usual bedtime to assess the dim-light melatonin onset (DLMO). For many years, attempts have been made to find a reliable objective determination of melatonin onset time either by fixed or dynamic threshold approaches. The here-developed hockey-stick algorithm, used as an interactive computer-based approach, fits the evening melatonin profile by a piecewise linear-parabolic function represented as a straight line switching to the branch of a parabola. The switch point is considered to reliably estimate melatonin rise time. We applied the hockey-stick method to 109 half-hourly melatonin profiles to assess the DLMOs and compared these estimates to visual ratings from three experts in the field. The DLMOs of 103 profiles were considered to be clearly quantifiable. The hockey-stick DLMO estimates were on average 4 minutes earlier than the experts' estimates, with a range of ?27 to +13 minutes; in 47% of the cases the difference fell within ±5 minutes, in 98% within ?20 to +13 minutes. The raters' and hockey-stick estimates showed poor accordance with DLMOs defined by threshold methods. Thus, the hockey-stick algorithm is a reliable objective method to estimate melatonin rise time, which does not depend on a threshold value and is free from errors arising from differences in subjective circadian phase estimates. The method is available as a computerized program that can be easily used in research settings and clinical practice either for salivary or plasma melatonin values.     

Effect of exogenous melatonin on the ovary, the embryo and the establishment of pregnancy in sheep

Administration of melatonin to advance the breeding season in sheep has been widely used, since this hormone conveys the photoperiodic signal to the reproductive neuroendocrine axis. An increased lambing percentage has been reported following such treatment during anoestrus, which could be mediated through a higher rate of embryonic survival, either by an improvement in luteal function or by a reduction in the antiluteolytic mechanisms. The aim of this article is to review the body of knowledge on the effect of melatonin on the ovine ovary, the embryo and the establishment of pregnancy. Some studies using synchronized ewes have found that melatonin treatments during anoestrus do not improve ovulation rate by modifying the timing of follicle emergence, but increasing the number of ovulatory follicles by decreasing the atresia of medium and large follicles. On the other hand, the addition of melatonin to the in vitro maturation medium does not improve oocyte maturation rate in oocytes from sheep ovaries recovered either in anoestrus or in the breeding season. However, a luteotrophic effect of melatonin at either short or medium term has been reported. We have recently observed that melatonin implants tend to improve the survival of embryos collected from ewes after superovulation in anoestrus. More specifically, melatonin induced a significant reduction of the number and rate of non-viable (degenerate and retarded) embryos. Preliminary data from our laboratory suggest that the uterine sensitivity to progesterone - in terms of progesterone receptor expression - of superovulated ewes could be reduced by melatonin treatment. It can be concluded that the success of exogenous melatonin as a means to improve lamb production of sheep is due, at least in part, to an improvement of luteal support and embryonic survival.     

Effects of pinealectomy, melatonin injections and melatonin antibody production on the mean duration of individual hibernation bouts in Spermophilus tridecemlineatus

1. 1.|Pinealectomy did not significantly alter the length of the mean durations of individual hibernation bouts in adult Spermophilus tridecemlineatus.

2. 2.|Similarly, no statistically-significant differences in mean durations of individual hibernation bouts between pinealectomized and sham-pinealectomized group squirrels were observed for young-of-the year S. tridecemlineatus

3. 3.|Some apparent differences of pinealectomy are suggested as they relate to age, stage of hibernation cycle and time of year during which pinealectomies are performed.

4. 4.|Daily s.c. injections of 50 μg of melatonin late in the subjective photophase induced no statistically-significant differences in mean durations of individual hibernation bouts between either adult, female pinealectomized or intact S. tridecemlineatus and their respective controls.

5. 5.|Adult, female S. tridecemlineautus with reduced titres of endogenous melatonin displayed on statistically-significant difference in the mean duration of initial hibernation bouts compared with controls.

6. 6.|It is possible that melatonin per se or that pinealectomy or melatonin titre alterations are generally failing to induce any significant changes in the torpor/arousal cycles of seasonal hibernators; or that a parameter other than the duration of torpor/arousal cycles is being modulated by the pineal complex.

Hormonal Modulation of Cyclic Melatonin Production in the Pineal Gland of Rats and Syrian Hamsters: Effects of Thyroidectomy or Thyroxine Implant

Night-time pineal levels of tryptophan, 5-hydroxytryptophan, serotonin, N-acetylserotonin, melatonin, 5-hydroxyindoleacetic acid and the activities of the two enzymes N-acetyltransferase and hydroxyindole-O-methyltransferase involved in the cyclic production of melatonin were determined in male albino rats and Syrian hamsters that were implanted with thyroxine or thyroidectomized two weeks earlier. Both treatments depressed nocturnal pineal melatonin content in rats and hamsters. The cause of this depression is not known, although minor alterations in the substrates and the enzymes involved in melatonin production were observed. The data suggest that alterations in thyroid hormone levels may increase the release of nocturnal melatonin from the pineal, thereby allowing less to accumulate in the gland.     

Expression of melatonin synthesis genes is controlled by a circadian clock in the pike pineal organ but not in the trout

The photosensitive teleost pineal organ exhibits a daily rhythm in melatonin production. In most teleosts, including the pike, this is driven by an endogenous pineal clock. An exception is the trout, in which the pineal melatonin rhythm is a direct response to darkness. This fundamental difference in the regulation of melatonin production in two closely related species provides investigators a novel opportunity to study the molecular mechanisms of vertebrate clock function. We have studied the circadian regulation of mRNA encoding two melatonin synthesis enzymes by Northern blot analysis. These two enzymes are serotonin N-acetyltransferase (AA-NAT), the penultimate enzyme in melatonin synthesis, and tryptophan hydroxylase (TPH), the first enzyme in melatonin synthesis. A clock controls expression of both AA-NAT and TPH mRNAs in the pineal organ of pike, but not that of trout, in which the levels of these mRNAs are tonically elevated. A parsimoneous explanation of this is that a single circadian system regulates the expression of both AA-NAT and TPH genes in most teleosts, and that in trout this system has been disrupted, perhaps by a single mutation.     

Melatonin synchronizes protein synthesis rhythm in hepatocyte cultures as an agonist of intercellular calcium and protein kinases

Melatonin in nanomolar concentrations synchronizes protein synthesis in primary cultures of hepatocytes through calcium-dependent activation of protein kinases. The synchronizing effect of melatonin is blocked by the cytoplasmic calcium chelating agent BAPTA-AM (20 μM) as well as by the inhibitor of protein kinases 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine dihydrochloride (40 μM). Thus, protein phosphorylation is the key event in hepatocyte synchronization by melatonin, as we have demonstrated previously for gangliosides and biogenic amines. The antagonist of melatonin receptors luzindole (20 nM) blocks the synchronizing function of melatonin.