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Seasonal postembryonic maturation of the diurnal rhythm of serotonin in the chicken pineal gland

机译:鸡松果体中5-羟色胺昼夜节律的季节性胚后成熟

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Previously, we have demonstrated the postembryonic development of chicken (Gallus gallus domesticus L.) pineal gland functions expressed as changes in melatonin (MEL) biosynthesis. Pineal concentrations of MEL and its precursor serotonin (5-HT) were shown to increase between the 2nd and 16th day of life. We also found that levels of the mRNAs encoding the enzymes participating in the final two steps of MEL biosynthesis from 5-HT: arylalkylamine-N-acetyltransferase (AANAT) and hydroxyindole-O-methyltransferase (HIOMT), as well as their enzymatic activities, were raised during postembryonic development. Moreover, the manner of these changes was season-of-hatch dependent, even in animals kept under constant laboratory conditions (L:D 12:12). The most pronounced changes were seen in the concentrations of 5-HT and MEL, as well as in Aanat mRNA level and its enzymatic activity. The high daily variability in 5-HT content suggested that season-and age-dependent changes in the activity of the chicken pineal gland might rely on the availability of 5-HT, i.e. it may be limited by changes in pineal tryptophan (TRP) and/or 5-hydroxytryptophan (5-HTP) levels as well as by the activity of tryptophan hydroxylase (TPH) and aromatic L-amino acid decarboxylase (AADC): two enzymes participating in the conversion of TRP to 5-HT. The present study was undertaken with the following objectives: (1) to examine whether the pineal concentration of the 5-HT precursors TRP and 5-HTP exhibit age-and season-related changes; (2) to look for season-related differences in the transcription of the Tph1 and Ddc genes encoding enzymes TPH and AADC; (3) to identify the step(s) in postembryonic development in which these season-related variations in pineal gland function are most pronounced. Male Hy-line chickens hatched in the summer or winter, from eggs laid by hens held in L: D 16: 8 conditions were kept from the day of hatch in L: D 12: 12 conditions. At the age of 2 or 9 days, animals were sacrificed every 2 or 4 h over a 24-h period and their pineal glands were isolated under dim red light and processed for the measurement of (i) the pineal content of TRP, 5-HTP and 5-HT, and (ii) the level of Tph1 and Ddc mRNAs. Circadian rhythmicity of all the measured parameters was evaluated by the cosinor method. The pineal levels of TRP and 5-HT as well as the Tph1 and Ddc transcripts changed during postembryonic development in a season-related way. Whereas, the 5-HTP concentration did not vary between animals from both age groups, regardless of the season. Circadian rhythmicity of all the measured parameters was dependent on both the age and the season of hatch, and was greatest in older animals in the summer. These findings indicated that the efficiency of season-related MEL biosynthesis, reported previously, is limited by 5-HT availability and this limitation depends on the transcription of both the Tph1 and Ddc genes. Moreover, Ddc mRNA level in 9-d-old birds changed rhythmically, even though this gene is generally considered to be arrhythmic.
机译:以前,我们已经证明了鸡(Gallus gallus domesticus L.)松果体的胚后发育,以褪黑素(MEL)生物合成的变化表示。 MEL和其前体5-羟色胺(5-HT)的松果浓度显示在生命的第2天和第16天之间增加。我们还发现,编码从5-HT参与MEL生物合成的最后两个步骤的酶的mRNA水平:芳基烷基胺-N-乙酰基转移酶(AANAT)和羟基吲哚-O-甲基转移酶(HIOMT),以及它们的酶促活性,在胚胎后发育过程中生长。而且,即使在实验室条件不变的情况下,这些变化的方式也取决于孵化季节(L:D 12:12)。在5-HT和MEL的浓度以及Aanat mRNA水平及其酶活性中观察到最明显的变化。 5-HT含量的日变化较大,表明松果腺活性随季节和年龄的变化可能取决于5-HT的可用性,即它可能受到松果氨酸(TRP)和/或5-羟色氨酸(5-HTP)的水平以及色氨酸羟化酶(TPH)和芳族L-氨基酸脱羧酶(AADC)的活性:这两种酶参与了TRP向5-HT的转化。进行本研究的目的是:(1)检查5-HT前体TRP和5-HTP的松果浓度是否表现出与年龄和季节相关的变化; (2)寻找与季节相关的编码TPH和AADC酶的Tph1和Ddc基因的转录差异; (3)确定胚胎后发育的步骤,其中与季节相关的松果腺功能变化最为明显。从L:D 12:12条件下孵化之日起,在夏季或冬季从在L:D 16:8条件下饲养的母鸡所产的卵孵化的雄性Hy-line鸡。在2或9天大的时候,在24小时内每2或4 h处死动物,并在暗红色光下分离其松果体,并处理以测量(i)TRP的松果含量,即5- HTP和5-HT,以及(ii)Tph1和Ddc mRNA的水平。通过余弦法评估所有测量参数的昼夜节律。胚后发育过程中,TRP和5-HT的松果水平以及Tph1和Ddc转录本的变化与季节相关。然而,无论哪个季节,两个年龄段的动物之间的5-HTP浓度均没有变化。所有测得参数的昼夜节律都取决于年龄和孵化季节,并且在夏季的较大动物中最大。这些发现表明,以前报道的与季节相关的MEL生物合成的效率受到5-HT可用性的限制,而这种限制取决于Tph1和Ddc基因的转录。而且,即使通常认为该基因是心律不齐的,在9日龄的家禽中Ddc mRNA的水平也会有节奏地变化。

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