Institute of Developmental Biology, Russian Academy of Sciences, Moscow, Russia.
Dokl Biochem Biophys. 2020 Jan;490(1):34-37. doi: 10.1134/S1607672920010123. Epub 2020 Apr 27.
One of the features of the developing suprachiasmatic nucleus (SCN), the "biological clock" of the body, is the early expression of dopamine (DA) receptors in the absence of dopaminergic neurons as a source of DA. Only recently we showed that DA in SCN is synthesized together by nerve fibers containing only tyrosine hydroxylase (TH) and neurons containing only aromatic L-amino acid decarboxylase (AADC). This study was aimed to assess specific characteristics of the phenotype of TH-fibers in ontogenesis. For this purpose, PCR and immunohistochemical analysis of the expression of genes and proteins such as TH, AADC, vesicular monoamine transporter (VMAT), and receptors for DA (D1, D2) was performed. We have detected numerous TH-immunoreactive fibers in SCN of young and adult rats. VMAT was observed in some of them, which suggests vesicular storage of L-DOPA. Considering the key role of TH-fibers in cooperative synthesis of DA, we assumed the presence of their dopamine regulation. Using double immunolabeling, we showed that D1 and D2 are present in TH-fibers in adult rats, and only D1 in young rats. According to PCR, D1 and D2 are also expressed in neurons of SCN in adult rats and only D1 in young rats. Thus, it was shown for the first time that VMAT and D1 are coexpressed in TH-fibers synthesizing L-DOPA in SCN in young and adult rats, and also D2 receptors in adult rats, which suggests vesicular storage and dopamine regulation of L-DOPA secretion, respectively.
视交叉上核(SCN)是身体的“生物钟”,其发育的一个特征是,在没有多巴胺能神经元作为多巴胺(DA)来源的情况下,早期表达多巴胺(DA)受体。直到最近我们才发现,SCN 中的 DA 是由仅含有酪氨酸羟化酶(TH)的神经纤维和仅含有芳香族 L-氨基酸脱羧酶(AADC)的神经元共同合成的。本研究旨在评估 TH 纤维在发生过程中的表型的特定特征。为此,进行了 PCR 和免疫组织化学分析,以评估 TH、AADC、囊泡单胺转运体(VMAT)和 DA 受体(D1、D2)等基因和蛋白的表达。我们在幼鼠和成年大鼠的 SCN 中检测到大量的 TH 免疫反应性纤维。在其中一些纤维中观察到 VMAT,这表明 L-DOPA 的囊泡储存。鉴于 TH 纤维在 DA 协同合成中的关键作用,我们假设它们存在多巴胺调节。通过双重免疫标记,我们表明 D1 和 D2 存在于成年大鼠的 TH 纤维中,而仅存在于幼鼠的 TH 纤维中。根据 PCR,D1 和 D2 也存在于成年大鼠的 SCN 神经元中,而仅存在于幼鼠的 SCN 神经元中。因此,首次表明 VMAT 和 D1 在合成 L-DOPA 的 SCN 中的 TH 纤维中共同表达,并且在成年大鼠中还表达 D2 受体,这分别表明 L-DOPA 分泌的囊泡储存和多巴胺调节。