MACRO Domain Containing 2 (MacroD2) is a neurodevelopmental disorder-related mono-ADP-ribosylhydrolase. Molecular features of this protein in neural tissues are largely unknown. In this study, we generated a specific antibody against MacroD2, and carried out expression and morphological analyses of the molecule during mouse brain development. In Western blotting, 2 MacroD2 isoforms with molecular masses of ∼70 and ∼75 kDa started to be expressed at embryonic day 16.5, reached the maximal level at postnatal day 8, and then gradually decreased through P30. In contrast, other isoforms with molecular masses of ∼110 and ∼140 kDa gradually increased during embryonic to postnatal development. In immunohistochemical analyses, MacroD2 was strongly detected in cortical neurons in layer II-V at P0 and P7, while the protein expression decreased significantly in the neurons at P30. Immunofluorescence analyses revealed that MacroD2 was mainly distributed in the soma and to a lesser extent in the axon and dendrite of immature primary cultured mouse hippocampal neurons. On the other hand, in the matured hippocampal neurons, while MacroD2 was detected in the soma, it displayed in dendrites a punctate distribution pattern with a partial colocalization with synaptic markers, synaptophysin, and PSD95. The obtained results indicate that MacroD2 is expressed and may have a physiological role in the central nervous system during brain development.

中文翻译：

---

神经发育障碍相关的单ADP核糖基水解酶的MACRO结构域的生化和形态学表征，包含2。

包含2的MACRO域（MacroD2）是与神经发育障碍相关的单ADP-核糖基水解酶。这种蛋白质在神经组织中的分子特征在很大程度上是未知的。在这项研究中，我们生成了针对MacroD2的特异性抗体，并在小鼠大脑发育过程中对该分子进行了表达和形态分析。在蛋白质印迹法中，分子质量分别约为70和75 kDa的2个MacroD2亚型在胚胎第16.5天开始表达，在出生后第8天达到最高水平，然后通过P30逐渐降低。相反，在胚胎到出生后的发育过程中，分子量约为110和140 kDa的其他同工型逐渐增加。在免疫组化分析中，在P0和P7的II-V层的皮质神经元中强烈检测到MacroD2，而在P30时，神经元中的蛋白表达显着下降。免疫荧光分析表明，MacroD2主要分布在未成熟的原代培养的小鼠海马神经元的轴突和树突中，程度较小。另一方面，在成熟的海马神经元中，虽然在体细胞中检测到MacroD2，但它在树突中显示出点状分布模式，并与突触标记物，突触素和PSD95部分共定位。获得的结果表明MacroD2在大脑发育过程中在中枢神经系统中表达并可能具有生理作用。另一方面，在成熟的海马神经元中，虽然在体细胞中检测到MacroD2，但它在树突中显示出点状分布模式，并与突触标记物，突触素和PSD95部分共定位。获得的结果表明MacroD2在大脑发育过程中在中枢神经系统中表达并可能具有生理作用。另一方面，在成熟的海马神经元中，虽然在体细胞中检测到MacroD2，但它在树突中显示出点状分布模式，并与突触标记物，突触素和PSD95部分共定位。获得的结果表明MacroD2在大脑发育过程中在中枢神经系统中表达并可能具有生理作用。