Jeong Y-G, Rosales J L, Marzban H, Sillitoe R V, Park D-G, Hawkes R, Lee K-Y
Department of Cell Biology and Anatomy, The University of Calgary, Alberta, Calgary, Canada T2N 4N1.
Neuroscience. 2003;118(2):323-34. doi: 10.1016/s0306-4522(03)00002-2.
Cyclin-dependent kinase 5 (Cdk5) activity is required for CNS development. The Cdk5 activator, p35, is well characterized but its isoform, p39, has been less studied. Previously, p39 mRNA expression in rat brain was shown to peak at 3 weeks postnatal, and the level remains high in the adult cerebellum [Neurosci Res 28 (1997) 355]. However, p39 protein expression and specific localization in the cerebellum, where p39 mRNA level significantly exceeds that of p35, have not been examined. Here, we explored the specific cerebellar localization of the p39 protein in the developing and adult mice. Adult cerebellar Purkinje cell somata and dendritic arbors were strongly positive for p39 but only rare and barely detectable p39 was observed in Purkinje cell axons. Cdk5 also localized in Purkinje cell somata and dendrites of the adult cerebellum, but p35 localized only in Purkinje cell somata, further suggesting a functional difference between p35 and p39. During development, cerebellar p39 was first noted at P10. Primary cultures of a developing cerebellum also showed strong p39 immunoreactivity in Purkinje cell somata and dendrites, but weak p39 immunoreactivity in Purkinje cell axons. Starting from P10, p39 was observed in a subset of Purkinje cells that form parasagittal bands throughout the vermis and hemispheres. These bands were bilaterally symmetrical and continuous from one lobule to another. Conversely, Cdk5 and p35 showed a uniform staining pattern. The pattern of p39 closely resembled that of zebrin II/aldolase C, suggesting that p39 may play a role in the adult cerebellum rather than in pattern development. This premise is consistent with the normal pattern of zebrin II/aldolase C zones and stripes in mutant p39-/- mice. The alternating p39 parasagittal band pattern may reflect a role for p39 or Cdk5/p39 in the functional compartmentation of the cerebellum.
细胞周期蛋白依赖性激酶5(Cdk5)活性对于中枢神经系统发育是必需的。Cdk5激活剂p35已得到充分研究,但其同种型p39的研究较少。先前研究表明,大鼠脑中p39 mRNA表达在出生后3周达到峰值,且在成年小脑内该水平仍维持在高位[《神经科学研究》28(1997)355]。然而,在小脑内p39 mRNA水平显著超过p35的情况下,p39蛋白表达及特异性定位尚未得到研究。在此,我们探究了p39蛋白在发育中和成年小鼠小脑中的特异性定位。成年小脑浦肯野细胞的胞体和树突分支对p39呈强阳性,但在浦肯野细胞轴突中仅观察到罕见且难以检测到的p39。Cdk5也定位于成年小脑的浦肯野细胞胞体和树突,但p35仅定位于浦肯野细胞胞体,这进一步表明p35和p39之间存在功能差异。在发育过程中,小脑p39最早在出生后10天被发现。发育中小脑的原代培养物在浦肯野细胞胞体和树突中也显示出强p39免疫反应性,但在浦肯野细胞轴突中显示出弱p39免疫反应性。从出生后10天开始,在贯穿蚓部和半球形成矢状旁带的一部分浦肯野细胞中观察到p39。这些带双侧对称且从一个小叶连续到另一个小叶。相反,Cdk5和p35呈现均匀的染色模式。p39的模式与zebrin II /醛缩酶C的模式非常相似,表明p39可能在成年小脑中发挥作用,而不是在模式发育中发挥作用。这一前提与突变型p39-/-小鼠中zebrin II /醛缩酶C区和条纹的正常模式一致。交替的p39矢状旁带模式可能反映了p39或Cdk5 / p39在小脑功能分区中的作用。