Yan C, Bentley J K, Sonnenburg W K, Beavo J A
Department of Pharmacology, University of Washington, Seattle 98195.
J Neurosci. 1994 Mar;14(3 Pt 1):973-84. doi: 10.1523/JNEUROSCI.14-03-00973.1994.
Based on their relative abundance and regulation by Ca2+ and by phosphorylation in vitro, it is thought that the Ca2+/calmodulin-dependent phosphodiesterases (CaM-PDEs) are important modulators of cyclic nucleotide function in the brain. Two of the most abundant CaM-PDEs in the brain are the 61 kDa and 63 kDa isozymes. In this study, the regional and cellular expression of mRNA encoding these two different isoforms in mouse brain has been determined by in situ hybridization. The 63 kDa CaM-PDE mRNA has a wide-spread but uneven distribution. Very strong hybridization signals are present in the caudate-putamen, nucleus accumbens, olfactory tubercle, and dentate gyrus of the hippocampus. Somewhat lesser amounts of 63 kDa CaM-PDE mRNA are present in the olfactory bulb and piriform cortex. Weaker but still easily discernible hybridization signals are seen in several layers of the cerebral cortex, CA1 and CA3 regions of the hippocampus, amygdaloid nuclear complex, thalamus, hypothalamus, midbrain, brainstem, cerebellum, and spinal cord. A weak hybridization signal was detected in the globus pallidus of the basal ganglia. In general, the distribution of the 63 kDa CaM-PDE is very similar to that of dopamine receptors, suggesting that it may modulate dopamine function. In contrast, the 61 kDa CaM-PDE mRNA has a more limited and much different distribution, with the highest level of expression in the cerebral cortex and in the pyramidal cells of the hippocampus. A moderate hybridization signal was detected in the medial habenula and amygdaloid nuclear complex. In addition, small subsets of neurons in several other regions showed specific hybridization. Both PDE mRNAs appear to be localized exclusively in neuronal cell bodies. Their distinct distribution suggests important but different physiological roles for these two isozymes in the regional regulation of cyclic nucleotides in the CNS. Since these two isozymes are differentially phosphorylated by cAMP-dependent and Ca2+/CaM-dependent protein kinases, the differential expression also provides a potential mechanism by which these PDEs can differentially regulate cAMP and cGMP in different brain areas. The high expression levels in specific subsets of neurons also suggest that agents increasing Ca2+ in these neurons will increase the rate of cyclic nucleotide degradation.
基于它们的相对丰度以及在体外受Ca2+和磷酸化的调节,人们认为钙/钙调蛋白依赖性磷酸二酯酶(CaM-PDEs)是大脑中环状核苷酸功能的重要调节因子。大脑中最丰富的两种CaM-PDEs是61 kDa和63 kDa的同工酶。在本研究中,通过原位杂交确定了小鼠脑中编码这两种不同同工型的mRNA的区域和细胞表达。63 kDa CaM-PDE mRNA分布广泛但不均匀。在尾状核-壳核、伏隔核、嗅结节和海马齿状回中存在非常强的杂交信号。嗅球和梨状皮质中63 kDa CaM-PDE mRNA的含量略少。在大脑皮质的几层、海马的CA1和CA3区域、杏仁核复合体、丘脑、下丘脑、中脑、脑干、小脑和脊髓中可见较弱但仍易于分辨的杂交信号。在基底神经节的苍白球中检测到微弱的杂交信号。总体而言,63 kDa CaM-PDE的分布与多巴胺受体的分布非常相似,表明它可能调节多巴胺功能。相比之下,61 kDa CaM-PDE mRNA的分布更有限且差异很大,在大脑皮质和海马锥体细胞中的表达水平最高。在内侧缰核和杏仁核复合体中检测到中等强度的杂交信号。此外,其他几个区域的一小部分神经元显示出特异性杂交。两种PDE mRNA似乎都仅定位于神经元细胞体。它们不同的分布表明这两种同工酶在中枢神经系统中环状核苷酸的区域调节中具有重要但不同的生理作用。由于这两种同工酶被cAMP依赖性和Ca2+/CaM依赖性蛋白激酶差异磷酸化,这种差异表达也提供了一种潜在机制,通过该机制这些PDEs可以在不同脑区差异调节cAMP和cGMP。特定神经元亚群中的高表达水平还表明,增加这些神经元中Ca2+的试剂将提高环状核苷酸的降解速率。