Durham D, Woolsey T A
J Comp Neurol. 1978 Apr 15;178(4):629-44. doi: 10.1002/cne.901780403.
The autoradiographic 2-deoxy-D glucose (2-DG) method has been used to map relative changes in metabolic activity in the CNS during various functional states (Plum et al., '76). Here we describe the application of the 2-DG method to assay regional activity in the posteromedial barrel subfield (PMBSF) region of the mouse SmI cortex after acute removal of mystacial vibrissae. One day prior to isotope injection, various combinations of vibrissae (e.g., all vibrissae, row-C only, rows-B and -D only) were plucked from adult male Swiss Webster mice under anesthetic. The next day, 5 muCi of 14C-2-DG were injected into a tail vein, and the mice were allowed to actively explore an empty cage for 45 minutes. The animals were then sacrificed, the brains quickly removed, frozen, and sectioned either parallel or perpendicular to the pia at 80 mum in a cryostat. The sections were mounted, dried on coverslips, and were used to expose X-ray film, after which the sections were stained with thionin and the X-ray film developed. The tissue sections and matching autoradiograms were compared directly from photomicrographs of each. The autoradiograms showed areas of higher activity in barrels for which corresponding vibrissae were present and lower activity in barrels for which appropriate vibrissae were missing. In tangential sections from animals with all vibrissae intact, the PMBSF was uniformly and consistently higher in activity than in cases with all vibrissae missing. The removal of row-C or rows-B and -D resulted in strips of decreased activity in the corresponding PMBSF rows. The same patterns of increased or decreased activity were also seen in sections normal to the pia, but the changes in activity, while greatest in layer IV, extended through all layers of the cortex. Finally, in a number of the autoradiograms, density patterns could be recognized which later were shown to relate directly to sides of individual barrels. The results indicate: (1) Acute removal of the peripheral vibrissal hairs is sufficient to deprive the related contralateral cortical barrel neurons of normal activity. Thus in the mouse somatosensory system it may be possible to determine the relative importance of sensory deprivation and neonatal peripheral lesions in normal cortical development. (2) The barrels are part of a functional cortical columnar organization similar to that in other sensory systems. And, (3) the 14C-2-DG-X-ray technique is sufficiently sensitive to reveal parts of individual barrels in autoradiograms and thus, with some modification, may be suitable for the study of small populations of neurons.
放射自显影2-脱氧-D-葡萄糖(2-DG)法已被用于绘制中枢神经系统在各种功能状态下代谢活动的相对变化(普拉姆等人,1976年)。在此,我们描述2-DG法在急性去除触须后对小鼠初级体感皮层(S1)后内侧桶状亚区(PMBSF)区域局部活动的检测应用。在注射同位素前一天,在麻醉状态下从成年雄性瑞士韦伯斯特小鼠身上拔除各种组合的触须(例如,所有触须、仅C排、仅B排和D排)。第二天,将5微居里的14C-2-DG注入尾静脉,让小鼠在空笼子里主动探索45分钟。然后处死动物,迅速取出大脑,冷冻,并在低温恒温器中以80微米的厚度平行或垂直于软脑膜切片。将切片固定在盖玻片上干燥,用于曝光X射线胶片,之后用硫堇对切片染色并冲洗X射线胶片。直接从每个切片的显微照片中比较组织切片和匹配的放射自显影图。放射自显影图显示,有相应触须的桶状区域活动较高,而缺少相应触须的桶状区域活动较低。在所有触须完整的动物的切线切片中,PMBSF的活动始终均匀且一致地高于所有触须缺失的情况。去除C排或B排和D排会导致相应PMBSF排的活动条带减少。在垂直于软脑膜的切片中也观察到了相同的活动增加或减少模式,但活动变化虽然在IV层最大,但延伸至皮层的所有层。最后,在一些放射自显影图中,可以识别出密度模式,后来证明这些模式与各个桶状区域的侧面直接相关。结果表明:(1)急性去除外周触须足以使相关的对侧皮层桶状神经元丧失正常活动。因此,在小鼠体感系统中,有可能确定感觉剥夺和新生儿外周损伤在正常皮层发育中的相对重要性。(2)桶状区域是类似于其他感觉系统的功能性皮层柱状组织的一部分。并且,(3)14C-2-DG-X射线技术足够灵敏,能够在放射自显影图中显示各个桶状区域的部分,因此,经过一些改进,可能适用于研究少量神经元群体。