Chinestra P, Leinekugel X, Ben Ari Y, Pollard H
INSERM U 29, Laboratoire de Neurobiologie et Physiopathologie du Développement, Hôpital de Port Royal, Paris, France.
Neuroreport. 1994 Jul 21;5(12):1461-5. doi: 10.1097/00001756-199407000-00013.
We have developed a method allowing suitable morphological conservation combined with in situ hybridization, on hippocampal slices used in conventional electrophysiological studies. After a bath application of kainate (KA, 750 nM, 2 min 15 s), electrical stimulation of the mossy fibre zone evoked epileptiform activity for up to 2 h. In situ hybridization performed on these slices showed a marked increased in expression of the transcription factor Zif/268 over the pyramidal and the granule cells and the surrounding neuropils. Bath application of tetraethylammonium (TEA, 25 mM, 10 min) elicited long-term potentiation in CA1 lasting up to 4 h. This was associated with enhanced expression of Zif/268 which returned to control values after 2 h 30 min. These observations suggest that slice preparations are suitable for the study of the role of neuronal activity in the regulation of gene expression.
我们已经开发出一种方法,可在常规电生理研究中使用的海马切片上,实现合适的形态学保存并结合原位杂交。在用红藻氨酸(KA,750 nM,2分15秒)进行浸浴处理后,对苔藓纤维区进行电刺激可诱发长达2小时的癫痫样活动。对这些切片进行原位杂交显示,转录因子Zif/268在锥体细胞、颗粒细胞以及周围神经毡中的表达显著增加。用四乙铵(TEA,25 mM,10分钟)进行浸浴处理可在CA1区诱发长达4小时的长时程增强。这与Zif/268表达增强有关,2小时30分钟后其表达恢复到对照值。这些观察结果表明,切片标本适合用于研究神经元活动在基因表达调控中的作用。