Popratiloff Anastas, Pollack Seth M, Giaume Christian, Peusner Kenna D
Department of Anatomy and Cell Biology, and Neuroscience Program, George Washington University Medical Center, Washington, DC, USA.
J Neurosci Res. 2003 Mar 1;71(5):617-28. doi: 10.1002/jnr.10535.
The chick tangential nucleus is a major vestibular nucleus whose principal cells receive convergent inputs from primary vestibular and nonvestibular fibers and participate in the vestibular reflexes. During development, the principal cells gradually acquire the mature firing pattern in part by losing a specific potassium current around hatching (H). Here we focus on characterizing the expression of connexin 43 (Cx43), a gap junction protein found mainly between astrocytes in the mature brain. The astrocytic syncytium plays an important role in maintaining extracellular potassium ion balance in the brain. Accordingly, it is important to characterize the potential of this syncytium to communicate during the critical developmental age of hatching. Using fluorescence immunocytochemistry, we investigated whether Cx43 staining was concentrated in specific cellular compartments at H1 by applying well-known markers for astrocytes (glial fibrillary acidic protein; GFAP), oligodendrocytes (antimyelin), neurons (microtubule-associated protein 2), and synaptic terminals (synaptotagmin). GFAP-positive astrocytes and GFAP-negative nonneuronal cells around the principal cell bodies were labeled with Cx43, suggesting that Cx43 was expressed exclusively by nonneuronal cells near the neuronal elements. Next, the developmental pattern of expression of Cx43 was studied at embryonic day 16 (E16), H1, and H9. At E16, Cx43 was present weakly as random small clusters in the tangential nucleus, whereas, at H1, overall staining became localized, with increases in size, brightness, and number of immunostained clusters. Finally, at H9, Cx43 staining decreased, but cluster size and location remained unchanged. These results suggest that Cx43 is developmentally regulated with a peak at birth and is associated primarily with astrocytes and nonneuronal cells near the principal cell bodies.
鸡胚切线核是一个主要的前庭核,其主要细胞接收来自初级前庭纤维和非前庭纤维的汇聚输入,并参与前庭反射。在发育过程中,主要细胞部分通过在孵化(H)前后失去一种特定的钾电流,逐渐获得成熟的放电模式。在这里,我们重点研究连接蛋白43(Cx43)的表达特征,Cx43是一种主要存在于成熟大脑中星形胶质细胞之间的缝隙连接蛋白。星形胶质细胞合体在维持大脑细胞外钾离子平衡中起重要作用。因此,在关键的孵化发育阶段,表征这种合体的通讯潜力很重要。我们使用荧光免疫细胞化学方法,通过应用星形胶质细胞(胶质纤维酸性蛋白;GFAP)、少突胶质细胞(抗髓磷脂)、神经元(微管相关蛋白2)和突触终末(突触结合蛋白)的知名标志物,研究了在H1时Cx43染色是否集中在特定的细胞区室。主要细胞体周围的GFAP阳性星形胶质细胞和GFAP阴性非神经元细胞被Cx43标记,这表明Cx43仅由神经元元件附近的非神经元细胞表达。接下来,我们研究了在胚胎第16天(E16)、H1和H9时Cx43的表达发育模式。在E16时,Cx43在切线核中以随机的小簇形式微弱存在,而在H1时,整体染色变得局部化,免疫染色簇的大小、亮度和数量增加。最后,在H9时,Cx43染色减少,但簇的大小和位置保持不变。这些结果表明,Cx43在发育过程中受到调控,在出生时达到峰值,并且主要与主要细胞体附近的星形胶质细胞和非神经元细胞相关。