Damak Sami, Mosinger Bedrich, Margolskee Robert F
Department of Physiology and Biophysics, Mount Sinai School of Medicine, 1425 Madison Avenue, Box 1677, New York, New York, 10029, USA.
BMC Neurosci. 2008 Oct 2;9:96. doi: 10.1186/1471-2202-9-96.
Anatomical tracing of neural circuits originating from specific subsets of taste receptor cells may shed light on interactions between taste cells within the taste bud and taste cell-to nerve interactions. It is unclear for example, if activation of type II cells leads to direct activation of the gustatory nerves, or whether the information is relayed through type III cells. To determine how WGA produced in T1r3-expressing taste cells is transported into gustatory neurons, transgenic mice expressing WGA-IRES-GFP driven by the T1r3 promoter were generated.
Immunohistochemistry showed co-expression of WGA, GFP and endogenous T1r3 in the taste bud cells of transgenic mice: the only taste cells immunoreactive for WGA were the T1r3-expressing cells. The WGA antibody also stained intragemmal nerves. WGA, but not GFP immunoreactivity was found in the geniculate and petrosal ganglia of transgenic mice, indicating that WGA was transported across synapses. WGA immunoreactivity was also found in the trigeminal ganglion, suggesting that T1r3-expressing cells make synapses with trigeminal neurons. In the medulla, WGA was detected in the nucleus of the solitary tract but also in the nucleus ambiguus, the vestibular nucleus, the trigeminal nucleus and in the gigantocellular reticular nucleus. WGA was not detected in the parabrachial nucleus, or the gustatory cortex.
These results show the usefulness of genetically encoded WGA as a tracer for the first and second order neurons that innervate a subset of taste cells, but not for higher order neurons, and demonstrate that the main route of output from type II taste cells is the gustatory neuron, not the type III cells.
对源自特定味觉受体细胞亚群的神经回路进行解剖学追踪,可能有助于揭示味蕾内味觉细胞之间的相互作用以及味觉细胞与神经之间的相互作用。例如,目前尚不清楚II型细胞的激活是否会直接激活味觉神经,或者信息是否通过III型细胞进行传递。为了确定表达T1r3的味觉细胞中产生的WGA是如何运输到味觉神经元中的,我们构建了由T1r3启动子驱动表达WGA-IRES-GFP的转基因小鼠。
免疫组织化学显示转基因小鼠的味蕾细胞中WGA、GFP和内源性T1r3共表达:唯一对WGA有免疫反应的味觉细胞是表达T1r3的细胞。WGA抗体也使味蕾内神经染色。在转基因小鼠的膝状神经节和岩神经节中发现了WGA免疫反应性,但未发现GFP免疫反应性,这表明WGA是通过突触运输的。在三叉神经节中也发现了WGA免疫反应性,这表明表达T1r3的细胞与三叉神经元形成突触。在延髓中,在孤束核中检测到了WGA,但在疑核、前庭核、三叉神经核和巨细胞网状核中也检测到了WGA。在臂旁核或味觉皮层中未检测到WGA。
这些结果表明,基因编码的WGA作为一种示踪剂,对于支配一部分味觉细胞的一级和二级神经元是有用的,但对于高级神经元则不然,并且表明II型味觉细胞的主要输出途径是味觉神经元,而不是III型细胞。