Expressions of VGLUT1/2 in the inspiratory interneurons and GAD65/67 in the inspiratory Renshaw cells in the neonatal rat upper thoracic spinal cord.
Expressions of VGLUT1/2 in the inspiratory interneurons and GAD65/67 in the inspiratory Renshaw cells in the neonatal rat upper thoracic spinal cord.
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DOI：
10.1016/j.ibror.2018.08.001
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发表时间：
2018-12
期刊：
影响因子：
2.6
通讯作者：
Izumizaki M
中科院分区：
文献类型：
--
作者：
Iizuka M;Ikeda K;Onimaru H;Izumizaki M
第三胸段腹内侧区的吸气中间神经元中约有一半是交感神经能的。这些交感神经能中间神经元可能增强吸气肋间运动活动。吸气Renshaw细胞存在于第三胸段腹内侧区。这些Renshaw细胞中的大多数是GABA能的，并且在新生期引起单个锋电位，随后是腹根刺激。虽然吸气脊髓中间神经元被认为提供了一个主要部分的兴奋性突触电位吸气肋间运动神经元，这还没有得到证实。为了阐明某些吸气脊髓中间神经元是否是神经元能的，我们从新生大鼠的脑干-脊髓分离标本中获得了第三胸段腹内侧区的全细胞记录，并将记录的细胞填充荧光黄用于以后的可视化。然后，我们研究了囊泡谷氨酸转运蛋白1和/或2（VGLUT 1/2）的mRNA的存在进行原位杂交。为了区分中间神经元和运动神经元，我们电刺激记录侧的第三胸前根，结果证实没有诱发逆向锋电位或兴奋性突触后电位。在腹根刺激诱发去极化突触后电位的情况下，我们研究了谷氨酸脱羧酶65和/或67（GAD 65/67）mRNA的存在，使用混合探针，以验证该细胞是否是真正的Renshaw细胞。记录到的中间神经元长径为22 ± 8 μm，短径为13 ± 4 μm。中间神经元的输入电阻为598 ± 274 MΩ。Renshaw电池具有相似的尺寸和输入电阻。11个中间神经元中有6个表达VGLUT 1/2，5个Renshaw细胞中有4个表达GAD 65/67。我们的研究结果表明，在新生大鼠胸脊髓腹内侧区的吸气中间神经元中约有一半是神经元，这些中间神经元可能会增强吸气肋间运动活动。
About half of the inspiratory interneurons in the ventromedial area of the third thoracic segment are glutamatergic. These glutamatergic interneurons may enhance the inspiratory intercostal motor activity. Inspiratory Renshaw cells exist in the ventromedial area of the third thoracic segments. Most of these Renshaw cells are GABAergic, and cause a single spike followed by ventral root stimulation at neonatal stage. Although the inspiratory spinal interneurons are thought to provide a major fraction of the excitatory synaptic potentials to the inspiratory intercostal motoneurons, this has not been confirmed. To clarify whether some inspiratory spinal interneurons are glutamatergic, we obtained whole-cell recordings from the ventromedial area of the third thoracic segments in an isolated brainstem-spinal cord preparation from neonatal rat, and the recorded cells were filled with Lucifer Yellow for later visualization. We then examined the existence of mRNA of vesicular glutamate transporters 1 and/or 2 (VGLUT1/2) by performing in situ hybridization. To discriminate the interneurons from motoneurons, we electrically stimulated the third thoracic ventral root on the recorded side, and the results verified that the antidromic spike or excitatory postsynaptic potential was not evoked. In cases in which the ventral root stimulation evoked depolarizing postsynaptic potentials, we examined the existence of glutamic acid decarboxylase 65 and/or 67 (GAD65/67) mRNA using a mixed probe to verify whether the cell was truly a Renshaw cell. The long diameter of the recorded interneurons was 22 ± 8 μm; the short diameter was 13 ± 4 μm. The interneurons' input resistance was 598 ± 274 MΩ. The Renshaw cells had similar sizes and input resistance. Six of 11 interneurons expressed VGLUT1/2, and four of five Renshaw cells expressed GAD65/67. Our findings suggest that approximately one-half of the inspiratory interneurons in the ventromedial area of the neonatal rat thoracic spinal cord are glutamatergic, and these interneurons might enhance the inspiratory intercostal motor activity.
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