Li Yang, Su Shanchu, Yu Jiaqi, Peng Minjing, Wan Shengjun, Ke Changbin
Hubei Key Laboratory of Embryonic Stem Cell Research, Department of Anesthesiology, Institute of Anesthesiology & Pain (IAP), Taihe Hospital, Hubei University of Medicine, Shiyan, China.
Front Aging Neurosci. 2021 Mar 10;13:640265. doi: 10.3389/fnagi.2021.640265. eCollection 2021.
A patch-clamp recording in slices generated from the brain or the spinal cord has facilitated the exploration of neuronal circuits and the molecular mechanisms underlying neurological disorders. However, the rodents that are used to generate the spinal cord slices in previous studies involving a patch-clamp recording have been limited to those in the juvenile or adolescent stage. Here, we applied an N-methyl-D-glucamine HCl (NMDG-HCl) solution that enabled the patch-clamp recordings to be performed on the superficial dorsal horn neurons in the slices derived from middle-aged rats. The success rate of stable recordings from substantia gelatinosa (SG) neurons was 34.6% (90/260). When stimulated with long current pulses, 43.3% (39/90) of the neurons presented a tonic-firing pattern, which was considered to represent γ-aminobutyric acid-ergic (GABAergic) signals. Presumptive glutamatergic neurons presented 38.9% (35/90) delayed and 8.3% (7/90) single-spike patterns. The intrinsic membrane properties of both the neuron types were similar but delayed (glutamatergic) neurons appeared to be more excitable as indicated by the decreased latency and rheobase values of the action potential compared with those of tonic (GABAergic) neurons. Furthermore, the glutamatergic neurons were integrated, which receive more excitatory synaptic transmission. We demonstrated that the NMDG-HCl cutting solution could be used to prepare the spinal cord slices of middle-aged rodents for the patch-clamp recording. In combination with other techniques, this preparation method might permit the further study of the functions of the spinal cord in the pathological processes that occur in aging-associated diseases.
对取自大脑或脊髓的脑片进行膜片钳记录,有助于探索神经回路以及神经系统疾病背后的分子机制。然而,在以往涉及膜片钳记录的研究中,用于制备脊髓切片的啮齿动物仅限于幼年或青少年阶段的动物。在此,我们应用了一种盐酸N-甲基-D-葡萄糖胺(NMDG-HCl)溶液,使得能够在源自中年大鼠的脑片的浅表背角神经元上进行膜片钳记录。从胶状质(SG)神经元获得稳定记录的成功率为34.6%(90/260)。当用长电流脉冲刺激时,43.3%(39/90)的神经元呈现紧张性放电模式,这被认为代表γ-氨基丁酸能(GABA能)信号。推测的谷氨酸能神经元呈现38.9%(35/90)的延迟放电模式和8.3%(7/90)的单峰模式。两种神经元类型的内在膜特性相似,但与紧张性(GABA能)神经元相比,延迟(谷氨酸能)神经元的动作电位潜伏期和阈电流值降低,似乎更易兴奋。此外,谷氨酸能神经元是整合的,接受更多的兴奋性突触传递。我们证明了NMDG-HCl切割溶液可用于制备中年啮齿动物的脊髓切片以进行膜片钳记录。结合其他技术,这种制备方法可能有助于进一步研究脊髓在衰老相关疾病发生的病理过程中的功能。