Rehabilitation Science PhD Program, University of Florida, Gainesville, Florida.
Department of Physical Therapy, University of Florida, Gainesville, Florida.
J Neurophysiol. 2022 Nov 1;128(5):1133-1142. doi: 10.1152/jn.00026.2022. Epub 2022 Aug 17.
Pompe disease is a lysosomal storage disease resulting from absence or deficiency of acid α-glucosidase (GAA). Tongue-related disorders including dysarthria, dysphagia, and obstructive sleep apnea are common in Pompe disease. Our purpose was to determine if designer receptors exclusively activated by designer drugs (DREADDs) could be used to stimulate tongue motor output in a mouse model of Pompe disease. An adeno-associated virus serotype 9 (AAV9) encoding an excitatory DREADD (AAV9-hSyn-hM3D(Gq)-mCherry, 2.44 × 10 vg) was administered to the posterior tongue of 5-7-wk-old Gaa null () mice. Lingual EMG responses to intraperitoneal injection of saline or a DREADD ligand (JHU37160-dihydrochloride, J60) were assessed 12 wk later during spontaneous breathing. Saline injection produced no consistent changes in lingual EMG. Following the DREADD ligand, there were statistically significant ( < 0.05) increases in both tonic and phasic inspiratory EMG activity recorded from the posterior tongue. Brainstem histology confirmed mCherry expression in hypoglossal (XII) motoneurons in all mice, thus verifying retrograde movement of the AAV9 vector. Morphologically, XII motoneurons showed histological characteristics that are typical of Pompe disease, including an enlarged soma and vacuolization. We conclude that lingual delivery of AAV9 can be used to drive functional expression of DREADD in XII motoneurons in a mouse model of Pompe disease. In a mouse model of Pompe disease, lingual injection of adeno-associated virus (AAV) serotype 9 encoding DREADD was histologically verified to produce transgene expression in hypoglossal motoneurons. Subsequent intraperitoneal delivery of a DREADD ligand stimulated tonic and phase tongue motor output.In a mouse model of Pompe disease, lingual injection of adeno-associated virus (AAV) serotype 9 encoding DREADD was histologically verified to produce transgene expression in hypoglossal motoneurons. Subsequent intravenous delivery of a DREADD ligand stimulated tonic and phase tongue motor output.
庞贝病是一种溶酶体贮积病，由酸性α-葡萄糖苷酶（GAA）缺失或缺乏引起。言语障碍包括构音障碍、吞咽困难和阻塞性睡眠呼吸暂停在庞贝病中很常见。我们的目的是确定设计受体是否可以专门被设计药物激活（DREADDs）用于刺激庞贝病小鼠模型中的舌运动输出。腺相关病毒血清型 9（AAV9）编码一种兴奋性 DREADD（AAV9-hSyn-hM3D（Gq）-mCherry，2.44×10 vg）被给予 5-7 周龄 Gaa 缺失（）小鼠的舌后。在自主呼吸期间，在腹腔内注射盐水或 DREADD 配体（JHU37160-二盐酸盐，J60）12 周后评估舌肌电图反应。盐水注射未引起舌肌电图的一致变化。在 DREADD 配体之后，记录的后舌强直和相呼吸肌电图活动均有统计学意义（<0.05）增加。脑干组织学证实，所有小鼠的舌下神经（XII）运动神经元中均表达 mCherry，从而验证了 AAV9 载体的逆行运动。形态学上，XII 运动神经元表现出典型的庞贝病的组织学特征，包括体增大和空泡化。我们得出结论，AAV9 舌内给药可用于驱动 Pompe 病小鼠模型中 XII 运动神经元的 DREADD 功能表达。在 Pompe 病小鼠模型中，腺相关病毒（AAV）血清型 9 编码的 DREADD 的舌内注射在组织学上被证实可在舌下运动神经元中产生转基因表达。随后腹腔内给予 DREADD 配体刺激了强直和相舌运动输出。在 Pompe 病小鼠模型中，腺相关病毒（AAV）血清型 9 编码的 DREADD 的舌内注射在组织学上被证实可在舌下运动神经元中产生转基因表达。随后静脉内给予 DREADD 配体刺激了强直和相舌运动输出。
