Two-photon imaging of cerebral hemodynamics and neural activity in awake and anesthetized marmosets.
Two-photon imaging of cerebral hemodynamics and neural activity in awake and anesthetized marmosets.
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DOI：
10.1016/j.jneumeth.2016.07.003
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发表时间：
2016-09-15
影响因子：
3
通讯作者：
Silva AC
中科院分区：
文献类型：
--
作者：
Santisakultarm TP;Kersbergen CJ;Bandy DK;Ide DC;Choi SH;Silva AC
绒猴是一种强大的，新兴的人类行为和神经系统疾病的模型。然而，纵向成像模式，可视化细胞结构和功能的皮质内是不可用的，在这个动物模型。因此，我们实施了一种方法来量化血管拓扑结构，血流动力学和神经活动在清醒的绒猴使用双光子显微镜（2PM）。使绒猴适应定制的立体定位系统。将AAV 1-GCaMP 5G注射到躯体感觉皮层中以光学指示神经活动，并植入颅室。纵向2PM显示皮质表面下500 µm的血管和神经元。血管反应和感觉刺激过程中的神经活动分别保存超过5个月和3个月，光学质量恶化之前。对血管重塑（包括迂曲度和分支增加）进行定量。然而，从小动脉到小静脉的毛细血管连通性保持不变。此外，手术前后的行为评估表明对认知和运动功能没有影响。免疫组织化学证实星形胶质细胞活化最小，无局灶性损伤。6个月后，可见的总皮质深度下降。在麻醉状态下，最显著的异氟醚诱导的血管舒张发生在毛细血管和较小的小动脉。这些结果证明了在清醒的绒猴中重复观察皮层生理学数月的能力。这项工作提供了一种新的和有见地的技术，调查清醒的绒猴神经系统疾病的关键机制，而不会引入麻醉混淆。
Marmosets are a powerful, emerging model for human behavior and neurological disorders. However, longitudinal imaging modalities that visualize both cellular structure and function within the cortex are not available in this animal model. Hence, we implemented an approach to quantify vascular topology, hemodynamics, and neural activity in awake marmosets using two-photon microscopy (2PM). Marmosets were acclimated to a custom stereotaxic system. AAV1-GCaMP5G was injected into somatosensory cortex to optically indicate neural activity, and a cranial chamber was implanted. Longitudinal 2PM revealed vasculature and neurons 500 µm below the cortical surface. Vascular response and neural activity during sensory stimulation were preserved over 5 and 3 months, respectively, before optical quality deteriorated. Vascular remodeling including increased tortuosity and branching was quantified. However, capillary connectivity from arterioles to venules remained unchanged. Further, behavioral assessment before and after surgery demonstrated no impact on cognitive and motor function. Immunohistochemistry confirmed minimal astrocyte activation with no focal damage. Over 6 months, total cortical depth visualized decreased. When under anesthesia, the most prominent isoflurane-induced vasodilation occurred in capillaries and smaller arterioles. These results demonstrate the capability to repeatedly observe cortical physiology in awake marmosets over months. This work provides a novel and insightful technique to investigate critical mechanisms in neurological disorders in awake marmosets without introducing confounds from anesthesia.
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影响因子：
3
作者：
Chen, Yu;Aguirre, Aaron D.;Ruvinskaya, Lana;Devor, Anna;Boas, David A.;Fujimoto, James G.
通讯作者：
Fujimoto, James G.
影响因子：
5.7
作者：
Chen BR;Bouchard MB;McCaslin AF;Burgess SA;Hillman EM
通讯作者：
Hillman EM
影响因子：
5.7
作者：
Liu, Junjie V.;Hirano, Yoshiyuki;Nascimento, George C.;Stefanovic, Bojana;Leopold, David A.;Silva, Afonso C.
通讯作者：
Silva, Afonso C.
DOI：
10.1073/pnas.95.26.15741
发表时间：
1998-12-22
影响因子：
11.1
作者：
Kleinfeld, D;Mitra, PP;Denk, W
通讯作者：
Denk, W
影响因子：
5.7
作者：
FRISTON, KJ;HOLMES, AP;TURNER, R
通讯作者：
TURNER, R