Awake Mouse Imaging: From Two-Photon Microscopy to Blood Oxygen Level-Dependent Functional Magnetic Resonance Imaging
Awake Mouse Imaging: From Two-Photon Microscopy to Blood Oxygen Level-Dependent Functional Magnetic Resonance Imaging
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DOI：
10.1016/j.bpsc.2018.12.002
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发表时间：
2019-06-01
影响因子：
5.9
通讯作者：
Devor, Anna
中科院分区：
文献类型：
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作者：
Desjardins, Michele;Kilic, Kivilcim;Devor, Anna
背景技术背景：清醒行为小鼠的功能性磁共振成像（fMRI）能够很好地桥接大脑活动的详细细胞水平视图，由于显微光学成像和遗传学的最新进展，该视图已成为可用的，以人类非侵入性可观测的宏观尺度。然而，尽管微观（例如，尽管在许多实验室中，在行为小鼠中的双光子成像）研究已经成为现实，但清醒小鼠的fMRI仍然是一个挑战。由于动物之间的行为的变异性，在同一主题内进行所有类型的测量是非常可取的，可以导致更高的科学严谨性。方法：我们证明了血氧水平依赖性功能磁共振成像在清醒的小鼠植入长期颅窗，允许光学显微成像方式和光遗传学刺激。我们从单血管直径变化的双光子成像开始（n = 1）。接下来，我们实施了血氧和血流的固有光学成像，并结合血流的激光散斑成像，获得了血液动力学反应的介观图像（n = 16）。然后获得相应的血氧水平依赖的fMRI数据（n = 5）。所有的测量可以在相同的小鼠在响应相同的感觉和optogenetic stimulations.RESULTS：颅窗没有恶化的fMRI的质量，并允许在每个subjects.CONCLUSIONS成像模式之间的交替：这份报告提供了一个证据，在清醒的小鼠多尺度成像方法的可行性。在未来，该协议可以扩展到包括可翻译为人类的复杂认知行为，例如感觉辨别或注意力。
BACKGROUND: Functional magnetic resonance imaging (fMRI) in awake behaving mice is well positioned to bridge the detailed cellular-level view of brain activity, which has become available owing to recent advances in microscopic optical imaging and genetics, to the macroscopic scale of human noninvasive observables. However, though microscopic (e.g., two-photon imaging) studies in behaving mice have become a reality in many laboratories, awake mouse fMRI remains a challenge. Owing to variability in behavior among animals, performing all types of measurements within the same subject is highly desirable and can lead to higher scientific rigor.METHODS: We demonstrated blood oxygenation level-dependent fMRI in awake mice implanted with long-term cranial windows that allowed optical access for microscopic imaging modalities and optogenetic stimulation. We started with two-photon imaging of single-vessel diameter changes (n = 1). Next, we implemented intrinsic optical imaging of blood oxygenation and flow combined with laser speckle imaging of blood flow obtaining a mesoscopic picture of the hemodynamic response (n = 16). Then we obtained corresponding blood oxygenation level-dependent fMRI data (n = 5). All measurements could be performed in the same mice in response to identical sensory and optogenetic stimuli.RESULTS: The cranial window did not deteriorate the quality of fMRI and allowed alternation between imaging modalities in each subject.CONCLUSIONS: This report provides a proof of feasibility for multiscale imaging approaches in awake mice. In the future, this protocol could be extended to include complex cognitive behaviors translatable to humans, such as sensory discrimination or attention.