Dai Min, Nuttall Alfred, Yang Yue, Shi Xiaorui
Oregon Hearing Research Center (NRC04), Department of Otolaryngology/Head & Neck Surgery, Oregon Health & Science University, 3181 SW Sam Jackson Park Road, Portland, OR 97239, USA.
Hear Res. 2009 Aug;254(1-2):100-7. doi: 10.1016/j.heares.2009.04.018. Epub 2009 May 5.
Pericytes, mural cells located on microvessels, are considered to play an important role in the formation of the vasculature and the regulation of local blood flow in some organs. Little is known about the physiology of cochlear pericytes. In order to investigate the function of cochlear pericytes, we developed a method to visualize cochlear pericytes using diaminofluorescein-2 diacetate (DAF-2DA) and intravital fluorescence microscopy. This method can permit the study of the effect of vasoactive agents on pericytes under the in vivo and normal physiological condition. The specificity of the labeling method was verified by the immunofluorescence labeling of pericyte maker proteins such as desmin, neural proteoglycan (NG2), and thymocyte differentiation antigen 1 (Thy-1). Superfused K(+) and Ca(2+) to the cochlear lateral wall resulted in localized constriction of capillaries at pericyte locations both in vivo and in vitro, while there was no obvious change in cochlear capillary diameters with application of the adrenergic neurotransmitter noradrenaline. The method could be an effective way to visualize cochlear pericytes and microvessels and study lateral wall vascular physiology. Moreover, we demonstrate for the first time that cochlear pericytes have contractility, which may be important for regulation of cochlear blood flow.
周细胞是位于微血管上的壁细胞,被认为在脉管系统的形成以及某些器官局部血流的调节中发挥重要作用。关于耳蜗周细胞的生理学知之甚少。为了研究耳蜗周细胞的功能,我们开发了一种使用二氨基荧光素-2二乙酸酯(DAF-2DA)和活体荧光显微镜观察耳蜗周细胞的方法。该方法能够在体内和正常生理条件下研究血管活性药物对周细胞的影响。通过对周细胞标记蛋白如结蛋白、神经蛋白聚糖(NG2)和胸腺细胞分化抗原1(Thy-1)进行免疫荧光标记,验证了标记方法的特异性。在体内和体外,向耳蜗外侧壁灌注钾离子(K⁺)和钙离子(Ca²⁺)均导致周细胞所在位置的毛细血管局部收缩,而应用肾上腺素能神经递质去甲肾上腺素时,耳蜗毛细血管直径无明显变化。该方法可能是观察耳蜗周细胞和微血管并研究外侧壁血管生理学的有效途径。此外,我们首次证明耳蜗周细胞具有收缩性,这可能对耳蜗血流调节具有重要意义。