Visualization and contractile activity of cochlear pericytes in the capillaries of the spiral ligament.
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.
Dai
Min
M
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.
Nuttall
Alfred
A
Yang
Yue
Y
Shi
Xiaorui
X
D000818:Animals; D002196:Capillaries / Q000473:pathology / Q000502:physiology*; D003051:Cochlea / Q000473:pathology / Q000502:physiology*; D005260:Female; D019793:Fluorescein / Q000494:pharmacology; D006168:Guinea Pigs; D007202:Indicators and Reagents / Q000494:pharmacology; D008297:Male; D008856:Microscopy, Fluorescence / Q000379:methods; D008953:Models, Anatomic; D020286:Pericytes / Q000473:pathology / Q000502:physiology*; D011509:Proteoglycans / Q000378:metabolism; D055493:Spiral Ligament of Cochlea / Q000473:pathology / Q000502:physiology*; D018800:Thy-1 Antigens / Q000096:biosynthesis; D013997:Time Factors
C414689:4,5-diaminofluorescein diacetate; D007202:Indicators and Reagents; D011509:Proteoglycans; D018800:Thy-1 Antigens; D019793:Fluorescein