Evaluation of three tracers for labeling distal cerebrospinal fluidcontacting neurons  

作　　者：Fang Zhou Jiayou Wang Hongxing Zhang He Liu Guangping Zhao Cuihua Zu Xiaoxing Lu Licai Zhang 

机构地区：[1]Department of Anesthesiology, The First Clinical College, China Medical University [2]Jiangsu Province Key Laboratory of Anesthesiology, Xuzhou Medical College [3]Jiangsu Key Laboratory of Brain Disease Bioinformation, Xuzhou Medical College [4]NursingSchool, Xuzhou Medical College [5]Jiangsu Province Key Laboratory of Anesthesiology,Xuzhou Medical College

出　　处：《Neuroscience Bulletin》2013年第5期576-580,共5页神经科学通报（英文版）

基　　金：supported by grants from the National Natural Science Foundation of China (30871307, 81200862);the Fund of Science and Technology Innovation Team in Jiangsu Province, China (2009), Open Project Funds of the Jiangsu Key Laboratory of Brain Disease Bioinformation (Jsbl1107);the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (12KJD320004)

摘　　要：It has been reported that distal cerebrospinal fluidcontacting neurons（dCSF-CNs）can be detected by immunohistochemical assay using cholera toxin subunit B-conjugated horseradish peroxidase（CBHRP）.In the present study,another two methods with CB alone or CB-conjugated FITC（CB-FITC）were used,and the results from the three methods were compared.Adult Sprague-Dawley rats were randomly divided into three groups with CB-HRP,CB or CB-FITC.Tracers were diluted to 30%in artificial cerebrospinal fluid and injected separately（in a volume of 3μL）into the lateral ventricle.Animals from the CB-HRP and CB groups were perfused 48 h after surgery while animals from the CB-FITC group were perfused 1,3,6,12,24 or 48 h after surgery.The brain was sectioned（40μm）for immunofluorescence and five sections with positive neurons were selected from each rat for neuron counting.Three clusters of positive neurons in a＇Y-like＇distribution were detected ventral to the cerebral aqueduct of rats from the three groups.No significant difference was observed among the quantitative data.In the CB-FITC group,stable staining was detected even at 6 h after injection.Taken together,lateral ventricle injection of CB/CB-FITC is a useful method for labeling dCSFCNs in rats.The CB-FITC method makes dCSF-CNs labeling much simpler and more convenient.It has been reported that distal cerebrospinal fluidcontacting neurons（dCSF-CNs）can be detected by immunohistochemical assay using cholera toxin subunit B-conjugated horseradish peroxidase（CBHRP）.In the present study,another two methods with CB alone or CB-conjugated FITC（CB-FITC）were used,and the results from the three methods were compared.Adult Sprague-Dawley rats were randomly divided into three groups with CB-HRP,CB or CB-FITC.Tracers were diluted to 30%in artificial cerebrospinal fluid and injected separately（in a volume of 3μL）into the lateral ventricle.Animals from the CB-HRP and CB groups were perfused 48 h after surgery while animals from the CB-FITC group were perfused 1,3,6,12,24 or 48 h after surgery.The brain was sectioned（40μm）for immunofluorescence and five sections with positive neurons were selected from each rat for neuron counting.Three clusters of positive neurons in a＇Y-like＇distribution were detected ventral to the cerebral aqueduct of rats from the three groups.No significant difference was observed among the quantitative data.In the CB-FITC group,stable staining was detected even at 6 h after injection.Taken together,lateral ventricle injection of CB/CB-FITC is a useful method for labeling dCSFCNs in rats.The CB-FITC method makes dCSF-CNs labeling much simpler and more convenient.

关 键 词：distal cerebrospinal fluid-contacting neurons cholera toxin subunit B-conjugated horseradish peroxidase cholera toxin subunit B cholera toxin subunit B-conjugated FITC label 

分 类 号：R614[医药卫生—麻醉学]