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Evaluation of three tracers for labeling distal cerebrospinal fluidcontacting neurons
Alternative TitleEvaluation of three tracers for labeling distal cerebrospinal fluidcontacting neurons
Zhou Fang1; Wang Jiayou2; Zhang Hongxing1; Liu He1; Zhao Guangping2; Zu Cuihua2; Lu Xiaoxing2; Zhang Licai1
2013
Source PublicationNEUROSCIENCE BULLETIN
ISSN1673-7067
Volume29Issue:5Pages:576-580
AbstractIt has been reported that distal cerebrospinal fluid-contacting 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 3 mu L) 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 (40 mu m) 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.
Other AbstractIt has been reported that distal cerebrospinal fluid-contacting 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 3 μL) 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 (40 μm) 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
KeywordFLUID-CONTACTING NEURONS RAT-BRAIN SYSTEM CELLS HRP distal cerebrospinal fluid-contacting neurons cholera toxin subunit B-conjugated horseradish peroxidase cholera toxin subunit B cholera toxin subunit B-conjugated FITC label
Indexed ByCSCD
Language英语
Funding Project[National Natural Science Foundation of China] ; [Fund of Science and Technology Innovation Team in Jiangsu Province, China] ; [Jiangsu Key Laboratory of Brain Disease Bioinformation] ; [Natural Science Foundation of the Jiangsu Higher Education Institutions of China]
CSCD IDCSCD:4975289
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Document Type期刊论文
Identifierhttp://ir.imr.ac.cn/handle/321006/150514
Collection中国科学院金属研究所
Affiliation1.中国科学院金属研究所
2.Xuzhou Med Coll, Jiangsu Prov Key Lab Anesthesiol, Xuzhou 221002, Peoples R China
3.Xuzhou Med Coll, Jiangsu Key Lab Brain Dis Bioinformat, Xuzhou 221002, Peoples R China
4.Xuzhou Med Coll, Sch Nursing, Xuzhou 221002, Peoples R China
Recommended Citation
GB/T 7714
Zhou Fang,Wang Jiayou,Zhang Hongxing,et al. Evaluation of three tracers for labeling distal cerebrospinal fluidcontacting neurons[J]. NEUROSCIENCE BULLETIN,2013,29(5):576-580.
APA Zhou Fang.,Wang Jiayou.,Zhang Hongxing.,Liu He.,Zhao Guangping.,...&Zhang Licai.(2013).Evaluation of three tracers for labeling distal cerebrospinal fluidcontacting neurons.NEUROSCIENCE BULLETIN,29(5),576-580.
MLA Zhou Fang,et al."Evaluation of three tracers for labeling distal cerebrospinal fluidcontacting neurons".NEUROSCIENCE BULLETIN 29.5(2013):576-580.
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