Evaluation of three tracers for labeling distal cerebrospinal fluidcontacting neurons

Fang Zhou, Jiayou Wang, Hongxing Zhang, He Liu, Guangping Zhao, Cuihua Zu, Xiaoxing Lu, Licai Zhang

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

It 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/CB-FITC is a useful method for labeling dCSFCNs in rats. The CB-FITC method makes dCSF-CNs labeling much simpler and more convenient.

Original languageEnglish
Pages (from-to)576-580
Number of pages5
JournalNeuroscience Bulletin
Volume29
Issue number5
DOIs
StatePublished - Oct 2013
Externally publishedYes

Keywords

  • cholera toxin subunit B
  • cholera toxin subunit B-conjugated FITC
  • cholera toxin subunit B-conjugated horseradish peroxidase
  • distal cerebrospinal fluid-contacting neurons
  • label

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