Immunohistochemical detection of wheat germ agglutinin-horseradish peroxidase (WGA-HRP)

Suzhen Gong, Mark Ledoux

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Traditional histochemical detection of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) can impose substantial technical limitations on studies requiring co-localization of neurotransmitters, receptors and other neural antigens. The goal of our experiments was to establish the ideal conditions and reagents for immunohistochemical detection of WGA-HRP. WGA-HRP was injected into the tongues and vibrissae pads of adult rats to characterize labeling of somas and synapses, respectively. Rats were perfused with either 4% paraformaldehyde (for light microscopy, LM) or 4% paraformaldehyde/0.15% glutaraldehyde (for electron microscopy, EM) after survival times of 2, 3, 4, 5 or 6 days. For LM, brainstem tissue was cut on a cryostat at 20 μm and collected onto glass slides. For EM, tissue was sectioned with a vibratome at 50 μm and processed free floating. For LM, WGA-HRP was detected with goat anti-HRP, goat anti-WGA, biotinylated goat anti-HRP or biotinylated goat anti-WGA antibodies. For EM, WGA-HRP was detected with biotinylated goat anti-WGA and anti-HRP antibodies. Survival intervals of 3 days were ideal for staining of hypoglossal neurons, whereas an interval of 4 days produced the strongest staining of synapses within the spinal trigeminal nucleus. For LM, the biotinylated antibodies resulted in better signal-to-noise ratios than the unconjugated antibodies. At both the LM and EM levels, the biotinylated antibody to WGA produced better quality staining than the biotinylated antibody to HRP.

Original languageEnglish (US)
Pages (from-to)25-34
Number of pages10
JournalJournal of Neuroscience Methods
Volume126
Issue number1
DOIs
StatePublished - Jun 15 2003

Fingerprint

Wheat Germ Agglutinins
Horseradish Peroxidase
Goats
Microscopy
Electron Microscopy
Light
Antibodies
Staining and Labeling
Synapses
Anti-Idiotypic Antibodies
Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate
Spinal Trigeminal Nucleus
Vibrissae
Neurotransmitter Receptor
Glutaral
Signal-To-Noise Ratio
Carisoprodol
Tongue
Brain Stem
Glass

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

Cite this

Immunohistochemical detection of wheat germ agglutinin-horseradish peroxidase (WGA-HRP). / Gong, Suzhen; Ledoux, Mark.

In: Journal of Neuroscience Methods, Vol. 126, No. 1, 15.06.2003, p. 25-34.

Research output: Contribution to journalArticle

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