Pronase treatment increases the staining intensity of GABA-immunoreactive structures in the paravertebral sympathetic ganglia.

Histochemistry
E DobóJ R Wolff

Abstract

A novel tissue preparation technique for improving gamma-aminobutyric acid (GABA) immunocytochemistry has been developed. The influence of the glutaraldehyde concentration in the fixative and the effect of pronase treatment on the GABA immunostaining were tested. This method includes fixation with a high concentration of glutaraldehyde, gelatin embedding and treatment of the sections with pronase. In sympathetic (paravertebral) ganglia and their connectives, the most intense and specific immunoreaction was obtained with the following procedure: immersion fixation in 5% glutaraldehyde, infiltration and embedding in 15% gelatin, secondary fixation of the samples with 4% formaldehyde, floating frozen sections and digestion with 0.1% pronase for 15-20 min. With this technique, the GABA-containing structures (cells and nerve fibers with varicosities forming basket-like networks around some principal neurons) were selectively labeled. The data presented suggest that (1) a high concentration (5%) of glutaraldehyde in the primary fixative is necessary to preserve a large proportion of the GABA content; (2) this glutaraldehyde fixation partly masks the GABA immunoreactivity; and (3) this masking may be overcome by a proteolytic treatmen...Continue Reading

References

Mar 1, 1985·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·P SomogyiA Erdei
Jan 1, 1989·Anatomy and Embryology·S DavangerJ Storm-Mathisen
Aug 13, 1986·Brain Research·R J WentholdR A Altschuler
Nov 1, 1986·Journal of the Autonomic Nervous System·S L Kenny, M A Ariano
Jan 2, 1987·Neuroscience Letters·S DavangerJ Storm-Mathisen

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Citations

Aug 1, 1995·Neuroscience·M Rickmann, J R Wolff
May 1, 1991·Zentralblatt für Veterinärmedizin. Reihe B. Journal of veterinary medicine. Series B·T Segall, A A Lindberg

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