High-pressure freezing followed by cryosubstitution as a tool for preserving high-quality ultrastructure and immunoreactivity in the Xenopus laevis pituitary gland.

Details

Serval ID
serval:BIB_A0D15D3F1CBF
Type
Article: article from journal or magazin.
Collection
Publications
Title
High-pressure freezing followed by cryosubstitution as a tool for preserving high-quality ultrastructure and immunoreactivity in the Xenopus laevis pituitary gland.
Journal
Brain Research Protocols
Author(s)
Wang L., Humbel B.M., Roubos E.W.
ISSN
1385-299X (Print)
ISSN-L
1385-299X
Publication state
Published
Issued date
2005
Volume
15
Number
3
Pages
155-163
Language
english
Abstract
Subcellular localisation of proteins and peptides yields fundamental information about cell functioning. Immunoelectron microscopy is a powerful tool to achieve this goal, but combining good tissue preservation with strong immunoreactivity is a great challenge in electron microscopy. We have applied a novel approach, using high-pressure freezing (HPF) followed by cryosubstitution, to prepare the pituitary gland of the amphibian Xenopus laevis for immunogold-electron microscopy. In this way, we investigated the subcellular distribution of brain-derived neurotrophic factor and the amphibian neurohormone mesotocin in the pituitary neural lobe, and the peptide hormone alpha-melanophore-stimulating hormone and its protein precursor proopiomelanocortin in melanotrope cells of the pituitary intermediate lobe. In contrast to conventional chemical fixation (followed by cryosubstitution), HPF not only revealed strong immunoreactivity of the secretory products, but also provided excellent ultrastructural preservation of cell organelles, including secretory granule subtypes. We conclude that HPF followed by cryosubstitution provides a preparation technique of choice when both optimal tissue ultrastructure and strong immunoreactivity are required.
Keywords
Animals, Freezing, Organ Preservation/methods, Organ Preservation Solutions, Pituitary Gland/immunology, Pituitary Gland/ultrastructure, Xenopus laevis
Pubmed
Web of science
Create date
28/02/2012 18:45
Last modification date
20/08/2019 15:06
Usage data