Comparison of high-pressure freezing/freeze substitution and chemical fixation of catfish barbel taste buds.

Abstract

The barbel taste buds of catfish are widely used as a model system for investigating the structure and function of vertebrate taste buds. We have examined the ultrastructure of the taste buds of the channel catfish, Ictalurus punctatus, as part of a comparative study of the morphology of taste buds in various mammalian and non-mammalian vertebrates. Since conventional chemical fixation methods have limited usefulness for certain kinds of ultrastructural studies (i.e., localization of diffusible substances or labeling techniques requiring retention of biological activity), we have developed methods for fixing catfish barbel taste buds by high-pressure freezing followed by freeze-substitution (HPF/FS) and have compared the ultrastructure of taste buds fixed by this technique and by chemical fixation procedures. The morphological details of the cells within taste buds are significantly affected by the method of fixation employed. In general, membrane contours are smoother and intracellular organelles more regular in shape in HPF/FS samples as compared with the chemically fixed specimens. Absolute and relative electron-densities of various tissue components are also affected by the fixation method employed. Certain ultrastructural features are more clearly visualized by one or the other of the fixation procedures. Fixation of barbel taste buds by HPF/FS not only provides an alternative view of the ultrastructure of taste bud cells but also offers a method of tissue preparation that may prove to be preferable to chemical methods for ultrastructural investigations involving procedures such as immunohistochemical labeling.