In Situ Localization of F-Actin in the Normal and Injured Guinea-pig Tympanic Membrane

Abstract

Although cell migration is an important function of the epithelial cells of the tympanic membrane (TM), little is known about the distribution of the F-actin cytoskeleton, a contractile protein important in cell motility. The purpose of this experiment was to study the in situ localization of F-actin in the epithelial cells of the TM. F-actin, localized using Rhodamine-phalloidin, was present as a thin cortical band at the margin of both the mucosal cells on the inner side of the drum, and the suprabasal cells of the epidermis. The basal cells showed diffuse circumferential F-actin staining sometimes appearing as short microfilaments. Following a full thickness injury, changes in the distribution of F-actin could be observed with in situ localization. While the diffuse F-actin staining of the basal cells was reduced, both long F-actin microfilament bundles extending parallel to the long axis of the cell and focal aggregates of F-actin were prominent. The suprabasal cells became elongated, and while the F-actin remained localized to the cell margin, faint central F-actin microfilaments were observed. The staining of the mucosal cells remained unchanged. This study showed that the guinea pig TM is a useful model to study the distribution of epithelial F-actin in situ under normal and repair conditions, and that the basal cell layer may be important in regulating migration in the epidermis.

Key Words:

• epithelium
• cytoskeleton
• microfilaments
• rhodamine-phalloidin
• cell migration