Quantitative changes and ultrastructural alterations of the cornea in response to ultraviolet light. II. Effects of amphibia; elucidation of desmosomal structure and basement membrane synthesis.

The effects of far ultraviolet light irradiation upon an amphibian cornea were studied to compare the effects observed both quantitatively and ultrastructurally with data obtained after UV irradiation of mammalian corneas. The ultimate goal of this series of investigations is the elucidation of the alterations and the regeneration mechanisms, which might reflect existing morphological diversities among the species, observed in vertebrate corneas following exposure to UV light. It was found that while the epithelial cells undergo oedema after low dose exposures and are gradually damaged after high doses of UV light, 2-4 days leter a new epithelium has been formed. Intercellular permeability is increased by low dose exposure as was detected by the penetration of Ruthenium Red into the intercellular clefts. Under these conditions desmosomal structure revealed a 21-laminar configuration. The basement membrane of the amphibian, unlike that of the mammal, does not dissolve away upon exposure but shows localized disruptions which are thought to accommodate the passage of leucocytes from stroma to epithelium. That a new basement membrane is subsequently formed is evident by the existence of extracellular and intracellular secretion granules. In comparison to irradiated rabbit corneas, this stroma remains remarkably at the same thickness following a high dose exposure although a noticeable disorganization of collagen arrangement is apparent. Finally, as in the case of the rabbit corneas, a secondary degeneration of endothelium was observed 4 days after a moderate dose exposure.     

UV spectra of indoles in strong sulphuric acid

Measurement of the UV spectra in 12·7 M H₂SO₄ at 30° and 70° provides a simple, quantitative and specific method for the identification of variously substituted, naturally occurring plant indoles in μg quantities.     

Isolation from earthworms of a proteinaceous chemoattractant to garter snakes

We have isolated a potent proteinaceous chemoattractant from aqueous washes of earthworm (Lumbricus terrestris) for garter snakes (Thamnophis sirtalis) by means of a covalent chromatography. It contained free sulfhydryl groups and showed an apparent mass of 20 kDa. The chemoattractive activity of this protein could be destroyed by heating as well as by proteolysis. Its activity could also be reversibly blocked by mixed disulfide formation with dithiodipyridine, suggesting that the free sulfhydryl(s) was essential for its function as a chemoattractant. This bioactive material had a tendency to form intermolecular crosslinked aggregates during isolation, if reducing agents were not included. Some of the high-molecular-weight aggregates cochromatographed with earthworm cuticle collagen on Ultragel AcA 34 or 44 columns. In contrast to an earlier report by D. M. Kirschenbaum, N. Schulman, and M. Halpern [1986) Proc. Natl. Acad. Sci. USA 83, 1213-1216) the purified earthworm collagen showed no chemoattractive activity to garter snakes.