Three-step monitoring of glycan and galectin profiles in the anterior segment of the adult chicken eye.

A histochemical three-step approach is applied for processing a panel of sections that covers the different regions of fixed anterior segment of the adult chicken eye. This analysis gains insight into the presence of binding partners for functional pairing by galectin/lectin recognition in situ. Glycophenotyping with 11 fungal and plant lectins (step 1) revealed a complex pattern of reactivity with regional as well as glycan- and cell-type-dependent differences. When characterizing expression of the complete set of the seven adhesion/growth-regulatory chicken galectins immunohistochemically (step 2), the same holds true, clearly demonstrating profiles with individual properties, even for the CG-1A/B paralogue pair. Testing this set of labeled tissue lectins as probes (step 3) detected binding sites in a galectin-type-dependent manner. The results of steps 2 and 3 reflect the divergence of sequences and argue against functional redundancy among the galectins. These data shape the concept of an in situ network of galectins. As consequence, experimental in vitro studies will need to be performed from the level of testing a single protein to work with mixtures that mimic the (patho)physiological situation, a key message of this report.