Postnatal development and localization of an N-acetylgalactosamine containing glycoconjugate associated with...

We have analyzed the appearance of N-acetylgalactosamine containing glycoconjugates by staining sections of fixed cat visual cortex with the lectins from Vicia villosa (VVA) and Glycine max (SBA) conjugated to fluorescent labels, horseradish peroxidase, or biotin. The appearance of lectin staining during postnatal development followed an inside-out gradient starting in the deep cortical layers at the fourth postnatal week, successively including more of the superficial layers during the first three postnatal months until labelled cells were present throughout layers II-VI at the seventh postnatal month. Staining was associated with the soma and proximal dendrites of bipolar and multipolar nonpyramidal neurons. Outside layer IV large neurons with basket cell and neurogliaform morphology predominated, whereas small multipolar cells were stained in layer IV. High power observation revealed a lattice-like staining on neuronal surfaces. In the cortical white matter staining was found at the nodes of Ranvier. The ultrastructural localization of lectin binding sites was assessed by a pre-embedding histochemical procedure with biotinylated lectin visualized with HRP-conjugated avidin and diaminobenzidine as the chromogen. The reaction product was found exclusively in close association with synaptic terminals on somata and proximal dendrites of nonpyramidal neurons. There was no preference with respect to the morphology of the synaptic structure, i.e., symmetric synapses with ovoid or asymmetric synapses with round vesicles. In the vicinity of synaptic contacts the staining was associated with membranes of astrocytic processes ensheathing the synapses. In the cortical white matter astrocytic processes running along myelinated axons were strongly labelled at the nodes of Ranvier. Comparison of Western blots from 4-week-old and adult cat cortex membranes revealed one lectin-positive protein band with an apparent molecular weight of about 24 kD in conjunction with the histochemical expression of perisynaptic staining in the adult tissue. It is concluded that the employed lectins selectively visualize distal astroglial processes ensheathing synapses on a subpopulation of cortical interneurons and nodes of Ranvier. On the basis of the developmental appearance and localization we consider and discuss the possibility that the N-acetylgalactosamine containing glycoconjugate is involved in the stabilization of synaptic contacts on GABA-ergic interneurons.