Macromolecular markers for glial cells have been sought for a variety of reasons. One of the earliest was the need for a means of assessing the purity of cell and subcellular fractions prepared from nervous tissue. While there is still a requirement for this kind of tool, emphasis has shifted towards seeking information on biochemical differentiation among cells and their functional interactions. A brief general review will be made of glial markers and two of these, 2′,3′-cyclic nucleotide 3′-phosphohydrolase (CNP) and glutamine synthetase (GS), will be considered in detail. Until recently studies of markers have been concentrated on the higher vertebrates and those on lower vertebrates and invertebrates have hardly begun. However, such comparative studies may lead to fresh insight into old problems. For example, CNP has long been regarded as a marker for myelin and oligodendrocytes but it has not been possible to attribute a functional role to it and its relation to myelination has remained obscure. The finding that it is present in the glia of a moth Manduca sexta which lacks myelin provides a stimulus for a fresh approach to the problem. Another example is provided by studies on GS. This enzyme is found in astrocyte feet and preliminary results indicate that it is localized also in the perineurial glia of Aplysia ganglia. These results lead to a reconsideration of the perennial question of the possible role of astrocyte feet in barrier mechanisms. Extension of comparative studies may not only raise new questions but also provide some answers.