Developmental alterations in membrane organization of rat subpial astrocytes

Subpial astrocytic processes were examined in developing rats, mainly with complementary replicas, to see how orthogonal arrays of particles (OAs) are formed and become numerous in membranes covered by basal lamina. Only a few (4.2%) endfeet in the membranes contacting the basal lamina (subpial membranes) had acquired OAs by the 19-day foetal stage. The number of endfeet provided with OAs increased drastically in the prenatal period, continued to increase at birth (P0), and somewhat more slowly in the early postnatal period (P0-P3), reaching 100% at P10. There were neuronal processes as well abutting on the basal lamina at the pial surface but they were easy to distinguish from astrocytic endfeet because of their larger intramembrane particles (IMPs), which are sparsely distributed and in patch-like aggregations. The distribution density of OAs in differentiated astrocytic endfeet also increased very gradually with age until P0, a little faster in the early postnatal period, and drastically from P10 to adult. Ordinary globular IMPs increased in number with age and continued to increase in the lateral membrane where OAs were still very few, though less rapidly in the subpial membrane as OAs became numerous. With maturation, larger IMPs became conspicuous in the lateral membrane but not in the subpial, suggesting that larger IMPs were predominantly used to form OAs. We have proposed the idea that relatively large IMPs line up to form single linear arrays (SLs), appearing as grooves on the E face, and that occasionally some SLs line up in multiple rows [multiple linear arrays (MLs)] and that SLs or MLs fuse with one another to become rod-like strands, then divide into squares to become OAs. SLs and MLs appeared ontogenetically earlier than OAs, and continued to appear in membranes provided with OAs. In areas where membranes were bent, transition of these three structures was observable and the proportion of OAs increased with age. Further, in such areas, alignment of OAs was different according to membrane curvature: concentric in and around protrusions, perpendicular to the edge of invaginations. This unique association of OA alignment with membrane curvature suggests that OAs contribute to some membrane stability in the area covered by the basal lamina and provide the membrane with special resistance to bending.