Accumulation of laminin and microglial cells at sites of injury and regeneration in the central nervous system of the leech

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Abstract
Profuse sprouting of leech neurons occurs in culture when they are plated on a substrate consisting of laminin molecules extracted from extracellular matrix that surrounds the central nervous system (CNS). To assess the role of laminin as a potential growth-promoting molecule in the animal, its distribution was compared in intact and regenerating CNS by light and electronmicroscopy, after it had been labelled with an anti-leech-laminin monoclonal antibody (206) and conjugated second antibodies. In frozen sections and electron micrographs of normal leeches the label was restricted to the connective-tissue capsule surrounding the connectives that link ganglia. Immediately after the connectives had been crushed the normal structure was disrupted but laminin remained in place. Two days after the crush, axons began to sprout vigorously and microglial cells accumulated in the lesion. At the same time, labelled laminin molecules were no longer restricted to the basement membrane but appeared within the connectives in the regions of neurite outgrowth. The distribution of laminin at these new sites within the CNS was punctate at two days, but changed over the following two weeks: the laminin became aggregated as condensed streaks running longitudinally within the connectives beyond the lesion. The close association of regenerating axons with laminin suggests that it may promote axonal growth in the CNS of the animal as in culture.