ABSTRACT
Rat brain vitamin D-dependent calcium-binding protein (D-CaBP) was assessed for vitamin D dependency, calcium binding and ultrastructural localization within neurons. No evidence of vitamin D dependency could be derived from the experiments on vitamin D-deficient rats. A 95% pure extract of the 27-kDa brain D-CaBP was shown to bind 45Ca on nitrocellulose membrane after sodium dodecyl sulphate-electrophoresis, specifically on the 27-kDa CaBP band. Immunogold staining with electron microscopy allowed detection of D-CaBP into Purkinje cells and climbing fibers of the cerebellum. The immunoreactivity was found to be hyaloplasmic and never membrane-bound. It was present in neuronal soma, neurites and postsynaptic as well as presynaptic terminals. These findings rule out D-CaBP as a possible neurotransmitter and bring further support to the hypothesis that the protein functions as a cytosolic calcium buffer. Immunohistochemical detection of D-CaBP is proposed as a means for morphologic detection of neurons with high calcium metabolism.
Subject(s)
Cerebellum/metabolism , S100 Calcium Binding Protein G/metabolism , Animals , Calcium/metabolism , Cell Membrane/metabolism , Cerebellum/ultrastructure , Female , Microscopy, Electron , Purkinje Cells/metabolism , Rats , Rats, Inbred Strains , Vitamin D Deficiency/metabolismABSTRACT
Using immunohistochemistry, vitamin D-dependent calcium binding protein (D-CaBP) has been detected in human retina. In the photoreceptor layer the cones are positive but the rods are negative. In the inner nuclear layer, horizontal cells and some bipolar cells are D-CaBP. In the ganglion cell layer both small and large somata are immunoreactive for D-CaBP. Beaded fibres from the outer plexiform, inner plexiform and fibre layers are also positive.
