4B2: A monoclonal antibody to ganglion and bipolar cells in the retina of the cat, generated by intrasplenic immunization

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Abstract
Using tissue from the inner layers of the retina as the immunogen, we have prepared a monoclonal antibody which is selective for two classes of neuron in the cat retina. The antibody, termed 4B2, was generated by intrasplenic injection, demonstrating that neuron-specific antibodies can be elicited by this technique, which requires only micrograms of immunogen. The 4B2 binds to the somas, dendrites and axons of ganglion cells. Double labelling with 4B2 and with a retrograde tracer injected into the retino-recipient nuclei of the brain demonstrates that, of the cell classes in the ganglion cell layer, 4B2 labels only ganglion cells, and that, amongst ganglion cells, 4B2 is strongly selective for large (α-) and medium-sized (β- and perhaps γ-) cells. Double labelling with 4B2 and with markers for amacrine cells confirms tht 4B2 does not label amacrine cells. Double labelling with 4B2 and with anti-GFAP confirms that 4B2 does not label the macroglia of the retina. The 4B2 also labels bipolar cells, showing their somas, dendrites and axons. The axons of the labelled cells terminate in large boutons, in the innermost part of the inner plexiform layer and in the ganglion cell layer. This level of termination suggests that 4B2 labels rod bipolars, and perhaps a subgroup of cone bipolars. Double labelling with 4B2 and with markers for amacrine, horizontal and Müller cells indicates that other cell classes with somas in the inner nuclear layer remain unlabelled. In addition, we have examined the ontogeny of 4B2 labelling in the cat retina. The developmental sequence of labelling with 4B2 follows the sequence in which the cells are born, first ganglion and then bipolar cells.