Alignment of EphA4 and ephrin‐B2 expression patterns with developing modularity in the lateral cortex of the inferior colliculus

In the multimodal lateral cortex of the inferior colliculus (LCIC), there are two neurochemically and connectionally distinct compartments, termed modular and extramodular zones. Modular fields span LCIC layer 2 and are recipients of somatosensory afferents, while encompassing extramodular domains receive auditory inputs. Recently, in developing mice, we identified several markers (among them glutamic acid decarboxylase, GAD) that consistently label the same modular set, and a reliable extramodular marker, calretinin, (CR). Previous reports from our lab show similar modular–extramodular patterns for certain Eph–ephrin guidance members, although their precise alignment with the developing LCIC neurochemical framework has yet to be addressed. Here we confirm in the nascent LCIC complementary GAD/CR‐positive compartments, and characterize the registry of EphA4 and ephrin‐B2 expression patterns with respect to its emerging modular–extramodular organization. Immunocytochemical approaches in GAD67‐GFP knock‐in mice reveal patchy EphA4 and ephrin‐B2 domains that precisely align with GAD‐positive LCIC modules, and are complementary to CR‐defined extramodular zones. Such patterning was detectable neonatally, yielding discrete compartments prior to hearing onset. A dense plexus of EphA4‐positive fibers filled modules, surrounding labeled ephrin‐B2 and GAD cell populations. The majority of observed GABAergic neurons within modular boundaries were also positive for ephrin‐B2. These results suggest an early compartmentalization of the LCIC that is likely instructed in part through Eph–ephrin guidance mechanisms. The overlap of developing LCIC neurochemical and guidance patterns is discussed in the context of its seemingly segregated multimodal input–output streams. Multiple‐labeling immunocytochemical experiments in the lateral cortex of the inferior colliculus in GAD67‐GFP mice reveal patchy EphA4 and ephrin‐B2 expression that overlaps with its modular–extramodular neurochemical compartments. This registry is apparent at birth and throughout the early postnatal period, providing a framework for its multimodal input–output arrays.