Distinct deep short-axon cell subtypes of the main olfactory bulb provide novel intrabulbar and extrabulbar GABAergic connections

A universal feature of neuronal microcircuits is the presence of GABAergic interneurons that control the activity of glutamatergic principal cells and each other. In the rat main olfactory bulb (MOB), GABAergic granule and periglomerular cells innervate mitral and tufted cells, but the source of their own inhibition remains elusive. Here, we used a combined electrophysiological and morphological approach to investigate a rather mysterious cell population of the MOB. Deep short-axon cells (dSACs) of the inframitral layers are GABAergic and have extensive and characteristic axonal ramifications in various layers of the bulb, based on which unsupervised cluster analysis revealed three distinct subtypes. Each dSAC subtype exhibits different electrical properties but receives similar GABAergic and glutamatergic inputs. The local axon terminals of all dSAC subtypes selectively innervate GABAergic granule and periglomerular cells and evoke GABA(A) receptor-mediated IPSCs. One subpopulation of dSACs (GL-dSACs) creates a novel intrabulbar projection from deep to superficial layers. Another subpopulation (GCL-dSACs) is labeled by retrogradely transported fluorescent microspheres injected into higher olfactory areas, constituting a novel projection-cell population of the MOB. Our results reveal multiple dSAC subtypes, each specialized to influence MOB activity by selectively innervating GABAergic interneurons, and provide direct evidence for novel intrabulbar and extrabulbar GABAergic projections.