Layer II of the ventral-intermediate entorhinal area (VIE) is narrow and organized in clumps that are very close to each other, rendering a continuous appearance when studied with low magnification. In contrast, in the dorsal-intermediate entorhinal area (DIE), layer II contains rather big, rounded neurons that stand out in regular Nissl and NeuN stains since they are darkly stained. A very narrow, relatively acellular band separates layer II from layer III in much of the DIE extent. Layer III of VIE is made up of densely packed, medium-size pyramids, whereas in DIE this layer is wide, and clearly presents a narrow, more densely packed outer zone with its neurons arranged in cell clusters, and a less densely and irregularly packed inner zone.
No major differences between the two areas are apparent in deep layers V and VI, with the exception that the border between layer V and the lamina dissecans is very much obscured in DIE compared to VIE and the latter area also contains much more large, darkly stained pyramidal cells than DIE does.
Calbindin is strongly expressed in deep layer I, layer II and III in
VIE, both in neurons and neuropil. In DIE, the staining in layers I and
III seems to be less dense, but the staining of individual neurons in
layer II is more intense in DIE than in VIE. Whereas we observed only
very weak staining in VIE for parvalbumin. DIE shows an overall higher
intensity when stained for parvalbumin, particularly in layers II and
III.
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