Extracellular diffusion is fast and isotropic in the stratum radiatum of hippocampal CA1 region in rat brain slices

Abstract

Molecular transport in brain extracellular space (ECS) is hindered by the structure of the tissue. Diffusion analysis of small extracellular markers quantifies tissue hindrance, expressed as tortuosity lambda = (D/D*)(1/2), where D is the free diffusion coefficient and D* is the effective diffusion coefficient in tissue. In healthy brain, lambda is approximately 1.6, but the nature of this parameter is poorly understood. We report that the stratum radiatum of the hippocampal CA1 region in vitro, previously shown to be anisotropic (i.e., different along the x-, y-, and z-axes) in in vivo study, is isotropic like somatosensory neocortex but has a reduced lambda. Diffusion of fluorophore-labeled dextran (f-dex, M(r) 3,000) and tetramethylammonium (TMA(+), M(r) 74) was measured in rat brain slices (400 mum) using integrative optical imaging (IOI) and real-time iontophoresis (RTI), respectively. In the stratum radiatum, diffusion of f-dex was similar along the x-, y-, and z-axes (lambda(x), lambda(y); lambda(z) were 1.55, 1.53, and 1.55), but the tortuosity was significantly lower than in the neocortex, where lambda = 1.81. This finding was confirmed by the RTI method, which measured lambda with TMA(+), a much smaller molecule, and determined volume fraction alpha, the proportion of tissue occupied by the ECS. In stratum radiatum, lambda(x), lambda(y), and lambda(z) were 1.47, 1.44, and 1.46, while in neocortex, lambda was 1.65. The ECS volume fraction was similar (0.24) in both regions. It is proposed that in the hippocampus, low lambda reflects a reduced occurrence of concave extracellular microdomains, referred to as dead spaces, which increase tortuosity by transient trapping of markers. Functionally, a low lambda may permit structural plasticity and facilitate extrasynaptic communication. It may also enhance the spread of neuroactive substances and thus contribute to the sensitivity of the hippocampal CA1 region to ischemia and epilepsy.