Culture condition | Number of co-cultures | |||||
---|---|---|---|---|---|---|
wt | wt(−) | wt/rl | rl(−) | rl | Total | |
7 DIV | ||||||
E+/+/H+/+ | 12 | 3 | 0 | 0 | 0 | 15 |
Erl/+/Hrl/+ | 11 | 1 | 0 | 0 | 0 | 12 |
Erl/rl/Hrl/rl | 0 | 0 | 0 | 6 | 13 | 19 |
Erl/rl/Hrl/+ | 30 | 11 | 0 | 0 | 0 | 41 |
Erl/+/Hrl/rl | 0 | 0 | 0 | 13 | 17 | 30 |
15 DIV | ||||||
E+/+/H+/+ | 12 | 0 | 0 | 0 | 0 | 12 |
Erl/+/Hrl/+ | 13 | 0 | 0 | 0 | 0 | 13 |
Erl/rl/Hrl/rl | 0 | 0 | 0 | 3 | 18 | 21 |
Erl/rl/Hrl/+ | 14 | 1 | 0 | 0 | 0 | 15 |
Erl/+/Hrl/rl | 0 | 0 | 0 | 3 | 11 | 14 |
Entorhino-hippocampal co-cultures were prepared from +/+,rl/+, and rl/rl tissue, and the entorhino-hippocampal pathway was traced with biocytin and fixed after 7 and 15 DIV. Co-cultures were classified according to the pattern of entorhinal innervation in the SLM/ML. wt, Wild-type pattern: entorhinal fibers target to the stratum lacunosum-moleculare and molecular layer occupying the entire thickness of these layers, with a high density of innervation (>10 fibers/75 μm), and single fibers exhibit abundant collateral branches and axonal varicosities. wt−, Scarce wild-type pattern: similar to the pattern described in wt, but with a decreased density of innervation (<10 fibers/75 μm). rl,reeler-like pattern: the entorhinal termination in the stratum lacunosum-moleculare is very scarce and restricted to the vicinity of the hippocampal fissure, with fibers following straight courses and exhibiting very few axonal collaterals and varicosities; projection to the dentate gyrus appears densely packed in the outer molecular layer (>20 fibers/75 μm), with abundance of collateral branches and axonal boutons; presence of misrouted fibers in stratum radiatum and stratum pyramidale of the CA1–CA3 regions and in the hilus. rl−, Scarce reeler-like pattern: similar to the pattern described in rl, with a decreased density of innervation (<10 fibers/75 μm) in the dentate gyrus. wt/rl, Wild-type/reeler pattern: co-cultures showing intermediate (rl) or (wt) pattern of termination.