Analysis Tools
nervous system
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• the characteristic lamination pattern of mossy fibers emanating from the dentate gyrus is altered; at rostral levels, mossy fibers fail to bifurcate and prematurely invade the stratum pyramidale, forming thick, fasciculated bundles that aberrantly navigate between CA3 somata
• more caudally, the suprapyramidal bundle is discernable but appears less compact and reduced in width, especially in CA3c
• the infrapyramidal bundle is absent or misplaced in the stratum pyramidale
• CB+ mossy fiber density in the stratum pyramidale of CA3 is increased
• the length of both the suprapyramidal bundle and infrapyramidal bundle and the distance between them are decreased
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• the organization of the suprapyramidal bundle and stratum pyramidale is intact initially at P0-P5, however numerous axons penetrate the stratum pyramidale in between dispersed CA3 neurons from P7 onwards indicating aberrant pattern of mossy fiber innervation
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• growth cones of dentate gyrus neurons show an increase in F-actin in the central domain and in large bundles emanating from this region towards the growth cone membrane
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• mice show ectopic synapses in the stratum pyramidale; numerous synaptoporin+ mossy fiber terminals are seen around CA3 pyramidal neuron somata in the stratum pyramidale and these aberrantly located terminals form synapse-like junctions containing numerous synaptoporin+ vesicles directly on the somata of CA3 pyramidal neurons or on large and branched spine-like structures emanating from the soma, often forming multiple postsynaptic densities
• 21 days in vitro (DIV) hippocampal neurons show an increase in the number of synaptoporin+;vGlut1+ puncta on CA3 somata indicating increased synapse formation on CA3 somata
• 21 DIV co-cultures of dentate gyrus neurons from mutant mice make more synaptic contacts onto wild-type CA3 somata as compared with wild-type dentate gyrus neurons
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• the number of Rab6 secretory vesicles in the growth cone of dentate gyrus neurons grown in culture is increased and many vesicles accumulate in areas of abnormal F-actin expression
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cellular
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• mouse embryonic fibroblasts (MEFs) display a compact, round morphology instead of the normal fibroblast-like irregular shape and are smaller
• MEFs exhibit strongly bundled F-actin and increased amount of peripheral F-actin
• MEFs show an increase in the number of Rab6 vesicles at the cell periphery in areas of dense F-actin bundles, however overall density of Rab6 vesicles is unchanged
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