Comparison of neuroplastin and synaptic marker protein expression in acute and cultured organotypic hippocampal slices from rat.
Organotypic hippocampal slice cultures can be used to study hippocampal biochemistry and physiology over a chronic period on the days to weeks timescale. In order to validate the organotypic hippocampal slice culture for our ongoing studies of synaptic function, we have compared, using Western blotting, the levels of a number of synaptic proteins from in vitro organotypic hippocampal slice cultures with those from in vivo hippocampal slices prepared from age-matched controls. We chose to follow the developmental expression of the neuroplastin (np) family of immunoglobulin related cell adhesion molecules (CAMs), np65, a brain specific isoform highly expressed in hippocampal neurones and np55 a more widely expressed isoform and two synaptic marker proteins, synaptophysin, a pre-synaptic marker and post-synaptic density protein-95, PSD95, a post-synaptic marker. All showed increasing expression over the developmental time period, both in vivo and in vitro. The level of both neuroplastins was also consistent between the in vivo and in vitro preparations, whereas the level of PSD95 was markedly increased in the organotypic hippocampal slice cultures while the level of synaptophysin was slightly decreased. Whilst these findings may indicate some differences in the composition and organisation of synapses, the developmental expression profiles of these synaptic proteins within organotypic hippocampal slice cultures suggests they are a valid model for the study of synapse function and development in vitro.
Buckby
Lucy E
LE
School of Biological Sciences, Royal Holloway University of London, Egham Hill, Egham, Surrey, TW20 0EX, UK.
Mummery
Rosemary
R
Crompton
Mark R
MR
Beesley
Philip W
PW
Empson
Ruth M
RM
D000375:Aging / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers; D015153:Blotting, Western; D015815:Cell Adhesion Molecules / Q000378:metabolism; D015816:Cell Adhesion Molecules, Neuronal / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D006624:Hippocampus / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009434:Neural Pathways / Q000166:cytology / Q000378:metabolism*; D009924:Organ Culture Techniques / Q000379:methods*; D017729:Presynaptic Terminals / Q000378:metabolism* / Q000648:ultrastructure; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D016708:Synaptophysin / Q000378:metabolism; D013997:Time Factors
D015415:Biomarkers; D015815:Cell Adhesion Molecules; D015816:Cell Adhesion Molecules, Neuronal; D009419:Nerve Tissue Proteins; D016708:Synaptophysin; C033652:postsynaptic density proteins