Culture of mature hippocampus slices for 4 days in a newly developed medium: preservation of transmitter release and leucine incorporation into protein.
A procedure and a medium for sustaining mature hippocampus slices in vitro for 4 days are described. The ionic composition of the medium, in which the slices were incubated for 1 h of recovery following preparation, strongly affected their ability, 4 days later, to take up and to release D-[3H]aspartate and [14C]GABA. A medium deficient in Na+ and Ca2+ proved best for recovery of the fresh slices prior to transfer to culture medium. The newly developed CSF-like culture medium was the best among several media tested in maintaining the potential of the slices for uptake and for induced release of D-[3H]aspartate and [14C]GABA. Glutamine, present in most culture media, appeared to be particularly toxic. Relative to fresh slices, the slices after 4 days in culture maintained 118% and 97% of the uptake of D-[3H]aspartate and of [14C]GABA respectively. K(+)-induced release of D-[3H]aspartate and of [14C]GABA was 104% and 82% of the respective values in fresh slices. Under the optimal culture conditions worked out, the slices also regained a considerable capacity for incorporation of labelled leucine into protein, which was low in fresh slices.
Kleinberger-Doron
N
N
Department of Biological Chemistry, Hebrew University of Jerusalem, Israel.
Schramm
M
M
D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D002250:Carbon Radioisotopes; D002412:Cations / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology*; D046508:Culture Techniques; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism*; D007930:Leucine / Q000378:metabolism*; D008297:Male; D009419:Nerve Tissue Proteins / Q000096:biosynthesis*; D018377:Neurotransmitter Agents / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013997:Time Factors; D014316:Tritium; D014701:Veratridine / Q000494:pharmacology; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D002250:Carbon Radioisotopes; D002412:Cations; D003470:Culture Media; D009419:Nerve Tissue Proteins; D018377:Neurotransmitter Agents; D014316:Tritium; D001224:Aspartic Acid; D005680:gamma-Aminobutyric Acid; D014701:Veratridine; D007930:Leucine