Cortical Neuronal Cell Cultures Research Articles

Comparison of rates of neuronal development and survival in human and rat cerebral cortical cell cultures

A problem that has not been addressed directly at the cellular and molecular levels concerns the basic mechanisms governing rates of development and aging in the nervous system. We have begun to examine this problem by employing parallel cell cultures of cerebral cortical neurons taken from rat and man, two species with markedly different developmental periods and life spans. Cortical cultures were established from rat and human fetuses and were maintained under identical conditions. Long-term neuronal survival in human cortical cultures was strikingly greater than in rat cortical cultures, and axonal outgrowth was significantly slower in the human neurons. These differences were consistently observed in a variety of culture media. Exchanges of culture medium between cultures of the two species did not alter the differences in neuronal survival and axon outgrowth suggesting that the differences between human and rat neurons were probably not due to factors released into the culture medium. Furthermore, the differences were not related to developmental stage at the time of culture initiation since similar results were obtained in cultures established from fetuses of different gestational ages. These initial data indicate that differences in the developmental time courses and life spans of human and rat cerebral cortex are reflected at the level of the individual neuron, and provide a system in which to explore the mechanisms responsible for these differences.

Cortical neurons containing somatostatin‐ or parvalbumin‐like immunoreactivity are atypically vulnerable to excitotoxic injury in vitro

We exposed murine cortical neuronal cell cultures for 24 hours to defined concentrations of N-methyl-D-aspartate (NMDA), kainate, or alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), and assessed the resultant neuronal degeneration quantitatively by the efflux of lactate dehydrogenase to the bathing medium. The small subpopulations of neurons that stained immunohistochemically for either somatostatin- or parvalbumin-like reactivity were atypically affected by these excitotoxins. Limited exposure to kainate or AMPA did little damage to the general neuronal population, but destroyed nearly all somatostatin- or parvalbumin-reactive cells. Conversely, these immunoreactive cells were more resistant to NMDA-induced injury than the general population. In view of reports suggesting that somatostatin- and parvalbumin-reactive cortical neurons may be preferentially damaged in Alzheimer's disease, these observations support a hypothesis that the overactivation of non-NMDA receptors could be involved in Alzheimer's disease pathogenesis.

Effect of ferric nitrilotriacetate on predominantly cortical neuronal cell cultures.

Predominately neuronal cell cultures were produced as described in previous communications. Neuronal cells were exposed to ferric nitrilotriacetate (Fe-NTA) at varying concentrations. Studies of the neuronal cells were performed at 13 and 20 days in culture. In addition to morphologic studies, biochemical assays including choline acetyltransferase (ChAT) activity, specific [3H]flunitrazepam (FLU) binding, clonazepam (CLO)-displaceable [3H]FLU binding, Ro5-4864-displaceable [3H]FLU binding, high-affinity [3H]GABA uptake, and protein determinations were performed. The data demonstrate that chelated ferric iron has an adverse effect on predominately neuronal cultures after 7 days of exposure as measured by choline acetyltransferase activity, while other measures remained unaffected; however, after 14 days of exposure all measures were significantly decreased. The effects of Fe-NTA exposure appear to be both concentration and duration-of-exposure related.

Arachidonic acid inhibits uptake of glutamate and glutamine but not of GABA in cultured cerebellar granule cells

The effects of arachidonic acid (20:4) on the uptake of glutamate were studied in primary cultures of cerebellar granule cells and were compared to cortical neurons and astrocytes. At a dose of 0.005 mM, the glutamate uptake was significantly inhibited in cerebellar granule cells. This inhibition was dose and time dependent. The uptake of glutamate was equally sensitive to 20:4 in primary cell cultures of cortical neurons, whereas the uptake in astrocytes was much less sensitive to 20:4. Glutamine uptake was inhibited by 20:4 in cultured cerebellar granule cells and cerebral cortical astrocytes but was not affected in cerebral cortical neurons. Furthermore, the uptake of gamma-aminobutyric acid was not affected by 20:4 in cerebellar granule cells.