M. Palestini

Cytoplasmic calcium buffer, calbindin-D28k, is regulated by excitatory amino acids.

Excessive intracellular calcium in neurones is thought to underlie the pathophysiology of several neurodegenerative diseases. An extensively studied animal model is the neurotoxic increases in intracellular Ca2+ induced by excitatory amino acid. We report here that the calcium-binding protein, calbindin-D28k, increases rapidly in Purkinje cells of rat cerebellar slices superfused with excitatory and excitotoxic concentrations of glutamate or its analogue, kainic acid. The increase is reversible and reproducible, is blocked by CNQX and is independent of Ca2+ influx. These results indicate that calbindin containing neurones can regulate their Ca2+ buffering capacity in response to a specific agonist and this regulation is not mediated by cytosolic calcium increases.

Copper suppresses hippocampus LTP in the rat, but does not alter learning or memory in the morris water maze.

The objective of our study was to determinate the effect of copper on long-term potentiation (LTP) in hippocampus slices and a learning test in the Morris Water Maze (MWM). A group of adult Wistar rats received intraperitoneal (ip) injections of 1 mg/kg of CuSO(4) dissolved in saline for 30 consecutive days (Cu.R). A group of control rats (Sal.R), received saline by the same routes and duration. After this period, every individual of both groups was submitted to learning in MWM. Once the learning was completed, the LTP was studied in slices of hippocampus of both groups. The statistical assessment shows that the rats in both groups did not show significant differences in their progressive learning, notwithstanding that group Cu.R had 14.2 times more copper in their hippocampus and 16.7 times more in the visual cortices than in those of group Sal.R. On the other hand, the neurons of CA1 in hippocampus slices of Sal.R showed a significant development of LTP, but this was not observed in group Cu.R. In a second situation, 13 rats received training in MWM. Then, a group of 6 animals were injected with copper i.p. at the dose and time previously described. The 7 other animals were administered saline. Afterward, both groups were retrained in the MWM. The results obtained in Cu.R were similar to those obtained in Sal.R. Both groups maintained the concentrations of copper in the hippocampus indicated above, nonetheless, only the hippocampus slices of Cu.R did not show LTP. The spatial learning behavior of the rats was not affected by high copper concentration.

Interference of chronically ingested copper in long-term potentiation (LTP) of rat hippocampus

The objective of our study was to find the evidence of copper interaction in LTP, motivated by copper involvement in neurodegenerative illness, like Parkinson, Alzheimer and Amyotrophic Lateral Sclerosis, and we initiated the study of this element in the LTP. For this purpose we used hippocampus slices of rats chronically consuming copper dissolved in water (CuDR; n=26) and non-copper-consuming rats (CR; n=20). The CuDR rats received 8--10 mg/day during 20--25 days. Electrophysiological tests showed absence of LTP in CuDR slices, contrary to CR slices. The stimulus-response test applied before and after LTP showed significant increases of synaptic potential in the CR group. This did not occur in the CuDR group, except for the initial values, which probably seem associated to an early action of copper. The paired-pulse (PP) test, applied to CR and CuDR prior to tetanic stimulation, showed a significant reduction in PP, for the 20-, 30- and 50-ms intervals in CuDR. At the end of the experiments, copper concentration was 54.2 times higher in CuDR slices, compared to the concentration present in CR slices. Our results show that copper reduces synaptic sensibility and also the facilitation capability. These effects represent a significant disturbance in the plasticity phenomenon associated with learning and memory.

Glutamate, GABA, calbindin-D28k and parvalbumin immunoreactivity in the pulvinar-lateralis posterior complex of the cat : relation to the projection to the Clare-Bishop area

Neurons of the pulvinar-lateralis posterior complex (Pul-LP) containing glutamate (Glu) and GABA, as presumed neurotransmitters, and calbindin- D28k (calbindin) and parvalbumin (PV), as Ca-binding proteins, were identified in the cat by using immunohistochemical methods. In vibratome sections, neurons immunoreactive (IR) to each of the four antibodies were observed throughout the Pul-LP. In semithin sections, GABA-IR neurons were also PV-IR but not calbindin-IR and some of them also co-localized Glu. The Glu-IR neurons which were negative for GABA co-localized calbindin but not PV. The neurons of the Pul-LP projecting to the Clare-Bishop area (CB) in the suprasylvian gyrus were identified with a retrogradely transported tracer and the sections were then immunostained for Glu, GABA, calbindin and PV. Only Glu- and calbindin-IR neurons were retrogradely labeled. These results show that, if calbindin and PV have a Ca-binding role, the presumably excitatory Glu-IR neurons projecting to the CB are use calbindin whereas the presumably inhibitory GABA-IR neurons are intrinsic and use PV. This relationship implies that these proteins probably have other roles specifically related to the kind of agonist to be released at the neuron.