Learning-induced alterations in hippocampal PKC-immunoreactivity: a review and hypothesis of its functional significance.
1. To localize protein kinase C (PKC) in the hippocampus, PKC activity measures, mRNA in situ hybridization, and [3H]phorbol ester binding techniques were used until in the 1980s antibodies became available for in situ immunocytochemistry. In the late 1980s, PKC-isoform-specific antibodies were first used to map hippocampal PKC at the cellular and subcellular level. The mammalian hippocampus contains all four Ca(2+)-dependent PKC isoforms, but the (sub)cellular localization is both isoform- and species-specific. 2. Hippocampally-dependent spatial and associative learning in rat, mice and rabbit induce an increase in PKC immunoreactivity (ir) in hippocampal principal cells studied 24 hours after the animals had learned the task. Among the four Ca(2+)-dependent PKC subtypes, this increase is selective for the gamma-isoform. The presence of the gamma-isoform in dendritic spines (the most likely site for synaptic plasticity and information storage), in contrast to PKC alpha, beta 1, and beta 2, may underlie the isoform-selectivity. 3. Compared to fully trained animals, subjects halfway training showed intermediate levels of increased PKC gamma-ir. Poor learners that were not able to learn the task showed considerably less enhanced PKC gamma-ir as compared to good learners. 4. Associative learning induced a decrease in astroglial PKC beta 2 and gamma-ir in those regions where a simultaneous increase in neuronal PKC gamma-ir was observed. This decrease most likely reflects PKC down-regulation, enabling the astrocytes to maintain their K+ buffering capacity necessary to support neuronal activity such as accompanying learning and memory. 5. Western blot analyses revealed that the increase in PKC gamma-ir was not due to an increase in total amount of PKC gamma, translocation, or the proteolytic generation of the fragment PKM. The increase in PKC gamma-ir must therefore reflect a learning-induced conformational change in the PKC gamma molecule that results in the exposure of the antigenic site(s). 6. Although a large number of hippocampal pyramidal cells display learning-induced enhancement of PKC gamma-ir at the 24 hours post-training time point, this does not indicate, however, that all synapses in these neurons are used, or that the maximal PKC signal transduction capacity per call has been reached. 7. The enhanced PKC gamma-ir may reflect a form of activated PKC, since PKC stimulation by phorbol esters (both in hippocampal slices and mildly aldehyde fixed sections) mimicked the increase in PKC gamma-ir similar as seen after learning. 8. The most likely transmitter systems which may have induced the altered PKC gamma-ir are acetylcholine and glutamate. Their contribution and interaction at the cellular level are depicted in a schematic circuit terminating on a CA1 pyramidal cell (Fig. 4). 9. Several functional roles for PKC gamma in learning and memory are discussed, and a hypothetical model is proposed based on an endogeneous PKC inhibitor protein that may explain altered antibody-binding to PKC gamma after learning (Fig. 6). 10. The immunocytochemical approach can contribute significantly to the ongoing attempts to decipher part of the cellular and biochemical mechanism of learning and memory. The development of ever more specific and better characterized antibodies reactive with different sites of proteins like PKC gamma will offer the necessary tools for further immunocytochemical research to help unravel complex brain functions.
Van der Zee
E A
EA
Dept. of Cell & Molecular Biology, Northwestern University Medical School, Chicago, IL, USA.
Luiten
P G
PG
Disterhoft
J F
JF
D000818:Animals; D001245:Association Learning; D006624:Hippocampus / Q000502:physiology*; D007527:Isoenzymes / Q000378:metabolism; D007858:Learning / Q000502:physiology*; D051379:Mice; D008959:Models, Neurological; D009474:Neurons / Q000502:physiology; D015240:Phorbol 12,13-Dibutyrate / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism*; D011817:Rabbits; D051381:Rats; D013028:Space Perception
D007527:Isoenzymes; D015240:Phorbol 12,13-Dibutyrate; D011493:Protein Kinase C