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In the present study, we examined changes in immunoreactivity (ir) for the gamma-isoform of protein kinase C (PKC gamma) in mouse hippocampus in relation to spatial memory processes employing the monoclonal antibody 36G9 raised against purified PKC gamma. Learning and memory were assessed by performance in a free-choice spatial pattern paradigm in a hole board in which the animals learned the pattern of 4 baited holes out of 16 holes. Adult male house mice were used, divided in four groups. Three control groups were formed: group N, naive (blank controls); group H, habituated (animals were for 5 consecutive days introduced to the hole board with all holes baited); and group PT, pseudotrained (animals were for 13 consecutive days introduced to the hole board with all holes baited). The T (trained) group was for 5 consecutive days introduced to the hole board with all holes baited (similar to the H and PT groups) followed by 8 successive days with only four holes baited in a fixed pattern. Behaviorally, following the first 5 d, the PT group crossed the hole board randomly, whereas the T group gradually learned to orientate in the hole board. The mice were killed 24 hr after the last performance. A shift in 36G9-ir appeared from the cell somata to the dendrites of hippocampal principal neurons when comparing the H and PT group, respectively. In contrast, the T group showed strong PKC gamma-ir in both cell somata and dendrites, which clearly exceeded that of the H and PT mice. In this way, 36G9-ir reveals the physiologically activated neurons involved in hole board learning.(ABSTRACT TRUNCATED AT 250 WORDS)

This study describes changes in the immunoreactivity for muscarinic acetylcholine receptors (mAChRs) in the hippocampus of mice in relation to spatial discrimination behavior, employing the monoclonal antibody M35 raised against purified bovine mAChR protein. Performance in a hole board in which the animals learned the pattern of 4 baited holes out of 16 holes served as the measure of spatial discrimination learning and memory. Twenty-six adult male house mice were used, divided into four groups. Three groups served as various controls: group N (naive; blank controls); group H (habituated; animals were introduced to the hole board with all holes baited for 5 consecutive days), and group P (pseudo-trained; the animals were admitted to the hole board for 13 consecutive days with all holes baited). The T group (trained) was subjected to the hole board for 5 consecutive habituation days with all holes baited (similar to the H and P groups), followed by 8 successive training days with only four holes baited in a fixed pattern. During the 8 training days, the T group gradually acquired a pattern to visit the baited holes, whereas the P group continued visiting holes in a random fashion. The mice were killed 24 h after the last behavioral session. All principal cells in teh cornu ammonis (CA) and dentate gyrus (DG) of the habituated animals revealed increased levels of mAChR immunoreactivity (mAChR-ir) over the naive mice. A minor increase in mAChR-ir was found in the apical dendrites of the CA1 pyramidal cells. Pseudotraining resulted in a CA1-CA2 region with a low level of mAChR-ir, resembling naive animals, whereas the trained mice showed a further increase in mAChR-ir in the CA1-CA2 pyramidal cell bodies and apical dendrites. Optical density measures of the mAChR-ir in the CA1 region revealed a significant (P < 0.05) increase in the pyramidal cell bodies of the H and T group over the N and P group, and a significant (P < 0.05) increase in the apical dendrites of the T group over all other groups. In contrast to the CA1-CA2 region, both pseudotrained and trained mice revealed high mAChR staining in the CA3-CA4 region and the DG. These results indicate that prolonged exposure to the hole board is sufficient for an enhanced mAChR-ir in the CA3-CA4 and DG, whereas the increase in CA1-CA2 pyramidal cells is a training-specific feature related to spatial orientation. Nonpyramidal neurons within the CA1-CA2 region with enhanced mAChR-ir in the pyramidal cells, however, revealed a decreased level of mAChR-ir. The opposing effect of pyramidal and nonpyramidal cells suggests a shift in the excitability of the hippocampal microcircuitry. Previously we demonstrated an increase and redistribution of hippocampal protein kinase C gamma-immunoreactivity (PKC gamma-ir) induced by hole board learning in mice (Van der Zee et al., 1992, J Neurosci 12:4808-4815). Immunofluorescence double-labeling experiments conducted in the present study in naive and trained animals revealed that the principal cells and DG interneurons co-express mACh...

Arginine-vasopressin (AVP) is a homeostatic modulator of body temperature during fever and may also be involved in normal body temperature control. In the present study the hypothalamus of mice bi-directionally selected for thermoregulatory nest-building behavior was immunocytochemically labeled for AVE The low-selected mice had a 1.5-fold higher number of AVP-immunoreactive neurons in the suprachiasmatic nuclei (SCN) compared to the unselected control and the high-selected mice. No differences between the selected lines could be detected in the number of AVP-immunoreactive neurons in the paraventricular nuclei (PVN). The neuroanatomical data suggest a possible role of AVP in the SCN and control of thermoregulatory nest-building behavior. Our selected mice may prove to be a model system to study the role of AVP in the SCN.

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