Abnormal axonal guidance and brain anatomy in mouse mutants for the cell recognition molecules close homolog of L1 and NgCAM-related cell adhesion molecule.

Cell recognition molecules of the L1 family serve important functions in the developing and the mature nervous system. Mutations in genes encoding the L1 family members close homolog of L1 (CHL1) and NgCAM-related cell adhesion molecule (NrCAM) have been found to alter connectivity and morphology of several brain regions. In order to emphasize similarities and differences of these two structurally related molecules, null mutants for CHL1 and NrCAM were directly compared with respect to axonal guidance in the hippocampus and the olfactory bulb and the sizes of the ventricular system and the cerebellar vermis using a combined structural magnetic resonance imaging (MRI) and histological approach. The results demonstrate that the absence of CHL1 leads to aberrant hippocampal mossy fiber projections whereas in both mutants, CHL1 and NrCAM, the guidance of the olfactory nerve projections is disturbed. Both mutations also alter the size of the ventricular system and the vermis with a specific profile of changes and partially opposite effects in each of the mutants. CHL1/NrCAM double-mutant mice do not show any enhancement of the single mutant's phenotype but balance the opposing effects on the ventricular system. In summary, the results show that CHL1 and NrCAM both affect axonal guidance and the anatomy of the ventricular system and the cerebellar vermis but act differently on these processes.

Severe cognitive and motor coordination deficits in tenascin-R-deficient mice.

The extracellular matrix molecule tenascin-R (TN-R), predominantly expressed in the central nervous system, has been implied in a variety of functions, e.g. during myelination, cerebellar neurite fasciculation and hippocampal long-term potentiation. In this study, we investigated in detail the impact of TN-R deficiency on the living animal by analyzing the behavior of TN-R-deficient mice. The general state, gross sensory functions, reflexes and motoric capabilities appeared normal. In contrast, motor coordination on the rota-rod was compromised in these mice, indicating a deficit in cerebellar functions. In the open field and the hole board, the mutants interact differently with their environment, probably due to differences in their exploratory behavior. TN-R-deficient mice were able to learn a reference memory task in the Morris water maze. In contrast to wild-type mice, the mutants displayed an alternative strategy; swimming around the pool using a stereotypical circling pattern, crossing all possible platform positions after relocation of the escape platform (reversal). These results, confirmed by relocating the platform in the center of the pool, suggest that TN-R-deficient mice may be impaired in constructing a goal-independent representation of space. In addition, a two-way active avoidance test (shuttle box) revealed a severe deficit in associative learning in TN-R-deficient mice. Our results support important functions of TN-R in vivo in the central nervous system, in particular in the cerebellum and the hippocampus.

Altered processing of novel information in N-CAM-deficient mice.

N-CAM-deficient mice display anatomical and electrophysiological abnormalities in the CNS and behavioral deficits. Here, we address the question whether information processing is altered in these mice by analysis of the expression of c-fos and arg 3.1/arc in N-CAM-deficient mice after presentation of saccharin as novel or familiar and water as neutral gustatory stimulus. When compared to their wild-type control littermates, increased expression of c-fos mRNA in the amygdala after the novel taste and of arg 3.1/arc mRNA in the dentate gyrus 4.5 h after the neutral taste was detected in the absence of N-CAM. Furthermore, the novelty-induced increase in arg 3.1/arc expression in the cingulate cortex 4.5 h after the novel taste was not observed in N-CAM-deficient mice. These data suggest that information processing mediated by immediate-early gene expression is altered in N-CAM-deficient mice.

Misguided axonal projections, neural cell adhesion molecule 180 mRNA upregulation, and altered behavior in mice deficient for the close homolog of L1.

Cell recognition molecules are involved in nervous system development and participate in synaptic plasticity in the adult brain. The close homolog of L1 (CHL1), a recently identified member of the L1 family of cell adhesion molecules, is expressed by neurons and glia in the central nervous system and by Schwann cells in the peripheral nervous system in a pattern overlapping, but distinct from, the other members of the L1 family. In humans, CHL1 (also referred to as CALL) is a candidate gene for 3p- syndrome-associated mental impairment. In the present study, we generated and analyzed CHL1-deficient mice. At the morphological level, these mice showed alterations of hippocampal mossy fiber organization and of olfactory axon projections. Expression of the mRNA of the synapse-specific neural cell adhesion molecule 180 isoform was upregulated in adult CHL1-deficient mice, but the mRNA levels of several other recognition molecules were not changed. The behavior of CHL1-deficient mice in the open field, the elevated plus maze, and the Morris water maze indicated that the mutant animals reacted differently to their environment. Our data show that the permanent absence of CHL1 results in misguided axonal projections and aberrant axonal connectivity and alters the exploratory behavior in novel environments, suggesting deficits in information processing in CHL1-deficient mice.

Altered odor-induced expression of c-fos and arg 3.1 immediate early genes in the olfactory system after familiarization with an odor.

In adult rats, repeated exposure to an odorant, in absence of any experimentally delivered reinforcement, leads to a drastic decrease in mitral/tufted (M/T) cell responsiveness, not only for the familiar odor but also for other novel odors. In the present study, using two different and complementary in situ hybridization methods, we analyzed the effect of familiarization with an odorant on c-fos and arg 3.1 mRNA expression levels, and we examined the odor specificity of this effect. Odor exposure induces a specific increase in c-fos and arg 3.1 expression in some particular olfactory bulb quadrants. Previous familiarization with the test odor results in a decreased expression of both IEGs in these quadrants, leading to the alteration of the odor-specific pattern of c-fos and arg 3.1 expression. In contrast, this odor-specific pattern is not affected when different odors are used for familiarization and test. Similarly, an odor-specific familiarization effect leading to a reduced c-fos and arg 3.1 expression was also detected in the cingulate cortex and in the anterior piriform cortex. These results support our hypothesis that the decrease in M/T cell responsiveness following a preceding familiarization with an odorant may be related to a particular form of synaptic plasticity involving changes at the genomic level, and reveals further insight in olfactory information processing and the cellular mechanisms underlying familiarization in the olfactory system.

Novelty-induced increased expression of immediate-early genes c-fos and arg 3.1 in the mouse brain.

The detection of novel stimuli is a memory-dependent process. The presented stimulus has to be compared with memory contents to judge its novelty. In addition, the novelty of stimuli activates attention-related processes that facilitate memory formation. To determine the involvement of limbic and neocortical brain structures in novelty detection, we exposed mice to a novel gustatory stimulus (0.5% saccharin) added to their drinking fluid. We then compared the novelty-induced expression of the two immediate-early genes (IEGs) c-fos and arg 3.1, with their expression in mice familiarized with the same stimulus or mice not exposed to that stimulus. Exposure to taste novelty increased expression of c-fos and arg 3.1 mRNA in the cingulate cortex and deep layers of the parietal cortex. In addition, c-fos mRNA expression was increased in the amygdala and arg 3.1 mRNA was increased in the dentate gyrus. Expression of c-fos and arg 3.1 was elevated 30 min after the exposure to novelty. For arg 3.1, a second peak of expression was found 4.5 h after presentation of the novel stimulus. Our results indicate that the amygdala, the dentate gyrus, and the cingulate and parietal cortices may be involved in novelty detection and associated cognitive events, and suggest that c-fos and arg 3.1 play distinct roles in these processes.

Odour-induced c-fos expression in the rat olfactory bulb: involvement of centrifugal afferents.

Expression of the proto-oncogene c-fos is known to increase in granule cells of the olfactory bulb following a sustained olfactory stimulation. Most granule cells displaying high levels of Fos accumulation are located in the bulbar columns defined by the odour-induced foci of high 2-deoxyglucose glomerular uptake. The present studies were undertaken in order to assess the possible involvement of centrifugal afferents in the modulation of odour-induced patterns of either 2-deoxyglucose accumulation or c-fos expression in the olfactory bulb. A unilateral olfactory peduncle section had no effect on the odour-induced 2-deoxyglucose foci but induced a significant decrease in the number of Fos-containing neurons in odour-selective areas of both olfactory bulbs, ipsilateral and contralateral to the lesion. This suppressive effect was much more pronounced in the side ipsilateral to the peduncle section. It is concluded that c-fos expression induced by a sustained stimulation with propionic acid vapours is not only determined by the olfactory peripheral input but also by afferents of central origin. In order to estimate the contingent involvements of the cholinergic and noradrenergic afferents in this control of c-fos expression, we attempted to mimic the effects of the surgical deafferentation on odour-induced c-fos expression by using a pharmacological approach with selective cholinergic and noradrenergic antagonists. The beta-adrenergic antagonist propanolol induced a suppression of the odour-related patterns of Fos accumulation similar to the one caused by the surgical deafferentation of the olfactory bulb. The muscarinic antagonist scopolamine did not alter c-fos expression in the odour-selective area but increased significantly Fos labelling in the other bulbar aspects. Pharmacological investigations indicate that the noradrenergic and cholinergic centrifugal systems are likely involved in the central modulation of c-fos expression in the OB. The Fos protein could be expressed as an early nuclear signal triggering further long-term modifications of the neuronal phenotype, in certain conditions of sensory stimulation involving the activation of centrifugal systems.

C-fos expression and 2-deoxyglucose uptake in the olfactory bulb of odour-stimulated awake rats.

Using a selective monoclonal Fos antibody, we have studied the expression of the c-fos proto-oncogene in the olfactory bulb of awake rats stimulated with propionic acid vapours. We have perfected a method which allows the comparison of patterns of either cellular c-fos immunoreactivity or glomerular 2-deoxyglucose uptake in the same olfactory bulb. C-fos was expressed in local bulbar interneurones, principally granule cells. We observed that the low level of basal c-fos expression raised significantly under olfactory stimulation in the bulbar column defined by the foci of high 2-deoxyglucose glomerular uptake. This is the first demonstration that c-fos expression can be triggered by afferent olfactory input in the olfactory bulb in normally breathing awake rats. These data support the assumption that the olfactory bulb is a suitable model to elucidate the priming conditions and functional involvement of Fos protein synthesis triggered by physiological sensory stimulation.

Apomorphine disrupts odour-induced patterns of glomerular activation in the olfactory bulb.

The olfactory bulb of adult male rats stimulated with propionic acid vapours displays a characteristic focus of high metabolic activity in the dorso-medial glomeruli, as revealed by the 2-deoxyglucose (2DG) method. Injection of the dopaminergic agonist apomorphine (1.5 mg kg-1) prior to odour stimulation completely abolishes this selective pattern of glomerular activation, while the metabolic activity of other bulbar areas is not significantly altered. This effect is abolished by a previous injection of the dopamine antagonist haloperidol (0.1 mg kg-1). These observations emphasize the probable involvement of dopamine and dopamine receptors in the bulbar processing of olfactory information.