Effect of hypoxia on the morphology of mouse striatal neurons.

Symptoms of high altitude sickness including headache and neuropsychological dysfunction are thought to result from prolonged exposure to hypoxia. In order to explain how the brain adapts to lower oxygen pressure at high altitude, CD1 mice were exposed to 3 weeks of hypobaric hypoxic conditions. Analyses of the neuronal morphology of striatal medium spiny neurons (MSNs) revealed a significant decrease in dendritic length, yet no change in dendritic volume, in hypoxic mice relative to normoxic mice. Vascular data indicated an increase in blood vessel area in the striatum of mice exposed to prolonged hypoxia. A mouse model of high altitude exposure may assist in elucidating the mechanisms of cerebral adaptation to high altitudes in humans, and therefore aid in developing successful prevention techniques and treatment of problems associated with high altitude disease.

Long-term potentiation is associated with changes in synaptic ultrastructure in the rat neocortex.

Long-term potentiation (LTP) in the sensorimotor cortex of freely moving rats has been associated with changes in dendritic morphology and dendritic spine density. The current research examined changes in synaptic number and ultrastructure associated with LTP in this cortical region. LTP was induced over a 1 h period and the animals were sacrificed 2 h after the initial stimulation of the LTP group. Synapses within the terminal area of the apical dendrites from layer III pyramidal neurons were quantified by determining the total number of synapses per neuron, the number of excitatory and inhibitory contacts, number of synapses with different curvature subtypes, number of perforated synapses, and synaptic length. Several changes in synaptic morphology of excitatory synapses were revealed but no overall increase in the number of synapses per neuron was evident. Specifically, the induction of LTP was associated with an increased number of excitatory perforated and concave shaped synapses. Increased numbers of perforated concave synapses were also found to be significantly correlated with the degree of potentiation in the LTP animals. These and previous results suggest similar synaptic changes in both the cortex and hippocampus during the early phases of LTP maintenance and distinct synaptic changes during later phases of LTP maintenance.

Sequential changes in the synaptic structural profile following long-term potentiation in the rat dentate gyrus: III. Long-term maintenance phase.

LTP has been associated with changes in synaptic morphology but the nature of these changes over the time course of the enhanced electrophysiological response has not been fully determined. The current research involved an examination of synaptic structure in the rat hippocampus during the long-term maintenance phase of LTP. Synapses were examined in the middle third of the molecular layer (MML) of the rat dentate gyrus following repeated high frequency tetanization of the perforant path. Synapses from both the ipsilateral inner third of the dentate molecular layer (IML), which was not directly stimulated during the induction of LTP, as well as implanted, nonstimulated animals, served as controls. LTP was induced over a 4-h period, and the animals were sacrificed 5 days after the final stimulation of the LTP group. Ultrastructural quantification included the total number of synapses per neuron, synaptic curvature, the presence of synaptic perforations, and the maximum length of the synapses. No overall changes in the number of synapses per neuron, shape, or synaptic perforations were observed. There was, however, a significant increase in the length of synapses in the directly stimulated LTP tissue. This increase in synaptic length was particularly evident in the concave-shaped synapses which were also more perforated. These results, together with previous findings, describe a sequence of changes in synaptic morphology that accompany LTP in a structure that is associated with learning and memory.

Morphology of layer III pyramidal neurons is altered following induction of LTP in sensorimotor cortex of the freely moving rat.

The organization of specific cortical connections can be altered by sensory and motor experience. These changes are believed to result from activity-dependent changes in synaptic connectivity, similar to those induced in the hippocampus by high-frequency stimulation in long-term potentiation (LTP) experiments. If similar mechanisms are involved, then neocortical LTP induction may induce some of the same morphological changes that are seen following learning. We induced LTP in the contralateral sensorimotor cortex by repeated, daily tetanization of the corpus callosum in chronically implanted, freely moving rats. Anatomical results showed that the LTP induction was associated with alterations in dendrite morphology and increased spine density. These changes are qualitatively and quantitatively similar to those commonly observed in studies in which rats are housed in complex environments. The similarity of results following exposure to complex environments and after LTP induction in the neocortex may indicate a reliance on the same cellular mechanisms in both situations.

Specificity and efficiency of Cre-mediated recombination in Emx1-Cre knock-in mice.

Emx1 is a mouse homologue of the Drosophila homeobox gene empty spiracles and its expression is restricted to the neurons in the developing and adult cerebral cortex and hippocampus. We reported previously the creation of a line of transgenic mice in which the cre gene was placed directly downstream of the putative Emx1 promoter using ES cell technology. We showed that Cre protein was present in the cerebral cortex of the transgenic mice and was able to mediate loxP-specific recombination in vitro. In the present study, the specificity and efficiency of the cre-mediated recombination were determined using three independent lines of reporter mice and a combination of histochemical staining, neuronal culture, and Southern detection of the genomic DNA. Our results showed that the recombination was highly efficient in all three lines of reporter mice tested and confirmed that the deletion was restricted to the neurons in the cerebral cortex and hippocampus. Furthermore, we have determined that the recombination efficiency in the cerebral cortex was 91%. Our results suggest that Emx1 is not expressed in every neuron in the developing and adult cerebral cortex. This line of cre mice should contribute to the studies of cortical development and plasticity.

Sequential changes in the synaptic structural profile following long-term potentiation in the rat dentate gyrus. II. Induction/early maintenance phase.

Long-term potentiation (LTP), one of the most compelling models of learning and memory, has been associated with changes in synaptic morphology. In this study, LTP was induced and animals were sacrificed 1 h after the stimulation of the LTP group (induction / early maintenance phase). Synapses in the directly stimulated middle third of the dentate gyrus molecular layer (MML) were examined while synapses from the inner third of the dentate molecular layer (IML) of the LTP animals and both the MML and the IML of implanted animals served as controls. The total number of synapses per neuron, synaptic curvature, the presence of synaptic perforations, and the maximum length of the synaptic contact and active zone were examined. No overall change in the number of synapses per neuron was observed in the LTP tissue. LTP was associated with a significant increase in the proportion of perforated and irregular-shaped synapses compared to controls. The increase in perforated synapses was particularly apparent in the proportion of concave perforated synapses. Nonperforated concave synapses were found to be significantly larger in potentiated tissue. The total synaptic length per neuron of synapses in a concave configuration was also significantly higher following potentiation. These results suggest that the specific structural profile associated with 1-h post-LTP induction, which differed from the profile observed at 24 h post-induction, may represent a unique early phase of synaptic remodeling in a series of changes observed during LTP induction, maintenance, and decay.

Long-term potentiation in the reciprocal corticohippocampal and corticocortical pathways in the chronically implanted, freely moving rat.

The hippocampus and adjacent cortical structures, including the entorhinal, perirhinal, and parahippocampal cortices, appear to serve as an integrated memory system. This extended hippocampal system is believed to influence memory and consolidation through an extensive set of reciprocal connections with widespread areas of the neocortex. Long-term potentiation (LTP) has been well-examined in the intrinsic connections of the hippocampus and neocortex. However, LTP in the pathways and structures thought to convey information between the hippocampus and neocortex has received little attention. If these pathways and structures are involved in information storage, and if LTP reflects a general synaptic encoding mechanism, then these systems are also likely to support LTP. In this paper we discuss a series of experiments aimed at investigating LTP in the efferents between the hippocampus and neocortex in chronically implanted animals. In the first experiment, the efferents of the perirhinal cortex were stimulated. LTP in the dentate gyrus (DG) reached asymptote more slowly than is typically seen following perforant path stimulation, whereas the frontal area (M1) reached asymptote more quickly than reported following corticocortical stimulation. The DG and M1 LTP was long-lasting, but entorhinal cortex LTP had decayed to baseline levels after a week. In the second experiment, the hippocampal efferents were stimulated. The perirhinal, entorhinal, and frontal cortex showed a similar slow potentiation, with only the perirhinal cortex levels returning to baseline after a week. In the third experiment, the projections from M1 were tested. The perirhinal cortex and hippocampus showed a long-lasting LTP. Although LTP was found in all pathways examined, there were differences in the induction and decay rate, and these properties may correspond to differences in learning rate and longevity of information storage.

Physiological consequences of morphologically detectable synaptic plasticity: potential uses for examining recovery following damage.

A growing literature indicates that brain structure is modified in various ways with experience. In this paper we briefly survey evidence that the brain retains the capacity to modify its organization in response to demands, including demands resulting from learning, throughout the lifetime. We attempt to address whether these experience-induced changes are accompanied by physiological changes that indicate a functional reorganization of the brain. The kinds of morphological changes that have been observed following brain injury appear to be very similar to those seen after learning. The similarity suggests that many of the basic mechanisms of synaptic change in the brain may be utilized for both functions. This suggests that we can take advantage of some of the methods used to test the changes in physiology with behavioral manipulations to examine the damaged brain. We advocate utilizing electrophysiological techniques to measure functional recovery from brain injury as these may be useful in evaluating both spontaneous recovery from damage and the therapeutic benefits of training, or other therapies.

Sequential changes in the synaptic structural profile following long-term potentiation in the rat dentate gyrus: I. The intermediate maintenance phase.

Changes in synaptic structure have been reported following the induction of long-term potentiation (LTP). The structure of synapses during the intermediate maintenance of LTP has yet to be fully characterized in chronically implanted freely moving animals. The present study examined synapses in the middle third of the molecular layer (MML) of the rat dentate gyrus following repeated high frequency tetanization of the perforant path. Synapses from both 1) the ipsilateral inner third of the dentate molecular layer (IML), which was not directly stimulated during the induction of LTP, as well as 2) implanted, nonstimulated animals, served as controls. LTP was induced over a 4-h period, and the animals were sacrificed 24 h after the final stimulation of the LTP group. Ultrastructural quantification included the total number of synapses, synaptic curvature, the presence of synaptic perforations, and the maximum length of the synaptic contact. Although LTP was not associated with an overall increase in synaptic number, there was a significant increase in the proportion of presynaptically concave-shaped synapses. Further, the concave synapses in the LTP tissue were found to be significantly smaller than control concave synapses. There was also a significant increase in the number of perforated concave synapses which exceeded the overall increase in concave synapses, and occurred despite the lack of a general increase in perforated synapses. It was concluded that this specific structural profile, observed at 24 h postinduction, may help support the potentiated response observed at this stage of LTP maintenance.

Changes in field potentials and membrane currents in rat sensorimotor cortex following repeated tetanization of the corpus callosum in vivo.

Repeated, daily tetanization of the corpus callosum induces lasting changes in sensorimotor cortex field potential responses, but the synaptic populations that mediate these responses and support long-term potentiation (LTP) have not been characterized. Current source density analyses of field responses were compared between control animals and those in which LTP was induced by 10 daily series of tetanizations. Tetanization and paired-pulse stimulation (100 ms interval) enhanced the duration of initial (approximately 3 ms onset) deep-negative population spike activity generated by a current sink in layer V that peaked repeatedly at a frequency of approximately 400 Hz. The early (approximately 10 ms to peak) surface-negative component of field responses was generated by a current sink in upper layer V and a source in layer VI. This monosynaptic component followed high stimulation frequencies, recovered quickly from the effects of anaesthesia, and was enhanced by both tetanization and paired-pulse stimulation. The late (approximately 20 ms to peak) surface-negative component was generated by a sink in upper layer V and a source deep in layer V, and was greatly enhanced by tetanization and paired-pulse stimulation. The late component did not follow high-frequency stimulation and recovered slowly from anaesthesia, suggesting that it is driven polysynaptically. Potentiation of monosynaptic thalamic and cortico-cortical afferents probably mediates enhancements of the early component and population spikes, while potentiation of polysynaptic afferents to layer V may contribute to growth in the late component.

The degree of potentiation is associated with synaptic number during the maintenance of long-term potentiation in the rat dentate gyrus.

There is a considerable degree of variation in the amount of potentiation induced in different animals following the induction of long-term potentiation (LTP). This variation provided us with the opportunity to determine what types of synaptic changes were dependent upon the degree of induced potentiation. To examine possible 'degree of potentiation' effects on synapses, we conducted a multiple regression analysis examining the relationship between the degree of potentiation in LTP animals and a series of synaptic structural measures. We examined synapses in the middle third of the molecular layer (MML) of the rat dentate gyrus following repeated high frequency tetanization of the perforant path. LTP was induced over a 4 h period, and the animals were sacrificed 24 h after the final stimulation. Synapses from the ipsilateral inner third of the dentate molecular layer (IML) and from implanted only animals were also examined for comparison. Ultrastructural quantification included the total number of synapses per neuron, synaptic curvature, the presence of synaptic perforations, and the maximum length of the synaptic apposition. The only structural change that was significantly associated with the degree of potentiation was a positive correlation between the degree of LTP and the number of synapses per neuron. Therefore, synaptic number, while not appearing to be significantly associated with the induction of LTP, appears to be important for the degree of LTP expressed.

Skilled forelimb movements in prey catching and in reaching by rats (Rattus norvegicus) and opossums (Monodelphis domestica): relations to anatomical differences in motor systems.

Traditional anatomical/behavioral classifications suggest that rats and opossums have simple motor systems and are impoverished with respect to their ability to make prehensile movements. Nevertheless, the motor system in rats and opossums represent extremes in relative size and complexity suggesting that a behavioral analysis of the movement competencies of these species will provide insights into the significance of such anatomical differences. This paper examines the movements that the two species use in catching crickets and in reaching for food items. Both species could use a single limb to reach out and grasp prey during prey catching and both could use a single limb to take food from a shelf. Both species could transport the food to the mouth by using a single paw. The food handling behavior of the rats was more complex than that of the opossums, however. They used a variety of prey catching movements and extensively manipulated the prey to remove the legs and wings before eating only the head and body. Additionally the rats made rotatory limb movements of aiming, pronation, and supination, when reaching. For both cricket catching and reaching, they used their digits more than did the opossums. The suggestion also emerged from the results that the movements of the opossums were more fixed and species-typical whereas those of the rats were more plastic and individualistic. Thus, the skilled movements of both species are more complex than is generally recognized and the greater complexity of the rat movements parallels their more complex motor system. These results are discussed in relation to anatomical differences in the motor system and, specifically, to differences in the terminal fields of the pyramidal tract. It is concluded that the motor abilities of nonprimate mammals have been vastly underrated.

Mouse NGF promoter upstream sequences do not affect gene expression in mouse fibroblasts.

The expression of nerve growth factor (NGF) is tightly controlled in a tissue-specific manner during development and in response to injury. In fibroblasts and in other cell types, expression of NGF is regulated at the transcriptional level. In order to elucidate the mechanism of this regulation, we have undertaken the analysis of the mouse NGF promoter in a mouse fibroblast cell line (LTA), using transient transfection of NGF promoter-human growth hormone (hGH) reporter gene plasmids. We find that sequences between +8bp and +120bp, containing an AP-1 site, confer increased levels of expression from the full length and truncated NGF promoters. When this region is deleted, a significant decrease in expression is observed from both the full length promoter and truncated versions thereof. A gradual increase in expression is observed with successive 5' deletions of both the AP-1 containing and AP-1 deleted promoters; this effect results from the juxtapositioning of adjacent plasmid sequences closer to the transcription initiation site and not from deletion of promoter sequences as was previously reported. When the NGF promoter is analyzed using a luciferase reporter plasmid, these 5' promoter deletions have no significant effect on reporter gene expression in fibroblasts. Thus, sequences downstream of the transcription start site influence NGF promoter activity in fibroblasts, but sequences upstream of the TATA box fail to affect promoter activity in these cells.