Differential effects of aging on hippocampal ultrastructure in male vs. female rats.

Age-related decline in physical and cognitive functions are facts of life that do not affect everyone to the same extent. We had reported earlier that such cognitive decline is both sex- and context-dependent. Moreover, age-associated ultrastructural changes were observed in the hippocampus of male rats. In this study, we sought to determine potential differences in ultrastructural changes between male and female rats at various stages of life. We performed quantitative electron microscopic evaluation of hippocampal CA1 region, an area intimately involved in cognitive behavior, in both male and female adolescent, adult and old Wistar rats. Specifically, we measured the number of docking synaptic vesicles in axo-dendritic synapses, the length of active zone as well as the total number of synaptic vesicles. Distinct age- and sex-dependent effects were observed in several parameters. Thus, adult female rats had the lowest synaptic active zone compared to both adolescent and old female rats. Moreover, the same parameter was significantly lower in adult and old female rats compared to their male counterparts. On the other hand, old male rats had significantly lower number of total synaptic vesicles compared to both adolescent and adult male rats as well as compared to their female counterparts. Taken together, it may be suggested that age- and sex-dependent ultrastructural changes in the hippocampus may underlie at least some of the differences in cognitive functions among these groups.

Propionic acid affects the synaptic architecture of rat hippocampus and prefrontal cortex.

It is well documented that propionic acid (PPA) produces behavioral, morphological, molecular and immune responses in rats that are characteristic of autism spectrum disorder in humans. However, whether PPA affects the ultrastructure and synaptic architecture of regions of autistic brain has not been adequately addressed. Earlier we show that single intraperitoneal (IP) injection of PPA (175 mg/kg) produces superficial changes in the spatial memory and learning of adolescent male Wistar rats. However, in neurons, synapses and glial cells of hippocampal CA1 area and medial prefrontal cortex transient (mainly) or enduring alterations were detected. In this study, we used electron microscopic morphometric analysis to test the effect of PPA on different structural parameters of axodendritic synapses of the hippocampus and prefrontal cortex. The animals were treated with a single IP injection of PPA (175 mg/kg). The length and width of synaptic active zone, the area of presynaptic and postsynaptic mitochondria, the distance between presynaptic mitochondria and the synapse active zone, the distance between postsynaptic mitochondria and postsynaptic density and the depth and opening diameter of neuronal porosome complex were evaluated. Our results show that synaptic mitochondria of the hippocampus and prefrontal cortex are the most vulnerable to PPA treatment: in both regions, the area of postsynaptic mitochondria were increased. In general, our results show that even small dose of PPA, which produces only superficial effects on spatial memory and learning is able to alter the synapse architecture in brain regions involved in cognition and autism pathogenesis. Therefore, the microbiome may be involved in the control of neurotransmission in these regions.

Short- and long-term effects of chronic toluene exposure on spatial memory in adolescent and adult male Wistar rats.

Addiction to toluene-containing volatile inhalants is of significant medical and social concern, particularly among youth. These concerns are underscored by the fact that the majority of adult abusers of toluene started as teenagers. Surprisingly, however, the lasting effects of chronic toluene exposure, especially in various age groups, have not been well investigated. Recently, we reported that adolescent and adult male Wistar rats show differential responses to chronic toluene exposure in recognition memory tasks. Since different cognitive functions may be differentially affected by drugs of abuse, we used the same model to evaluate the short- and long-term effects of chronic toluene on spatial learning and memory using Morris water maze. Daily exposure to toluene (2000 ppm) for 40 days (5 min/day) resulted in age-dependent behavioral changes. For example, only adolescent animals showed a decrease in time and distance travelled to find the hidden platform 24 h after the last toluene exposure. In contrast, only adult rats exhibited a decrease in acquisition time and distance travelled at 90 days' post toluene exposure. Our data provide further support for the contention that age-dependent responses should be taken into consideration in interventional attempts to overcome specific detrimental consequences of chronic toluene exposure.

Anxiety and ultrastructural consequences of chronic mild stress in rats.

Detrimental consequences following exposure to severe stress, either acute or chronic are well recognized. Chronic mild stress (CMS) is also a leading cause of emotional distress and neuropsychiatric conditions such as anxiety disorders. However, the neurobiological substrates of the latter, particularly at the ultrastructural levels have not been adequately investigated. In this study, adult male Wistar rats were subjected to 4 h daily mild restraint for 20 days and their behavior in open field and elevated plus maze (EPM) were evaluated 24 h after the last restraint. Anxiety-like behavior was evident in CMS exposed rats by increases in rearing and grooming in the open field and the avoidance of open arms in the EPM. Concomitant ultrastructural alterations such as chromatolysis, agglutination of synaptic vesicles or mitochondrial damage were also observed in the central nucleus of amygdala (CNA), an area intimately involved in emotional and fear response, in CMS exposed rats. These results while confirming detrimental consequences of CMS, also suggest that ultrastructural alterations in CNA may be a basis for CMS-induced anxiety.

Short- and long-term effects of chronic toluene exposure on recognition memory in adolescent and adult male Wistar rats.

Abuse of toluene-containing volatile inhalants, particularly among youth, is of significant medical and social concern worldwide. Teenagers constitute the most abundant users of toluene and the majority of adult abusers of toluene started as teenagers. Although the euphoric and neurotoxic effects of acute toluene have been widely studied, lasting effects of chronic toluene exposure, especially in various age groups, have not been well investigated. In this study, we used adolescent and adult male Wistar rats to evaluate the short- and long-term effects of chronic toluene on various behaviors including cognitive function. Daily exposure to toluene (2000 ppm) for 40 days (5 min/day) resulted in age-dependent behavioral impairments. Specifically, adolescent animals showed recognition memory impairment the day after the last exposure, which had normalized by day 90 post- exposure, whereas such impairment in adult animals was still evident at day 90 post-exposure. Our data suggest that age-dependent responses should be taken into consideration in interventional attempts to overcome specific detrimental consequences of chronic toluene exposure.

Aging affects cognition and hippocampal ultrastructure in male Wistar rats.

It is now well established that aging is associated with emotional and cognitive changes. Although the basis of such changes is not fully understood, ultrastructural alterations in key brain areas are likely contributing factors. Recently, we reported that aging-related anxiety in male Wistar rats is associated with ultrastructural changes in the central nucleus of amygdala, an area that plays important role in emotional regulation. In this study, we evaluated the cognitive performance of adolescent, adult, and aged male Wistar rats in multi-branch maze (MBM) as well as in Morris water maze (MWM). We also performed ultrastructural analysis of the CA1 region of the hippocampus, an area intimately involved in cognitive function. The behavioral data indicate significant impairments in few indices of cognitive functions in both tests in aged rats compared to the other two age groups. Concomitantly, a total number of presynaptic vesicles as well as vesicles in the resting pool were significantly lower, whereas postsynaptic mitochondrial area was significantly higher in aged rats compared to the other age groups. No significant differences in presynaptic terminal area or postsynaptic mitochondrial number were detected between the three age groups. These results indicate that selective ultrastructural changes in specific hippocampal region may accompany cognitive decline in aging rats.

Electron microscopy demonstrating noise exposure alters synaptic vesicle size in the inferior colliculus of cat.

CONTEXT: White noise is known to have detrimental effects on different brain regions, especially auditory regions, including inferior colliculus. Although the basis for such alterations has been hypothesized to result from abnormalities in neurotransmitter release, the mechanism is unclear. The final step in neurotransmission is the docking and transient fusion of synaptic vesicles at the base of cup-shaped lipoprotein structures called porosomes at the presynaptic membrane and the consequent release of neurotransmitters. Earlier studies in cat brain document altered morphology of the secretory portal the porosome at nerve terminals in the inferior colliculus following white noise exposure. The current study was performed to test the hypothesis of possible changes to synaptic vesicle size in the colliculus, following white noise exposure. MATERIAL AND METHODS: Electron microscopic morphometry of synaptic vesicles size in axo-dendritic synapses at the colliculus region of the cat brain was performed. RESULTS: We report, for first time, decreased size of both docked and undocked vesicles in high-intensity white noise-exposed animals. In both control and experimental animals, docked vesicles are demonstrated to be smaller than undocked vesicles, suggesting fractional discharge of vesicular contents via porosome-mediated kiss-and-run mechanism. CONCLUSION: These studies advance our understanding of neurotransmitter release and the impact of white noise on brain function.

Age-related cognitive decline in rats is sex and context dependent.

Previously, we had observed age-related cognitive decline in male rats compared to adolescent and adult rats. This was shown in both a multi-branched maze test (MBM), as well as in the Morris water maze test (MWM). In the present study, we compared the behavior of similar age groups in both male and female rats using the same paradigms. The results confirmed the increase in errors and time spent in MBM in aged male rats compared to other age groups. However, no such differences were observed in female rats. In the acquisition phase of MWM, aged male rats did not differ significantly from the other two groups in terms of time spent in quadrants, whereas aged female rats spent significantly more time in quadrants compared to the other 2 age groups. Aged male rats also travelled significantly more than the other 2 age groups during the acquisition phase, whereas no such differences were observed in female rats. In both short term (30 min post acquisition) and long term (24 h after acquisition) retrieval phases of MWM, significant gender-related differences were also observed in all age groups. These findings suggest gender- and context-dependent alterations in cognitive functions during aging.

Age-related behavioral and ultrastructural changes in the rat amygdala.

Although the relationships between brain structure and emotions may alter across the life span, this relationship is of particular importance during aging when significant alterations in emotions may be manifested. Understanding the structural-behavioral relationship could not only provide a neurobiological basis of these changes, but could also suggest potential intervention. Since anxiety is commonly observed in aging population, we undertook this study to determine the extent of this behavioral manifestations as well as the associated ultrastructural changes in the amygdala. Rats of various age groups, adolescent, adult, and aged were tested for anxiety-like behavior and the ultrastructure/presynaptic architecture of the central nucleus of amygdala (CNA) were evaluated using transmission electron microscopy (EM). Aged rats were consistently more anxious than the other groups as evidenced by their scores in the elevated plus maze. Morphometric EM analysis of axodendritic synapses revealed that the aged rats had a lower presynaptic area as well as number of synapses, but unexpectedly a higher number of presynaptic mitochondria in CNA. Since presynaptic mitochondria are known to provide the energy for neurotransmission, it may be concluded that compensatory mechanisms are still operational during aging, and hence, may be a target for therapeutic intervention at this stage of life span.