Barbara Fischer

Complement C1q and C3 mRNA expression in the frontal cortex of Alzheimer's patients

The levels and cellular localization of mRNA for complement C1q and C3 were examined by RNA gel blot and nonradioactive in situ hybridization in the frontal cortex of patients with Alzheimers disease (AD) and age-matched controls. We found that the hybridization signal for C1q mRNA was markedly increased (approx. 3.5-fold) in the frontal cortex of AD patients compared to that in age-matched controls. In contrast to previous reports we also found that the levels of C3 mRNA, although well expressed, did not differ significantly between AD cases and age-matched controls. Nonradioactive in situ hybridization using digoxigenin-labeled riboprobes revealed that transcripts coding for both C1q and C3 were closely associated with neurons. These results support the hypothesis that complement could play a role in neuronal degeneration which has been observed in the brain of AD patients.

Sarcopenic obesity: molecular clues to a better understanding of its pathogenesis?

Abstract An age-dependent decline in skeletal muscle mass, strength, and endurance during the aging process is a physiological development, but several factors may exacerbate this process, leading to the threatening state of sarcopenia, frailty, and eventually higher mortality rates. Obesity appears to be such a promoting factor and has been linked in several studies to sarcopenia. The reason for this causal association remains poorly understood. Notwithstanding the fact that a higher body mass might simply lead to diminished physical activity and therefore contribute to a decline in skeletal muscle, several molecular mechanisms have been hypothesized. There could be an obesity derived intracellular lipotoxicity (i.e., elevated intramuscular levels of lipids and their derivatives), which induces apoptosis by means of an elevated oxidative stress. Paracrine mechanisms and inflammatory cytokines, such as CRP and IL-6 could be confounders of the actual underlying pathological mechanism. Due to a cross-talk of the hypothalamo-pituitary axis with nutritional status, obese subjects are more in a catabolic state of metabolism, with a higher susceptibility to muscle wasting under energy restriction. Obesity induces insulin resistance in the skeletal muscle, which consequently leads to perturbed metabolism, and misrouted signaling in the muscle cells. In obesity, muscle progenitor cells could differentiate to an adipocyte-like phenotype as a result of paracrine signals from (adipo)cytokines leading to a reduced muscular renewal capacity. The present review outlines current knowledge concerning possible pathways, which might be involved in the molecular pathogenesis of sarcopenic obesity.

Seizure induced c-fos mRNA in the rat brain: comparison between young and aging animals

The molecular mechanisms associated with age-related alterations in the pharmacological and physiological properties of hippocampal and cortical neurons in response to chemically induced seizure are largely unknown. Administration of pentylenetetrazole (PTZ) (50 mg/kg body weight) to rats of various ages evoked tonic-colonic seizures. Using RNA gel blot analysis we found that 1 h after the onset of seizure, the mRNA for the protooncogene c-fos was increased in the hippocampus and cortex of 3-month-old rats. The levels of c-fos mRNA in the hippocampus and cortex of 3-month-old rats returned to control levels by 3 h after PTZ administration. The levels of c-fos mRNA in the hippocampus and cortex of 20-month-old and 30-month-old rats peaked at 3 h and returned to basal levels by 15 h following PTZ treatment. These results suggest that the induction of immediate-early gene expression, as exemplified by c-fos, is not impaired in the aged rat brain. However, the aged rat brain responded more slowly to chemically induced seizure and the levels of c-fos mRNA induction are decreased by about 49% in the cortex and by 27% in the hippocampus of 30-month-old rats, as compared to the levels expressed by 3-month-old rats.

Impaired Postprandial Response of Active Ghrelin and Prolonged Suppression of Hunger Sensation in the Elderly

BACKGROUND The role of the orexigenic hormone ghrelin is of major interest in the altered appetite regulation of the elderly. METHODS Basal and postprandial levels of active and total ghrelin were measured in 15 younger (mean age 35.4 years) and 19 older (80.7 years) participants following a carbohydrate-rich test meal. RESULTS Our results showed that older participants felt postprandially less hungry and more full. Although basal levels were not significantly different, active and total ghrelin levels declined postprandially only in the younger study participants. Highly significant differences between the two age groups were shown for the changes of the area under the curve for active ghrelin (p = .024). CONCLUSIONS Our study demonstrates for the first time that differences in hunger and satiety sensations in relation to age are paralleled by a substantially different response of acylated and total ghrelin, that is, the absence of a postprandial decline in ghrelin levels.

Gender-specific differences in the development of sarcopenia in the rodent model of the ageing high-fat rat

Sarcopenia is linked to functional impairments, loss of independence, and mortality. In the past few years, obesity has been established as being a risk factor for a decline in muscle mass and function. There are several molecular pathological mechanisms, which have been under discussion that might explain this relationship. However, most studies were conducted using male animals and for a short period of time.

Pentylenetetrazole-induced seizure up-regulates levels of microtubule-associated protein 1B mRNA and protein in the hippocampus of the rat

Abstract: Stimuli that evoke seizure are capable of inducing structural changes in the hippocampus. However, late‐acting genes related to these changes have not been described. Administration of pentylenetetrazole (PTZ; 50 mg/kg) to rats of various ages evoked tonic‐clonic seizures. Using RNA gel blot analysis we found that the level of the mRNA for microtubule‐associated protein 1B (MAP1B) was robustly increased in the hippocampus of 3‐month‐old rats. The levels of MAP1B mRNA in hippocampus peaked at 40 h and began to decline by 72 h following PTZ treatment. Immunoblotting with anti‐MAP1B antibody demonstrates the increase in content of immunoreactive proteins 40–72 h after seizure onset in the hippocampus of PTZ‐treated rats. These results indicate that MAP1B is a sensitive indicator of hippocampal structural changes occurring in response to PTZ‐induced seizure activity.

Anomalous expression of microtubule-associated protein 1B in the hippocampus and cortex of aged rats treated with pentylenetetrazole

The aim of the present study was to assess the age-dependent response of microtubule-associated protein 1B, a plasticity-associated protein deriving from a late gene, following administration of an epileptogenic stimulus. The effect of a single administration of the convulsant pentylenetetrazole on microtubule-associated protein 1B expression in the hippocampal formation and cortex of three-, 18- and 28-month-old rats was assessed using northern blot analysis, in situ hybridization and immunohistochemistry. In three-month-old rats, we detected initial increases in microtubule-associated protein 1B messenger RNA at 15 h following pentylenetetrazole administration in the granule cells of the dentate gyrus, in the CA3 region of the hippocampus and in layers II/III of the entorhinal cortex, and these reached a maximum at 44 h. However, in the hippocampus and cortex of 18-month-old rats, the peak occurred at 15 h, and in the brains of 28-month-old rats a blunted peak was reached at 3 h. Pentylenetetrazole treatment in young rats resulted in a robust induction of microtubule-associated protein 1B immunoreactivity in the granule cells of the dentate gyrus and in layers II/III of the entorhinal cortex, but also produced a large decrease in the retrosplenial cortex. However, following pentylenetetrazole treatment in older rats, the granule cells of the dentate gyrus were nearly devoid of microtubule-associated protein 1B immunoreactivity, whereas the retrosplenial cortex showed no changes at all, and the entorhinal cortex had an expression pattern similar to that of young rats. Aberrant immunolabeling of microtubule-associated protein 1B occurred in cortical layer VI of the aged rats where, unlike in young rats, there was heavy staining of neuronal somata. These results suggest that the regulation of the plasticity-associated protein microtubule-associated protein 1B is altered in the ageing rat brain, with the peak of expression shifted to earlier times in 18-month-old rats and blunted, variable increases at even earlier times in 28-month-old rats.

Dynamics of gene expression for immediate early- and late genes after seizure activity in aged rats

The ability of the rodent brain to support plasticity-related phenomena declines with increasing age. A decreased coordination of genes implicated in brain plasticity may be one factor contributing to this decline. Synaptic rearrangement that occurs after seizure activity is regarded as a model of brain plasticity. In a rat model of seizure-related brain plasticity, we found that the induction of immediate-early genes, as exemplified by c-fos and tissue plasminogen activator ( tPA), is not impaired in the aged rat brain. However, the aged rat brain responded more slowly to chemically induced seizure, and the levels of c-fos and tPA mRNAs induction are decreased in the cortex and in the hippocampus of 30 month old rats, as compared to the levels expressed by 3 month old rats. In addition, at the peak induction, the TPA transcripts were restricted to certain cortical layers of the older rats. Surprisingly, in applying the same experimental paradigm to late genes, we found that there was a shift toward earlier times in the maximum expression of growth-related molecules, the microtubule-associated protein 1B (MAP1B) mRNA, which was very evident in 18 month old rats. Aberrant immunolabeling of MAP1B occurred in cortical layer VI of the aged rats where, unlike in young rats, there was heavy staining of neuronal somata. These results suggest that (1) one consequence of aging, besides decreases in the levels of mRNA, is a progressive loss of coordination in gene activity following the administration of a stimulus; (2) since c-fos, TPA and MAP1B have been implicated in neuronal plasticity, these findings could explain, in part, the limited plasticity of the aging brain.

V+ fibronectin mRNA is increased in the brains of aged rats: effect of food restriction

Food restriction increases life-span in rodents. With regard to the central nervous system, underfeeding has been shown to have beneficial effects on synaptic transmission in old rats. However, the molecular events underlying functional changes in the brains of food-restricted rats are largely unknown. In the present study the levels of fibronectin mRNA containing the alternatively spliced segment V (FN-V+) as well as the levels of FN mRNA containing the alternatively spliced segment EIIIB (FN-EIIIB+), were examined by RNA gel blot hybridization in the brains of 3-day-old, 24-day-old, 10-month-old, 18-month-old, 30-month-old ad libitum (AL) fed, 30-month-old food-restricted (FR), and 35-37-month-old, FR rats. The hybridization signal for the FN-EIIIB+ mRNA was relatively abundant at early postnatal stages but very few transcripts were detected in the brains of adult, AL rats. The transcripts coding for FN-V+ mRNA were moderately expressed in the brains of 3-day-old, 24-day-old, 10-month-old, and 18-month-old rats. However, the FN-V+ mRNA signal was then prominently increased (approx. 3-fold) in the brains of the 30-month-old, AL rats vs. 10-month-old, AL rats, and further increased (approx. 2-fold) in the brains of 30-month-old, FR, as compared to 30-month-old, AL rats. However, the levels of FN-V+ mRNA were slightly decreased in the brains of very old (35-37-month) FR rats vs. 30-month-old FR rats. The distribution of fibronectin messenger RNA and protein was also investigated by non-radioactive in situ hybridization and immunohistochemistry, respectively. The most prominent expression of FN-V+ messenger RNA was seen in neurons of the hippocampus, including the granule cells of the dentate gyrus, and in layers III and V of the cortex of 30-month-old, FR rats. FN immunostaining closely paralleled the distribution of FN mRNA and was confined to the neuronal cell periphery. The upregulation of fibronectin gene expression upon exposure to glucocorticoids is well documented. Prolonged food restriction, acting as a stress factor, combined with decreased plasticity of glucocorticoid regulatory responses in the aged rats could cause an increase in the levels of FN mRNA and protein in the brains of old, FR rats. Since FN has been shown to provide an adhesive substrate for extending neurites, we conclude that food restriction may potentiate synaptic plasticity, via glucocorticoid receptor binding elements of the FN gene, in the brains of old rats.

Altered expression of microtubule-associated protein 1B in cerebral cortical structures of pentylenetetrazole-treated rats

Using Northern blot, immunoblotting, immunocytochemistry, and in situ hybridization, we show that a single administration of the convulsant pentylenetetrazole leads to robust, long‐term changes in microtubule‐associated protein 1B and its mRNA, in the adult rat brain. The first increases in MAP1B mRNA were detected at 15 hr following pentylenetetrazole administration in the temporal (Te2) and perirhinal cortex followed by increases in microtubule‐associated protein 1B immunoreactivity at 72 hr postseizure. In contrast, the levels of microtubule‐associated protein 1B mRNA and protein in layers I–II of the retrosplenial and parietal cortex (Par2) declined visibly by 24 hr and 72 h, respectively, post‐seizure. The changes included loss of staining in layers I–II and development of structures resembling “strings‐of‐beads” along the fibers of projection neurons of layer V. The levels of microtubule‐associated protein 1B mRNA in the entorhinal cortex peaked at later times (72 h), especially in layers II–III, and returned to control levels by 10 days. Whereas the levels of microtubule‐associated protein 1B immunoreactivity in the retrosplenial and parietal cortex recovered by 5–10 days, it persisted at high levels through day 35 in layer V of the temporal cortex (Te2), layers II–III of the perirhinal cortex and layers I–II of the lateral entorhinal cortex. These results indicate that seizure activity leads to long‐term upregulation of genes coding for structural elements that are characteristic of the immature brain such as microtubule‐associated protein 1B. J. Neurosci. Res. 51:646–657, 1998. © 1998 Wiley‐Liss, Inc.