Decreased expression of ErbB4 and tyrosine hydroxylase mRNA and protein in the ventral midbrain of aged rats

Decreased availability or efficacy of neurotrophic factors may underlie an increased susceptibility of mesencephalic dopaminergic cells to age-related degeneration. Neuregulins (NRGs) are pleotrophic growth factors for many cell types, including mesencephalic dopamine cells in culture and in vivo. The functional NRG receptor ErbB4 is expressed by virtually all midbrain dopamine neurons. To determine if levels of the NRG receptor are maintained during aging in the dopaminergic ventral mesencephalon, expression of ErbB4 mRNA and protein was examined in young (3 months), middle-aged (18 months), and old (24–25 months) Brown Norway/Fischer 344 F1 rats. ErbB4 mRNA levels in the substantia nigra pars compacta (SNpc), but not the adjacent ventral tegmental area (VTA) or subtantia nigra pars lateralis (SNl), were significantly reduced in the middle-aged and old animals when compared to young rats. Protein expression of ErbB4 in the ventral midbrain was significantly decreased in the old rats when compared to the young rats. Expression of tyrosine hydroxylase (TH) mRNA levels was significantly reduced in the old rats when compared to young animals in the SNpc, but not in the VTA or SNI. TH protein levels in the ventral midbrain were also decreased in the old animals when compared to the young animals. These data demonstrate a progressive decline of ErbB4 expression, coinciding with a loss of the dopamine-synthesizing enzyme TH, in the ventral midbrain of aged rats, particularly in the SNpc. These findings may implicate a role for diminished NRG/ErbB4 trophic support in dopamine-related neurodegenerative disorders of aging such as Parkinson's disease.     

Vasopressin and renin response to dehydration in aged rats

Abnormalities in neurohypophyseal function have been postulated to contribute to the alterations in fluid and electrolyte balance observed during aging. In this study, parameters of fluid and electrolyte balance were evaluated during chronic water deprivation in old (30 months) and young (3 months) Fischer 344 rats. The increases in serum vasopressin (VP) and renin concentrations observed in the 3 month animals following chronic water deprivation were absent in the aged rats (p<0.05 and p<0.02, respectively). This occurred in spite of apparently comparable alterations in fluid volume and osmolality (assessed by changes in body weight, hematocrit and plasma osmolality). Relative to body weight, VP content of the neural lobe was significantly reduced and was more severely depleted by dehydration in aged rats than in young rats. Thus, inadequate neurohypophyseal hormone stores may contribute to the inability of the aged animals to attain elevated serum VP concentrations during chronic stimulation. Several parameters of renal function were examined in the aged rats. Although none of the old rats were in renal failure, they all showed some indication of reduced renal function. In spite of renal abnormalities including reduced concentrating capabilities, the old rats did demonstrate a significant antidiuretic response to dehydration. However, with prolonged fluid deprivation, they were unable to attain serum VP or renin concentrations comparable to that achieved by the young rats.     

Expression profiling identifies dysregulation of myosin heavy chains IIb and IIx during limb immobilization in the soleus muscles of old rats

Aged individuals suffer from multiple dysfunctions during skeletal muscle atrophy. The purpose of this study was to determine differential changes in gene expression in atrophied soleus muscle induced by hindlimb immobilization in young (3–4 months) and old (30–31 months) rats. The hypothesis was that differentially expressed mRNAs with age-atrophy interactions would reveal candidates that induce loss of function responses in aged animals. Each muscle was applied to an independent set of Affymetrix micoarrays, with 385 differentially expressed mRNAs with atrophy and 354 age-atrophy interactions detected by two-factor ANOVA (α of 0.05 with a Bonferroni adjustment). Functional trends were observed for 23 and 15 probe sets involved in electron transport and the extracellular matrix, respectively, decreasing more in the young than in the old. Other functional categories with atrophy in both ages included chaperones, glutathione-S-transferases, the tricarboxylic acid cycle, reductions in Z-line-associated proteins and increases in probe sets for protein degradation. Surprisingly, myosin heavy chain IIb and IIx mRNAs were suppressed in the atrophied soleus muscle of old rats as opposed to the large increases in the young animals (16- and 25-fold, respectively, with microarrays, and 61- and 68-fold, respectively, with real-time PCR). No significant changes were observed in myosin heavy chain IIb and IIx mRNA with micoarrays in the atrophied soleus muscles of old rats, but they were found to increase six- and fivefold, respectively, with real-time PCR. Therefore, deficiencies in pre-translational signals that normally upregulate myosin heavy chain IIb and IIx mRNAs during atrophy may exist in the soleus muscle of old animals.     

Vasopressin and renin response to dehydration in aged rats

Abnormalities in neurohypophyseal function have been postulated to contribute to the alterations in fluid and electrolyte balance observed during aging. In this study, parameters of fluid and electrolyte balance were evaluated during chronic water deprivation in old (30 months) and young (3 months) Fischer 344 rats. The increases in serum vasopressin (VP) and renin concentrations observed in the 3 month animals following chronic water deprivation were absent in the aged rats (p<0.05 and p<0.02, respectively). This occurred in spite of apparently comparable alterations in fluid volume and osmolality (assessed by changes in body weight, hematocrit and plasma osmolality). Relative to body weight, VP content of the neural lobe was significantly reduced and was more severely depleted by dehydration in aged rats than in young rats. Thus, inadequate neurohypophyseal hormone stores may contribute to the inability of the aged animals to attain elevated serum VP concentrations during chronic stimulation. Several parameters of renal function were examined in the aged rats. Although none of the old rats were in renal failure, they all showed some indication of reduced renal function. In spite of renal abnormalities including reduced concentrating capabilities, the old rats did demonstrate a significant antidiuretic response to dehydration. However, with prolonged fluid deprivation, they were unable to attain serum VP or renin concentrations comparable to that achieved by the young rats.     

Vasopressin and oxytocin excretion in the Brown-Norway rat in relation to aging, water metabolism and testosterone

There is considerable disagreement in the literature on changes in the hypothalamo-neurohypophyseal system (HNS) with aging: some reports support HNS degeneration, whereas others claim an activation of this system in senescence. In order to study age-related changes in vasopressin (VP) and oxytocin (OT) excretion in relation to water metabolism, six young (4 months) and 12 aged (34 months) male Brown-Norway rats were placed in metabolism cages. Since plasma testosterone levels have been reported to affect HNS activity and to decline progressively with age, half of the aged animals were given subcutaneous testosterone implants. Urine volume and water intake were significantly increased in aged animals, while urine osmolality was significantly reduced. These changes could not be attributed to diminished VP secretion, since 24-h urinary excretion of this peptide was elevated in the aged animals. In addition, 24-h OT excretion was elevated in the aged animals, indicating an overall activation of the HNS in senescence. VP excretion was significantly correlated with urine osmolality, urine volume and urinary VP concentration. No significant differences were observed between testosterone- and sham-implanted aged rats. It is concluded that the moderate polyuria/polydipsia in the senescent Brown-Norway rat is probably due to renal changes and is accompanied by a compensatory rise in both VP and OT secretion. Testosterone does not affect these changes.     

老年大鼠血液白细胞基因表达谱的生物信息学分析

目的了解老年大鼠血液白细胞基因表达水平的变化。方法将雄性Wister大鼠分为老年组（18月龄）和年轻组（4月龄），用illumina大鼠全基因组芯片研究其分子水平的变化。结果生物信息学分析显示．造成老年组和年轻组显著生物学差异的基因功能及所属基因是：28条凋亡基因和22条炎性基因上调，9条涉及氧与活性氧中间物代谢的基因下调。结论差异基因功能导致老年大鼠和年轻大鼠的生物学差异。     

Treatment with dehydroepiandrosterone (DHEA) stimulates oxidative energy metabolism in the cerebral mitochondria. A comparative study of effects in old and young adult rats

The content of the neurosteroids, dehydroepiandrosterone (DHEA) in the brain decreases with aging. Also the oxidative energy metabolism is known to decrease with aging. Hence we examined the effects of treatment with DHEA (0.2 or 1.0 mg/kg body weight for 7 days) on oxidative energy metabolism in brain mitochondria from old and young adult rats. State 3 respiration rates in brain mitochondria from old animals were considerably lower than those in young adults. Treatment with DHEA stimulated state 3 and state 4 respiration rates in both the groups of the animals in a dose-dependent manner. In the old rats following DHEA treatment, the state 3 respiration rates became comparable to or increased beyond those of untreated young adults. In contrast to the old rats, stimulatory effect of DHEA treatment was of greater magnitude in the young adults. However, at higher dose (1.0 mg) the effect declined. Cytochrome aa3 content in the brain mitochondria from old rats was significantly low but the content of cytochrome b was unchanged while the content of cytochromes c+c1 had increased. Treatment with DHEA increased the content of cytochrome aa3 and b in old as well as in young adult animals. Higher dose of DHEA (1.0 mg) had adverse effect on the content of cytochrome c+c1. DHEA treatment stimulated ATPase activity in a dose-dependent manner in young adult rats whereas in the old rats the effect on ATPase activity was marginal. Dehydrogenases activities were somewhat lower in the old rats. DHEA treatment stimulated mitochondrial dehydrogenases activities in both the groups. Results of our studies suggest that judicious use of DHEA treatment can improve oxidative energy metabolism parameters in brain mitochondria from young adult as well as old rats.     

Expression of c-Fos in response to stressogenic stimuli in the amygdala of old vs. young rats--a preliminary study

During aging many of the brain functions became deteriorated or altered. One of the most important age related changes is an increase of anxiety level, reported both in humans and in animals. Our study was intended to compare c-fos gene expression in amygdala, the key structure in anxiety/fear regulation, in old (24 months) and young (4 months) rats exposed to various behavioral stimulations. There were no differences between age groups in basal c-Fos expression. After social encounter c-Fos expression level in amygdala increased significantly, but still remained independent on age. Significant differences between both groups appeared after open field test and immobilization test. Contrary to the findings on young adults indicating the correlation between increased anxiety level and higher c-Fos expression, old rats showed increased anxiety together with significantly lower c-Fos expression.     

Androgenic regulation of oxidative stress in the rat prostate: involvement of NAD(P)H oxidases and antioxidant defense machinery during prostatic involution and regrowth

Little is known about the roles of androgens in the regulation of redox state in the prostate, a cellular process believed to profoundly influence normal and aberrant prostate functions. We demonstrate that castration induced discrete oxidative stress (OS) in the acinar epithelium of rat ventral prostate (VP), as evident from marked increases in 8-hydroxy-2'-deoxy-guanosine and 4-hydroxynonenal protein adducts in the regressing epithelium. Testosterone replacement partially reduced OS in VP epithelia of castrates, but the level remained higher than in intact rats. Quantification of steady-state mRNA levels of 14 genes involved in the anabolism and catabolism of reactive oxygen species (ROS) showed that castration resulted in dramatic increases of three ROS-generating NAD(P)H oxidases (Noxs) including Nox1, gp91(phox), and Nox4, significant reductions of key ROS-detoxifying enzymes (superoxide dismutase 2, glutathione peroxidase 1, thioredoxin, and peroxiredoxin 5), and unchanged levels of catalase, glutathione reductase, gamma-glutamyl transpeptidase, and glutathione synthetase. Testosterone replacement in castrated rats partially reduced expression of Noxs but restored expression of superoxide dismutase 2, glutathione peroxidase 1, thioredoxin, and peroxiredoxin 5 to complete normalcy and induced a compensatory increase in expression of catalase, glutathione reductase, gamma-glutamyl transpeptidase, and glutathione synthetase in the regenerating VP. Expression of superoxide dismutase 1, glutathione S-transferase-pi, and glucose-6-phosphate dehydrogenase was unaffected by castration and testosterone replacement. These findings indicate androgen-deprivation induces OS in the rat VP through elevation of ROS anabolism and diminution of antioxidant detoxification. Androgen replacement partially reduces OS in rat VP to precastration levels. Expression of Noxs remained high amid a broad-based recovery of antioxidant defense mechanism(s). These data might have implications on the use of androgen blockade for prostate cancer prevention and androgen therapy for andropause treatment in elderly men.     

Pituitary and hypothalamic concentration of met-enkephalin in young and old rats

Anterior pituitary and hypothalamic concentration of met-enkephalin was measured by a specific radioimmunoassay in old (22–24 months) and young (4–6 months) rats. The old rats were reproductively abnormal and based on their vaginal cytology, they were further classified under pseudopregnant, irregular cycle and constant estrus groups. All old rats had significantly higher concentration of met-enkephalin in the anterior pituitary gland when compared with young rats in estrus. Pseudopregnant rats had the highest concentration of met-enkephalin (7.38 ± 1.72 ng met-enkephalin/mg protein, mean ± SEM) followed by rats with irregular cycles (3.07 ± 1.54) and old rats exhibiting constant estrus (2.22 ± 0.47). Young rats in estrus had a mean concentration of 0.13 ± 0.08 ng met-enkephalin/mg protein in the anterior pituitary gland. However, there was no significant difference in the hypothalamic concentration of met-enkephalin of old animals when compared with young animals.     

Different pathways of neurohormonal hypothalamic control of the adrenal cortex function in young and old rats

The pathways of hypothalamic control of the adrenal cortex function were studied in the experiments on young and old male Wistar rats. Immunohistochemical, morphometric, densitometric and radioimmunoassay methods were used. It was shown that in young rats under stress vasopressin (VP) is released into the portal circulation and probably in this way stimulates ACTH and enhances secretion of corticosterone in the adrenal cortex. In old rats stress-reaction is delayed and less adequate compared to young animals. It seems likely that in old rats VP is secreted into the general circulation mainly via the posterior pituitary lobe (PP), exerting a direct influence on the adrenal cortex. The conclusion is made that in old rats regulation of the adrenal cortex function is realised via the pathway phylogenetically more ancient than in young animals.     

Age-specific changes in expression, activity, and activation of the c-Jun NH(2)-terminal kinase and p38 mitogen-activated protein kinases by methyl methanesulfonate in rats

The stress-activated protein kinases (SAPKs), c-Jun NH(2)-terminal kinases (JNKs) and p38 mitogen-activated protein kinases, were evaluated in the liver and brain of young and old rats in response to a direct-acting alkylating agent, methyl methanesulfonate (MMS). A slight but statistically significant increase in the baseline expression levels of JNK isoforms was detected in both the liver and brain of old as compared with young rats. In the liver of both young and old rats, no basal activities of JNKs were detected. In the brain, JNK activities were constitutively high and significantly increased in old rats compared with their young counterparts. Upon MMS treatment, JNKs were strongly activated in the liver, but not in the brain, of both young and old animals. The basal activity of p38 significantly increased in both the liver and brain of old rats as compared with young rats. An increase in the basal expression of p38 was detected in the brain but not in the liver of old rats. Upon treatment with MMS, p38 was activated in the liver of both young and old rats. In the brain, p38 was only activated in young but not in old rats. Taken together, these results demonstrate age-specific as well as organ-specific SAPKs signaling pathways in the rat in vivo. The possible implications of these findings in terms of resistance to endogenous and environmentally induced genotoxic stress during aging are discussed.     

Pituitary and hypothalamic concentration of met-enkephalin in young and old rats

Anterior pituitary and hypothalamic concentration of met-enkephalin was measured by a specific radioimmunoassay in old (22–24 months) and young (4–6 months) rats. The old rats were reproductively abnormal and based on their vaginal cytology, they were further classified under pseudopregnant, irregular cycle and constant estrus groups. All old rats had significantly higher concentration of met-enkephalin in the anterior pituitary gland when compared with young rats in estrus. Pseudopregnant rats had the highest concentration of met-enkephalin (7.38 ± 1.72 ng met-enkephalin/mg protein, mean ± SEM) followed by rats with irregular cycles (3.07 ± 1.54) and old rats exhibiting constant estrus (2.22 ± 0.47). Young rats in estrus had a mean concentration of 0.13 ± 0.08 ng met-enkephalin/mg protein in the anterior pituitary gland. However, there was no significant difference in the hypothalamic concentration of met-enkephalin of old animals when compared with young animals.     

5-HT5a receptors in the carotid body chemoreception pathway of rat

Due to the possible detrimental impact of local inflammatory responses in neurodegenerative disease, it was of interest to measure the expression of extracellular matrix-degrading enzymes, a group of proteases that are induced during an inflammatory response, in the brains of old and young animals in a model of neuronal death. Doses of kainic acid were administered that resulted in comparable hippocampal pyramidal neuron loss in young and old F344/BN hybrid rats, even though each age group received widely differing doses. Two matrix metalloproteinases (MMPs), MMP-2 and MMP-9, were differentially induced with respect to time after kainic acid in sensitive brain regions in both young and old rats. However, the elevation of MMP-9 in the temporal lobe 12 h after injection in old rats was significantly greater than that observed in young animals. These results suggest that early and late induction of MMPs may play a role in neuronal death and repair mechanisms, respectively, and that inflammatory mechanisms in the central nervous system (CNS) of old rats are exaggerated compared to young rats.     

Age-related excretion of six glycosidases in rat urine.

The activity of beta-N-acetylglucosaminidase (NAG), beta-galactosidase, alpha-L-fucosidase, beta-glucuronidase, beta-glucosidase and alpha-mannosidase was determined in the urine of rats at progressive ages from newborn to old animals. The age-dependence of urinary creatinine, protein and pH values was also studied. Enzyme activity, related to urinary creatinine, was significantly higher in the newborn group than other ages. The excretion of NAG increased significantly in adult rats (3-6 months old) compared to young rats (1 month old). Most of the enzyme activities were diminished in old rats (25 months old). Increased proteinuria and creatinine excretion were observed in rats since 3 months of age. Age-related differences among enzyme activities therefore should be considered when these urinary glycosidases are to be studied in rats.     

Age-dependent changes in recovery sleep after 48 hours of sleep deprivation in rats

To characterize possible changes in homeostatic regulation of sleep with aging, we have examined sleep stages during recovery sleep after 48 h of sleep deprivation in young (3 months), middle aged (12 months), and old (24 months) rats. It was found that young and middle aged, in contrast to old rats, had large (21-24%) increases in total sleep time during recovery sleep; the old rats experienced a quantitatively small (8%) but significant rise in total sleep. NREM sleep increased significantly during the recovery period in young and middle aged, but not older rats. High voltage NREM sleep (HS2) declined by 30% during recovery in the young animals, but remained unchanged compared to baseline in the middle aged and old animals. The young and middle aged rats had increases in REM sleep during recovery compared to their baseline by 96% and 93%, respectively, which was significantly greater than a 65% increase during recovery in the old rats. Increases in total sleep and REM sleep during recovery were largely confined to the first 6 h in young and middle aged rats, but maxima for the old rats occurred in the second 6 h.     

Increased carrageenan-induced acute lung inflammation in old rats

Ageing is associated with increased susceptibility to lung infections and delayed resolution of pulmonary infiltrates. The purpose of this study was to investigate the effect of age on the onset of carrageenan-induced lung inflammation. When compared with carrageenan-treated young rats (3 months old), old rats (>18 months old) exhibited a preponderance of pleural exudation and polymorphonuclear cell infiltration. Lung myeloperoxidase activity, an index of neutrophil infiltration and activation, was significantly increased in old rats in comparison with young rats. Consistent with the biochemical markers of inflammation, increased lung damage, as assessed by nitrosative stress and lipid peroxidation, was observed in carrageenan-treated old rats. In the lung exudate obtained from old rats, a significant reduction in interleukin-10 (IL-10) was observed, while similar expression of monocyte chemotactic protein-1 was induced, suggesting that a decrease in IL-10 rather than increased chemotaxis may account for the preponderance of the inflammatory cellular infiltrate in old rats. Similar to the in vivo situation, freshly isolated alveolar macrophages obtained from old rats produced less IL-10. This defective IL-10 production could be explained by a reduction in the cAMP-dependent signalling pathway, which mediates IL-10 production. Indeed, we found decreased cAMP-responsive element binding protein (CREB) and phosphorous-CREB (P-CREB) expression in old rats, which may account for reduced IL-10 production in old rats.     

Age-dependent changes of the sympathetic innervation of the rat kidney

The influence of aging on the sympathetic innervation of the kidney was studied in 3- (considered to be young), 12- (considered to be adult) and 24- (considered to be old) month-old male Sprague-Dawley rats by means of high pressure liquid chromatography with electrochemical detection, catecholamine histofluorescence and acetylcholinesterase (AChE) histochemistry. Body and kidney weights were significantly increased in adult in comparison with young rats. No further increase of either body or kidney weight was appreciated in old rats. Noradrenaline levels were increased by about 48% in adult rats and were decreased in old rats (by approx. 22% vs. young and 60% vs. adult). The density of perivascular noradrenergic fibres was significantly increased in adult rats and decreased in old animals. The percentage of kidney glomeruli supplied by AChE-positive nerve fibres is also remarkably increased in 12-month old rats and decreased in 24-month-old rats. The present data indicate that there is a striking increase in the expression of sympathetic innervation of rat kidney at 12 months of age followed by a significant decrease in the expression of innervation in old age. These changes are discussed in relation to the age-dependent impairment of renal function.