Age-dependent changes of nucleic acid labeling in different rat brain regions

The effects of aging on in vivo DNA and RNA labeling and on RNA content in various brain regions of 4-, 12-, and 24-month-old rats were investigated. No difference in [methyl-¹⁴C]thymidine incorporation into DNA of cerebral cortex and cerebelllum during aging was observed.The ratio of RNA/DNA content significantly decreased from 4 to 24 months of age in cerebral cortex, cerebellum and striatum. RNA labeling decreased by 15% in cerebral cortex of 24-month-old animals while in the other brain areas examined (cerebellum, hippocampus, hypothalamus, brainstem, striatum) did not change during aging.In the cerebral cortex, the ratio of the specific radioactivity of microsomal RNA to that of nuclear RNA, determined by in vivo experiments, was not affected by the aging process. A significant decrease of total, poly(A)+ RNA and poly(A)- RNA content was observed in the same brain area of 24-month-old rats compared to 4-month-old ones. Moreover, densitometric and radioactivity patterns obtained by gel electrophoresis of labeled RNA after in vitro experiments (tissue slices of cerebral cortex) showed a different ribosomal RNA processing during aging. In vivo chronic treatment with CDP-choline was able to increase RNA labeling in corpus striatum of 24-month-old animals.     

Choline acetyltransferase activity and muscarinic binding in brain regions of aging fischer-344 rats

Muscarinic receptor binding and choline acetyltransferase (EC 2.3.1.6.) activity were assayed in three brain regions of 4-, 12- and 24-month-old Fischer-344 rats. Statistically significant age differences in cholinergic parameters were observed in each region. The affinity for [³H]quinuclidinyl benzilate increased in the cortex (24 vs 12 and 4 months), but B_max decreased in the cortex (24 vs 12 vs 4 months), striatum (24 vs 12 vs 4 months) and hippocampus (24 vs 12 and 24 vs 4). Assays of carbamylcholine inhibition of [³H]quinuclidinyl benzilate binding in the hippocampus showed that high affinity agonist binding increased with age (24 vs 12 and 4 months), and the percentage of muscarinic binding to high affinity agonist sites decreased (24 vs 12 vs 4 months). In addition, the affinity of the agonist oxotremorine for muscarinic binding sites also increased in the hippocampus (12 and 24 vs 4 months). Although the K_m of choline acetyltransferase for choline chloride did not change in any region tested, the K_m for acetyl coenzyme A decreased in the hippocampus (24 vs 12 months), but increased (4 vs 12 months) and then decreased (12 vs 24 months) in the striatum. Statistically significant age-related declines in V_max for choline acetyltransferase were noted in the striatum (24 < 12 < 4 months), but no age differences in this parameter were observed in the cortex or the hippocampus. Statistically significant positive correlations between V_max for choline acetyltransferase and B_max for [³H]quinuclidinyl benzilate binding were observed in each of the brain regions of 4-, 12- and 24-month-old rats.

The findings have implications for use of the Fischer-344 male rat as an animal model of aging and age-related disorders of the human brain, including dementia of the Alzheimer type.     

Decrease of Choline Acetyltransferase Activity of Rat Cortex Capillaries with Aging

Choline acetyltransferase (ChAT) activity was estimated in brain cortex capillaries isolated from 3-, 12-, 18-, and 24-month-old rats. Maximum enzymatic activity was found at 12 months (55 +/- 0.3 pmol X mg-1 protein X min-1; mean +/- SEM) and then it decreased to reach a minimum at 24 months (34 +/- 3.1 pmol X mg-1 protein X min-1). A less marked decrease of enzymatic activity was also found in cortex homogenate and in a synaptosomal fraction obtained from the same groups of rats. Loss of ChAT of brain capillaries with aging could be related to a general phenomenon of cortical cholinergic deficit in that condition.     

The Lumped Constant in the Deoxyglucose Procedure Declines with Age in Fischer-344 Rats

Concentrations of [14C]2-deoxy-D-glucose ([14C]DG) and of glucose were measured in plasma of arterial and sagittal sinus venous blood from awake Fischer-344 rats at 3, 12, and 24 months of age, during continuous intravenous infusion of [14C]DG and after a steady-state arterial plasma concentration of [14C]DG was reached. Brain extraction, i.e., the difference between arterial and venous plasma concentrations divided by the arterial plasma concentration, was calculated for both [14C]DG and glucose. Because exchange of both substances between rat plasma and erythrocytes is slow, the ratio of the brain extraction of [14C]DG to that of glucose is identical to the lumped constant in the deoxyglucose procedure of Sokoloff et al. [J. Neurochem. 28, 897-916. (1977)]. This ratio equaled 0.502 +/- 0.015 (SEM) at 3 months, 0.456 +/- 0.007 at 12 months, and 0.418 +/- 0.006 at 24 months of age (n = 15); the means differed significantly from each other (p less than 0.05). The results indicate that the lumped constant declines between 3 and 24 months of age in awake rats, and suggest that many reported age reductions in regional cerebral glucose utilization, of 15-25%, are artifactual.     

Noradrenaline content of the brain of rats of various ages following adaptation to a new situation and to the development of a passive avoidance conditioned reflex

Noradrenaline (NA) levels in cortico-striatal (including cerebral cortex, hippocamp, striatum) and hypothalamo-brainstem (including hypothalamus, thalamus, tectum + tegmentum) regions were determined by fluorometry in I- and 2-month-old male rats after 7-day adaptation to experimental conditions and passive avoidance learning by single electric foot shock. Neither the new environment nor a week's adaptation to it resulted in any significant alteration of NA content in both brain regions of 1- and 2-month-old rats. No considerable differences in NA levels were found in rats of both age groups with and without passive avoidance responses. But 24 hours after the exposure to foot shock NA basal levels markedly decreased in both brain regions of 1-month-old rats, while in 2-month-old ones NA basal levels markedly increased in hypothalamo-brainstem region.     

NGF restores decrease in catalase activity and increases superoxide dismutase and glutathione peroxidase activity in the brain of aged rats.

The effects of ageing on the activity of copper-zinc superoxide dismutase (SOD), selenium-dependent and independent glutathione peroxidase (GSH-Px) and catalase in several areas of the brain in 3-, 12-, and 24-month-old rats were studied. In addition, the effects of a subacute intracerebroventricular treatment of NGF (1 microgram daily for 28 consecutive days) on SOD, GSH-Px, and catalase activity in the same areas of the brain were assessed. The effects of ageing on the activities of antioxidant enzymes varied considerably in the different brain areas studied. Copper-zinc SOD was alone in being unaffected by ageing. Intraventricular infusion of NGF significantly increased SOD activity in the prefrontal cortex, hypothalamus, caudate nucleus, and mesencephalon of 24-month-old rats. Selenium-dependent GSH-Px activity did not significantly change in 12-month-old rats but it increased in the lower brain stem of 24-month-old animals. In comparison to vehicle-treated rats, NGF significantly increased selenium-dependent GSH-Px activity in all brain areas studied in 12- and 24-month-old rats. Catalase activity decreased significantly in the majority of the brain areas studied in 12- and 24-month-old rats. NGF completely restored the fall in catalase activity in 12- and 24-month-old animals to levels similar to those occurring in young rats. In conclusion, the present experiments show, for the first time, that long-term intraventricular administration of NGF significantly increases in old animals the activity of key enzymes involved in the metabolic degradation of superoxide radicals and hydrogen peroxide.     

Local cerebral glucose utilization in the brain of old, learning impaired rats

The local cerebral glucose utilization (LCGU) was measured in 63 different cortical areas and nuclei of the telencephalon, diencephalon and rhombencephalon of young adult (3 to 4-month-old) rats and of 27-month-old Wistar rats, in which learning impairments had been proven by a water maze test. The LCGU was determined by [14C]2-deoxyglucose autoradiography. In the old rats the mean LCGU of all brain regions was significantly reduced by about 10% compared with the young control group; the mean LCGU was 74.2 mumol glucose/(100 g x min) in the young and 66.7 in the old rats. Different degrees of LCGU decrease were found in the different regions. Most of the brain regions with significantly reduced LCGU values in the aged, learning impaired rats were associated with auditory and visual functions, the dopaminergic system, and structures known to be involved in learning and memory processes. Therefore, the regional pattern of LCGU reduction found in the aged, learning impaired rats did not resemble any known pattern found after lesions of a single transmitter system or systemic administration of transmitter agonists or antagonists.     

Cerebral Blood Flow and Oxidative Metabolism in Conscious Fischer-344 Rats of Different Ages

Abstract: The cerebral metabolic rates for O₂ and for glucose were measured in conscious, fasted male Fischer-344 rats at the ages of 3, 12, and 24 months, and cerebral blood flow was determined with ¹⁴C-iodoantipyrine. The metabolic rates for oxygen and glucose were obtained by multiplying blood flow by the O₂ and glucose concentration differences, respectively, between blood in the femoral artery and in the superior sagittal sinus. Mean cerebral blood flow and the metabolic rates for oxygen and glucose did not differ significantly (p > 0.05) between 3 and 12 or between 12 and 24 months. Nor did the arteriovenous differences for O₂ and for glucose change significantly with age. Because the superior sagittal sinus drains blood mainly from the cerebral cortex, the results indicate that average cerebral cortical oxidative metabolism, and the coupling ratios between the cerebral metabolic rate for oxygen and cerebral blood flow and between the cerebral metabolic rate for glucose and cerebral blood flow, do not change significantly with age in the Fischer-344 rat.     

Lack of Effects of Inhibition of Nitric Oxide Synthesis on Local Glucose Utilization in the Rat Brain

Abstract: The effects of chronic treatment with N ^G-nitro- l -arginine methyl ester, a potent inhibitor of nitric oxide synthase activity, on local cerebral glucose utilization were examined in conscious rats. Intraperitoneal injections of 50 mg/kg of the nitroarginine twice daily for 4 days have been found to result in almost complete inhibition of nitric oxide synthase activity in brain. Local cerebral glucose utilization was determined by means of the quantitative autoradiographic [¹⁴C]deoxyglucose method in an experimental group (n = 7) of rats that were treated with the nitroarginine according to this schedule and in a normal control group (n = 7) treated similarly with saline. The rats were conscious but partially restrained during the determinations of local cerebral glucose utilization. The nitroarginine treatment raised mean arterial blood pressure statistically significantly to 147 ± 3 mm Hg (mean ± SEM) from a level of 120 ± 5 mm Hg in the saline controls ( p < 0.001 by grouped t test), but there were no statistically significant effects on glucose utilization in any of 39 brain structures examined. It is concluded that nitric oxide normally exerts no significant influence on energy metabolism in the rat brain.     

No Effect of Hyperketonemia on Local Cerebral Glucose Utilization in Conscious Rats

Abstract: Local cerebral glucose utilization was measured by the [¹⁴C]2-deoxy- d -glucose method in conscious control and hyperketonemic rats. Hyperketonemia was induced by 3 days of starvation or by infusion of 3- hydroxybutyrate in fed rats. These treatments produced combined blood ketone body concentrations (acetoacetate + 3-hydroxybutyrate) of from 1.2 to 2.4 mM. Neither treatment significantly affected glucose utilization in any of the 15 brain regions studied. These observations indicate that hyperketonemia in resting, conscious rats does not interfere with brain uptake and phosphorylation of glucose.     

Regional Cerebral Glucose Utilization During Insulin-Induced Hypoglycemia in Unanesthetized Rats

Regional cerebral glucose utilization (rCMRgl) was studied during insulin-induced hypoglycemia in unanesthetized rats. Rats were surgically prepared using halothane and nitrous oxide anesthesia and allowed 5 h to recover from the anesthesia before rCMRgl was measured. The rCMRgl was measured using [6-14C]glucose in a normoglycemic control group and two hypoglycemic groups, A (30 min after insulin injection) and B (2 h after insulin injection). The mean plasma glucose level was 7.03 mumol/ml in the normoglycemic group, 1.96 mumol/ml in hypoglycemic group A, and 1.40 mumol/ml in hypoglycemic group B. The rCMRgl in hypoglycemic group A decreased 8-18% in 17 brain regions measured; five changes were statistically significant. The rCMRgl in hypoglycemic group B decreased significantly in all but one of the brain regions measured; the decrease ranged from 15% in the pyramidal tract to 36% in the motor and auditory cortices. The rCMRgl in every brain region decreased when the plasma glucose level fell below 1.5-2.5 mumol/ml. No brain region could maintain rCMRgl at plasma glucose concentrations lower than predicted by regional glucose influx described in previous studies. Glucose utilization in all brain regions appears to be limited by the influx of glucose.     

Onset and evolution of nephropathy in rats with spontaneous diabetes mellitus

The occurrence of renal diabetic complications was studied in diabetic nonobese IIM/FmeSS (eSS) rats. The results were compared with eumetabolic Wistar rats paired by sex and age. Between 6 and 12 months of age, eSS male rats had higher fructosamine values and glucose intolerance as well as increasing proteinuria and uremia. Enhancement in water, calcium and phosphorus fractional excretion with a concomitant lower sodium excretion, was observed from 12 months of age on. 18- and 21-month-old eSS rats exhibited fasting hyperglycaemia and rising values of fructosamine, glucose intolerance and glycosuria. Simultaneously, a notorious worsening of proteinuria as well as alterations in glomerular filtration were verified. Optic microscopy of 12-month-old eSS rat kidneys showed areas of tubular dilatation with protein cylinders. In 21-month-old eSS animals, kidneys appeared overtly damaged. Increased capsular, glomerular and Henle's thin loop diameters were verified in 12- and 21-month-old eSS rats. Glomeruli showed diffuse hypertrophy of mesangial tissue and thickening of the basement membrane. Areas of markedly atrophic and dilated tubules containing acidophilic proteinaceous material were observed. At age of 21 months, kidneys of eumetabolic Wistar control rats presented foci of interstitial and pielic inflammatory infiltrates.     

Local cerebral glucose utilization in the hippocampus of old rats

The local cerebral glucose utilization (LCGU) was measured in the different areas and layers of the Ammon's horn and dentate gyrus of young adult (3 to 4-month-old) rats, and of 27-month-old rats with proven cognitive deficits. The LCGU was determined by quantitative [14C]2-deoxyglucose autoradiography. Compared to young animals, in the old rats the LCGU was significantly reduced by 12% to 15% in the oriens layers of CA1 and CA2, the pyramidal layers of the CA sectors 1-3, the radiatum and lacunosum-molecular layers of CA2 and CA3 and in the lucidum layer of CA3. The LCGU values of all the other layers of the Ammon's horn and the dentate gyrus did not differ significantly between young and old rats. The pattern of the LCGU reduction found in the old rats roughly resembles changes found after fimbra-fornix lesions or systemic administration of scopolamine, suggesting a functionally important deficit in the cholinergic innervation of the old rats' hippocampi.     

Extracellular Glucose Concentrations in the Rat Hippocampus Measured by Zero-Net-Flux

Abstract : The concentration of glucose in the brain's extracellular fluid remains controversial, with recent estimates and measurements ranging from 0.35 to 3.3 m M . In the present experiments, we used the method of zero-net-flux microdialysis to determine glucose concentration in the hippocampal extracellular fluid of awake, freely moving rats. In addition, the point of zero-net-flux was measured across variations in flow rate to confirm that the results for glucose measurement were robust to such variations. In 3-month-old male Sprague-Dawley rats, the concentration of glucose in the hippocampal extracellular fluid was found to be 1.00 ± 0.05 m M , which did not vary with changes in flow rate. Three-month-old and 24-month-old Fischer-344 rats both showed a significantly higher hippocampal extracellular fluid glucose concentration, at 1.24 ± 0.07 and 1.21 ± 0.04 m M , respectively ; there was no significant difference between the two age groups. The present data demonstrate variation in extracellular brain glucose concentration between rat strains. When taken together with previous data showing a striatal extracellular glucose concentration on the order of 0.5 m M , the data also demonstrate variation in extracellular glucose between brain regions. Traditional models of brain glucose transport and distribution, in which extracellular concentration is assumed to be constant, may require revision.     

Local cerebral glucose utilization in the brain of old,learning impaired rats

Summary The local cerebral glucose utilization (LCGU) was measured in 63 different cortical areas and nuclei of the telencephalon, diencephalon and rhombencephalon of young adult (3 to 4-month-old) rats and of 27-month-old Wistar rats, in which learning impairments had been proven by a water maze test. The LCGU was determined by [¹⁴C]2-deoxyglucose autoradiography. In the old rats the mean LCGU of all brain regions was significantly reduced by about 10% compared with the young control group; the mean LCGU was 74.2 mol glucose/(100 g × min) in the young and 66.7 in the old rats. Different degrees of LCGU decrease were found in the different regions. Most of the brain regions with significantly reduced LCGU values in the aged, learning impaired rats were associated with auditory and visual functions, the dopaminergic system, and structures known to be involved in learning and memory processes. Therefore, the regional pattern of LCGU reduction found in the aged, learning impaired rats did not resemble any known pattern found after lesions of a single transmitter system or systemic administration of transmitter agonists or antagonists.     

The Effects of Apomorphine upon Local Cerebral Glucose Utilization in Conscious Rats and in Rats Anesthetized with Chloral Hydrate

The effects of the dopaminergic agonist apomorphine (1 mg . kg-1 i.v.) upon local cerebral glucose utilization in 43 anatomically discrete regions of the CNS were examined in conscious, lightly restrained rats and in rats anesthetized with chloral hydrate by means of the quantitative autoradiographic [14C]2-deoxyglucose technique. In animals anesthetized with chloral hydrate, glucose utilization was reduced throughout all regions of the CNS from the levels observed in conscious animals, although the magnitude of the reductions in glucose use displayed considerable regional heterogeneity. With chloral hydrate anesthesia, the proportionately most marked reductions in glucose use (by 40-60% from conscious levels) were noted in primary auditory nuclei, thalmaic relay nuclei, and neocortex, and the least pronounced reductions in glucose use (by 15-25% from conscious levels) were observed in limbic areas, some motor relay nuclei, and white matter. In conscious, lightly restrained rats, the administration of apomorphine (1 mg . kg-1) effected significant increased in glucose utilization in 15 regions of the CNS (e.g., subthalamic nucleus, ventral thalamic nucleus, rostral neocortex, substantia nigra, pars reticulata), and significant reductions in glucose utilization in two regions of the CNS (lateral habenular nucleus and anterior cingulate cortex). In rats anesthetized with chloral hydrate, the effects of apomorphine upon local glucose utilization were less widespread and less marked than in conscious animals. In only two of the regions (the globus pallidus and septal nucleus), which displayed increased glucose use following apomorphine in conscious rats, were significant increases in local glucose utilization observed with this agent in chloral hydrate-anesthetized rats. In the pars compacta of the substantia nigra, in which apomorphine increased glucose utilization in conscious animals, significant reductions in glucose utilization were observed following apomorphine in rats anesthetized with chloral hydrate. The profound effects of chloral hydrate anesthesia upon local cerebral glucose use, and the modification by this anesthetic regime of the local metabolic responses to apomorphine, emphasize the difficulties which exists in the extrapolation of data from anesthetized animals to the conditions which prevail in the conscious animal.     

Effects of dexamethasone on insulin receptor in aging

The aim of this study was to examine the effects of dexamethasone (Dex) on functional properties of the rat insulin receptor (IR). Male Mill Hill hooded rats, 3, 6, 12, 18 and 21 months old, were injected with Dex (4 mg/kg) and rat liver and erythrocytes were used for experiments 18 h after Dex administration. Treatment with Dex lowered the specific binding (SB) of insulin (INS) in the liver of 3- and 18-month-old rats and concentration of INS binding sites (N1, N2) and the dissociation constant of low-affinity binding sites (Kd2) in the liver of 6- and 18-month-old rats. In addition, Dex treatment lowered the liver IR protein level in all analyzed groups, except 21-month-old rats where it remained unchanged, but raised the IR mRNA level in 18-month-old rats. In erythrocytes, treatment with Dex decreased SB and Kd2 (in animals 3 and 6 months old) and N1 (in ones 3 and 18 months old). Following Dex treatment, the INS plasma level increased (in rats 3, 18 and 21 months old), while glucose (Glu) concentration increased in 3 and 12 months old, but decreased in 6- and 21-month-old rats. In summary, Dex exerts the strongest effect on the erythrocyte IR of 3- and 6-month-old rats and the hepatic IR of 18-month-old rats. IR in both tissues is almost insensitive to Dex in 12- and 21-month-old rats. The pattern of age-related changes of IR induced by Dex does not correlate with changes of plasma Glu and INS.     

大鼠脑损伤后非损伤区域缺氧诱导因子与乳酸的变化研究

目的：研究大鼠脑损伤后非损伤区域缺氧诱导因子（hypoxia-inducible factor-1α,HIF-1α）与乳酸的表达变化。方法：取雄性SD大鼠36只,体重200-300g,参照统计学随机数字表将大鼠随机平均分为正常对照组（6只）、假手术组（6只）、造模组（24只）,3组,造模组分四个时间点12h、72h、1w、2w处死动物（每时间点6只）。使用立体定位仪和液压打击装置,靶向打击大脑中动脉,造大鼠脑外伤模型。采用免疫组织化学法检测脑外伤后不同时间点损伤临近区域脑组织中HIF-1α蛋白表达及乳酸含量的变化。结果：正常组和假手术组脑组织神经细胞HIF-1α表达和乳酸含量无明显变化,而模型组损伤临近区域HIF-1α的表达及乳酸含量的变化规律基本一致,12 h时增多,72h时达到高峰,1w表达下降至2w时恢复正常。造模组12h、72h、1w3个亚组与正常对照组比较差异具有统计学意义p〈0.01,造模组2w亚组与正常对照组比较差异无统计学意义p〉0.01。结论：脑外伤后非损伤区域也有缺血、缺氧的改变,可能与脑外伤后的脑萎缩有相关性。     

Age-related changes in albumin elimination in female WAG/Rij rats.

Albumin elimination rates were determined in 3-, 12-, 24- and 36-month-old female WAG/Rij rats. No change in elimination half-life was found with age. However, as there was an increase in the whole-body albumin pool, a concomitant increase in albumin clearance was observed at between 12 and 36 months of age. It was concluded that the increase in clearance between 12 and 24 months of age was only due to a change in the animal's physiology, whereas between 24 and 36 months of age it was also due to changes in the albumin molecule. The age-related changes in albumin clearance were thought not to be caused by changes in the albumin excretion via the urine or via the gastrointestinal tract.     

Changes in the cytochrone C and A (A3) content in the liver mitochondria of rats with hyperthyroidism

Thyroxin was administered daily to male Wistar rats aged 1, 3, 12 and 24 months in a dose of 250 g per 100 g body weight. After 9 days there was a significant increase in the content of cytochromes c and a (a(3)). An appreciable rise in the cytochrome c content was recorded as long as after one day, whereas the content of cytochromes a (a(3)) did not exceed normal even after 2 days. Following one day the content of cytochromes a (a(3)) in 3-, 12-, and 24-month-old rats was sightly lower than in normal rats. A significant temporary increase in the c/a (a(3)) ratio was observed after 1-2 days only in 12- and 24-month-old rats. The c/a (a(3)) ratio increased with age. It is suggested that application of thyroid hormones may be promising in studies on the regulation of mitochondrial biogenesis.