Acute effects of acephate and methamidophos and interleukin-1 on corticotropin-releasing factor (CRF) synthesis in and release from the hypothalamus in vitro

Acute effects of Ace, Meth and IL-1 on AChE activity, ACh and CRF mRNA levels in, and CRF-release from the hypothalamus were studied in vitro. The hypothalamus samples were dissected from the rat brain and were incubated in vitro with IL-1, Ace or Meth in the presence or absence of Dex, Atrop, PTL, PROP and GABA. Ace and Meth, but not IL-1, inhibited AChE activity, while all three compounds; (1) increased ACh and CRF mRNA levels in and CRF release from; (2) activated the CRE promoter region of CRF-gene in: and (3) increased cFos binding to the AP-1 region of the CRF-gene in the hypothalamus. Dex suppressed the effects of IL-1, possibly by inducing the nGRE regulatory sites of the CRF-gene. Dex, however, did not modulate the effects of Ace and Meth on the hypothalamus, which may be attributed to the failure of Dex to modulate the CRF-gene's nGRE regulatory sites. Atrop caused 80-90% inhibition of the effects of IL-1, but caused only 50-65% inhibition of the effects of Ace or Meth on CRF mRNA levels in and CRF release from the hypothalamus. PTL did not affect, while PROP slightly attenuated the effects of IL-1 and the insecticides on the hypothalamus. GABA attenuated the effects of the insecticides but not the effects of IL-1 on the hypothalamus. This suggests that the IL-1-induced augmentation of CRF synthesis in and release from the hypothalamus is mediated through a cholinergic pathway, while the insecticide-induced augmentation of CRF synthesis in and release from the hypothalamus is mediated through the cholinergic and GABAergic pathways. The insecticides, but not IL-1, disrupt feedback regulation of CRF synthesis in and release from the hypothalamus.     

The effect of insulin deficiency, dietary protein intake, and plasma amino acid concentrations on brain amino acid levels in rats

The effect of diabetes (streptozotocin, 65 mg/kg ip), dietary protein intake (15-60%), and plasma amino acid concentrations on brain large neutral amino acid levels in rats was examined. After 20 days, the plasma concentrations of methionine and the branched chain amino acids (BCAA), valine, isoleucine, and leucine were increased in diabetic rats. In brain tissue, methionine and valine levels were increased but threonine, tyrosine, and tryptophan concentrations were depressed. Increased protein consumption promoted a diabetic-like plasma amino acid pattern in normal rats while enhancing that of diabetic animals. However, with the exception of threonine, glycine, valine, and tyrosine, there was little effect on brain amino acid levels. A good association was found between the calculated brain influx rate and the actual brain concentration of threonine, methionine, tyrosine, and tryptophan in diabetic animals. There was no correlation, however, between brain influx rate and brain BCAA levels. Thus, the brain amino acid pattern in diabetes represents the combined effects of insulin insufficiency and composition of the diet ingested on plasma amino acid levels as well as metabolic adaptation within the brain itself.     

The hepatic amino acid system A transport activity, is up-regulated in obese Zucker rats

The utilization of L-alanine by liver is dependent on amino acid uptake from blood. This uptake, mainly mediated by the A transport system, may be regulated by different nutritional and physiologic conditions. The regulation of this transport system by diets with different protein content was tested in lean and obese Zucker rats. High-protein (HP) and low-protein (LP) diets led to changes in the rats’ growth patterns, especially in lean animals. However, homeostasis was relatively well maintained, as seen in plasma values, in spite of the increased urea production in the HP groups and increased triacylglycerides in the LP groups. The obese animals took up L-alanine at a higher rate than the lean animals. Obesity led to the emergence of a high-affinity component (K_M approximately 0.1–0.2 mM) in the transport system, which was not dependent on the protein content of the diet. This component has a 10-fold increase in affinity for L-alanine, but with an approximately 3- to 5-fold reduction in maximal velocity of transport.     

Correlations between amino acid concentrations in brains of seizure-susceptible and seizure-resistant rats

We have examined the relationship between free amino acid concentrations in the brains of genetically seizure-susceptible and seizure-resistant rats. The concentrations of free amino acids in the two strains do not differ significantly in the inferior colliculus or the cortex. However, animal-to-animal variations in the concentrations of numerous amino acid pairs are highly correlated. Glutamic acid decarboxylase activities did not vary between the two strains.We conclude that the strong correlations reported between glutamate and taurine levels in several species are not unique to this amino acid pair. Furthermore, unlike the situation with some experimentally-induced epilepsies, genetic epilepsy is not associated with major disturbances in free amino acid concentrations. The high correlations between amino acid pairs in some cases may reflect variations in cellular and subcellular compartment sizes that are shared by several amino acids.     

Nature of stress: differential effects on brain acetylcholinesterase activity and memory in rats

Effect of acute, chronic-predictable and chronic-unpredictable stress on memory and acetylcholinesterase (AChE) was investigated in rats. The animals were subjected to 3 type of stressors--(1) acute immobilization stress, (2) chronic-predictable stress i.e., immobilization daily for 5 consecutive days and (3) chronic-unpredictable stress that included reversal of light/dark cycle, over-night fasting, forced-swimming, immobilization and forced exercise in random unpredictable manner daily for 5 consecutive days. Learning and memory function was studied by single trial Passive avoidance test. AChE activity was assayed spectrophotometrically in the detergent (DS) and salt (SS) soluble fractions in different brain regions. Learning was obtained in acute and chronic-predictable stress groups but not in chronic-unpredictable group. Acute, chronic-predictable and chronic-unpredictable stress caused significant decrease in AChE activity in the DS fraction of cortex, hippocampus and hypothalamus as compared to control. Results indicate that AChE in DS fraction is predominantly affected in stressed and stressed-trained group but cognition is affected only by chronic-unpredictable stress. In acute and chronic-predictable groups the decreased AChE activity in the hippocampal DS fraction during learning may be responsible to maintain cognitive function by enhancing the cholinergic activity.     

Plasma amino acid concentrations in pregnant rats and in 21-day foetuses.

Plasma amino acid concentrations were determined in virgin female rats, in pregnant rats (12 and 21 days after impregnation) and in 21-day foetuses. The total amino acid concentration in plasma decreases significantly with pregnancy, being lower at 12 than at 21 days. Alanine, glutamine+glutamate and other 'gluconeogenic' amino acids decrease dramatically by mid-term, but regain their original concentrations at the end of the pregnancy. With most other amino acids, mainly the essential ones, the trend is towards lower concentrations which are maintained throughout pregnancy. These data agree with known nitrogen-conservation schemes in pregnancy and with the important demands on amino acids provoked by foetal growth. In the 21-day foetuses, concentrations of individual amino acids are considerably higher than in their mothers, with high plasma foetal/maternal concentration ratios, especially for lysine, phenylalanine and hydroxy-proline, suggesting active protein biosynthesis and turnover. All other amino acids also have high concentration ratios, presumably owing to their requirement by the foetuses for growth. Alanine, glutamine+glutamate, asparagine+aspartate, glycine, serine and threonine form a lower proportion of the total amino acids in foetuses than in the virgin controls or pregnant rats, probably owing to their role primarily in energy metabolism in the adults. The results indicate that at this phase of foetal growth, the placental amino acid uptake is considerable and seems to be higher than immediately before birth.     

The effects of starvation on plasma free amino acid and glucose concentrations in lake sturgeon

The effect of starvation on the metabolism of the lake sturgeon Acipenser fulvescens was examined by measuring haematocrit, plasma glucose concentrations, and plasma free amino acids. Plasma was sampled on day 0, 10, 20, 45 and 60 of a 60-day starvation period. Haematocrit was observed to decrease with starvation indicating a decreased oxygen carrying capacity of the blood. Plasma glucose levels differed only at day 10, with a decrease in blood glucose level in the starved group. No differences were detected between groups for alanine, aspartate, and serine, while elevated levels were observed for glutamine throughout the experiment. An increase in arginine, tyrosine, valine, methionine, tryptophan, phenylalanine, glutamate, glycine, isoleucine, histidine and leucine, concentrations were observed after 45 days of starvation. The maintenance, or increased plasma levels, of glucogenic amino acids in combination with the maintenance of blood glucose concentrations indicates active gluconeogenic processes in the liver supported by muscle proteolysis.     

Acute effects of caffeine injection on neutral amino acids and brain monoamine levels in rats

The injection of caffeine (100 mg/kg, i.p.) into male rats acutely increased brain levels of trytophan, serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA). Blood levels of glucose, nonesterified fatty acids (NEFA) and insulin also increased, while those of the aromatic and branched-chain amino acids fell. Serum tryptophan levels either did not fall, or increased. Consequently, the serum ratio of trypthopahn to the sum of other large neutral amino acids (LNAA) increased. Less consistently noted were increases in serum free tryptophan levels. Brain tyrosine levels were not appreciably altered by caffeine, nor was the serum tyrosine ratio. In dose-response studies, 25 mg/kg of caffeine was the minimal effective dose needed to raise brain tryptophan, but only the 100 mg/kg dose elevated all three indoles in brain. In no experiments did caffeine, at any time or dose, alter brain levels of dopamine or norepinephrine. Caffeine thus probably raises brain tryptophan levels by causing insulin secretion, and thereby changing plasma amino acid levels to favor increased tryptophan uptake into brain. The rises in brain 5-HT and 5-HIAA may follow from the increase in brain tryptophan, although further data are required clearly to establish such a mechanism.     

Regulation of uterine and umbilical amino acid uptakes by maternal amino acid concentrations

We tested the hypothesis that decreased fetal amino acid (AA) supply, produced by maternal hypoaminoacidemia (low AA) during hyperglycemia (HG), is reversible with maternal AA infusion and regulates fetal insulin concentration ([I]). We measured net uterine and umbilical AA uptakes during maternal HG/low AA concentration ([AA]) and after maternal intravenous infusion of a mixed AA solution. After 5 days HG, all maternal [AA] except glycine were decreased >50%, particularly essential [AA] (P < 0.00005). Most fetal [AA] also were decreased, especially branched-chain AA (P < 0.001). Maternal AA infusion increased net uterine uptakes of Val, Leu, Ile, Met, and Ser and net umbilical uptakes of Val, Leu, Ile, Met, Phe, and Arg but did not change net uteroplacental uptake of any AA. Fetal [I] increased 55 +/- 14%, P < 0.001, with correction of fetal [AA], despite the lack of change in fetal glucose concentration. Thus generalized maternal hypoaminoacidemia decreases uterine and umbilical uptakes of primarily the essential AA and decreases fetal branched-chain [AA]. These changes are reversed with correction of maternal [AA], which also increases fetal [I].     

Acute effects of ethanol on brain,plasma and adrenal monoamine concentrations in virgin and pregnant rats and their fetuses

The dose-response relationship in brain, plasma, and adrenal monoamine changes after acute oral ethanol administration (1, 2, 4 g/kg body wt) was studied in virgin rats to determine whether the response to the highest dose differed in 21-day pregnant animals, and to assess the potential consequences of ethanol on the neurotransmitter systems of their fetuses. Blood ethanol and acetaldehyde concentrations in blood increased progressively with the ethanol dose in virgin rats, and values in pregnant animals were very similar. Ethanol concentration in fetal blood and amniotic fluid did not differ from that in mother's blood whereas fetal acetaldehyde concentrations were negligible. In a dose-related manner, ethanol decreased brain DA, DOPAC and 5HT concentrations did not affect those of NA and 5HIAA, or adrenal A and NA concentrations, whereas it enhanced plasma NA levels. Basal levels of monoamines and their changes after ethanol intake did not differ in pregnant and virgin rats. Monoamine and metabolite concentrations were much lower in fetal than in maternal brains whereas plasma and adrenal catecholamine concentrations were very similar and maternal ethanol intake did not modify these fetal parameters in the fetus. Results are in agreement with the known similar metabolic response to ethanol in fed pregnant and virgin rats. The lack of fetal monoamine response to maternal ethanol intake may be a consequence of the incapacity of fetal liver to form acetaldehyde and the ability of the placenta to oxidize maternal acetaldehyde which protects the fetus from maternal alcohol intake at late gestation.     

Pharmacological effects on acetylcholinesterase activity of hypothalamic and cortical structures

The kinetics of cholinesterase activity in the sensomotor cortex, hippocampus, and anterior hypothalamus was determined in 93 rats by an electrometric method using an automatic recording system. The localization of acetylcholinesterase in the cell structures was revealed histochemically. In the control animals acetylcholinesterase was distributed unevenly among these structures. In rats aged 1 month its activity was higher in the sensomotor cortex. At later age periods (6, 12, 24 months) the acetylcholinesterase activity was highest in the posterior hypothalamus. While the unevenness of distribution of this substance continued to be observed, the level of its activity fell with age. A redistribution of acetylcholinesterase activity compared with the control was found 15 min after subcutaneous injection of adrenalin (300 µg/kg). Its activity was maximal in the anterior hypothalamus. The acetycholinesterase activity in all these structures was lowered after subcutaneous injection of chloral hydrate (300 mg/kg). Meanwhile the differences in the character of its distribution between the individual structures disappeared. These results indicate definite correlation between the functional state of the CNS and the distribution of acetylcholinesterase activity in the brain structures studied.A. A. Bogomolets' Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 5, No. 1, pp. 47–53, January–February, 1973.     

Acute endocrine effects of a single administration of methylmercury chloride (MMC) in rats

The acute effects of methylmercury chloride (MMC) on the endocrine functions were investigated with doses too small to cause any typical neurological dysfunctions. The hormones included PRL, LH, TSH, ACTH, corticosterone (Bk), testosterone (TLI), total thyroxine (T4) and free thyroxine (free T4). The changes in serum hormone levels from 1 hour through 10 days after a single injection of MMC (12 mg/kg s.c.) (Exp. 1), and dose-response relationships between MMC doses (2 to 16 mg/kg s.c.) and the serum hormone levels at 25 hours after MMC injection (Exp. 2) were examined. The acute effects revealed, which were all reversible, are summarized as follows; MMC might directly inhibit thyroxine synthesis; MMC could affect only stimulatively the pituitary-adrenal axis and PRL synthesis/release, the primary action site for which may be the CNS; and the effects of the pituitary-gonadal axis were inconsistent and, therefore, this axis seems to be relatively resistant to MMC. On the other hand, the responses of PRL and TSH to TRH loading, which were examined for both groups in Exp. 3, suggested that MMC could not affect the metabolizing activity for serum PRL and TSH. The hormone levels of the MMC group enhanced by TRH recovered very rapidly as in the control group. Thus, these acute and reversible endocrine effects seem to indicate relatively earlier development of possible chronic and irreversible effects on the endocrine functions when exposed to methylmercury chronically, and these should be examined further.     

Serum amino acid concentrations in aging men and women

The age and gender related differences in serum amino acid concentrations have been assessed in 72 (23-92 years) medically screened healthy men and women who were divided into three male and three female groups according to age. Free-time physical activity and food intake were analysed from the 5-day diaries. The subjects were instructed to eat according to their normal dietary habits and to avoid any clinical complementary nutritional products or other products that could increase protein or energy intake. The blood samples (5 ml) taken from the antecubital vein after an over-night fast were analysed for their amino acid contents by chromatography. In total nutrient intake of energy (P < 0.001), protein (P < 0.001), alcohol (P < 0.05), water (P < 0.01), sodium (P < 0.001) and fiber P < 0.001) decreased significantly with age. The concentration of total amino acids (P < 0.01), essential amino acids (P < 0.001), non-essential amino acids (P < 0.05) and branched-chain amino acids (P < 0.05) decreased, whereas citrulline (P < 0.001) and cysteine (P < 0.001) were the only amino acids, which increased with aging. In addition, men had significantly higher concentrations than women of essential amino acids (P < 0.001), branched-chain amino acids (P < 0.001), and 10 of the 22 individual amino acids assayed (P < 0.01). Women had significantly higher concentrations of aspartate (P < 0.05), glycine (P < 0.01), serine (P < 0.001) and taurine (P < 0.01) than men. It is concluded that the decrease in serum total amino acid concentration is associated with decreased energy and protein intake with aging and men have higher essential amino acid concentration in serum than women.     

Hypothalamic excitatory amino acid system during sexual maturation in female rats

The present results indicate that during sexual maturation the APOA-MBH from rats of 30 days of age released significantly higher quantities of GnRH than the tissue from 16-day-old rats (P < 0.01). The addition of NMDA, an agonist of the excitatory amino acids system (EAAs), to the medium after 30 min of incubation significantly increased (P < 0.01) the GnRH release in normal rats of both ages and this increase was significantly (P < 0.01) higher in 30-day-old rats (to 661%) than in rats of 16 days of age (to 273%). The administration of estrogen-progesterone (EP) to rats of 16 days of age did not modify the GnRH release response to NMDA. On the contrary, at 30 days of age EP administration significantly potentiated the GnRH release response to NMDA since while in the control group NMDA increased the GnRH release to 630%, in the EP-pretreated group this was to around 4700% (P < 0.01). EP pretreatment of prepubertal rats decreases the hypothalamic release of aspartate and glutamate, the excitatory amino acids involved in NMDA neurotransmission and glycine but increases EAAs release in peripubertal rats. On the basis of these results it is proposed that the increase in EAAs release by the hypothalamus is directly connected with the onset of puberty and that the maturation of the positive feedback effect of ovarian hormones on gonadotropin secretion is related to the maturation of the capacity of EP to increase hypothalamic EAAs. Before this maturational event EP inhibits EAAs release as well as gonadotropin release (prepubertal rats). NMDA receptor stimulation leads to a positive mechanism which increases the release of Asp and Glu from APOA-MBH both in prepubertal and peripubertal rats, but EP potentiates this mechanism only in peripubertal rats. This could be an additional neuroendocrine mechanism involved in the increase of gonadotropin during sexual maturation which induces the onset of puberty and the preovulatory discharge of these pituitary hormones.     

Effects of chronic lithium administration on brain weights, acetylcholinesterase activity and learning ability in rats

In order to test the effects of chronic lithium (Li) administration on learning and memory, 21 day old rats were subjected to different degrees of environmental stimulation (enriched condition, EC and impoverished condition, IC) with and without Li for 144 days. Li was administered with food (2.18 mEq/Kg weight/day). Average plasma Li concentration at the end of the experiment was 0.41 +/- 0.04 mu Eq/ml. Both Li treatment and the environmental condition showed an overall significant effect on the cortex/subcortex weight ratio and learning ability index, but not on AChE activity in occipital cortex. A similar pattern of brain Li distribution was observed in both EC-Li and IC-Li, with occipital cortex having the highest levels. Li tissue/protein/plasma ratio was higher in EC than in IC, in all the brain areas studied. Other organs (liver and kidney) did not show EC-IC differences in the tissue/protein/plasma Li ratio.     

Different effects of hypothermia on amino acid incorporation and on amino acid uptake in the brainin vivo

The temperature dependence of the incorporation of amino acids into cerebral proteins and that of the transport of amino acids through the blood-brain barrier were studied. We measured the protein synthesis rate in vivo over a wide temperature range (14°C–38°C) in male Sprague-Dawley rats using a flooding dose of labeled valine. There was a linear dependence of the protein synthesis rate on temperature. The temperature quotient expressed as per cent decrease per 1°C was somewhat lower at the lower temperatures, a decrease from 7.8% in the 37.7–32.5°C range to 6.7% in the 25.5–14°C range. The transport of the three amino acids phenylalanine, lysine, and alanine, representing there transport systems, through the blood-brain barrier showed no temperature dependence in vivo. The results show that in hypothermia cerebral metabolic rates are lowered to a great extent, while some aspects of metabolic transport are not affected.