Central administration of leucine, but not isoleucine and valine, stimulates feeding behavior in neonatal chicks

Branched-chain amino acids (BCAAs) are essential amino acids that play a major role in brain energy metabolism. This study was done to elucidate whether central injection of BCAAs influences feeding behavior in chicks. We found that the intracerebroventricular injection of leucine (200 microg) significantly stimulated food intake in neonatal chicks during 30 min postinjection. Additionally, the starting time of feeding and pecking rhythm after injection were significantly accelerated by leucine. In contrast, isoleucine and valine had no effect on ingestive response during experiment periods. Moreover, a metabolite of leucine (alpha-ketoisocaproic acid) at an equimolar concentration of leucine also did not increase food intake in chicks. These results suggest that leucine induces hyperphagia of neonatal chicks and it may be due to the synthesized glutamate by exogenous leucine.     

Atypical amino acids inhibit histidine, valine, or lysine transport into rat brain

Transport of histidine, valine, or lysine into rat brain slices and across the blood-brain barrier (BBB) was determined in the presence of atypical nonprotein amino acids. Competitors of histidine and valine transport in slices were large neutral amino acids including norleucine, norvaline, alpha-aminooctanoate, beta-methylphenylalanine, and alpha-aminophenylacetate. Less effective were aromatic amino acids with ring substituents; ineffective were basic amino acids and omega-amino isomers of norleucine and aminooctanoate. Lysine transport was moderately depressed by homoarginine or ornithine plus arginine; large neutral amino acids were also similarly inhibitory. Histidine or valine transport across the BBB was also strongly inhibited by large neutral amino acids that were the most effective competitors in the slices (norvaline, norleucine, alpha-aminooctanoate, and alpha-aminophenylacetate); homoarginine and 8-aminooctanoate were ineffective. Homoarginine, ornithine, and arginine almost completely blocked lysine transport, but the large neutral amino acids were barely inhibitory. When rats were fed a single meal containing individual atypical large neutral amino acids or homoarginine, brain pools of certain large neutral amino acids or of arginine and lysine, respectively, were depleted.     

Microbiological assay of amino acids in serum: valine, leucine, and methionine

A microbiological method for the assay of valine, leucine, and methionine in serum is described and its accuracy and reproducibility are assessed. Under the conditions specified the L-isomers only of the three amino acids are measured. The serum levels of the three amino acids in 60 normal subjects are presented.     

The effects of paradoxical sleep deprivation and valine on spatial learning and brain 5-HT metabolism

We have previously reported that rapid eye movement sleep deprivation (REMSD), induced by the flower pot technique, causes a deficit in reference spatial memory and increases rates of serotonin (5-HT) metabolism in the brain. In this study we used increased concentrations of dietary valine to inhibit tryptophan (TRP) transport across the blood-brain barrier in an attempt to modify the REMSD-induced increase of 5-HT metabolism. Rats were fed either a control diet or the same diet supplemented to 2% by weight valine, and were allocated to one of three experimental groups: cage control (CC), stress tank control (TC), or REMSD. Reference and working spatial memory of all rats was tested in a Morris water maze on Days 2, 3, and 4. REMSD produced a significant decrement in reference memory on Days 2 and 4, independent of dietary condition. The valine diet had a detrimental effect on the reference memory of TC rats on Day 2 but not Day 4. Measurements made on Day 4 indicated that the valine diet decreased brain TRP only in the CC rats. In contrast, the valine diet did not prevent increases in brain TRP or 5-HT metabolism in REMSD rats, and increased hypothalamic and brain stem TRP concentrations and the hippocampal 5-HIAA/5-HT ratio in TC rats. These results indicate that dietary valine does not prevent REMSD-induced changes in spatial memory or serotonin metabolism, although it does reduce brain TRP in nonstressed rats.     

Leptin access into the brain: a saturated transport mechanism in obesity.

In spite of extensive research in the last few years, we still do not have a clear understanding of how does leptin access its targets in the brain, and whether there is postreceptor defect in the brain of obese individuals. Recent data have shown that leptin is produced in the pituitary, where its receptor is also present. A better understanding of how leptin reaches the brain and how it modulates the release of hypothalamic neuropeptides and pituitary hormones is needed. This information is crucial in order to better understand the role that leptin may play in the pathophysiology of diseases, such as obesity, delayed puberty, or growth defects.     

Unidirectional transport of glucose and lactate into brain of fetal sheep and guinea-pig

The first-passage multiple-indicator dilution method was used to measure blood to brain transport of D- and L-glucose, D- and L-lactate and sucrose relative to 22Na, an impermeable reference tracer, in fetal sheep. Fractional extraction for D-glucose was 0.315 +/- 0.051 (S.E.M.) at normal glucose levels and fell to 0.198 +/- 0.041 at 5.2 +/- 0.4 mM-glucose. Fractional extractions for L-glucose, D- and L-lactate and sucrose were not different from zero. No specific blood-brain transport system was detected for L-lactate in fetal sheep in vivo (fractional extraction = -0.024 +/- 0.019). Uptake of L-lactate into isolated microvessels from fetal sheep cerebrum in vitro showed a slightly higher rate (32.2 +/- 8.9 pmol min-1 (mg protein)-1) than that for D-lactate (22.6 +/- 5.6). In fetal guinea-pigs, the carotid arterial injection method with tritiated water as the permeable reference was used to measure the brain uptake index (BUI). BUI was determined for D-glucose (0.304 +/- 0.065) sucrose (0.008 +/- 0.001), L-lactate (0.418 +/- 0.112) and D-lactate (0.071 +/- 0.024). Unidirectional influx calculated from these measurements and estimates of cerebral blood flow showed that transport would be rate-limiting for cerebral glucose utilization at arterial glucose levels below 0.5 mM in fetal sheep and 1.7 mM in fetal guinea-pig. In fetal sheep, but not in fetal guinea-pigs, lactate efflux may be limited by brain-blood transport.     

Ricin transport into cells: studies of endocytosis and intracellular transport

The plant toxin ricin binds to both glycoproteins and glycolipids with terminal galactose, and the toxin will therefore be endocytosed by the different mechanisms operating in a given cell. After endocytosis the toxin is transported to the Golgi apparatus by a process that differs from the Rab9-dependent transport of mannose-6-phosphate receptors. Retrograde toxin transport from the Golgi apparatus to the endoplasmic reticulum (ER) seems to be a requirement for subsequent toxin translocation to the cytosol where the toxin inhibits protein synthesis enzymatically. By using ricin we have characterized different types of endocytosis and the transport steps used by this toxin.     

Chronic loss of ovarian function decreases transport of leptin into mouse brain

Cannabinoid use is known to disrupt learning and memory in a number of species. cholecystokinin (CCK) release and CCK receptors have been implicated in spatial memory processes in rodents. Rat hippocampal CCK interneurons express cannabinoid 1 receptors (CB1). The CB1 agonist R(+)WIN 55,212-2 (WIN+), at 1 and 10 micromol, strongly inhibited potassium-evoked CCK release from rat hippocampal slices, while the inactive isomer S(-)WIN,55,212-3 (WIN-) had no effect. CCK release from cerebral cortical slices was not altered by WIN+.     

Low temperature inhibition on binding,transport, and incorporation of leucine,arginine, methionine,and histidine in Escherichia coli

A multiple amino acid auxotroph and a wild type of Escherichia coli K12 were used to study the effects of near minimum growth temperatures on the binding, transport, and cellular incorporation of selected amino acids. Both strains of the bacterium showed the same minimum growth temperature (8°C) when previously grown at 15°C. At 8°C and above, the auxotroph exhibited an overall greater ability to bind and transport amino acids than did the wild type. Below the minimum growth temperature, transport and cellular incorporation including respiration (uptake) were significantly lower for either organism. The NEU and HEPPEL osmotic shock treatment indicated the removal of the specific histidine-binding protein and the ability to bind histidine was not recovered by further incubation below 8°C. At 8°C and above, the cells recovered their ability to bind histidine within one hour. The evidence presented indicates a direct relationship between the auxotroph's minimum growth temperature and its ability to bind amino acids, specifically methionine.     

Talc injury: changes in glucosamine and leucine incorporation into liver and plasma proteins

Injury was induced in mice by sterile subcutaneous injection of talc. The host response to this injury was studied by measuring the incorporation of radioactive leucine and glucosamine into liver and plasma proteins, and the talc granuloma at various intervals between 2 and 528 hr following talc injection. Incorporation into liver proteins showed a marked transient increase at 21–45 hr following injury. Incorporation into plasma proteins indicated a biphasic response: a marked increase incorporation into the PCA-insoluble fraction at 21 hr with a return to normal values at 45 hr, and a similar marked increase into the PCA-soluble fraction at 45 hr with a gradual return towards normal values. This response was dependent upon the amount of talc injected. A biphasic pattern of incorporation was observed in the talc granuloma also, but this may have been related to the local healing processes.     

Mapping Pavlovian conditioning effects on the brain: blocking, contiguity, and excitatory effects

Pavlovian conditioning effects on the brain were investigated by mapping rat brain activity with fluorodeoxyglucose (FDG) autoradiography. The goal was to map the effects of the same tone after blocking or eliciting a conditioned emotional response (CER). In the tone-blocked group, previous learning about a light blocked a CER to the tone. In the tone-excitor group, the same pairings of tone with shock US resulted in a CER to the tone in the absence of previous learning about the light. A third group showed no CER after pseudorandom presentations of these stimuli. Brain systems involved in the various associative effects of Pavlovian conditioning were identified, and their functional significance was interpreted in light of previous FDG studies. Three conditioning effects were mapped: 1) blocking effects: FDG uptake was lower in medial prefrontal cortex and higher in spinal trigeminal and cuneate nuclei in the tone-blocked group relative to the tone-excitor group. 2) Contiguity effects: relative to pseudorandom controls, similar FDG uptake increases in the tone-blocked and -excitor groups were found in auditory regions (inferior colliculus and cortex), hippocampus (CA1), cerebellum, caudate putamen, and solitary nucleus. Contiguity effects may be due to tone-shock pairings common to the tone-blocked and -excitor groups rather than their different CER. And 3) excitatory effects: FDG uptake increases limited to the tone-excitor group occurred in a circuit linked to the CER, including insular and anterior cingulate cortex, vertical diagonal band nucleus, anterior hypothalamus, and caudoventral caudate putamen. This study provided the first large-scale map of brain regions underlying the Kamin blocking effect on conditioning. In particular, the results suggest that suppression of prefrontal activity and activation of unconditioned stimulus pathways are important neural substrates of the Kamin blocking effect.     

Comparison of behavioral effects of cortexin and cerebrolysin injected into brain ventricles

We compared central effects of polypeptide preparations cortexin and cerebrolysin injected into brain ventricles of Wistar rats in doses of 1, 10, and 100 μg. Both drugs exhibited moderate psychoactivating effect, the effects cortexin were more pronounced compared to those of cerebrolysin in all tests. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 143, No. 4, pp. 414–418, April, 2007     

Acid-evoked Ca2+ signalling in rat sensory neurones: effects of anoxia and aglycaemia

Ischaemia excites sensory neurones (generating pain) and promotes calcitonin gene-related peptide release from nerve endings. Acidosis is thought to play a key role in mediating excitation via the activation of proton-sensitive cation channels. In this study, we investigated the effects of acidosis upon Ca²⁺ signalling in sensory neurones from rat dorsal root ganglia. Both hypercapnic (pH_o 6.8) and metabolic–hypercapnic (pH_o 6.2) acidosis caused a biphasic increase in cytosolic calcium concentration ([Ca²⁺] _i ). This comprised a brief Ca²⁺ transient (half-time approximately 30 s) caused by Ca²⁺ influx followed by a sustained rise in [Ca²⁺] _i due to Ca²⁺ release from caffeine and cyclopiazonic acid-sensitive internal stores. Acid-evoked Ca²⁺ influx was unaffected by voltage-gated Ca²⁺-channel inhibition with nickel and acid sensing ion channel (ASIC) inhibition with amiloride but was blocked by inhibition of transient receptor potential vanilloid receptors (TRPV1) with (E)-3-(4-t-butylphenyl)-N-(2,3-dihydrobenzo[b][1,4] dioxin-6-yl)acrylamide (AMG 9810; 1 μM) and N-(4-tertiarybutylphenyl)-4-(3-cholorphyridin-2-yl) tetrahydropryazine-1(2H)-carbox-amide (BCTC; 1 μM). Combining acidosis with anoxia and aglycaemia increased the amplitude of both phases of Ca²⁺ elevation and prolonged the Ca²⁺ transient. The Ca²⁺ transient evoked by combined acidosis, aglycaemia and anoxia was also substantially blocked by AMG 9810 and BCTC and, to a lesser extent, by amiloride. In summary, the principle mechanisms mediating increase in [Ca²⁺] _i in response to acidosis are a brief Ca²⁺ influx through TRPV1 followed by sustained Ca²⁺ release from internal stores. These effects are potentiated by anoxia and aglycaemia, conditions also prevalent in ischaemia. The effects of anoxia and aglycaemia are suggested to be largely due to the inhibition of Ca²⁺-clearance mechanisms and possible increase in the role of ASICs.     

Intracellular content of thiol compounds, thiobarbituric acid reactive substances and gamma-glutamyl transpeptidase in rat brain during anoxia

Intracellular thiol compounds like glutathione play an important role in scavenging free radicals. Free radical reactions occur in the respiratory chain, and free radicals are known to accumulate in hypoxia and ischaemia. The content of thiobarbituric acid (TBA) reactive substances, thiol and non-protein thiol (reduced glutathione) concentrations along with the activity of gamma-glutamyl transpeptidase (GGTP), a membrane enzyme, were estimated in three regions of the brain in rats exposed to anoxia. It was observed that no change occurred in the content of TBA-reactive substances in any one of the three brain regions, although a significant increase in the non-protein thiol concentration occurred uniformly during anoxia. A significant increase in GGTP occurred in all the three brain regions during anoxia. The increase in the activity of GGTP is suggestive of an alteration in the function of blood-brain barrier during anoxia. These results are suggestive of an absence of lipid peroxidative damage during anoxia.