Desensitization of the Neurokinin 1 Receptor Is Mediated by the Receptor Carboxy-Terminal Region, but Is Not Caused by Receptor Internalization

Abstract: The carboxy-terminal cytoplasmic regions of the rat neurokinin 1 (substance P) and neurokinin 2 (neurokinin A) receptors have been exchanged to determine if this region of the neurokinin 1 receptor is involved in its desensitization. When expressed at similar levels in stably transfected Chinese hamster ovary (CHO) cell lines, receptors containing the carboxy-terminal region of the neurokinin 1 receptor desensitized significantly more (as measured by reduction of the inositol 1,4,5-trisphosphate response) when preexposed for 1 min to 1 µ M neurokinin, indicating a role for the carboxy-terminal region of the neurokinin 1 receptor in its desensitization. Measurement of receptor internalization using radiolabeled neurokinins (0.3 n M ) indicated that ∼75–80% of the receptors were internalized in each cell line after 10 min at 37°C, with no observable correlation between neurokinin receptor desensitization and internalization. Measurement of loss of receptor surface sites for cell lines CHO NK1 and CHO NK1NK2 following exposure to 1 µ M substance P also indicated no obvious relationship between the percent desensitization and percent of receptors internalized. Also, two inhibitors of neurokinin 1 receptor internalization, phenylarsine oxide and hyperosmolar sucrose, did not inhibit neurokinin 1 receptor desensitization. The protein kinase inhibitors Ro 31-8220, staurosporine, and Zn²⁺ had no effect on neurokinin 1 receptor desensitization, indicating that the kinases affected by these agents are not rate-limiting in neurokinin 1 receptor desensitization in this system.     

Developmental and Regional Expression of NMDA Receptor Subtypes Containing the NR2D Subunit in Rat Brain

Abstract: The regional and developmental expression of NMDA receptors containing the NR2D subunit was analyzed on the level of the subunit mRNA and protein in rat brain. RNase protection experiments indicated that among two proposed splice variants of the NR2D subunit, only the NR2D-2 subunit is expressed. The regional distribution of the NR2D subunit protein was visualized with a newly developed NR2D-2 subunit-specific antiserum on brain sections using the histoblot technique. In adult brain, NR2D immunoreactivity was mainly restricted to diencephalic, mesencephalic, and brainstem structures. During postnatal development, the NR2D subunit was detected transiently in certain regions, such as the ventro-basal complex of the thalamus, hippocampus, inferior colliculus, and brainstem reticular formation, suggesting that NR2D subunit-containing receptors play a role in these brain areas only during development. The level of NR2D subunit mRNA and protein decreased during late postnatal development. However, significant levels of NR2D subunit mRNA and protein were present in adulthood, in particular, in the globus pallidus, thalamus, subthalamic nuclei, and superior colliculus. These results indicate a functional relevance for NMDA receptors containing the NR2D subunit in the developing and adult brain, although its expression in the adult brain is less prominent and restricted to a few brain areas.     

Kinetics of tert-[35S]Butylbicyclophosphorothionate Binding in the Cerebral Cortex of Newborn and Adult Rats: Effects of GABA and Receptor Desensitization

Abstract: The effects of GABA on the kinetics of tert -[³⁵S]butylbicyclophosphorothionate ([³⁵S]TBPS) binding to the convulsant site of GABA_A receptors were studied in membrane suspensions from the cerebral cortex of newborn (1-day-old) and adult (90-day-old) rats. TBPS dissociation was biphasic in neonates and adults, indicating that more than one interconvertible state of [³⁵S]TBPS binding sites may be present in the cerebral cortex. In the absence of GABA, the fast ( t _1/2, 11 min) and slow ( t _1/2, 77 min) components of TBPS dissociation in newborn rats were approximately fourfold slower than in adults. The acceleration of the dissociation rates caused by 2 µ M GABA, however, was more robust in neonates than in adults (six- to ninefold vs. twofold increase, respectively). Moreover, the dissociation rates of TBPS in membranes preincubated with 2 µ M GABA (dissociation started by adding 40 µ M picrotoxin) were two- to fourfold slower than in membranes preincubated without GABA (dissociation started by adding 40 µ M picrotoxin plus 2 µ M GABA). Taken together, these results suggest that (1) the closed state of GABA_A receptors is associated with a more effective steric barrier for the binding of TBPS in neonates compared with adults, (2) GABA produces a larger acceleration of the binding kinetics of TBPS in neonates than in adults, and (3) long incubations with GABA may cause receptor desensitization, which in turn slows down the dissociation rates of TBPS.     

Sensory characteristics of the P afferent neurone of the crab thoracic-coxal muscle receptor organ

Intracellular recordings were made from the P fibre, the smallest of the three afferent neurones innervating the thoracic-coxal muscle receptor organ of the crab (Carcinus maenas). While the two larger afferents are nonspiking, the response of the P fibre to a trapezoidal change in receptor muscle length consists of a single action potential signalling the onset of stretch superimposed on a graded amplitude receptor potential. The P fibre is sensitive to the velocity of the applied stretch, but is insensitive to static joint position, stretch amplitude and the velocity of the release phase. The presence and amplitude of the action potential depends on the initial length of the receptor muscle, the tension caused by efferent activation of the receptor muscle prior to receptor stretch, and on the velocity of stretch. Length constant (1.9 mm) and specific membrane resistance (76 K · cm²) values obtained for the P fibre, together with its small diameter (7 m) suggest that this neurone is less well adapted to conveying passive signals to the thoracic ganglion than are the S and T fibres. It is likely that the P fibre complements the length sensitivity of the S fibre and the tension and velocity sensitivity of the T fibre by signalling the onset of receptor stretch via single action potentials.Abbreviations TCMRO thoracic-coxal muscle receptor organ - TTX tetrodotoxin     

Recent developments in vitamin E nutrition of turkeys

Research and field observations have shown that vitamin E status of poults is often inadequate during the 5 to 21-day posthatching period. Although overt consequences of this inadequacy may not be evident and clear-cut, subtle adverse effects on health and metabolic efficiency are probable. These latter effects can result in uneconomical performance of turkeys. It therefore seems advisable to supplement the first diet (starter or prestarter) of poults with 100 to 150 IU of vitamin E. The cost of doing so will contribute little to the total cost of production and may facilitate more efficient turkey production. In the meantime, other alternatives that may be effective and economical need to be evaluated.     

Degenerative Joint Disease and enthesopathies in a Bronze Age population from Sindou cave (Senaillac-Lauzes,Lot, France)

The anthropological characteristics of the people who lived during the cultural period of the Late Bronze Age in South West France still remain practically unknown because very few sites have provided skeletal remains which permit of an exhaustive study. The cave of Sindou is, in that sense, one of the scarce exceptions. Although the sample of Sindou cannot be considered as representative of the whole regional population (N=50), we studied the presence and severity of DJD and enthesopathies of microtraumatic origin with the aim of finding some data which contribute to the knowledge of several biological aspects of this human group. From the results of the comparisons of the Sindou remains with two different medieval samples, a great similarity is deduced for these skeletal markers, but the higher frequency and severity of Achilles tendon enthesopathy in Sindou is a probable index of a higher level of physical stress at this specific localisation.     

蔷薇属38个野生种果实的维生素含量及其与分组的关系

对蔷薇属 (Rosa) 38个野生种果实 (以下简称蔷薇果 )的经济性状进行了分析 ,并测定了 VC、VE 和胡萝卜素等重要维生素的含量。蔷薇果 VC 含量在该属种间差异很大 ,以秦岭蔷薇 (R.tsinglingensis)的含量为最高 (2 576mg/ 1 0 0 g) ,德钦蔷薇 (R.deqenensis)的含量为最低 (49mg/ 1 0 0 g)。胡萝卜素含量种间差异明显 ,以软条七蔷薇 (R.henryi)的含量为最高 (1 9.2 4 mg / 1 0 0 g) ,黄刺玫 (R.xanthina)的含量为最低 (0 .0 6 mg/ 1 0 0 g)。 VE 含量种间差异较小 ,在 1 .34 mg/ 1 0 0 g(黄刺玫 )至 3.86 mg/ 1 0 0 g(硕苞蔷薇R.bracteata)之间。对蔷薇亚属 54种野生种果实重要维生素含量的统计分析表明 ,维生素含量与分组具有一定相关性 ,尤以 VC 含量与分组的相关性最为明显 ,桂味组和小叶组 Vc含量很高 (均值都高于 1 80 0mg/ 1 0 0 g) ;合柱组、月季组、木香组和硕苞组含量很低 (均值都在 30 0 mg/ 1 0 0 g以下 ) ,芹叶组除宽刺蔷薇 (R.platyacantha) VC含量很高外 ,其余种类含量都很低 (均值为 1 90 mg/ 1 0 0 g)。胡萝卜素含量与分组也具有一定相关性 ,桂味组、芹叶组、合柱组和硕苞组的胡萝卜素含量较高 ,均值在 6 mg/ 1 0 0 g以上 ;月季组、小叶组和木香组含量较低 ,均值在 0 .4mg/ 1 0 0 g以下。V     

Regulation and functional significance of phospholipase D in myocardium

There is now clear evidence that receptor-dependent phospholipase D is present in myocardium. This novel signal transduction pathway provides an alternative source of 1,2-diacylglycerol, which activates isoforms of protein kinase C. The members of the protein kinase C family respond differently to various combinations of Ca²⁺, phosphatidylserine, molecular species of 1,2-diacylglycerol and other membrane phospholipid metabolites including free fatty acids. Protein kinase C isozymes are responsible for phosphorylation of specific cardiac substrate proteins that may be involved in regulation of cardiac contractility, hypertrophic growth, gene expression, ischemic preconditioning and electrophysiological changes. The initial product of phospholipase D, phosphatidic acid, may also have a second messenger role. As in other tissues, the question how the activity of phospholipase D is controlled by agonists in myocardium is controversial. Agonists, such as endothelin-1, atrial natriuretic factor and angiotensin 11 that are shown to activate phospholipase D, also potently stimulate phospholipase C- in myocardium. PMA stimulation of protein kinase C inactivates phospholipase C and strongly activates phospholipase D and this is probably a major mechanism by which agonists that promote phosphatidyl-4,5-bisphosphate hydrolysis secondary activate phosphatidylcholine-hydrolysis. On the other hand, one group has postulated that formation of phosphatidic acid secondary activates phosphatidyl-4,5-bisphosphate hydrolysis in cardiomyocytes. Whether GTP-binding proteins directly control phospholipase D is not clearly established in myocardium. Phospholipase D activation may also be mediated by an increase in cytosolic free Ca²⁺ or by tyrosine-phosphorylation.     

Vitamin A supplementation inhibits chemiluminescence and lipid peroxidation in isolated rat liver microsomes and mitochondria

In the present study we investigated if administration of vitamin A could protect rat liver microsomes and mitochondria from in vitro peroxidation. Appreciable decrease of chemiluminescence and lipid peroxidation was measured in microsomal membranes from rats receiving vitamin A, with respect to control animals. In membranes derived from control animals, the fatty acid composition was profoundly modified when subjected to in vitro peroxidation mediated by ascorbate-Fe⁺⁺, with a considerable decrease of 20:4 n6 and 22:6 n3 in mitochondria and 18:2 n6 and 20:4 n6 in microsomes. As a consequence the peroxidizability index, a parameter based on the maximal rate of oxidation of specific fatty acids was higher in supplemented animals than in control group when both kind of membranes were analyzed. These changes were less pronounced in membranes derived from rats receiving vitamin A. These results are in agreement with previous results that indicated that vitamin A may act as an antioxidant protecting membranes from deleterious effects.Abbreviations BHT butylated hydroxytoluene - BSA bovine serum albumin - CL chemiluminescence - PI peroxidizability index Member of Carrera del Investigador Científico, Consejo Nacional de Investigaciones Cientificas y Técnicas de la Republica Argentina     

Early degradation of photosynthetic membranes in carob and sunflower cotyledons

The assimilatory activity of cotyledons can play an essential role in the survival of seedlings with a slow and delayed development of primary leaves. Changes in the photosynthetic activity of the cotyledon, from the onset of greening through senescence, were studied in two such plants, carob and sunflower, in order to determine its efficiency and duration, also in connection with the achievement of assimilatory autonomy by the plantlet. Chlorophyll analyses showed that the cotyledon's chloroplasts reached maximal greening in plantlets with a pair of expanded leaves. In contrast, the cotyledon's photosynthetic activity, measured as the rate of oxygen release, started to decrease early, before expansion of primary leaves. The decrease was due to the inactivation of a number of photosystem II (PSII) units, as revealed by immunodetection of breackdown products of the reaction centre's D1 and D2 thylakoid proteins. No signals of PSII alteration were noticed in the primary leaf chloroplasts that differentiated under the same environmental conditions. The damage to the cotyledon PSII, occurring in a non-photoinhibitory situation, might be due to a slower rate of turnover of D1 polypeptide than in the leaf thylakoids. The differential turnover of this protein in cotyledons and in leaves might represent an organ-specific regulation of the photosynthetic activity. The peculiarity of the cotyledon thylakoids make these organs useful objects for studying the metabolic cycle of both D1 and D2 proteins in vivo, under non-photoinhibiting conditions.