NMDA Receptor Heterogeneity During Postnatal Development of the Rat Brain: Differential Expression of the NR2A, NR2B, and NR2C Subunit Proteins

Abstract: Changes in the expression of the NMDA receptor subunits (NRs) NR2A, 2B, and 2C were investigated in histo blots of the developing rat brain with subunit-specific antisera. At birth, the NR2B subunit was detected almost ubiquitously, the NR2A subunit staining was faint and restricted to the hippocampus, cerebral cortex, and striatum, and no NR2C subunit immunoreactivity was detected. During the first 3 postnatal weeks, the NR2B subunit became confined to forebrain structures, whereas the NR2A immunoreactivity became abundantly expressed throughout the brain. The NR2C immunoreactivity emerged 5 days after birth in the olfactory bulb, thalamus, and vestibular nuclei and became very intense after 10 days in cerebellar granule cells, its primary site of expression in adulthood. After 3 weeks, NR2A and NR2B immunoreactivity decreased to adult levels, whereas NR2C immunoreactivity remained unchanged. The patterns of distribution of the subunit proteins were in agreement with those of their corresponding mRNAs, as monitored by in situ hybridization histochemistry, although the mRNA translation appeared to be delayed by several days in certain areas. Our results reveal a progressive increase in the heterogeneity of NMDA receptors due to the comparably late onset of NR2A and NR2C subunit expression and by the area-specific rearrangement of NR2B subunit expression following birth.     

Comparison of techniques for measuring pulse-wave velocity in the rat

Mitchell, Gary F., Marc A. Pfeffer, Peter V. Finn, andJanice M. Pfeffer. Comparison of techniques for measuringpulse-wave velocity in the rat. J. Appl.Physiol. 82(1): 203-210, 1997.We evaluatedmethods for measuring average and regional pulse-wave velocity alongthe full length of the aorta in 18-mo-old ether-anesthetized malespontaneously hypertensive rats. Catheter-tip manometers were placed inthe ascending and descending thoracic aorta via the right carotid andleft femoral arteries, respectively. As the distal catheter waswithdrawn at 1-cm intervals, the relationship between distal catheterinsertion distance and distance between transducers was determined fromthe intercept of the insertion distance vs. transmission delayregression line. Methods that assessed the foot-to-foot time delaybetween pressures accurately predicted the separation between catheters(measured distance of 14.3 cm; intercept of 14.0 ± 0.5 cm;P = not significant) were highlyreproducible (coefficient of variation of 2.3% for repeated measurements) and showed minimal variability (range 509 ± 30 to 600 ± 29 cm/s) along the full length of the aorta. Methods that madeuse of the pressure-pressure transfer function were spatially (range ofvalues along the aorta 367 ± 17 to 722 ± 39 cm/s) and temporally more variable, especially during vasoconstriction with methoxamine, due to the effects of reflected waves.

     

Modeling the population dynamics of annual plants with seed bank and density dependent effects

A model is proposed for the population dynamics of an annual plant (Sesbania vesicaria) with a seed bank (i.e. in which a proportion of seeds remain dormant for at least one year). A simple linear matrix model is deduced from the life cycle graph. The dominant eigenvalue of the projection matrix is estimated from demographic parameters derived from field studies. The estimated values for population growth rate () indicates that the study population should be experiencing a rapid exponential increase, but this was not the case in our population.The addition of density dependent effects on seedling survivorship and adult fecundity, effects for which field studies provide evidence, considerably improves our model. Depending on the demographic parameters, the model leads to stable equilibrium, oscillations, or chaos. Study of the behaviour of this model in the parameter space shows that the existence of a seed bank allows higher among-year variation of adult fecundity, without leaving the region of demographic stability. Field data obtained over 3 years confirm this prediction.     

Influence of central command and ergoreceptors on the splanchnic circulation during isometric exercise

The splanchnic circulation can make a major contribution to blood flow changes. However, the role of the splanchnic circulation in the reflex adjustments to the blood pressure increase during isometric exercise is not well documented. The central command and the muscle chemoreflex are the two major mechanisms involved in the blood pressure response to isometric exercise. This study aimed to examine the behaviour of the superior mesenteric artery during isometric handgrip (IHG) at 30% maximal voluntary contraction (MVC). The pulsatility index (PI) of the blood velocity waveform of the superior mesenteric artery was taken as the study parameter. A total of ten healthy subjects [mean age, 21.1 (SEM 0.3) years] performed an IHG at 30% MVC for 90 s. At 5 s prior to the end of the exercise, muscle circulation was arrested for 90 s to study the effect of the muscle chemoreflex (post exercise arterial occlusion, PEAO). The IHG at 30% MVC caused a decrease in superior mesenteric artery PI, from 4.84 (SEM 1.57) at control level to 3.90 (SEM 1.07) (P = 0.015). The PI further decreased to 3.17 (SEM 0.70) (P = 0.01) during PEAO. Our results indicated that ergoreceptors may be involved in the superior mesenteric artery vasodilatation during isometric exercise.     

Specific immunotherapy with vaccinia oncolysates

Summary A live vaccinia virus-augmented tumor cell vaccine (vaccinia oncolysate) has been used as a specific active immune mechanism stimulator against certain advanced human cancers. This preliminary trial, in which the same dose of vaccinia oncolysate was used in 29 patients, showed the vaccine to be safe, non-toxic, and potentially effective. This study was designed as a toxicity/feasibility trial at a single vaccine dose. There was no effort to look at other dose levels. The heterogeneity of this early tumor group only reflects the type of patient available to the study during the early phase of vaccine testing. Randomized prospective trials have begun to make future analysis easier.     

A test for preference of association in a color polymorphic poeciliid fish: laboratory study

Synopsis The ecological and evolutionary forces maintaining genetic polymorphism within populations is of continuing interest to evolutionary biologists. Male pygmy swordtails,Xiphophorus pygmaeus, are polymorphic at a Y-linked locus controlling body color. Fish with the + and cp alleles have blue bodies; those with the con allele have bright gold bodies. Male and female fish were tested in the laboratory to determine if there were any preferences for association with groups of male fish based on color. Single test fish were presented with two groups of males of different phenotypes, and times in which the test fish spent in proximity to each group were recorded. Males of different phenotypes showed no differences in preference, nor did phenotype of the males in the groups affect the behavior of the test fish. Females also showed no preference for males of a particular phenotype. Finally, female test fish showed no preference for groups containing a male with the rare phenotype compared to a group containing all males of the same phenotype.     

Sperm motility in turbot,Scophthalmus marimus: initiation of movement and changes with time of swimming characteristics

Synopsis Turbot sperm motility is observed using dark field microscopy and stroboscopic illumination combined with video recording. Sperm motility is triggered by dilution of spermatozoa in sea water or in non ionic media (glucose or saccharose), presenting osmotic pressure ranging from 300 to 2100 mOsmol. The percentage of motile spermatozoa reaches 100% under conditions of osmotic pressure of 300 to 1100 mOsmol and pH close to 8.0. In full sea water, glucose or saccharose solutions an agglutination of spermatozoa is observed; this is prevented by addition of bovine serum albumin (5 mg ml^–1). Immediately after transfer in activation solutions, 100% spermatozoa are motile in most samples freshly stripped. This percentage drops suddenly between 15 and 30% after 70 to 100 sec. The beat frequency remains at a constant value of 50 Hz during 40 s post activation and then drops suddenly between 15 and 30 Hz. The spermatozoa velocity is about 200 micrometers s^–1 during 30 to 40 s and then declines to a stable value of 100 micrometers s^–1 at 50 s post activation. After 1.20 mn, more and more spermatozoa become motionless. The minimum calculated and averaged distance covered during 1.20 min, is about 12 mm. The high performances of turbot spermatozoa motility are interpreted as a compensatory mechanism for the low sperm production.     

Investigation of the folding pathway of the TEM-1 β-lactamase

The TEM-1 β-lactamase is a globular protein containing 12 proline residues. The folding mechanism of this enzyme was investigated by kinetic and equilibrium experiments with the help of fluorescence spectroscopy and circular dichroism. The equilibrium denaturation of the protein induced by guanidine hydrochloride occurs in two discrete steps, indicating the existence of a thermodynamically stable intermediate state. Thisstate is 5.2 ± 0.4 kcal/mol less stable than the native conformation and 5.7 ± 0.2 kcal/mol more stable than the fully denaturedprotein. This intermediate state exhibits a high content of native secondary structure elements but is devoid of specific tertiary organization; its relation to the “molten globule” is discussed. Refolding kinetic experimentsrevealed the existence of a transient intermediate conformation between thethermodynamically stable intermediate and the native protein. This transient intermediate appears rapidly during the folding reaction. It exhibits a secondary structure content very similar to that of the native protein and has also recovered a significant amount of tertiary organisation. The final refolding step of the TEM-1 β-lactamase, leading to the native enzyme, is dominated by two major slow kinetic phases which probablyreflect a very complex process kinetically limited by proline cis/transisomerization. © 1995 Wiley-Liss, Inc.     

An estimate of spin diffusion in a spin subset: Application to iterative distance calculation from 3D 15N NOESY-HMQC

Summary A method for quantification of distances between amide hydrogens using only the 3D NOESY-HMQC experiment recorded on a ¹⁵N-labelled protein is presented. This method is based on an approximate expression of the NOE intensities between amide hydrogens obtained from continuum modelling of the non-amide spins; this expression is used in a distance calculation algorithm. The algorithm has been named CROWD, standing for Continuum approximation of Relaxati On path Ways between Dilute spins. This approximation as well as the CROWD algorithm are tested on a simulated case; the CROWD algorithm is then applied to experimental data, measured on a fragment of bacteriorhodopsin.     

Tripeptides as selective inhibitors of src-SH2 phosphoprotein interactions

Summary This paper describes the synthesis of phosphorylated peptides of the general structural Ac-Tyr(PO₃H₂)-Glu-Xaa_NH₂, where Xaa represents a hydrophobic -amino acid of d-configuration. These peptides displayed activities in the micromolar range in inhibiting src-SH2 domain/epidermal growth factor receptor interactions.