FRET-FLIM Investigation of PSD95-NMDA Receptor Interaction in Dendritic Spines; Control by Calpain,CaMKII and Src Family Kinase

Little is known about the changes in protein interactions inside synapses during synaptic remodeling, as their live monitoring in spines has been limited. We used a FRET-FLIM approach in developing cultured rat hippocampal neurons expressing fluorescently tagged NMDA receptor (NMDAR) and PSD95, two essential proteins in synaptic plasticity, to examine the regulation of their interaction. NMDAR stimulation caused a transient decrease in FRET between the NMDAR and PSD95 in spines of young and mature neurons. The activity of both CaMKII and calpain were essential for this effect in both developmental stages. Meanwhile, inhibition of Src family kinase (SFK) had opposing impacts on this decrease in FRET in young versus mature neurons. Our data suggest concerted roles for CaMKII, SFK and calpain activity in regulating activity-dependent separation of PSD95 from GluN2A or GluN2B. Finally, we found that calpain inhibition reduced spine growth that was caused by NMDAR activity, supporting the hypothesis that PSD95-NMDAR separation is implicated in synaptic remodeling.     

Estrogen receptors in ovary and uterus of immature hamster and rat: effects of estrogens

Cytosolic and nuclear estrogen receptors in the ovary and uterus of immature rats and hamsters were determined to evaluate why exogenous estrogens were ineffective in stimulating follicular maturation in the hamster compared to the rat. Animals were injected sc with oil or single injection of 1 mg estradiol cyclopentylpropionate (ECP) on Day 23 or a daily injection of 2 mg diethylstilbestrol (DES) on Days 23-25 and killed on Day 26. Total binding sites for estrogen in ovarian cytosol of control hamsters were half the number in the rat ovary (28 fmole/mg protein) and about 50% of the receptors were occupied in the hamster. The apparent affinity of the estrogen-cytosol receptor complex was also lower in the hamster (Kd; 1.41 nM) than in the rat (Kd; 0.52 nM). After ECP treatment, there was a tendency for translocation in all 4 tissues examined even though some differences were not statistically significant. However, after DES treatment both cytosol and nuclear estrogen receptors decreased in both species. This discrepancy may be due to the difference in the time course of the nuclear translocation, the difference in metabolism and difference in the binding potencies of ECP and DES. The lack of ovarian responsiveness to estrogen in the hamster thus appears to be due to the reduced number of cytosol receptor sites which have a low affinity for estrogen and are already partially occupied.     

Cultural longevity: Morin on cultural lineages

Morin has written a rich and valuable book. Its main aim is to isolate the factors involved in maintaining behavioural lineages over time, and to understand how these factors might interact. In doing so, it takes issue with the abstract and idealised models and arguments of dual-inheritance theorists, which are alleged in this account to rely on an overly simplistic notion of imitative learning. Morin’s book is full of ethnographic, anthropological, and psychological research, and there is much to commend in it. However, Morin’s arguments against the dual-inheritance theorists are less convincing when put under scrutiny, and his positive picture which includes appeals to ostensive communication, intrinsic appeal and cultural attraction has some difficulties. I argue that when we contrast dual-inheritance theorists and Morin, we find that there may be fewer differences and greater commonalities than Morin’s book might suggest.     

Xenotransplantation: Biotechnological Aspects and Current Attitudes

Xenotransplantation of organs from the large domestic species will only be successful if the donor animals have been genetically modified, in particular regarding the α-Gal epitope, certain human complements (CD55 and CD59) and/or H-transferase. This requires, among other things, major embryo-technological efforts, and the rate of success is still far from an acceptable level in the domestic species. It is currently poor, but the progress is very good. In this brief review certain embryo-technological problems will be addressed with the focus on the pig as potential organ donor. In addition, certain views of the Danish ad hoc Committee on Gene Technology on xenotransplantation will be presented in this context as they are supposed to mirror the concern and the views of the issues important for the public and each individual.     

A rare hybridization event in two common Caribbean wrasses (genus <Emphasis Type="Italic">Halichoeres</Emphasis>; family Labridae)

Molecular tools were used to evaluate the hybrid status of a specimen with intermediate colour pattern between Halichoeres bivittatus and Halichoeres garnoti from Belize. Phylogenetic analyses of the two species, eight Halichoeres species from new and old world lineages and two outgroups showed that the study species are closely related and that H. garnoti is the maternal contributor to the putative hybrid specimen, based on partial mitochondrial COI data. Direct sequencing of Intron 1 of the nuclear ribosomal protein S7 identified H. bivittatus as sister to H. garnoti with the putative hybrid specimen in an intermediate position, due to heterozygosity at nucleotides alternatively fixed in the two putative parent species. This is consistent with the hybrid status of the specimen, with parental contributions from both H. garnoti and H. bivittatus. These results, combined with no evidence of introgression between the two parent species (based on the mtDNA and the single investigated nuclear marker) and the biogeography and ecology of these species suggests that this is a rare event with minimal evolutionary implications.     

Postexercise protein supplementation improves health and muscle soreness during basic military training in Marine recruits.

Elevated postexercise amino acid availability has been demonstrated to enhance muscle protein synthesis acutely, but the long-term impact of postexercise protein supplementation on variables such as health, muscle soreness, and function are unclear. Healthy male US Marine recruits from six platoons (US Marine Corps Base, Parris Island, SC; n = 387; 18.9 +/- 0.1 yr, 74.7 +/- 1.1 kg, 13.8 +/- 0.4% body fat) were randomly assigned to three treatments within each platoon. Nutrients supplemented immediately postexercise during the 54-day basic training were either placebo (0 g carbohydrate, 0 g protein, 0 g fat), control (8, 0, 3), or protein supplement (8, 10, 3). Subjects and observers making measurements and data analysis were blinded to subject groupings. Compared with placebo and control groups, the protein-supplemented group had an average of 33% fewer total medical visits, 28% fewer visits due to bacterial/viral infections, 37% fewer visits due to muscle/joint problems, and 83% fewer visits due to heat exhaustion. Recruits experiencing heat exhaustion had greater body mass, lean, fat, and water losses. Muscle soreness immediately postexercise was reduced by protein supplementation vs. placebo and control groups on both days 34 and 54. Postexercise protein supplementation may not only enhance muscle protein deposition but it also has significant potential to positively impact health, muscle soreness, and tissue hydration during prolonged intense exercise training, suggesting a potential therapeutic approach for the prevention of health problems in severely stressed exercising populations.     

Compensation of large motion sensor displacements during long recordings of limb movements

In motion capture applications using electromagnetic tracking systems the process of anatomical calibration associates the technical frames of sensors attached to the skin with the human anatomy. Joint centers and axes are determined relative to these frames. A change of orientation of the sensor relative to the skin renders this calibration faulty. This sensitivity regarding sensor displacement can turn out to be a serious problem with movement recordings of several minutes duration. We propose the “dislocation distance” as a novel method to quantify sensor displacement and to detect gradual and sudden changes of sensor orientation. Furthermore a method to define a so called fixed technical frame is proposed as a robust reference frame which can adapt to a new sensor orientation on the skin. The proposed methods are applied to quantify the effects of sensor displacement of 120 upper and lower limb movement recordings of newborns revealing the need for a method to compensate for sensor displacement. The reliability of the fixed technical frame is quantified and it is shown that trend and dispersion of the dislocation distance can be significantly reduced. A working example illustrates the consequences of sensor displacement on derived angle time series and how they are avoided using the fixed technical frame.