Zebrafish TARP Cacng2 is required for the expression and normal development of AMPA receptors at excitatory synapses

Fast excitatory synaptic transmission in the CNS is mediated by the neurotransmitter glutamate, binding to and activating AMPA receptors (AMPARs). AMPARs are known to interact with auxiliary proteins that modulate their behavior. One such family of proteins is the transmembrane AMPA receptor‐related proteins, known as TARPs. Little is known about the role of TARPs during development, or about their function in non‐mammalian organisms. Here we report the presence of TARPs, specifically the prototypical TARP, stargazin, in developing zebrafish. We find that zebrafish express two forms of stargazin, Cacng2a and Cacng2b from as early as 12‐h post fertilization (hpf). Knockdown of Cacng2a and Cacng2b via splice‐blocking morpholinos resulted in embryos that exhibited deficits in C‐start escape responses, showing reduced C‐bend angles, smaller tail velocities and aberrant C‐bend turning directions. Injection of the morphants with Cacng2a or 2b mRNA rescued the morphological phenotype and the synaptic deficits. To investigate the effect of reduced Cacng2a and 2b levels on synaptic physiology, we performed whole cell patch clamp recordings of AMPA mEPSCs from zebrafish Mauthner cells. Knockdown of Cacng2a results in reduced AMPA currents and lower mEPSC frequencies, whereas knockdown of Cacng2b displayed no significant change in mEPSC amplitude or frequency. Non‐stationary fluctuation analysis confirmed a reduction in the number of active synaptic receptors in the Cacng2a but not in the Cacng2b morphants. Together, these results suggest that Cacng2a is required for normal trafficking and function of synaptic AMPARs, while Cacng2b is largely non‐functional with respect to the development of AMPA synaptic transmission. © 2015 Wiley Periodicals, Inc. Develop Neurobiol 76: 487–506, 2016     

Activation of the PI3K/mTOR Pathway following PARP Inhibition in Small Cell Lung Cancer

Small cell lung cancer (SCLC) is an aggressive malignancy with limited treatment options. We previously found that PARP is overexpressed in SCLC and that targeting PARP reduces cell line and tumor growth in preclinical models. However, SCLC cell lines with PI3K/mTOR pathway activation were relatively less sensitive to PARP inhibition. In this study, we investigated the proteomic changes in PI3K/mTOR and other pathways that occur following PAPR inhibition and/or knockdown in vitro and in vivo. Using reverse-phase protein array, we found the proteins most significantly upregulated following treatment with the PARP inhibitors olaparib and rucaparib were in the PI3K/mTOR pathway (p-mTOR, p-AKT, and pS6) (p≤0.02). Furthermore, amongst the most significantly down-regulated proteins were LKB1 and its targets AMPK and TSC, which negatively regulate the PI3K pathway (p≤0.042). Following PARP knockdown in cell lines, phosphorylated mTOR, AKT and S6 were elevated and LKB1 signaling was diminished. Global ATP concentrations increased following PARP inhibition (p≤0.02) leading us to hypothesize that the observed increased PI3K/mTOR pathway activation following PARP inhibition results from decreased ATP usage and a subsequent decrease in stress response signaling via LKB1. Based on these results, we then investigated whether co-targeting with a PARP and PI3K inhibitor (BKM-120) would work better than either single agent alone. A majority of SCLC cell lines were sensitive to BKM-120 at clinically achievable doses, and cMYC expression was the strongest biomarker of response. At clinically achievable doses of talazoparib (the most potent PARP inhibitor in SCLC clinical testing) and BKM-120, an additive effect was observed in vitro. When tested in two SCLC animal models, a greater than additive interaction was seen (p≤0.008). The data presented here suggest that combining PARP and PI3K inhibitors enhances the effect of either agent alone in preclinical models of SCLC, warranting further investigation of such combinations in SCLC patients.     

Chemical Aspects of Santalin as a Histological Stain

Recent research on the chemical nature of the red dyes isolated from Pterocarpus santalinus and certain West African plants, viz., Baphia nitida, Pterocarpus osun and Pterocarpus soyauxii, have been reviewed. P. santalinus contains santalins A, B and C., but no santarubin. Santalins and santarubins have been found in P. osun, P. soyauxii and B. nitida. The structural formulae of the santalins are presented and their differences from santarubins indicated. Santalins A and B have some similarities in structure with hematein. This is probably responsible for their staining properties; the possible mechanism of staining is discussed.     

Effect of desiccation on mosquito oviposition site selection in Mediterranean temporary habitats

1. Hydroperiod duration has been identified as the main factor determining the faunal composition and structure of aquatic communities in temporary habitats. It is hypothesised that desiccation will positively affect mosquito oviposition habitat selection during the post-drought period due to the lack of antagonists. 2. An experiment was carried out in outdoor mesocosms to assess whether desiccation events have post-drought effects on community richness and mosquito oviposition. Three different treatments were randomly assigned to the mesocosms: (i) eight mesocosms were left dried for a week and then reflooded; (ii) eight mesocosms stayed dry for 4 weeks before they were reflooded; (iii) eight mesocosms were maintained at a constant water volume of 30 litres during the entire experiment as controls. Mosquito oviposition and invertebrate community richness were monitored in every mesocosm, along with environmental parameters [water temperature, pH, conductivity, dissolved oxygen, total suspended solids (TSS) and chlorophyll a concentration]. 3. Post-drought mosquito oviposition and larval abundance were higher in the short-drought and long-drought pools than in the control. Desiccation negatively affected the biomass of the filter feeder invertebrates in both desiccation treatments. Chlorophyll a concentrations were higher in the long-drought pools than in controls. The negative impact of desiccation on zooplankton led to a post-drought increase in algae, associated with an increase in mosquito oviposition. 4. Despite immediate negative effect on mosquitoes, pulsed disturbances can benefit mosquitoes as they favour oviposition during the post-disturbance recovery period due to a lower abundance of mosquito antagonists and higher food resources for their offspring.     

Embryonic characteristics in superovulated mouse strains. Comparative analyses of the incidence of chromosomal aberrations, morphological malformations, and mortality of embryos from two strains of superovulated mice

Superovulation was induced in an inbred (C3HeB/FeJ) and a random-bred (CD-1) strain of mice and pregnant females were serially sacrificed from 6-15 days of gestation. All embryos were examined chromosomally and by external morphology. A G-banding technique was used to detect specific chromosomal constitutions. The common types of chromosomal aberrations were monosomies and trisomies. Our results showed that the proportion of chromosomally abnormal embryos was significantly greater in the superovulated group of inbred mice, than in the spontaneously ovulating inbred group, an observation not made in the random-bred mice. In each strain, superovulation yielded a significantly higher proportion of morphologically malformed embryos, but no significant difference between strains with respect to this criterion was noted after superovulation. There was a significant increase in the post-implantation embryonic mortality in the random-bred strain but not in the inbred strain following superovulation. This study indicates that each strain has its intrinsic pattern with regard to the fate of embryos following superovulation.     

Genetic heterogeneity and population structure in eastern India: red cell enzyme variability in ten Assamese populations

This paper is a part of the genetic study of the people of Assam (eastern India), initiated by the Anthropometry and Human Genetics Unit, Indian Statistical Institute, Calcutta, India, and the Dept. of Human Biology/Physical Anthropology, University of Bremen, W. Germany. The results of 1. allele distribution of five red cell enzyme polymorphisms in ten Assamese populations, 2. heterogeneity of allele frequencies and extent of gene differentiation among these populations, and 3. standard genetic distances are presented here. A total of 1024 blood samples was screened for aP, E D, AK, ADA and LDH enzyme systems for Brahmins, Kalitas, Kaibartas, Rajbanshis, Muslims, Ahoms, Chutiyas, Kacharis, Karbis (Mikirs) and Sonowals, of which the latter three are tribes. The gene diversity (FST) is smallest (0.0035) for pa and highest (0.1604) for HbE. The total FST value (0.0399 +/- 0.0141) appears to be statistically significant. From distance analysis two major clusters with sub-clusters in each are visible, which are in conformity with the ethnohistory of these populations.     

Biological affinities between the migrant groups of fishermen of Puri Coast, Orissa, India

Biological affinities between the three endogamous groups of marine fishermen of Puri are examined with the help of nine anthropometric measurements and 22 dermatoglyphic traits of fingers and palms. The results are interpreted in the light of their ethnic, geographical, and migrational backgrounds. Multiple discriminant analysis and Mahalanobis's generalized distances suggest higher discrimination of populations with reference to anthropometric traits as compared to that in dermatoglyphic variables. While the nature of interpopulation distances conforms to the geographic pattern in anthropometrics, no clear picture emerges in the dermatoglyphic distances supporting either ethnic or geographic evidence.