Our current understanding of HSV latency is based on a variety clinical observations, and in vivo, ex vitro model systems, each with unique advantages drawbacks. The criteria for authentically modeling include the ability to easily manipulate host genetics biological pathways, as well mimicking immune response viral pathogenesis human infections. Although realistically necessary when choosing model, cost, time requirement, ethical constraints, reagent availability are also equally important....

Successful replication of the intracellular parasite Toxoplasma gondii within its parasitophorous vacuole necessitates a substantial increase in membrane mass. The possible diversion and metabolism host cell lipids lipid precursors by was therefore investigated using radioisotopic fluorophore-conjugated compounds. Confocal microscopic analyses demonstrated that is selective with regards to both acquisition compartmentalization lipids. Lipids were compartmentalized into endomembranes and,...

INTRODUCTION The moss Physcomitrella patens has been used as an experimental organism for more than 80 years. Within the last 15 years, its use a model to explore plant functions increased enormously. ability gene targeting and RNA interference methods study function, availability of many tools comparative functional genomics (including sequenced assembled genome, physical genetic maps, 250,000 expressed sequence tags [ESTs]), dominant haploid phase that allows direct forward analysis have...

INTRODUCTION This article includes a series of methods for culturing the moss Physcomitrella patens at all stages its life cycle. Gametophytes are axenically cultured on solid agar-based media and in shaken liquid cultures. For long-term storage gametophytes, cultures maintained medium 10°C very short day. Cryopreservation may also be used. Finally, sporophytes generated by self-fertilization sexual crossing.

Cystogenesis associated with autosomal dominant polycystic kidney disease (ADPKD) is characterized by perturbations in the polarized phenotype and function of cyst-lining epithelial cells. The polycystins, protein products genes mutated majority ADPKD cases, have been described recently, but pathological mechanism which causal mutations result mislocalization cell membrane proteins has remained unclear. This report documents dissociation from basolateral three molecules essential for spatial...

During invasion by Toxoplasma gondii, host cell transmembrane proteins are excluded from the forming parasitophorous vacuole membrane (PVM) tight apposition of and parasite cellular membranes. Previous studies suggested that basis for selective partitioning constituents may be a preference microdomains, this hypothesis was herein tested. The diverse group molecular reporters raft nonraft subdomains monitored during time-lapse video or confocal microscopy. Unexpectedly, both lipid probes, as...

Abstract: Myelin/oligodendrocyte glycoprotein (MOG) is a CNS‐specific integral membrane protein that an atypical member of the immunoglobulin (Ig) superfamily with two potential transmembrane domains based upon hydropathy analysis. With only one other exception, all Ig family members possess single or no spanning region. In order to analyze MOG topology, we prepared stably transfected cells express mouse and used three domain‐specific antisera ascertain localization these hydrophilic...

Polycystin-1, the product of major gene mutated in autosomal dominant polycystic kidney disease (ADPKD), has been shown to associate with multiple epithelial cell junctions. Our hypothesis is that polycystin-1 an important protein for initial establishment cell-cell junctions and maturation localization dependent on degree polarization. Using laser-scanning confocal microscopy two models polarization, desmosomes were found colocalize during contact when forming. However, colocalization was...

INTRODUCTION This protocol describes how to isolate individual protoplasts from young gametophyte tissue of the moss Physcomitrella patens and regenerate them into plants if desired. Protonemal is grown on cellophane-overlay plates, harvested, treated with 0.5% Driselase produce protoplasts. After filtration washing, are regenerated solid medium.

INTRODUCTION This protocol describes how to transform moss ( Physcomitrella patens ) protoplasts using polyethylene glycol (PEG)-mediated DNA uptake. The transformation rates for direct uptake by of with and without genomic sequence (a targeting construct) are typically 10 −5 −3 , respectively. (These the frequencies stable transformants among regenerants surviving procedure.)

ABSTRACT Herpes simplex virus (HSV) latency in neurons remains poorly understood, and the heterogeneity of sensory nervous system complicates mechanistic studies. In this study, we used primary culture adult trigeminal ganglion (TG) mouse microfluidic devices an vivo model to examine subtypes involved HSV latency. HSV-infected neurofilament heavy-positive (NefH + ) were more likely express latency-associated transcripts (LATs) than infected heavy-negative − neurons. This differential...

Epithelial cells explanted from autosomal dominant polycystic kidney disease (ADPKD) tissue exhibit impaired exocytosis, specifically between the Golgi and basolateral membrane (Charron A, Nakamura B, Bacallo R, Wandinger‐Ness A. Compromised cytoarchitecture polarized trafficking in cells. J Cell Biol 2000; 148: 111–124.). Here defect is shown to result accumulation of transport marker vesicular stomatitis virus (VSV) G protein complex. complex morphology consequently altered cells, evident...

Expression of the hominoid-specific TBC1D3 oncoprotein enhances growth factor receptor signaling and subsequently promotes cellular proliferation survival. Here we report that is degraded in response to signaling, suggesting expression regulated by a factor-driven negative feedback loop. To gain better understanding how regulated, studied effects on post-translational processing turnover. Using yeast two-hybrid screen, identified CUL7, scaffolding subunit CUL7 E3 ligase complex, as...

Alphaherpesviruses are ubiquitous DNA viruses and include the human pathogens herpes simplex virus 1 (HSV-1) HSV-2 significant causes of ulcerative mucosal sores, infectious blindness, encephalitis, devastating neonatal disease. Successful primary infection persistent coexistence with host immune defenses dependent on ability these to counter antiviral response. HSV-1 other primate within Simplexvirus genus encode US11, an antagonist that promotes production by preventing shutdown protein...

Expression of viral genes and activation innate antiviral responses during infection result in an increase reactive oxygen species (ROS) toxic by-products energy metabolism which can lead to cell death. The mitochondrion its associated proteins are crucial regulators these related pathways such as autophagy apoptosis. Through a mass spectrometry approach, we have shown that the herpes simplex virus 1 (HSV-1) neurovirulence- autophagy-modulating protein ICP34.5 interacts with numerous...

INTRODUCTION This protocol describes a series of procedures for isolating nucleic acids (DNA and RNA) proteins from moss ( Physcomitrella patens ) tissue. includes rapid, small-scale procedure DNA, which results in genomic DNA that is only suitable polymerase chain reaction (PCR), as well method obtaining much larger amounts Southern analysis. The latter makes use the Nucleon PhytoPure Genomic Extraction Kit (GE Healthcare). RNA extraction uses Plant Isolation Mini (Agilent) with some...

INTRODUCTION In this protocol, the transformation of moss ( Physcomitrella patens ) protoplasts is performed via Agrobacterium -mediated transfer T-DNA. Protoplasts are incubated with and acetoseringone in regeneration medium. They then washed plated on antibiotic-containing medium to select for T-DNA insertion stable transformants. The rate protocol typically 10 −4 (expressed as frequency transformants among regenerants surviving procedure).

INTRODUCTION As an alternative to sexual crossing, protoplasts from two strains of moss ( Physcomitrella patens ) can be hybridized using polyethylene glycol (PEG). Although the efficiency is low, it requires no sophisticated apparatus. Hybrids are readily obtained complementary auxotrophic mutants or with transgenic antibiotic resistance markers. It now routine obtain hybrids that hygromycin- G418-resistant by selecting on medium containing both antibiotics.

The microvascular wall is remarkably simple, consisting only of the endothelial lining, subjacent basal lamina, and underlying periendothelial cells. This study describes characterization a novel protein. 80,000-molecular weight protein was predominantly associated with electron-lucent amorphous material in capillary laminae therefore termed cablin (protein lamina). Consistent its immunolocalization to microvasculature, synthesized secreted by cultured cells vascular smooth muscle...

INTRODUCTION This method makes use of biolistic bombardment to propel gold particles coated with DNA into protonemal tissue the moss Physcomitrella patens . It is especially suitable for transient gene expression studies, but it can be used obtain stable transformants.