Organotypic Tissue Culture of Adult Rodent Retina Followed by Particle-Mediated Acute Gene Transfer In Vitro

BACKGROUND: Organotypic tissue culture of adult rodent retina with an acute gene transfer that enables the efficient introduction of variable transgenes would greatly facilitate studies into retinas of adult rodents as animal models. However, it has been a difficult challenge to culture adult rodent retina. The purpose of this present study was to develop organotypic tissue culture of adult rodent retina followed by particle-mediated acute gene transfer in vitro. METHODOLOGY/PRINCIPAL FINDINGS: We established an interphase organotypic tissue culture for adult rat retinas (>P35 of age) which was optimized from that used for adult rabbit retinas. We implemented three optimizations: a greater volume of Ames' medium (>26 mL) per retina, a higher speed (constant 55 rpm) of agitation by rotary shaker, and a greater concentration (10%) of horse serum in the medium. We also successfully applied this method to adult mouse retina (>P35 of age). The organotypic tissue culture allowed us to keep adult rodent retina morphologically and structurally intact for at least 4 days. However, mouse retinas showed less viability after 4-day culture. Electrophysiologically, ganglion cells in cultured rat retina were able to generate action potentials, but exhibited less reliable light responses. After transfection of EGFP plasmids by particle-mediated acute gene transfer, we observed EGFP-expressing retinal ganglion cells as early as 1 day of culture. We also introduced polarized-targeting fusion proteins such as PSD95-GFP and melanopsin-EYFP (hOPN4-EYFP) into rat retinal ganglion cells. These fusion proteins were successfully transferred into appropriate locations on individual retinal neurons. CONCLUSIONS/SIGNIFICANCE: This organotypic culture method is largely applicable to rat retinas, but it can be also applied to mouse retinas with a caveat regarding cell viability. This method is quite flexible for use in acute gene transfection in adult rodent retina, replacing molecular biological bioassays that used to be conducted in isolated cultured cells

Two novel proteins in the mitochondrial outer membrane mediate β-barrel protein assembly

Mitochondrial outer and inner membranes contain translocators that achieve protein translocation across and/or insertion into the membranes. Recent evidence has shown that mitochondrial β-barrel protein assembly in the outer membrane requires specific translocator proteins in addition to the components of the general translocator complex in the outer membrane, the TOM40 complex. Here we report two novel mitochondrial outer membrane proteins in yeast, Tom13 and Tom38/Sam35, that mediate assembly of mitochondrial β-barrel proteins, Tom40, and/or porin in the outer membrane. Depletion of Tom13 or Tom38/Sam35 affects assembly pathways of the β-barrel proteins differently, suggesting that they mediate different steps of the complex assembly processes of β-barrel proteins in the outer membrane

A direct proof of the weighted Pólya–Knopp inequality following Carleson’s method

The aim of the paper is to provide a direct proof of the weighted Pólya–Knopp inequality. This inequality (which is a limiting case of the Ariño–Muckenhoupt inequalities), involving non-increasing functions, was initially established by Sbordone–Wik, who proved its validity under the necessary and sufficient condition that the weight satisfies an appropriate doubling condition. Our main contribution is to use Carleson’s approach to Carleman’s inequality in conjunction with Hardy’s lemma and Sbordone–Wik’s doubling condition, in order to obtain the weighted Pólya–Knopp inequality

A barley PHD finger transcription factor that confers male sterility by affecting tapetal development

Controlling pollen development is of major commercial importance in generating hybrid crops and selective breeding, but characterized genes for male sterility in crops are rare, with no current examples in barley. However, translation of knowledge from model species is now providing opportunities to understand and manipulate such processes in economically important crops. We have used information from regulatory networks in Arabidopsis to identify and functionally characterize a barley PHD transcription factor MALE STERTILITY1 (MS1), which expresses in the anther tapetum and plays a critical role during pollen development. Comparative analysis of Arabidopsis, rice and Brachypodium genomes was used to identify conserved regions in MS1 for primer design to amplify the barley MS1 gene; RACE-PCR was subsequently used to generate the full-length sequence. This gene shows anther-specific tapetal expression, between late tetrad stage and early microspore release. HvMS1 silencing and overexpression in barley resulted in male sterility. Additionally, HvMS1 cDNA, controlled by the native Arabidopsis MS1 promoter, successfully complemented the homozygous ms1 Arabidopsis mutant. These results confirm the conservation of MS1 function in higher plants and in particular in temperate cereals. This has provided the first example of a characterized male sterility gene in barley, which presents a valuable tool for the future control of male fertility in barley for hybrid development

Further research on wavelet inversion formula

This note is an announcement of a continuation of the authors' paper “Some variations on wavelet inversion formula". The aim of the note is to replace the special orthogonal group by the spin group in the wavelet inversion formula. A matrix representation of the three dimensional rotation associated with a spin is given explicitly as an appendix.departmental bulletin pape

Widths of embeddings of 2-microlocal Besov spaces

We consider the asymptotic behaviour of the approximation, Gelfand and Kolmogorov numbers of compact embeddings between 2-microlocal Besov spaces with weights defined in terms of the distance to a $d$-set $U\subset \mathbb{R}^n$. The sharp estimates are shown in most cases, where the quasi-Banach setting is included.Comment: 28 page

An efficient experimental strategy for mouse embryonic stem cell differentiation and separation of a cytokeratin-19-positive population of insulin-producing cells

Embryonic stem cells are a potential source for insulin‐producing cells, but existing differentiation protocols are of limited efficiency. Here, the aim has been to develop a new one, which drives development of embryonic stem cells towards insulin producing cells rather than to neuronal cell types, and to combine this with a strategy for their separation from insulin‐negative cells.Fil: Naujok, O.. Institute of Clinical Biochemistry; AlemaniaFil: Francini, Flavio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Endocrinología Experimental y Aplicada. Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Endocrinología Experimental y Aplicada; Argentina. Institute of Clinical Biochemistry; AlemaniaFil: Jörns, A.. Hannover Medical School; Alemania. Institute of Clinical Biochemistry; AlemaniaFil: Lenzen, S.. Institute of Clinical Biochemistry; Alemani

3 可測関数

http://purl.org/coar/resource_type/c_e05

9 収束定理

http://purl.org/coar/resource_type/c_e05