ABSTRACT
The localization of the actin-monomer-binding protein profilin during the cell cycle of living Tradescantia virginiana stamen hair cells has been studied by microinjection of a fluorescently labeled analog of the protein. In contrast to previously published studies performed on chemically fixed animal cells, we do not find a specific colocalization of profilin with actin filament arrays. Our results show that, besides a general cytoplasmic distribution, profilin specifically accumulates in the nucleus in interphase and prophase cells. This nuclear localization was confirmed by means of electron microscopic immunolocalization of endogenous profilin (in Gibasis scheldiana stamen hair cells). During mitosis, as the nuclear envelope and nuclear matrix break down at the onset of prometaphase, the nuclear profilin redistributes equally into the accessible volume (cytosol) of the cell. During metaphase and anaphase no specific localization of profilin can be observed associated with the mitotic apparatus. However, during telophase, as nuclear envelopes and nuclear matrices re-form and the sister chromatids start to decondense, a subset of the microinjected profilin again localizes to the nucleus. No accumulation of profilin could be observed in the phragmoplast, where a distinct array of actin filaments exists. The function of profilin in the nucleus remains unclear.
Subject(s)
Cell Cycle , Cell Nucleus/metabolism , Contractile Proteins/metabolism , Hair Cells, Auditory/metabolism , Microfilament Proteins/metabolism , Tradescantia/metabolism , Actins/metabolism , Animals , Cell Nucleus/ultrastructure , Cells, Cultured , Chickens , Contractile Proteins/ultrastructure , Fluorescent Antibody Technique, Indirect , Hair Cells, Auditory/ultrastructure , Male , Microfilament Proteins/ultrastructure , Microinjections , Microscopy, Confocal , Phosphatidylinositol 4,5-Diphosphate/metabolism , Plant Proteins/metabolism , Plant Proteins/ultrastructure , Pollen/metabolism , Profilins , Tradescantia/ultrastructureABSTRACT
In animal cells and in fungi, small GTP-binding proteins of the Rho family have well-established roles in morphogenesis, cell-cycle progression, gene transcription and the generation of superoxide anions. The presence of these proteins in plant cells, however, has been established only recently, and the role of Rho GTPases in plants is now coming into view. Already, it is apparent that there are both striking similarities and fascinating differences in how Rho GTPases are regulated and used in plant versus animal and fungal cells. These new findings define certain core properties that might be common to members of this protein family in all eukaryotes.
Subject(s)
Plant Physiological Phenomena , rho GTP-Binding Proteins/metabolism , Actins/metabolism , Amino Acid Sequence , Animals , Guanine Nucleotide Exchange Factors , Humans , Molecular Sequence Data , Proto-Oncogene Proteins/metabolism , rac GTP-Binding Proteins/metabolism , rho GTP-Binding Proteins/physiologyABSTRACT
We report what we believe to be the first demonstration of a Kerr-lens mode-locked Cr(3+):LiSrAlF(6)laser that is pumped by a single narrow-stripe AlGaInP laser diode with a diffraction-limited output beam. A novel low-loss three-mirror laser cavity design is described in which strong, localized Kerr lensing was exploited such that 75-fs-duration pulses were obtained for only 36mW of incident pump power. This pump power was maintained for 18h by just three AA batteries as the electrical power source. We have shown that mode locking can be sustained for pump powers as low as 22mW.
ABSTRACT
We have examined the cytological effects of microinjecting recombinant birch profilin in dividing and interphase stamen hair cells of Tradescantia virginiana. Microinjection of profilin at anaphase and telophase led to a marked effect on cytokinesis; cell plate formation was often delayed, blocked, or completely inhibited. In addition, the initial appearance of the cell plate was wrinkled, thin, and sometimes fragmented. Injection of profilin at interphase caused a thinning or the collapse of cytoplasmic strands and a retardation or inhibition of cytoplasmic streaming in a dose-dependent manner. Confocal laser scanning microscopy of rhodamine-phalloidin staining in vivo revealed that high levels of microinjected profilin induced a degradation of the actin cytoskeleton in the phragmoplast, the perinuclear zone, and the cytoplasmic strands. However, some cortical actin filaments remained intact. The data demonstrate that profilin has the ability to act as a regulator of actin-dependent events and that centrally located actin filaments are more sensitive to microinjected profilin than are cortical actin filaments. These results add new evidence supporting the hypothesis that actin filaments play a crucial role in the formation of the cell plate and provide mechanical support for the cytoplasmic strands in interphase cells.
ABSTRACT
A tunable femtosecond solid-state amplifier system that uses only 3 W of 488-nm argon-ion pump power has been demonstrated to deliver microjoule pulses at repetition rates up to 20 kHz, with a maximum pulse energy of 14 mu;J obtained at 5 kHz. An all-solid-state, tunable, diode-pumped Cr:LiSrAlF(6) regenerative amplifier has been demonstrated, for the first time to our knowledge, that amplifies femtosecond pulses to energies exceeding 1 mu;mJ at up to a 16-kHz repetition rate.
