Analysis of distributions of mutants in clones of plant-cell aggregates

Retinal,cell aggregates develop in culture in a pattern similar to the in ovo retina, forming neurites first and then synapses. In the present study, we continuously exposed chick retinal cell aggregates to a high concentration (1 mM) of carbamylcholine (carbachol), an acetylcholine (ACh) analog that resists hydrolysis by acetylcholinesterase (AChE). This situation is similar to organophosphorus anticholinesterase poisoning in which the ACh level is elevated at synaptic junctions due to inhibition of AChE, Our objective was to determine whether continuous carbachol exposure either damaged cholino- ceptive neurites, cell bodies, and synaptic elements of the aggregates or influenced (hastened or retarded) their development.The retinal tissue was isolated aseptically from 11 day embryonic White Leghorn chicks and then enzymatically (trypsin) and mechanically (trituration) dissociated into single cells. After washing the cells by repeated suspension and low (about 200 x G) centrifugation twice, aggregate cell cultures (about l0 cells/culture) were initiated in 1.5 ml medium (BME, GIBCO) in 35 mm sterile culture dishes and maintained as experimental (containing 10-3 M carbachol) and control specimens.

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Sectioned rubisco crystals in TEM

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010016087x ◽

1990 ◽

Vol 48 (3) ◽

pp. 664-665

Author(s):

Gunnel Karlsson ◽

Jan-Olov Bovin ◽

Michael Bosma

Keyword(s):

Plant Cell ◽

Carbon Fixation ◽

Unit Cell ◽

Protein Crystals ◽

Unit Cell Parameters ◽

Cell Parameters ◽

Photosynthetic Carbon Fixation ◽

Photosynthetic Carbon

RuBisCO (D-ribulose-l,5-biphosphate carboxylase/oxygenase) is the most aboundant enzyme in the plant cell and it catalyses the key carboxylation reaction of photosynthetic carbon fixation, but also the competing oxygenase reaction of photorespiation. In vitro crystallized RuBisCO has been studied earlier but this investigation concerns in vivo existance of RuBisCO crystals in anthers and leaves ofsugarbeets. For the identification of in vivo protein crystals it is important to be able to determinethe unit cell of cytochemically identified crystals in the same image. In order to obtain the best combination of optimal contrast and resolution we have studied different staining and electron accelerating voltages. It is known that embedding and sectioning can cause deformation and obscure the unit cell parameters.

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Exploration of cell wall architecture with the rapid-freeze deep-etch technique

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100146485 ◽

1993 ◽

Vol 51 ◽

pp. 128-129

Author(s):

Béatrice Satiat-Jeunemaitre ◽

Chris Hawes

Keyword(s):

Plant Cell ◽

Cell Walls ◽

Cell Biology ◽

Molecular Model ◽

Three Dimensional ◽

Secretory Activity ◽

Wall Structure ◽

Specimen Preparation ◽

Molecular Architecture ◽

Plant Cell Walls

The comprehension of the molecular architecture of plant cell walls is one of the best examples in cell biology which illustrates how developments in microscopy have extended the frontiers of a topic. Indeed from the first electron microscope observation of cell walls it has become apparent that our understanding of wall structure has advanced hand in hand with improvements in the technology of specimen preparation for electron microscopy. Cell walls are sub-cellular compartments outside the peripheral plasma membrane, the construction of which depends on a complex cellular biosynthetic and secretory activity (1). They are composed of interwoven polymers, synthesised independently, which together perform a number of varied functions. Biochemical studies have provided us with much data on the varied molecular composition of plant cell walls. However, the detailed intermolecular relationships and the three dimensional arrangement of the polymers in situ remains a mystery. The difficulty in establishing a general molecular model for plant cell walls is also complicated by the vast diversity in wall composition among plant species.

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