The Organization of Actin Filaments in the Stereocilia of the Bird Cochlea

1980 ◽  
Vol 38 ◽  
pp. 554-555
Author(s):  
E.J. Battles ◽  
D. DeRosier ◽  
J.C. Saunders ◽  
L.G. Tilney
Keyword(s):  
Hair Cell ◽  
Diffraction Pattern ◽  
Sea Urchin ◽  
Actin Filaments ◽  
Optical Diffraction ◽  
Dense Material ◽  
Apical Surface ◽  
Structural Rigidity ◽  
High Degree ◽  
Degree Of Order

Extending from the apical surface of each hair cell of the chick cochlea are from 75 to 200 microvilli or stereocllia and one true cllium, the kinocilium. The stereocllia are arranged in rows of progressively increasing length (Fig. 1). Within each tapering sterocilium is a bundle of actin filaments with over 900 filaments near the tip yet only approximately 25 at the base where filaments are enmeshed in a dense material (Fig. 1); from here some of the filaments enter the apical surface of the cell (cuticular plate) as a rootlet. Examination of longitudinal sections of the stereocilia (Fig. 2) show that the filaments are aligned parallel to each other and show considerable order. Examination of an optical diffraction pattern of this bundle (Fig. 4) reveal that the actin filaments are packed such that the crossover points of adjacent actin filaments are inregister. A prominent reflection at 125Å−1 demonstrates that the filaments are cjossbridged by a macromolecular bridge situated at an average of 125Å−1 intervals (Fig. 4) in transverse sections the filaments appear hexagonally packed although there are regions where the filaments are less ordered (Fig. 3). In images processed in the computer to remove, noise and enhance detail periodic nature of the bridge can be clearly seen (see arrows Fig. 5). This image resembles that of an actin paracrystal formed from sea urchin extract composed of bundles of actin filaments crossbridged by a second protein. Thus the actin filaments in the bird stereocilia by being cross-bridged and packed with a high degree of order and produces a structure with considerable structural rigidity. Embryos were studied at various stages in development in an attempt to determine how the stereocilia form and how does the actin packing develops. These stages will be discussed.

scholarly journals Actin in triton-treated cortical preparations of unfertilized and fertilized sea urchin eggs.

1979 ◽  
Vol 82 (1) ◽  
pp. 212-226 ◽  
Author(s):  
A Spudich ◽  
J A Spudich
Keyword(s):  
Electron Microscopy ◽  
Sea Urchin ◽  
Actin Filaments ◽  
Cell Types ◽  
Optical Diffraction ◽  
Muscle Actin ◽  
Sea Urchin Eggs ◽  
Inner Surface ◽  
Low Ionic Strength ◽  
Helical Parameters

Triton-treated cortical fragments of unfertilized and fertilized sea urchin eggs prepared in the presence of greater than or equal to 5 mM EGTA contain 15-30% of the total egg actin. However, actin filaments are not readily apparent by electron microscopy on the cortical fragments of unfertilized eggs but are numerous on those of fertilized eggs. The majority of the actin associated with cortical fragments of unfertilized eggs is solubilized by dialysis against a low ionic strength buffer at pH 7.5. This soluble actin preparation (less than 50% pure actin) does not form proper filaments in 0.1 M KCl and 3 mM MgCl2, whereas actin purified from this preparation does, as judged by electron microscopy. Optical diffraction analysis reveals that these purified actin filaments have helical parameters very similar to those of muscle actin. Furthermore, the properties of the purified actin with regard to activation of myosin ATPase are similar to those of actin from other cell types. The possibility that actin is maintained in a nonfilamentous form on the inner surface of the unfertilized egg plasma membrane and is induced to assemble upon fertilization is discussed.

scholarly journals Actin filaments, stereocilia, and hair cells of the bird cochlea. I. Length, number, width, and distribution of stereocilia of each hair cell are related to the position of the hair cell on the cochlea.

1983 ◽  
Vol 96 (3) ◽  
pp. 807-821 ◽  
Author(s):  
L G Tilney ◽  
J C Saunders
Keyword(s):  
Hair Cell ◽  
Surface Area ◽  
Hair Cells ◽  
Actin Filaments ◽  
Hexagonal Lattice ◽  
Frequency Discrimination ◽  
Sensory Epithelium ◽  
Scanning Microscopy ◽  
Apical Surface ◽  
Length Width

Located on the sensory epithelium of the sickle-shaped cochlea of a 7- to 10-d-old chick are approximately 5,000 hair cells. When the apical surface of these cell is examined by scanning microscopy, we find that the length, number, width, and distribution of the stereocilia on each hair cell are predetermined. Thus, a hair cell located at the distal end of the cochlea has 50 stereocilia, the longest of which are 5.5 microns in length and 0.12 microns in width, while those at the proximal end number 300 and are maximally 1.5 microns in length and 0.2 micron in width. In fact, if we travel along the cochlea from its distal to proximal end, we see that the stereocilia on successive hair cells gradually increase in number and width, yet decrease in length. Also, if we look transversely across the cochlea where adjacent hair cells have the same length and number of stereocilia (they are the same distance from the distal end of the cochlea), we find that the stereocilia of successive hair cells become thinner and that the apical surface area of the hair cell proper, not including the stereocilia, decreases from a maximum of 80 microns2 to 15 microns2. Thus, if we are told the length of the longest stereocilium on a hair cell and the width of that stereocilium, we can pinpoint the position of that hair cell on the cochlea in two axes. Likewise, if we are told the number of stereocilia and the apical surface of a hair cell, we can pinpoint the location of that cell in two axes. The distribution of the stereocilia on the apical surface of the cell is also precisely determined. More specifically, the stereocilia are hexagonally packed and this hexagonal lattice is precisely positioned relative to the kinocilium. Because of the precision with which individual hair cells regulate the length, width, number, and distribution of their cell extensions, we have a magnificent object with which to ask questions about how actin filaments that are present within the cell are regulated. Equally interesting is that the gradient in stereociliary length, number, width, and distribution may play an important role in frequency discrimination in the cochlea. This conclusion is amplified by the information presented in the accompanying paper (Tilney, L.G., E.H. Egelman, D.J. DeRosier, and J.C. Saunders, 1983, J. Cell Biol., 96:822-834) on the packing of actin filaments in this stereocilia.

Light Optical Diffraction Studies of Macromolecular Ultrastructure

1970 ◽  
Vol 28 ◽  
pp. 258-259
Author(s):  
Glen B. Haydon
Keyword(s):  
High Resolution ◽  
Diffraction Analysis ◽  
Diffraction Pattern ◽  
Electron Micrographs ◽  
Optical Diffraction ◽  
Electron Micrograph ◽  
Its Analysis ◽  
Resolution Electron ◽  
Diffraction Patterns

Analysis of light optical diffraction patterns produced by electron micrographs can easily lead to much nonsense. Such diffraction patterns are referred to as optical transforms and are compared with transforms produced by a variety of mathematical manipulations. In the use of light optical diffraction patterns to study periodicities in macromolecular ultrastructures, a number of potential pitfalls have been rediscovered. The limitations apply to the formation of the electron micrograph as well as its analysis.(1) The high resolution electron micrograph is itself a complex diffraction pattern resulting from the specimen, its stain, and its supporting substrate. Cowley and Moodie (Proc. Phys. Soc. B, LXX 497, 1957) demonstrated changing image patterns with changes in focus. Similar defocus images have been subjected to further light optical diffraction analysis.

Feasibility of Molecular Structure Determination With a High Resolution Electron Microscope

1968 ◽  
Vol 26 ◽  
pp. 338-339
Author(s):  
T. A. Welton
Keyword(s):  
Electron Microscope ◽  
Substrate Surface ◽  
Helical Axis ◽  
Base Plane ◽  
Resolution Electron ◽  
High Resolution Electron Microscope ◽  
High Degree ◽  
Degree Of Order ◽  
Purine Pyrimidine

An ultimate design goal for an improved electron microscope, aimed at biological applications, is the determination of the structure of complex bio-molecules. As a prototype of this class of problems, we propose to examine the possibility of reading DNA sequence by an imaginable instrument design. This problem ideally combines absolute importance and relative simplicity, in as much as the problem of enzyme structure seems to be a much more difficult one.The proposed technique involves the deposition on a thin graphite lamina of intact double helical DNA rods. If the structure can be maintained under vacuum conditions, we can then make use of the high degree of order to greatly reduce the work involved in discriminating between the four possible purine-pyrimidine arrangements in each base plane. The phosphorus atoms of the back bone form in projection (the helical axis being necessarily parallel to the substrate surface) two intertwined sinusoids. If these phosphorus atoms have been located up to a certain point on the molecule, we have available excellent information on the orientation of the base plane at that point, and can then locate in projection the key atoms for discrimination of the four alternatives.

Comparison of TEM and STM observations of small silver clusters on different carbon supports

1990 ◽  
Vol 48 (4) ◽  
pp. 264-265
Author(s):  
Bernd Tesche ◽  
Tobias Schilling
Keyword(s):  
Vibration Isolation ◽  
Pyrolytic Graphite ◽  
Carbon Film ◽  
Silver Clusters ◽  
Carbon Supports ◽  
Stacking Order ◽  
System P ◽  
High Resolution Tem ◽  
High Degree ◽  
Degree Of Order

The objective of our work is to determine:a) whether both of the imaging methods (TEM, STM) yield comparable data andb) which method is better suited for a reliable structure analysis of microclusters smaller than 1.5 nm, where a deviation of the bulk structure is expected.The silver was evaporated in a bell-jar system (p 10−5 pa) and deposited onto a 6 nm thick amorphous carbon film and a freshly cleaved highly oriented pyrolytic graphite (HOPG).The average deposited Ag thickness is 0.1 nm, controlled by a quartz crystal microbalance at a deposition rate of 0.02 nm/sec. The high resolution TEM investigations (100 kV) were executed by a hollow-cone illumination (HCI). For the STM investigations a commercial STM was used. With special vibration isolation we achieved a resolution of 0.06 nm (inserted diffraction image in Fig. 1c). The carbon film shows the remarkable reduction in noise by using HCI (Fig. 1a). The HOPG substrate (Fig. 1b), cleaved in sheets thinner than 30 nm for the TEM investigations, shows the typical arrangement of a nearly perfect stacking order and varying degrees of rotational disorder (i.e. artificial single crystals). The STM image (Fig. 1c) demonstrates the high degree of order in HOPG with atomic resolution.

High-Resolution Cryo-Electron Microscopy of Biological Macromolecules

1990 ◽  
Vol 48 (1) ◽  
pp. 126-127
Author(s):  
Yoshinori Fujiyoshi
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Diffraction Pattern ◽  
Superfluid Helium ◽  
Copper Phthalocyanine ◽  
Optical Diffraction ◽  
Irradiation Damage ◽  
Biological Macromolecules ◽  
Cross Sectional ◽  
Instrumental Resolution

The resolution of direct images of biological macromolecules is normally restricted to far less than 0.3 nm. This is not due instrumental resolution, but irradiation damage. The damage to biological macromolecules may expect to be reduced when they are cooled to a very low temperature. We started to develop a new cryo-stage for a high resolution electron microscopy in 1983, and successfully constructed a superfluid helium stage for a 400 kV microscope by 1986, whereby chlorinated copper-phthalocyanine could be photographed to a resolution of 0.26 nm at a stage temperature of 1.5 K. We are continuing to develop the cryo-microscope and have developed a cryo-microscope equipped with a superfluid helium stage and new cryo-transfer device.The New cryo-microscope achieves not only improved resolution but also increased operational ease. The construction of the new super-fluid helium stage is shown in Fig. 1, where the cross sectional structure is shown parallel to an electron beam path. The capacities of LN2 tank, LHe tank and the pot are 1400 ml, 1200 ml and 3 ml, respectively. Their surfaces are placed with gold to minimize thermal radiation. Consumption rates of liquid nitrogen and liquid helium are 170 ml/hour and 140 ml/hour, respectively. The working time of this stage is more than 7 hours starting from full LN2 and LHe tanks. Instrumental resolution of our cryo-stage cooled to 4.2 K was confirmed to be 0.20 nm by an optical diffraction pattern from the image of a chlorinated copper-phthalocyanine crystal. The image and the optical diffraction pattern are shown in Fig. 2 a, b, respectively.

Enthalpien von Kupfer- und Silberchalkogeniden / Enthalpies of Copper and Silver Chalcogenides

1978 ◽  
Vol 33 (2) ◽  
pp. 190-196 ◽  
Author(s):  
Roger Blachnik ◽  
Paul-G. Gunia
Keyword(s):  
Thermodynamic Data ◽  
Transition Temperatures ◽  
Melting Points ◽  
Drop Calorimeter ◽  
Silver Chalcogenides ◽  
Increasing Temperature ◽  
High Degree ◽  
Degree Of Order ◽  
Bond Character

The enthalpy contents of Cu2Se, Cu2Te, Ag2Se and Ag2Te at temperatures between 350 K to 50 K above the melting points were measured in an isoperibolic drop-calorimeter. Thermodynamic data of the compounds were derived from these measurements. An analysis of the data leads to the conclusion that a high degree of order is retained in the melts of these compounds, and that the bond character of these melts is partially ionic. An unusual decrease of Cp with increasing temperature immidiately after the transition temperatures could be confirmed.

Differential localization of villin and fimbrin during development of the mouse visceral endoderm and intestinal epithelium

Development ◽  
1989 ◽  
Vol 106 (2) ◽  
pp. 407-419 ◽  
Author(s):  
R.M. Ezzell ◽  
M.M. Chafel ◽  
P.T. Matsudaira
Keyword(s):  
Intestinal Epithelium ◽  
Actin Filaments ◽  
Early Embryo ◽  
Actin Binding ◽  
Common Mechanism ◽  
Apical Surface ◽  
Visceral Endoderm ◽  
Apical Cytoplasm ◽  
Gut Epithelium ◽  
Microfilament Bundles

The apical surface of transporting epithelia is specially modified to absorb nutrients efficiently by amplifying its surface area as microvilli. Each microvillus is supported by an underlying core of bundled actin filaments. Villin and fimbrin are two actin-binding proteins that bundle actin filaments in the intestine and kidney brush border epithelium. To better understand their function in the assembly of the cytoskeleton during epithelial differentiation, we examined the pattern of villin and fimbrin expression in the developing mouse using immunofluorescence and immunoelectron microscopy. Villin is first detected at day 5 in the primitive endoderm of the postimplantation embryo and is later restricted to the visceral endoderm. By day 8.5, villin becomes redistributed to the apical surface in the visceral endoderm, appearing in the gut at day 10 and concentrating in the apical cytoplasm of the differentiating intestinal epithelium 2–3 days later. In contrast, fimbrin is found in the oocyte and in all tissues of the early embryo. In both the visceral endoderm and gut epithelium, fimbrin concentrates at the apical surface 2–3 days after villin; this redistribution occurs when the visceral endoderm microvilli first contain organized microfilament bundles and when microvilli first begin to appear in the gut. These results suggest a common mechanism of assembly of the absorptive surface of two different tissues in the embryo and identify villin as a useful marker for the visceral endoderm.

Sign in / Sign up

Export Citation Format

Share Document