Biomedical subjects

L G Tilney

Publications and source records attributed to L G Tilney.

At least 55 records · Page 3Linked to original sources

PubMed1992

How individual cells develop from a syncytium: merogony in Theileria parva (Apicomplexa).

The central problem for Theileria parva during merogony is how to form numerous individual, uninucleate merozoites from a syncytial schizont so that each merozoite contains a single nucleus and a prescribed assortment of organelles. The way T. parva packages all the requisite organelles into free merozoites is by binding these organelles to the nuclear envelope, which in turn becomes associated, both directly and through the rhoptry complex, with the schizont plasma membrane. Formation of the merozoites occurs in a synchronous manner by a budding process. The merozoites develop with the rhoptry complex at the apical end by the progressive, outward evagination of the schizont plasma membrane. This evagination of the plasma membrane is associated with, and presumably induced by, the development of an orderly array of tubules that originate from the apical end and progressively form a longitudinal basket enclosing first the rhoptry complex, then the mitochondria and ribosomes, and finally the nucleus. The process of merogony is compared to sporogony within the tick salivary gland and with the differentiation of the intra-erythrocytic piroplasm stage. Because all three processes occur by a morphologically similar mechanism, the possibility that the parasite uses a single cassette of genes to perform each of these similar processes is discussed.

Animals↗

PubMed1991

The structure of plasmodesmata as revealed by plasmolysis, detergent extraction, and protease digestion.

Plasmodesmata or intercellular bridges that connect plant cells are cylindrical channels approximately 40 nm in diameter. Running through the center of each is a dense rod, the desmotubule, that is connected to the endoplasmic reticulum of adjacent cells. Fern, Onoclea sensibilis, gametophytes were cut in half and the cut surfaces exposed to the detergent, Triton X 100, then fixed. Although the plasma membrane limiting the plasmodesma is solubilized partially or completely, the desmotubule remains intact. Alternatively, if the cut surface is exposed to papain, then fixed, the desmotubule disappears, but the plasma membrane limiting the plasmodesmata remains intact albeit swollen and irregular in profile. Gametophytes were plasmolyzed, and then fixed. As the cells retract from their cell walls they leave behind the plasmodesmata still inserted in the cell wall. They can break cleanly when the cell proper retracts or can pull away portions of the plasma membrane of the cell with them. Where the desmotubule remains intact, the plasmodesma retains its shape. These images and the results with detergents and proteases indicate that the desmotubule provides a cytoskeletal element for each plasmodesma, an element that not only stabilizes the whole structure, but also limits its size and porosity. It is likely to be composed in large part of protein. Suggestions are made as to why this structure has been selected for in evolution.

Cell Wall↗

PubMed1991

The entry of Theileria parva sporozoites into bovine lymphocytes: evidence for MHC class I involvement.

We have examined the process of Theileria parva sporozoite entry into susceptible bovine lymphocytes and have begun to identify one of the possible molecular interactions involved in the process. The entry process involves a defined series of events and we have used a number of experimental procedures in combination with a method of quantitation to examine various aspects of this process. T. parva sporozoites are nonmotile organisms and the initial sporozoite-lymphocyte interaction is a chance event which can occur at 0-2 degrees C. All subsequent stages in the process are temperature dependent, require the participation of live intact sporozoites and host cells, and involve some cytochalasin-inhibitable rearrangement of the host cell surface membrane or cytoskeleton. Sporozoite entry can be inhibited by antibodies (mAbs) reactive with major histocompatibility complex (MHC) class I molecules (IL-A 19, IL-A 88) and with beta 2 microglobulin (B1G6), whereas mAbs reactive with MHC class II molecules (IL-A 21, J 11), and a common panleucocyte surface antigen, (IL-A 87; a bovine equivalent of CD 11a) have no effect. These results indicate that MHC class I molecules play a role in the process of T. parva sporozoite entry into bovine lymphocytes although as yet the precise role has not been determined. Once internalized within the lymphocyte, a process that takes less than 3 min at 37 degrees C, the sporozoite rapidly escapes from the encapsulating host cell membrane; a process which occurs concurrently with the discharge of the contents of the sporozoite rhoptries and microspheres. The intracytoplasmic parasite is covered by a layer of sporozoite-derived fuzzy material to which host cell microtubules rapidly become associated.

Actins↗

PubMed1990

Actin filament nucleation by the bacterial pathogen, Listeria monocytogenes.

Shortly after Listeria is phagocytosed by a macrophage, it dissolves the phagosomal membrane and enters the cytoplasm. 1 h later, actin filaments coat the Listeria and then become rearranged to form a tail with which the Listeria moves to the macrophage surface as a prelude to spreading. If infected macrophages are treated with cytochalasin D, all the actin filaments associated with the Listeria break down leaving a fine, fibrillar material that does not decorate with subfragment 1 of myosin. This material is associated with either the surface of the Listeria (the cloud stage) or one end (the tail stage). If the cytochalasin-treated infected macrophages are detergent extracted and then incubated in nuclei-free monomeric actin under polymerizing conditions, actin filaments assemble from the fine, fibrillar material, the result being that each Listeria has actin filaments radiating from its surface like the spokes of a wheel (cloud form) or possesses a long tail of actin filaments formed from the fine, fibrillar material located at one end of the Listeria. Evidence that the fine fibrillar material is involved in nucleating actin assembly comes from a Listeria mutant. Although the mutant replicates at a normal rate in macrophages, actin filaments do not form on its surface (cloud stage) or from one end (tail stage), nor does the bacterium spread. Furthermore it does not form the fine fibrillar material. Evidence that the nucleating material is a secretory product of Listeria and not the macrophage comes from experiments using chloramphenicol, which inhibits protein synthesis in Listeria but not in macrophages. If chloramphenicol is applied 1 h after infection, a time before actin filaments are found attached to the Listeria in untreated macrophages, actin filaments never assemble on the Listeria even when fixed 3 h later. Furthermore the fine fibrillar material is absent, although there is a coat of dense granular material.

Actin Cytoskeleton↗

PubMed1990

The distribution of plasmodesmata and its relationship to morphogenesis in fern gametophytes.

Fern (Onoclea sensibilis) gametophytes when grown in the dark form a linear file of cells (one-dimensional) called a protonema. In the light two-dimensional growth occurs which results in a heart-shaped prothallus one cell thick. The objective of this paper is to relate the most common pattern of cell division observed in developing gametophytes to the formation of the plasmodesmatal network. Since the prothalli are only two dimensional, we can easily determine from thin sections the total number and the density (number per unit surface area) of plasmodesmata at each developmental stage. As the prothallus grows the number of plasmodesmata increases 50-fold in the apical or meristematic cell. This number eventually reaches a plateau even though the density continues to increase with each new cell division. What is particularly striking is that both the number and density of plasmodesmata between adjacent cells is precisely determined. Furthermore, the pattern of plasmodesmata distribution is predictable so that (1) we can identify the apical meristematic cells by their plasmodesmata number, or density, as well as by their size, shape and location, (2) we can predict, again from plasmodesmata number, the location of a future wall of the apical cell prior to its actual formation, (3) we can show that the density of plasmodesmata in the triangular apical cell of the prothallus (14 plasmodesmata microns-2) is comparable to those reported for secretory glands which are known to have high rates of plasmodesmatal transport and (4) we can show that once the plasmodesmata have been formed during division, no subsequent change in the number of plasmodesmata occurs following cell plate formation.

Cell Division↗

PubMed1989

Gamma interferon limits access of Listeria monocytogenes to the macrophage cytoplasm.

The effect of rIFN-gamma and rTNF on the fate of hemolytic and nonhemolytic (hly-) Listeria monocytogenes in cultured mouse peritoneal macrophages was investigated. In untreated macrophages, approximately 80% of the hemolytic bacteria were killed during the first 2 h of incubation, but the survivors doubled between two and three times. In rIFN-gamma-treated macrophages, although the bacterial killing was identical to the controls during the first 2 h, there was no subsequent bacterial growth, and bactericidal activity continued for the duration of the experiment. rTNF has no affect by itself, but acted synergistically with rIFN-gamma to promote bacterial killing. Infected macrophages with or without rIFN-gamma were examined by EM. The results clearly showed that the role of rIFN-gamma was to prevent access of L. monocytogenes to the macrophage cytoplasm, which would prevent cell-to-cell spread of the bacteria. In addition, rIFN-gamma-treated macrophages exhibited enhanced digestive capacity of the intracellular bacteria.

Animals↗

PubMed1989

Actin filaments and the growth, movement, and spread of the intracellular bacterial parasite, Listeria monocytogenes.

Listeria monocytogenes was used as a model intracellular parasite to study stages in the entry, growth, movement, and spread of bacteria in a macrophage cell line. The first step in infection is phagocytosis of the Listeria, followed by the dissolution of the membrane surrounding the phagosome presumably mediated by hemolysin secreted by Listeria as nonhemolytic mutants remain in intact vacuoles. Within 2 h after infection, each now cytoplasmic Listeria becomes encapsulated by actin filaments, identified as such by decoration of the actin filaments with subfragment 1 of myosin. These filaments are very short. The Listeria grow and divide and the actin filaments rearrange to form a long tail (often 5 microns in length) extending from only one end of the bacterium, a "comet's tail," in which the actin filaments appear randomly oriented. The Listeria "comet" moves to the cell surface with its tail oriented towards the cell center and becomes incorporated into a cell extension with the Listeria at the tip of the process and its tail trailing into the cytoplasm behind it. This extension contacts a neighboring macrophage that phagocytoses the extension of the first macrophage. Thus, within the cytoplasm of the second macrophage is a Listeria with its actin tail surrounded by a membrane that in turn is surrounded by the phagosome membrane of the new host. Both these membranes are then solubilized by the Listeria and the cycle is repeated. Thus, once inside a host cell, the infecting Listeria and their progeny can spread from cell to cell by remaining intracellular and thus bypass the humoral immune system of the organism. To establish if actin filaments are essential for the spread of Listeria from cell to cell, we treated infected macrophages with cytochalasin D. The Listeria not only failed to spread, but most were found deep within the cytoplasm, rather than near the periphery of the cell. Thin sections revealed that the net of actin filaments is not formed nor is a "comet" tail produced.

Actin Cytoskeleton↗

PubMed1989

Preliminary biochemical characterization of the stereocilia and cuticular plate of hair cells of the chick cochlea.

The sensory epithelium of the chick cochlea contains only two cell types, hair cells and supporting cells. We developed methods to rapidly dissect out the sensory epithelium and to prepare a detergent-extracted cytoskeleton. High salt treatment of the cytoskeleton leaves a "hair border", containing actin filament bundles of the stereocilia still attached to the cuticular plate. On SDS-PAGE stained with silver the intact epithelium is seen to contain a large number of bands, the most prominent of which are calbindin and actin. Detergent extraction solubilizes most of the proteins including calbindin. On immunoblots antibodies prepared against fimbrin from chicken intestinal epithelial cells cross react with the 57- and 65-kD bands present in the sensory epithelium and the cytoskeleton. It is probable that the 57-kD is a proteolytic fragment of the 65-kD protein. Preparations of stereocilia attached to the overlying tectorial membrane contain the 57- and 65-kD bands. A 400-kD band is present in the cuticular plate. By immunofluorescence, fimbrin is detected in stereocilia but not in the hair borders after salt extraction. The prominent 125 A transverse stripping pattern characteristic of the actin cross-bridges in a bundle is also absent in hair borders suggesting fimbrin as the component that gives rise to the transverse stripes. Because the actin filaments in the stereocilia of hair borders still remain as compact bundles, albeit very disordered, there must be an additional uncharacterized protein besides fimbrin that cross-links the actin filaments together.

Animals↗

PubMed1989

The structure of the cuticular plate, an in vivo actin gel.

The cuticular plate is a network of actin filaments found in hair cells of the cochlea. In the alligator lizard, it consists of rootlets, emanating from the stereocilia, and of cross-connecting actin filaments that anchor these rootlets. In thin sections, this network displays striking patches of 650 +/- 110-A striae. By quantitative analyses of the images, the mystery of the striae can be explained. They are due in part to the rootlets which are sets of flat ribbons of actin filaments. The ribbons in each set are separated by approximately 650 A. Numerous whiskers 30 A in diameter extend from each ribbon's face, interconnecting adjacent ribbons. The nonrootlet filaments, except at the margins of the cell, occur primarily as single filaments. Like the ribbons, they are bristling with whiskers. The patches of striae are explained by ribbons and filaments held at a 650-A separation by the whiskers that project from them. A simple model for regions of bewhiskered filaments is a box crammed full of randomly oriented test-tube brushes. A thin slice through the box will show regions of dark lines or striae due to the wire backbones of the brushes separated from one another by the bristle length. Using the computer instead of test-tube brushes, we have been able to model quantitatively the filament distribution and pattern of striae seen in the cuticular plate of the lizard. The organization of actin filaments we have deduced from our simulations differs from that found in macrophages or in the terminal web of intestinal epithelial cells.

Actin Cytoskeleton↗

PubMed1988

New observations on the stereocilia of hair cells of the chick cochlea.

The final step in the staircase of tall hair cells is longer than lower steps, whereas the final step of the staircase of short hair cells is about the same increment in height as the lower steps. Furthermore the stereocilia in the final step of the staircase in tall hair cells are thinner and contain fewer actin filaments than the stereocilia at this step in short hair cells. Thus, the disproportionate lengths and widths of stereocilia in tall hair cells make them sensitive 'antennae' to mechanical oscillations. In contrast, the stereocilia in short hair cells are more robust and the increment between the tallest rows and the next tallest row is not disproportionately longer than that of the lower rows which would make the whole bundle stiffer and less likely to be displaced mechanically, a morphology more consistent with a separate function. Apart from these differences between tall and short hair cells, there are two features common to all hair cells. First, as one goes directly up the staircase in the 1.0 lattice plane, a plane we refer to as the 1.00 lattice plane, each successive stereocilium is taller than the one lower down. If, instead of going directly up the staircase, one goes up at 60 degrees to the 1.00 lattice plane, successive stereocilia are not necessarily longer than those below. Second, in looking up the staircase on the 1.00 lattice plane we see that the stereocilia are ordered into parallel rows. Adjacent rows are offset from each other by 1/2 the width of a stereocilium. Thus, in order for all the next to tallest stereocilia in adjacent rows in the 1.00 lattice plane to contact a stereocilium on the final step, all of which are the same height, the final step must minimally contain twice as many stereocilia as lower steps in the staircase. This is what is observed. Both of these features are necessary if the number of tip linkages from taller to less tall stereocilia is to be maximized.

Animals↗

PubMed1988

Actin filaments, stereocilia, and hair cells of the bird cochlea. V. How the staircase pattern of stereociliary lengths is generated.

The stereocilia on each hair cell are arranged into rows of ascending height, resulting in what we refer to as a "staircase-like" profile. At the proximal end of the cochlea the length of the tallest row of stereocilia in the staircase is 1.5 micron, with the shortest row only 0.3 micron. As one proceeds towards the distal end of the cochlea the length of the stereocilia progressively increases so that at the extreme distal end the length of the tallest row of the staircase is 5.5 micron and the shortest row is 2 micron. During development hair cells form their staircases in four phases of growth separated from each other by developmental time. First, stereocilia sprout from the apical surfaces of the hair cells (8-10-d embryos). Second (10-12-d embryos), what will be the longest row of the staircase begins to elongate. As the embryo gets older successive rows of stereocilia initiate elongation. Thus the staircase is set up by the sequential initiation of elongation of stereociliary rows located at increased distances from the row that began elongation. Third (12-17-d embryos), all the stereocilia in the newly formed staircase elongate until those located on the first step of the staircase have reached the prescribed length. In the final phase (17-d embryos to hatchlings) there is a progressive cessation of elongation beginning with the shortest step and followed by taller and taller rows with the tallest step stopping last. Thus, to obtain a pattern of stereocilia in rows of increasing height what transpires are progressive go signals followed by a period when all the stereocilia grow and ending with progressive stop signals. We discuss how such a sequence could be controlled.

Actin Cytoskeleton↗

PubMed1988

Three-dimensional reconstruction of an actin bundle.

We present the three-dimensional structure of an actin filament bundle from the sperm of Limulus. The bundle is a motile structure which by changing its twist, converts from a coiled to an extended form. The bundle is composed of actin plus two auxiliary proteins of molecular masses 50 and 60 kD. Fraying the bundle with potassium thiocyanate created three classes of filaments: actin, actin plus the 60-kD protein, and actin plus both the auxiliary proteins. We examined these filaments by transmission electron microscopy and scanning transmission electron microscopy (STEM). Three-dimensional reconstructions from electron micrographs allowed us to visualize the actin subunit and the 60- and 50-kD subunits bound to it. The actin subunit appears to be bilobed with dimensions 70 X 40 X 35 A. The inner lobe of the actin subunit, located at 20 A radius, is a prolate ellipsoid, 50 X 25 A; the outer actin lobe, at 30 A radius, is a 35-A-diam spheroid. Attached to the inner lobe of actin is the 60-kD protein, an oblate spheroid, 55 X 40 A, at 50 A radius. The armlike 50-kD protein, at 55 A radius, links the 60-kD protein on one of actin's twin strands to the outer lobe of the actin subunit on the opposite strand. We speculate that the 60-kD protein may be a bundling protein and that the 50-kD protein may be responsible for the change in twist of the filaments which causes extension of the bundle.

Actin Cytoskeleton↗

PubMed1988

The actin filament content of hair cells of the bird cochlea is nearly constant even though the length, width, and number of stereocilia vary depending on the hair cell location.

By direct counts off scanning electron micrographs, we determined the number of stereocilia per hair cell of the chicken cochlea as a function of the position of the hair cell on the cochlea. Micrographs of thin cross sections of stereociliary bundles located at known positions on the cochlea were enlarged and the total number of actin filaments per stereocilium was counted and recorded. By comparing the counts of filament number with measurements of actin filament bundle width of the same stereocilium, we were able to relate actin filament bundle width to filament number with an error margin (r2) of 16%. Combining this data with data already published or in the process of publication from our laboratory on the length and width of stereocilia, we were able to calculate the total length of actin filaments present in stereociliary bundles of hair cells located at a variety of positions on the cochlea. We found that stereociliary bundles of hair cells contain 80,000-98,000 micron of actin filament, i.e., the concentration of actin is constant in all hair cells with a range of values that is less than our error in measurement and/or biological variation, the greatest variation being in relating the diameters of the stereocilia to filament number. We also calculated the membrane surface needed to cover the stereocilia of hair cells located throughout the cochlea. The values (172-192 micron 2) are also constant. The implications of our observation that the total amount of actin is constant even though the length, width, and number of stereocilia per hair cell vary are discussed.

Actins↗

PubMed1987

The distribution of hair cell bundle lengths and orientations suggests an unexpected pattern of hair cell stimulation in the chick cochlea.

A detailed analysis of the morphological polarity of the hair cell bundles on the chick cochlea was carried out. Although the pattern is identical from cochlea to cochlea, the morphological polarity of the bundles varies at different positions on the cochlea. More specifically, the hair cell bundles located immediately adjacent to the inferior and superior edges are oriented with their morphological polarity perpendicular to the margins. As we move across the cochlea (transect it), there is a gradual rotation in the polarity of the bundles so that in the center of the cochlea the hair cells are oriented at an angle to those at the edges. As we continue to the superior edge the polarity gradually rotates back again. The amount of rotation depends on the position of the transect such that at the extreme proximal end there is little rotation, while at the distal end the rotation is up to 90 degrees. The rotation is always in the same direction with the tallest rows of stereocilia nearest the distal end of the cochlea. Measurements of the length of the longest stereocilia in the hair cell bundles revealed that not only are the bundles systematically longer from the proximal to distal end of the cochlea, but also the hair cells on the superior edge are significantly longer than those on the inferior edge at the same distance from one end of the cochlea. If we draw on micrographs of the cochlea contour lines through hair cells whose stereocilia are the same height, these lines coincide with the morphological polarity of the hair cells included in these contours. Furthermore analysis of damage to the cochlea induced by pure tones of high intensity also roughly follows the same contour lines. We conclude that unlike what has been thought, the stimulation of hair cells by pure tones may not occur in a strictly transverse pattern, but instead may follow the oblique contours demonstrated here.

Acoustic Stimulation↗

PubMed1987

Hair cell damage produced by acoustic trauma in the chick cochlea.

Examination of pure-tone acoustic damage in the chick basilar papilla revealed that the location and extent of hair cell damage was a function of both the stimulus intensity and the age at which the chicks were exposed. Scanning electron microscopic evaluation of noise-exposed cochleae at post-hatching days 1, 10 and 30 permitted the identification of discrete regions of damage, including hair cells with stereocilia injuries as well as those lost from the epithelium. The hair cell damage was tonotopically distributed along the cochlea according to frequency. However, for each exposure frequency two distinct sites of damage were often produced, and their locations were correlated with stimulus intensity. At low intensities, a longitudinal strip of hair cell damage ran along the superior edge of the basilar papilla. As exposure intensity increased, a second damage site developed along the inferior edge of the basilar papilla, distal to the longitudinal strip. This second type of damage initially took the form of a series of laterally-oriented wedges, but at higher intensities, the wedges coalesced to form a large crescent-shaped patch of damage. The location of the damage sites for each frequency did not shift with age. However, there were differences in the extent and position of the damage which could be correlated with stimulus intensity and with changes in middle ear admittance during development [(1983) Development of Auditory and Vestibular Systems, pp. 3-25. Editor: R. Romand. Academic Press, New York]. These results suggest that developmental changes in the location and extent of hair cell damage depend on the effective stimulus intensity reaching the cochlea, rather than on alterations in the frequency coding of the hair cells.

Acoustic Stimulation↗

PubMed1987

Flagellar gyration and midpiece rotation during extension of the acrosomal process of Thyone sperm: how and why this occurs.

The midpiece of Thyone sperm contains a large mitochondrion and a centriolar pair. Associated with one of the pair, i.e., the basal body of the flagellum, are satellite structures which apparently anchor the flagellar axoneme to the mitochondrion and to the plasma membrane covering the midpiece. Immediately before and as the acrosomal process elongates, the flagellum and the midpiece begin to rotate at 1-2 rotations per second even though the head of the sperm, by being firmly attached on its lateral surfaces to the coverslip, does not rotate at all. This rotation is not observed in the absence of flagellar beating whose frequency is much greater than that of its gyration. To understand how the midpiece rotates relative to the sperm head, it is first necessary to realize that in Thyone the flagellar axoneme projects at an acute angle to the principal axis of the sperm and is bent towards one side of this axis. Thus movement of the flagellum induces the sperm to tumble or yaw in solution. If the head is stuck, the midpiece will rotate because all that connects the sperm head to the midpiece is the plasma membrane, a liquid-like layer. A finger-like projection extends from the proximal centriole into an indentation in the basal end of the nucleus. In contrast to the asymmetry of the flagellum, this indentation is situated exactly on the principal axis of the sperm and, along with the finger-like projection, acts as a biological bearing to maintain the orderly rotation of the midpiece. The biological purpose of flagellar gyration during fertilization is discussed.

Acrosome↗

PubMed1987

Movement of the actin filament bundle in Mytilus sperm: a new mechanism is proposed.

An actin filament bundle approximately 2-5 microns in length is present in the sperm of the blue mussel, Mytilus. In unfired sperm this bundle extends from the midpiece through a canal in the center of the nucleus to terminate on the membrane limiting the inside of the cone-shaped acrosomal vacuole. The bundle is composed of 45-65 actin filaments which are hexagonally packed and regularly cross-bridged together to form an actin paracrystal so well ordered that it has six nearly equal faces. Upon induction of the acrosomal reaction, a needle-like process is formed in a few seconds. Within this process is the actin filament bundle which appears unchanged in filament number and packing as determined by optical diffraction methods. Using fluorescein-conjugated phalloidin we were able to establish that the bundle does not change length but instead is projected anteriorly out of the midpiece and nuclear canal like an arrow. Existing mechanisms to explain this extension cannot apply. Specifically, the bundle does not increase in length (no polymerization), does not change its organization (no change in actin twist), does not change filament number (no filament sliding), and cannot move by myosin (wrong polarity). Thus we are forced to look elsewhere for a mechanism and have postulated that at least a component of this movement, or cell elongation, is the interaction of the actin filament bundle with the plasma membrane.

Actin Cytoskeleton↗

PubMed1986

Actin filaments, stereocilia, and hair cells of the bird cochlea. III. The development and differentiation of hair cells and stereocilia.

The cochleae of chick embryos of 8 days of incubation until hatching (21 days) were examined by scanning electron microscopy. Unlike what one would expect from the literature, the total number of hair cells per cochlea (10,405 +/- 529) is already determined and visible in a 10-day embryo and the growth of the cochlea is a result of the growth in size and surface area of the hair cells. We also find that the hair cells differentiate simultaneously throughout the cochlea and have followed the differentiation of individual hair cells throughout development. During development we find that the total number, hexagonal packing, and orientation of the stereocilia in each hair cell is determined early and accurately (9- to 10-day embryos). The stereocilia then begin to elongate in all the cells of the cochlea at approximately 0.5 micron/day. By Day 12 the tallest stereocilia in each cell are 1.5-1.8 micron long, the mature length for cells at the proximal end of the cochlea. At this point all stereocilia cease elongating, but those along the inferior edge gradually increase in width from 0.11 micron to maximally 0.19 micron in 17-day embryos. When the stereocilia on the inferior edge reach their mature width, widening ceases and the elongation of stereocilia in the distal hair cells begins again. When these stereocilia have attained their mature lengths, they stop growing. Thus elongation and widening of stereocilia are separated in time. During this period, 11 to 13 days, the shape of the tufts at the proximal end of the cochlea changes. This occurs because stereocilia in the front of each tuft are absorbed while others at the sides appear de novo. This rearrangement converts a circular bundle of stereocilia to a rectangular bundle.

Actins↗