Ciliary necklace

Primary cilia mediate sensory signaling in multiple organisms and cell types but have structures adapted for specific roles. Structural defects in them lead to devastating diseases known as ciliopathies in humans. Key to their functions are structures at their base: the basal body, the transition zone, the "Y-shaped links," and the "ciliary necklace." We have used cryo-electron tomography with subtomogram averaging and conventional transmission electron microscopy to elucidate the structures associated with the basal region of the "connecting cilia" of rod outer segments in mouse retina...

It is well known that dysfunction of primary cilia during embryonic development causes a range of developmental disorders such as cleft lip and palate, lung, kidney and heart disease. Both Pitchfork and Kinesin family member 3a (Kif3a) are associating with primary cilia, but whether there is a correlation between them are still inconclusive. Our research confirmed that Pitchfork over-expression induced lateral cleft palate and primary cilia disassembly during palate development. We also demonstrated that Sonic hedgehog (Shh) and Patched1 (Ptc1) expression levels were altering in the over-expressed Pitchfork group during palate development...

The use of cryosectioning facilitates the morphological analysis and immunocytochemistry of cells in tissues in atomic force microscopy (AFM). The cantilever can access all parts of a tissue sample in cryosections after the embedding medium (sucrose) has been replaced with phosphate-buffered saline (PBS), and this approach has enabled the production of a type of high-resolution image. The images resembled those obtained from freeze-etching replica electron microscopy (EM) rather than from thin-section EM. The AFM images showed disks stacked and enveloped by the cell membrane in rod photoreceptor outer segments (ROS) at EM resolution...

This essay records a voyage of discovery from the "cradle of cell biology" to the present, focused on the biology of the oldest known cell organelle, the cilium. In the "romper room" of cilia and microtubule (MT) biology, the sliding MT hypothesis of ciliary motility was born. From the "summer of love," students and colleagues joined the journey to test switch-point mechanisms of motility. In the new century, interest in nonmotile (primary) cilia, never lost from the cradle, was rekindled, leading to discoveries relating ciliogenesis to autophagy and hypotheses of how molecules cross ciliary necklace barriers for cell signaling...

Cilia and flagella are surface-exposed, finger-like organelles whose core consists of a microtubule (MT)-based axoneme that grows from a modified centriole, the basal body. Cilia are found on the surface of many eukaryotic cells and play important roles in cell motility and in coordinating a variety of signaling pathways during growth, development, and tissue homeostasis. Defective cilia have been linked to a number of developmental disorders and diseases, collectively called ciliopathies. Cilia are dynamic organelles that assemble and disassemble in tight coordination with the cell cycle...

Both the basal body and the microtubule-based axoneme it nucleates have evolutionarily conserved subdomains crucial for cilium biogenesis, function and maintenance. Here, we focus on two conspicuous but underappreciated regions of these structures that make membrane connections. One is the basal body distal end, which includes transition fibres of largely undefined composition that link to the base of the ciliary membrane. Transition fibres seem to serve as docking sites for intraflagellar transport particles, which move proteins within the ciliary compartment and are required for cilium biogenesis and sustained function...

The sensory and motility functions of eukaryotic cilia and flagella are essential for cell survival in protozoans and for cell differentiation and homoeostasis in metazoans. Ciliary biology has benefited early on from the input of electron microscopy. Over the last decade, the visualization of cellular structures has greatly progressed, thus it becomes timely to review the ultrastructure of cilia and flagella. Briefly touching upon the typical features of a 9+2 axoneme, we dwell extensively on the transition zone, the singlet zone, the ciliary necklace, cap and crown...

Eukaryotic cilia and flagella perform motility and sensory functions which are essential for cell survival in protozoans, and to organism development and homoeostasis in metazoans. Their ultrastructure has been studied from the early beginnings of electron microscopy, and these studies continue to contribute to much of our understanding about ciliary biology. In the light of the progress made in the visualization of cellular structures over the last decade, we revisit the ultrastructure of cilia and flagella...

Cilia and flagella are important organizing centers for signaling in both development and disease. A key to their function is a poorly characterized barrier at their base that allows the protein and lipid composition of the ciliary membrane to be distinct from that of the plasma membrane. We review current models of ciliary membrane biogenesis, highlighting several structures, including the ciliary necklace and ciliary pocket, that appear during biogenesis and that likely contribute to the barrier. The regulated movement of membrane proteins and lipids across this barrier is central to the sensory function of these organelles...

A primary cilium is an antenna-like structure extending from the surface of most vertebrate cells. It is structurally divided along its vertical axis into sub-compartments that include the ciliary tip, the shaft, the ciliary necklace segment, the transitional zone and the basal body. We recently discovered that the shaft of the primary cilia has a distinct molecular compartment, termed the "Inv compartment", which is characterized by the accumulation of Inv at the base of primary cilia. Inv was discovered as a causative gene in inv mutant mice...

As an introduction to a symposium of nematode chemoreception, the anatomy of nematode chemosensilla, their distribution on plant parasitic nematodes, and their possible functional roles is briefly reviewed. Comparison of nematode chemosensilla with those of other animals shows their greater resemblance to olfactory primary sense cells of vertebrates. Although the sensory process is obviously derived from a cilium, the absence of many ciliary features is noted. Retention of the ciliary necklace may be important functionally...

Tissues from the epidermis, alimentary tract and notochord of the cephalochordate Branchiostoma lanceolatum have been examined in both thin sections and freeze-fracture replicas to ascertain the nature of the intercellular junctions that characterize their cell borders. The columnar epithelial cells from the branchial chamber (pharynx), as well as from the anterior and posterior intestine, all feature cilia and microvilli on their luminal surfaces. However, their lateral surfaces exhibit zonulae adhaerentes only...

BACKGROUND: The airway cilia of patients with primary ciliary dyskinesia (PCD) exhibit several anomalies when studied by transmission electron microscopy, but little is known about the ultrastructural organisation of ciliary membranes in these patients. Freeze fracture replication of airway epithelium from patients with PCD provides a means of achieving high resolution views of cell membrane structure. Ciliary necklaces are a specialised structural feature of ciliary membranes thought to serve as a timing mechanism for ciliary beat, and their characterisation in the cilia of patients with PCD may contribute new insights into the pathophysiology of this syndrome...

Samples of tracheal mucosa were obtained from 10 healthy adult water buffaloes and 50 000 cilia were examined ultrastructurally. Ciliary abnormalities were found in all 10 subjects. Atypical cilia occurred as compound cilia (up to 1.5%), intracytoplasmic cilia (0.07%) and swollen cilia (0.05%). The microtubular pattern was determined in 5000 cross-sectioned cilia, with about 7% showing axonemal abnormalities in which peripheral defects prevailed. Some electron-dense plugs appeared inside the cylinder lumen of 2...

Specimens of tracheal mucosa were obtained from ten adult apparently healthy small ruminants (five goats and five sheep) both by a fibre optic endoscope and in a slaughterhouse. Ultrastructurally, a total of 50,000 cilia were examined. Pathological cilia were found in all examined subjects. The prevalence of compound cilia showed a range of 0.3% to 3%. Intracytoplasmic and swollen cilia ranged from 0.2% to 0.5%. The microtubular pattern was examined in 4,000 cross-sectioned cilia and an abnormal pattern was found in 5-7%...

Light microscopic immunohistochemistry coupled with freeze-substitution electron microscopic immunocytochemistry was used to localize alpha-subunits of G-proteins and type III adenylyl cyclase in developing rat olfactory epithelia. Some cilia immunoreacted with antibodies to GS alpha and type III adenylyl cyclase as early as prenatal day 15 (E15; E1 = sperm-positive), but immunolabelling with antibodies to Golf alpha was not observed until E16. From then on numbers of receptor cells with immunolabelled cilia increased for all three probes...

This article focusses on the structural development of human esophageal ciliated epithelium. A combination of transmission electron microscopic (TEM), scanning electron microscopic (SEM), radioautographic, and light microscopic (LM) analyses were carried out using intact fetal tissues between 8 and 20 weeks of gestation as well as cultured esophageal explants. Up to the age of 10 weeks, the stratified esophageal epithelium consisted of two longitudinal primary folds. The surface cells were undifferentiated and contained large glycogen aggregates...

The existence of a ciliary necklace in kinocilia, atypical cilia and mature and differentiating sensory cilia is described in rat and chicken. In addition to parallel horizontally oriented rows of the ciliary necklace, irregular forms of intramembranous particle (IMP) aggregations are also described. The function of the ciliary necklace is discussed, special attention being given to the appearance of circular rows of IMP prior to the outgrowth of olfactory cilia and to the existence of dynein arms in these...

This paper illustrates the structural diversity of the ciliary membrane of the somatic ciliature in 68 ciliate genera and describes the interior architecture of several presumptive sensory cilia. Freeze-fracture technique reveals a variety of intramembrane particle arrays known as ciliary necklace, ciliary plaques, and ciliary rosettes. While the function of these arrays is still largely unknown, their distribution among ciliates suggests a phylogenetic correlation. Whereas current systems of ciliate classification are primarily based on the character of the ciliature which surrounds the cytostome as well as on the morphogenesis of this oral ciliature, the data presented in this paper are well suited to emphasize the significance of the somatic ciliature for the reconstruction of ciliate phylogeny...

Cilia from seminal vesicles of Discoglossus (Anura, Amphibia) are partially covered with a fuzzy coat easily stained by the PATAg technique. The freeze-fractures of the ciliary membrane show large intramembrane particles (IMPs) not randomly distributed. At the base of the cilium, 3 or 4 rows of large IMPs form the ciliary necklace. Above the necklace, a 0.1 micrometer high region is devoid of particles after which the shaft membrane exhibits large IMPs coarsely lined up in 9 longitudinal rows 0.6 micrometer in height...