Ultrastructural changes in spermatozoa of the brush-tailed possum,Trichosurus vulpecula (Marsupialia), during epididymal transit

Summary The acrosome in spermatozoa from the caput epididymidis of the Australian Brush-tailed possum, Trichosurus vulpecula, typically forms a cup-like structure, sitting on the anterior third of the dorsal surface of the nucleus. The base of the acrosomal cup is narrowly separated from the nuclear surface, while the body of the cup projects voluminously away from the nucleus.During epididymal transit these pronounced marginal extensions of the acrosome are retracted towards the nucleus, and the electron dense acrosomal material undergoes a process of compaction within the plasma membrane of the head to produce the convex ovate form of the definitive acrosome. During this process a variety of bizarre forms of the acrosome are produced before its final configuration is attained.The authors would like to thank Dr. D.J.H. Cockayne, Director of the Electron Microscope Unit, University of Sydney, for the generous provision of transmission electron microscope facilities, and Dr. M.R. Dickson, Electron Microscopist in charge, Biomedical Electron Microscope Unit, University of New South Wales, for the use of other facilities. Thanks also are due to Miss Robin Arnold and Mrs. Eva Vassak of the Department of Histology and Embryology, University of Sydney, for their expert assistance. The assistance of the N.S.W. National Parks and Wildlife Service in the provision of permits to work on these native mammals is acknowledged     

Spermiogenesis in the brush-tailed possum,Trichosurus vulpecula (Marsupialia)

Summary Acrosome development in the Australian Brush-tailed possum, Trichosurus vulpecula, displays a number of extraordinary features. This is particularly evident in the later stages of spermiogenesis, when the area of the nuclear surface bounded by the nuclear ring, and covered by the acrosome, is reduced considerable. As a result, the acrosomal material becomes located over its definitive position on the anterior third of the dorsal nuclear surface; in this process it is thrown into a series of folds, and a wide subacrosomal space is formed.Further changes around the time of spermiation result in the release of a spermatozoon in which a thin layer of acrosomal material is closely applied to the nucleus over the area of the definitive location of the acrosome, whilst its margins are greatly extended and project freely away from the nucleus. The latter feature does not appear to have been reported for the sperm of other mammals.The authors would like to thank Dr. D.J.H. Cockayne, Director of the Electron Microscope Unit, University of Sydney, for the generous provision of transmission electron microscope facilities, and Dr. M.R. Dickson, Electron Microscopist in charge, Biomedical Electron Microscope Unit, University of New South Wales, for the use of other facilities. Also, thanks are due to Miss Robin Arnold and Mrs. Eva Vassak of the Department of Histology and Embryology, University of Sydney, for their expert assistance. The assistance of the N.S.W. National Parks and Wildlife Service in the provision of permits to work on these native mammals is acknowledged     

Ultrastructural changes in spermatozoa of the Brush-tailed possum,Trichosurus vulpecula (Marsupialia), during epididymal transit

Summary During epididymal transit, a fibre network and an array of vesicles develop in the posterior two-thirds of the midpiece in sperm of the Brush-tailed possum, Trichosurus vulpecula. The fibre network is developed by the time the sperm reach the corpus epididymidis, and is composed of evenly spaced, helically arranged fibres lying immediately beneath the plasma membrane. The angle of these fibrous helices is always counter to that of the underlying mitochondrial helix. Separating the fibre network from the mitochondria is a layer of granular material which develops at the same time, and over the same length of the midpiece as the fibre network.A somewhat tenuous fibre network is found between the fibrous sheath and plasma membrane in the anterior principal piece of sperm from all regions of the epididymis.The array of vesicles is developed by the time sperm reach the cauda epididymidis. The vesicles resemble pinocytotic vesicles; some appear as invaginations of the plasma membrane, and are open to the medium surrounding the spermatozoon by a narrow neck, while others are entirely enclosed within the spermatozoon, and lie at varying distances between the plasma membrane and the layer of granular material.The authors would like to acknowledge the assistance of Miss Dianne Higginbotham of the Electron Microscope Unit, University of Sydney for expert assistance with the scanning electron microscopy. Also to Dr. D.J.H. Cockayne, Director of the Electron Microscope Unit, for use of the transmission electron microscope facilities.     

A cytoarchitectonic study of the hypothalamus of the lizard,Calotes versicolor

Summary The hypothalamic nuclei of the lizard, Calotes versicolor, can be broadly divided into AF-positive and AF-negative. The AF-positive cell complexes include the nucleus supraopticus, nucleus paraventricularis, and a few interconnecting bridge cells. In addition, some AF-positive neurones are also observed in the median eminence. As many as 15 AF-negative nuclei-like accumulations of nerve cells can be identified in the hypothalamus. The nucleus periventricularis hypothalami of earlier authors is subdivided into eight circumscribed neuronal complexes. In addition, a few AF-negative nuclei, e.g. nucleus subfornicalis, nucleus ventralis tuberis, nucleus med. recessus infundibuli, nucleus lat. recessus infundibuli and nucleus praemamillaris, are regarded for the first time as anatomical entities. The distribution of the hypothalamic nuclei and their cytoarchitectonic features are described at the light microscopical level. An attempt has been made to interprete the nuclei identified in the present study on a comparative and phylogenetic basis.Dedicated to Professors W. Bargmann and B. Scharrer, the pioneers of neuroendocrinology, on the occasion of their birthday anniversariesSupported in part by research grant [No. F. 30-4 (6431)/76(SR-II)] by the University Grants Commission, New Delhi, India. The facilities provided by Dr. V. K. Thakare are hereby acknowledgedFormer scholar of the Alexander von Humboldt-Stuftung (1973–1975), Federal Republic of Germany. The present study was initiated by Professor A. Oksche, Department of Anatomy and Cytobiology, Justus Liebig University of GiessenThe facilities provided by Dr. A.K. Dorle are gratefully acknowledged

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Umsatz von Lymphozyten in Blut und Lymphknoten der Ratte

Summary Mature spermatozoa from the seminal vesicles of adult Agriolimax reticulatus have been studied by means of phase contrast and electron microscopy; sperm were either live, or sectioned or mounted whole and shadowed with gold. The cell is of the typical pulmonate sperm type with a simple acrosome and a spiral nucleus comprising the head, and a tail which is ensheathed along its entire length by mitochondrion. The 9 peripheral fibrils of the axial complex show no indication of a double nature. Within the spermatheca or gametolytic gland breakdown of sperm occurs; the nucleus and axial fibre bundle of the flagellum survive the longest. The complexity of the flagellum and the relative simplicity of the acrosome are discussed in light of the ecology of the spermatozoa. Many problems concerning the functional physiology of the sperm organelles remain to be investigated.     

Spermatogenesis revisited

Summary Gametogenesis is normal through meiosis and the earliest phases of spermiogenesis in male-sterile mice homozygous for the recessive, pleiotropic, mutation hpy (hydrocephalic-polydactyl). However, structurally complete sperm flagella were not encountered. Instead, partially assembled axonemal structures and/or poorly organized aggregates of other tail components (mitochondria, outer coarse fibers) were seen at the posterior poles of nuclei in older spermatids. The ultrastructure of centrioles in spermatids was normal, but that of axonemes associated with them was not. These findings suggest that the observed flagella dysgenesis results from defects in assembly rather than from defective intiation centers. Released gametes usually consisted of distorted nuclei and associated acrosome enclosed in a relatively close fitting plasma membrane. Perturbations of sperm head development were also encountered; they included extreme nuclear elongation, and distortion of the acrosome and underlying nuclear material by inpushings of finger-like processes of Sertoli cells. It is believed that sperm head anomalies are secondary consequences of the mutant condition. The findings support the view that the hpy locus represents one of a number of genes primarily involved in the mediation of flagella development.Aided by a grant from The National Foundation-March of DimesA preliminary account of some of the findings was presented elsewhere (Bryan, 1976)The author wishes to express his thanks to Mrs. Charla Danforth and Mr. James Barrows for capable technical assistance, to Dr. Walter J. Humphreys, Director, The Central Electron Microscope Laboratory, University of Georgia, for use of the facility, and to Dr. Willard F. Hollander, Iowa State University, for original breeding stock and helpful discussions     

Ultrastructure of paraspermatozoa,euspermatozoa and eusperm-like spermatozoa ofObtortio cf.fulva (Prosobranchia: Cerithiacea)

The cerithiaceanObtortio cf.fulva produces three distinct types of spermatozoa: (1) paraspermatozoa, (2) euspermatozoa and (3) eusperm-like spermatozoa. Like most mesogastropods, euspermatozoa ofObtortio are composed of a conical acrosome, short posteriorly invaginated nucleus, elongate midpiece and glycogen piece, and short terminal region. The midpiece, however, is distinctly cerithiacean in structure and is composed of four non-helical midpiece elements. Eusperm-like spermatozoa closely resemble euspermatozoa, but have a very short nucleus only one half to one third the length of the euspermatozoon nucleus. Paraspermatozoa of this species are composed of (1) head (mosaic sheath of dense blocks enveloping multiple axonemes which attach anteriorly to a long apical structure), (2) midpiece (multiple axonemes interspersed with elongate mitochondria), and (3) multiple tail tuft (axonemes each ensheathed by glycogen granules). The possible role of eusperm-like spermatozoa is briefly discussed together with the taxonomic implications of the structure of the three sperm types.     

Measurement of the electrical resistance of plasmodesmata and membranes of corn suspension-culture cells

Electrophysiological investigations of intercellular communication and membrane resistance in higher plants have been hampered by the difficulty in measuring these quantities independently. Uncertainty about the position of an electrode inserted into vacuolate tissue has further complicated such measurement. To overcome these problems sister cell pairs of a Zea mays L. Black Mexican Sweet suspension culture were used and dye was injected from the current-injecting electrode to determine the location of the electrode tip in each experiment. Of the impalements, 72% were cytoplasmic. The presence of plasmodesmata was fully incorporated into the electriccircuit model for the cell, and the resistance of the membrane of the current-injected cell was calculated, separate from the plasmodesmata resistance. This avoided some of the confusion resulting from work on multicellular tissue in which the position of the electrode and the extent of intercellular coupling is not determined. Using this technique, plasma-membrane resistivity was measured as 0.65 ·m², the resistivity of the tonoplast and plasma membrane in series was 1.35 ·m², and the resistance of a single plasmodesma was calculated to be 53 ± 11 G.Abbreviations BMS Black Mexican Sweet - PD potential difference - R_j resistance of the plasmodesmata in the junction between cells - R_m resistance of the plasma membrane of the current-injected cell - R_t resistance of the tonoplast - V₁, V₂ membrane PDs of sister cells This work was funded by an Australian Research Council grant to R.L.O. We are grateful to Dr. Maret Vesk (Electron Microscope Unit, The University of Sydney) for assistance with the preparation of EM sections, and to Dr. Richard Brettell (C.S.I.R.O. Division of Plant Industry) for assistance with the BMS culture.     

Ultrastructure of spermiogenesis in Plagioscion squamosissimus (Teleostei, Perciformes, Sciaenidae)

Spermiogenesis in Plagioscion squamosissimus occurs in cysts. It involves a gradual differentiation process of spermatids that is characterized mainly by chromatin compaction in the nucleus and formation of the flagellum, resulting in the spermatozoa, the smallest germ cells. At the end of spermiogenesis, the cysts open and release the newly formed spermatozoa into the lumen of the seminiferous tubules. The spermatozoa do not have an acrosome and are divided into head, midpiece, and tail or flagellum. The spermatozoa of P. squamosissimus are of perciform type with the flagellum parallel to the nucleus and the centrioles located outside the nuclear notch.     

Ultrastructural study of spermiogenesis in a rare desert amphisbaenian Diplometopon zarudnyi

Spermiogenesis, in particular the head differentiation of Diplometopon zarudnyi, was studied at the ultrastructural level by Transmission Electron Microscope (TEM). The process includes acrosomal vesicle development, nuclear elongation, chromatin condensation and exclusion of excess cytoplasm. In stage I, the proacrosomal vesicle occurs next to a shallow fossa of the nucleus, and a dense acrosomal granule forms beneath it. This step commences with an acrosome vesicle forming from Golgi transport vesicles; simultaneously, the nucleus begins to move eccentrically. In stage II, the round proacrosomal vesicle is flattened by projection of the nuclear fossa, and the dense acrosomal granule diffuses into the vesicle as the fibrous layer forms the subacrosomal cone. Circular manchettes surrounded by mitochondria develop around the nucleus, and the chromatin coagulates into small granules. The movement of the nucleus causes rearrangement of the cytoplasm. The nucleus has uniform diffuse chromatin with small indices of heterochromatin. The subacrosome space develops early, enlarges during elongation, and accumulates a thick layer of dark staining granules. In stage III, the front of the elongating nucleus protrudes out of the spermatid and is covered by the flat acrosome; coarse granules replace the small ones within the nucleus. One endonuclear canal is present where the perforatorium resides. In stage IV, the chromatin concentrates to dense homogeneous phase. The circular manchette is reorganized longitudinally. The Sertoli process covers the acrosome and the residues of the cytoplasmic lobes are removed. In stage V, the sperm head matures.     

Morphological aspects of fertilization in Phallusia (Ascidia) nigra (Ascidiacea,Tunicata)

The spermatozoa of Phallusia (Ascidia) nigra have an elongated head (approximately 5 m in length) in which a nucleus and a single mitochondrion are located side by side. There is no midpiece. The apex of the head is wedge-shaped. Acrosomal vesicles (approximately 55–65 nm in diameter) and moderately electron-dense material (MEDM) are present between the plasmalemma and the nuclear membranes in the anterior tip of the head. The MEDM occupies a central position and three or four acrosomal vesicles are seen in a line alongside it. The acrosomal vesicles disappear as the sperm makes contact with the surface of the chorion. Gamete fusion most likely occurs between a small process extending from the peripheral margin of the sperm apex and the egg surface, resulting in incorporation of the sperm into the egg from the anterior region of its head.     

Ultrastructure of the spermatozoa of<Emphasis Type="Italic"> Parenterodrilus taenioides</Emphasis> (Protodrilida: “Polychaeta”) and its phylogenetic significance

The interstitial annelid Parenterodrilus taenioides from coral sands of the island of Moorea (French Polynesia) is a mouthless species characterized by a vestigial non-functioning alimentary canal and a distinctive body shape. It is the only known annelid that has a non-functioning gut and no symbiotic bacteria. Parenterodrilus is thought to represent the sister group of the well-known taxon Protodrilus, generally recognized as Protodrilidae. An ultrastructural study revealed that, as in Protodrilus spp., the males of P. taenioides have lateral organs, i.e. paired epidermal, glandular and ciliated grooves, in the anterior body region where the sperm ducts are located. As in Protodrilus two types of spermatozoa were found, commonly called euspermatozoa and paraspermatozoa. Both types are filiform and the euspermatozoa, like those in Protodrilus spp., have a characteristic midpiece reinforced by symmetrically arranged supporting structures and reduced mitochondria. The ultrastructure of the spermatozoa in P. taenioides and Protodrilus spp. is basically the same but the species can be distinguished from each other on the basis of particular structural details of spermatozoa and the pattern of lateral organs. The two types of spermatozoa and the epidermal lateral organs are unique features of these two taxa; as they are not present in any other potentially related taxon, such as Saccocirrus and Protodriloides, they very likely represent autapomorphies of Protodrilidae. In Protodrilus spp. spermatophores are deposited on the epidermis of the females, whereupon sperm are released and penetrate the body wall. Since in polychaetes the structure of the sperm is correlated with the fertilization biology, the similar structure of sperm suggests that sperm transfer and fertilization are very probably the same in P. taenioides as in Protodrilus.This paper was presented at the symposium Morphology, molecules, evolution and phylogeny in Polychaeta and related taxa held in Haus Ohrbeck, Osnabrück, in September 2002, and is dedicated to Prof. Dr. Wilfried Westheide on the occasion of his 65th birthday     

Ultrastructural studies on the nucleoli of the follicle cells in the Lizards Acanthodactylus scutellatus hardyi and Eremias brevirostris

Summary In male mice homozygous for both p ^s and hpy, two recessive, pleiotrophic, mutations, gametogenesis is normal through meiosis but no functional spermatozoa are produced. Spermiogenesis is abnormal from the Golgi phase on. The types of abnormalities seen during the early and mid-stages of Spermiogenesis are characteristic of those associated with the presence of the p ^s mutation whereas those associated with the hpy mutation appear during the later stages of spermatid development. While centriolar ultrastructure was normal, axonemal structures were only rarely encountered and no late spermatids with recognizable flagella were seen. Some late spermatids showed head abnormalities of the type characteristic of the p^s mutation while others were recognizable as being of the hpy type. A released gamete usually consisted of a distorted nucleus and associated acrosome enclosed in a tightly fitting plasma membrane. No spermatids exhibiting a novel phenotype were encountered. The findings support the view that, despite their simultaneous presence in the double homozygote, each mutation acts autonomously. These studies also allow a similar inference to be made with respect to the extent of the interrelationship of the other major sub-processes of Spermiogenesis.The author wishes to express his thanks to Mr. Clifford S. Shuman III for capable technical assistance     

Electron microscopic studies on the palpi of Cybister fimbriolatus fimbriolatus (Say)

Summary The maxillary palpi of the predaceous diving beetle Cybister fimbriolatus fimbriolatus Say were observed by transmission and scanning electron microscopy. Scanning electron microscopy shows at least two types of sensilla at the tip of palp, which are referred to as circumvallate and naked sensilla. The former are innervated with outer segments of distal processes of sensory cells, but the latter are provided only with chitin and cuticular substances.This work was supported by Grant NB 04687 from the National Institutes of Health of the United States of America.We wish to express our gratitude to Prof. H. Stanley Bennett, Laboratories for Reproductive Biology, School of Medicine, University of North Carolina, Prof. G. Yasuzumi, Electron Microscope Research Laboratory, Department of Anatomy, Nara Medical University, and Prof. John E. Bardach, Department of Wildlife and Fisheries, School of Natural Resources, University of Michigan, for their valuable advice to the present work.     

The spermatozoon of Oikopleura dioica Fol (Larvacea,Tunicata)

Summary The spermatozoon of Oikopleura dioica is about 30 m long, with a spherical head, about 1 m wide, a 3 m long and 1 m wide midpiece, and a 25 m long tail with a tapered end piece. The head contains a nucleus with the chromatin volume limited to about 0.1 m³. A small acrosome is found in an anterior inpocketing, and a flagellar basal body in a posterior inpocketing of the nucleus. The midpiece contains a single mitochondrion with the flagellar axoneme embedded in a groove along its medial surface. The flagellar axoneme has the typical 9 + 2 substructure, and the basal body the typical 9+0 substructure. A second centriole and special anchoring fibres are absent.     

Fine structure of the spermatozoon of the sea cucumber,Leptosynapta clarki (Echinodermata: Holothuroidea)

Summary The spermatozoon of the holothurian Leptosynapta clarki has a small circular head measuring about 3.0 at the greatest diameter, a midpiece containing a single mitochondrion and a tail flagellum measuring between 35 and 45 in length. The acrosomal region contains a granule measuring 0.7 in diameter which consists of electron dense material arranged in concentric lamellae. Five concentric very electron dense lamellae alternate with areas of much less electron dense material in the central region of the granule. This granule rests in an anterior nuclear depression.The nucleus is circular in shape and contains one or two unbound vacuoles which frequently contain a fine granular material. Posteriorly the nucleus is bounded by a large mitochondrion and an occasional Golgi complex. The proximal centriole which contains a lateral arm of dense material lies in a deep fossa projecting into the nucleus. The distal centriole lies posteriorly in the mitochondrial mass and gives rise to nine satellite projections and their Y-shaped connective extensions.The tail contains the 9 + 2 tubule arrangement and tapers at its distal end.This investigation was supported by a National Research Council grant to F. S. Chia.     

Ultrastructure of spermatogenesis in Cerastoderma glaucum (Cardiacea) and Spisula subtruncata (Mactracea)

Summary

In Cerastoderma glaucum, Sertoli cells are rich in lipids, glycogen and lysosomes, and premeiotic cells exhibited nuage, a prominent Golgi complex and endoplasmic reticulum cisternae encircling the nucleus. The Golgi complex gives rise to proacrosomal vesicles during mid-spermiogenesis, and the round acrosomal vesicle, with a dense fibrillar core, migrates laterally while linked to the plasma membrane as it develops the subacrosomal material. In its final position, the vesicle becomes cap-shaped (0.6 μm) and differentiates into apical light and basal dense regions. The elongated and helicoidal nucleus (8–9.9 μm) has a thin tip (0.3 μm) that invades the subacrosomal space, and in the midpiece (0.8 μm) two of the four mitochondria extend laterally to the nucleus (1.5–2.1 μm). In Spisula subtruncata, Sertoli cells are rich in lipids, glycogen and phagocytosed sperm. Premeiotic cells exhibit nuage, a prominent Golgi complex that gives rise to proacrosomal vesicles from the leptotene stage and a flagellimi that is extruded at zygotene. The acrosomal vesicle forms during the round spermatid stage and differentiates into a large and dense basal region and an apical light region. It then migrates while linked to the plasma membrane by its apical pole. Development of the subacrosomal perforatorium is associated with nuage materials and endoplasmic reticulum vesicles. The mature cap-shaped (0.6 μm) acrosomal vesicle exhibits a large apical and irregular region with floccular contents and a basal dense region. The round nucleus becomes barrel-shaped (1.5 μm) and the midpiece (0.8 μm), with four mitochondria, contains a few glycogen particles.     

Spermatozoa and spermatogenesis in the northern quahaug <Emphasis Type="Italic">Mercenaria mercenaria</Emphasis> (Mollusca,Bivalvia)

We studied the ultrastructure of spermatogenesis and spermatozoa in the northern quahaug, the clam Mercenaria mercenaria. Spermatogenetic cells gradually elongate. Mitochondria gradually fuse and increase in size and electron density. During spermatid differentiation, proacrosomal vesicles migrate towards the presumptive anterior pole of the nucleus and eventually form the acrosome. The spermatozoon of M. mercenaria is of a primitive type. It is composed of head, mid-piece, and tail. The acrosome shows a subacrosomal space with a short conical contour. The slightly curved nucleus of the spermatozoon contains fine-grained dense chromatin. The middle piece consists of a centriolar complex which is surrounded by four mitochondria. The flagellum has a standard “9 + 2” microtubular structure. The ultrastructure of spermatozoa and spermatogenesis of M. mercenaria shares a number of features with other species of the family Veneridae. M. mercenaria may be a suitable model species for further investigations into the mechanisms of spermatogenesis in the Bivalvia.     

An ultrastructural analysis of the mature spermatozoon of Anguispira alternata (say) (pulmonata,stylommatophora)

Mature spermatozoa from the hermaphroditic duct of adult snails were examined using various techniques of light microscopy as well as scanning and transmission electron microscopy. The sperm are approximately 557 μm in length including a dextrally spiral head approximately 13 μm long. The head consists of an electron-dense nucleus sculptured into a double-ridged spiral and an acrosome projecting approximately 0.45 μm beyond the apex of the nucleus. The acrosome consists of a membrane-bound vesicle approximately 0.1 μm in diameter and a column of homogeneous material which extends along one side of the terminal spiral of the nucleus. This material is separated from the nucleus by the nuclear envelope. The neck region, though similar to that found in other pulmonates, possesses a unique coiled structure surrounding the central doublet of the axoneme. The midpiece axoneme possesses a 9+9+2 configuration anteriorly grading into a 9+2 pattern for the majority of its length. There are three mitochondrial helices – one primary and two secondary – in the midpiece. Only the primary helix persists throughout the midpiece.     

Ultrastructural observations of the testicular epithelium and spermatozoa of <Emphasis Type="Italic">Pseudochordodes bedriagae</Emphasis> (Gordiida,Nematomorpha)

Two questions are of interest concerning the male reproductive system in Gordiida: (1) is the epithelium surrounding the testis continuous or discontinuous and (2) is the type of spermatozoon as described at the transmission electron-microscopical level for the two species of Gordius typical for all Gordiida? An examination of the South American species Pseudochordodes bedriagae has allowed us to add new information to this poorly studied phylum. Testicular tubes are large, filled with spermatozoa, and surrounded by a continuous epithelium. The epithelial cells that line the posterior testes occasionally overlap, and their cytoplasm is narrow and contains dense granules, abundant endoplasmic reticulum, and vesicles. The plasma membrane possesses microvilli with many filaments. This epithelium rests on a basement membrane. The spermatozoa in P. bedriagae resemble the known spermatozoa of two Gordius species but differ in presenting a uniform halo layer of less dense chromatin that surrounds the dense chromatin in the nucleus. The finding that a similar type of spermatozoa occurs in both genera (Pseudochordodes and Gordius) makes it likely that it is present in all other Gordiida and is therefore an autapomorphy of the Gordiida.