Development of Specific Inclusion in the Mesophyll Cells of Ammopiptanthus mongolicus

The development of specific inclusion in the mesophyll cells of Ammopiptanthus mongolicus was observed by means of transmission electron microscopy. The inclusion is approximately oval-shaped with very high electron-density. It originates from outside of the central vacuole. During its early development it only contains small amount of vesicle-like elements and electron-dense materials. Then the two kinds of components gradually increase in amount and form a protuberance by endocytosis of the tonoplast. The protuberance becomes larger and larger, sometimes even occupies a great part of the vacuole. Later, the vesiclelike elements gradually decrease as the electron-dense materials rapidly increase, eventually filling up the protuberance. The protuberance gradually contracts and finally detaches from the tonoplast and immerses freely in the central vacuole. However, such inclusions were never found in small vacuoles. The inclusions usually appear in cold season and contain large quantity of lipid.     

Protist-like inclusions in amber, as evidenced by Charentes amber

The mid-Cretaceous amber of France contains thousands of protist-like inclusions similar in shape to some ciliates, flagellates and amoebae. The sheer abundance of these inclusions and their size variation within a single amber piece are not concordant with true fossil protists. French amber is coniferous in origin, which generally does not preserve well protists without cell walls. Thus, it would be surprising if French Cretaceous amber had preserved millions of protists. Here, we present a survey of the protist-like inclusions from French amber and attempt to elucidate their origins.Diverse Cretaceous ambers (from Spain, Germany and Lebanon), also derived from conifer resins, contain thousands of protist-like inclusions. In contrast, Tertiary ambers and modern resins are poor in protist-like fossils. This suggests these inclusions originated from early Cretaceous plant resins, probably secreted with the resin by trees that did not survive after the Cretaceous (such as the Cheirolepidiaceae). A review of the recent literature on amber microfossils indicates several protist-like inclusions that are unlikely to have a biological origin have already been described as real fossil protists. This is problematic in that it will bias our understanding of protist evolution.     

Morphogenesis of accessory nuclei during final stages of oogenesis in Cosmoconus meridionator (Hymenoptera: Ichneumonidae)

Morphogenesis of accessory nuclei (AN) in chorionated oocytes of Cosmoconus meridionator is described. Initially, each AN contains two dense, morphologically distinct inclusions. During the final stages of postvitellogenesis, these inclusions undergo characteristic transformation that is followed by the extrusion of some substances from AN to the surrounding periplasm. Histo- and cytochemical tests indicate that both inclusions contain RNA, although their precise composition is different. Our results support previous suggestions on the involvement of AN in the distribution of morphogenetic signals.     

沙冬青叶肉细胞中一种特殊内含物的发育

用透射电镜观察了沙冬青（Ａｍ ｍ ｏｐｉｐｔａｎｔｈｕｓｍ ｏｎｇｏｌｉｃｕｓ）叶肉细胞中一种电子密度很高的近似椭圆形的特殊内含物的发育． 它始于中央液泡外侧,起初只是少量的泡状成分和电子密度很高的物质,然后两种成分逐渐增多,并随液泡膜内吞形成突起,不断伸向液泡中央,有的突起占据了液泡很大一部分体积． 接着突起中的泡状成分开始解体,电子密度很高的物质越来越多,直至充满整个突起． 当突起继续内伸时,它的尾部不断收缩变小,最后完全脱离液泡膜而游离在中央液泡里面． 这种内含物一般只出现在严冬季节,里面含有大量的脂．     

Differential morphology and composition of inclusions in the R6/2 mouse and PC12 cell models of Huntington’s disease

The histological hallmark feature of Huntington’s disease (HD) and other polyglutamine repeat diseases is the presence of intracellular inclusions. Much work has been devoted to trying to determine the relationship between inclusion formation and neuronal injury. However, little attention has been paid to the variability and characteristics of inclusions themselves. Here, we characterize the morphological and biochemical composition of inclusions in both a transgenic mouse model (R6/2 line) and an inducible cell culture model of HD (iPC12Q74). We identified several morphologically distinct kinds of inclusions in different locations (nuclei, cytoplasm and cellular processes). Ubiquitin colocalized completely with all of these inclusions in both the iPC12Q72 and R6/2 models. In the inclusions in iPC12Q74 cells, the 20S and 11S proteasome subunits colocalized variably, and the 19S subunit did not colocalize at all. In inclusions in R6/2 mouse neurons, the 20S subunit colocalized completely, but neither the 11S nor the 19S subunits colocalized at all. While the role of inclusions in the pathogenesis of HD continues to be debated, we suggest that the content and structure of inclusions vary considerably, not only from cell to cell but even within individual cells. Their role in the pathogenesis of HD is likely to depend on their location as well as their composition.     

Accessory nuclei in insect oogenesis: in search of the function of enigmatic organelles

This review compiles present knowledge of the structure and molecular composition of the enigmatic cytoplasmic organelles called accessory nuclei. Most typically, they are found in the perinuclear cytoplasm in oocytes of insects and several other invertebrates. Accessory nuclei originate by budding of the oocyte nucleus (germinal vesicle) and are surrounded by an envelope identical to the nuclear envelope. They contain one or several dense inclusions called pseudonucleoli immersed in a translucent ground substance or matrix. Comparative analysis of the morphology, molecular composition and the ultimate fate of accessory nuclei and their inclusions revealed that there are two basic types of these organelles in insect oocytes. In mallophagans, accessory nuclei are associated with the oocyte nucleus throughout entire oogenesis and at least some of them are connected to the germinal vesicle by slender stems. Each accessory nucleus contains a single, dense, RNA-positive inclusion which is likely to correspond to a nucleolus. In hymenopterans, accessory nuclei initially surround the germinal vesicle but during oogenesis they separate from it and migrate toward the peripheral ooplasm. Within the accessory nucleus matrix usually two distinct inclusions develop. One of these is perfectly spherical, contains coilin and small nuclear ribonucleoproteins and is homologous to the Cajal body. In the light of recent discoveries, we discuss the role accessory nuclei play in insect oogenesis and early embryogenesis.     

The structure and composition of aleurone grains in the barley aleurone layer

Summary Cytochemical methods have been used in conjunction with light and electron microscopy to determine the nature of the inclusions in aleurone grains of barley aleurone layers. Two kinds of inclusions were found: (1) Globoids within globoid cavities which were not enclosed by a membrane: the globoids stained red with toluidin blue due to the presence of phytin, and with lipid stains; (2) Protein-carbohydrate bodies which stained green with toluidin blue. The characteristics of globoids and protein-carbohydrate bodies as seen in the electron microscope are described in detail using both glutaraldehyde- and permanganatefixed tissues. The protein-carbohydrate body was identified by silver-hexaminestaining; this was not caused by carbohydrate but by some component which stained green in toluidin blue and which also occurred in cell walls in a thin band adjacent to the cytoplasm. The characteristics of both bodies are discussed in relation to apparent confusion in their identities in previous electron-microscope studies.     

A survey on the intraclear inclusions in the Schrophulariaceae and their systematic significance

A survey of nuclear inclusions in chlorenchyma cells is given for 290 taxa of the Scrophulariaceae, covering 72 genera; 172 are new data and the remainder are from previous reports by the author. In addition to the 5 types of nuclear inclusions already determined by their different ultrastructural morphology — amorphous (A), lamellar (L), fibrillar (F), tubular (T), and crystalline (C) — and to subtypes C2, three new subtypes — A2, L2, T2 - specific to certain genera or small groups of genera are reported. These new subtypes are described in detail and their distribution in the family discussed. Although the proteinic nature was unquestionably demonstrated, the origin and function of plant nuclear inclusions is far from being understood. In the Scrophulariaceae they have been recognized in 242 out of 290 taxa studied (83.4%), in a total of 60 out of 72 genera (83.3%). On the basis of the occurrence and qualitative distribution of the nuclear inclusions, the family can be subdivided into four main group: 1) with lamellar inclusions, 2) with amorphous inclusions, 3) with inclusions other than those in the two former groups, and 4) with no nuclear inclusions. The relationship between the distribution on the nuclear inclusions throughout the family and the current systematic arrangement into subfamilies are analyzed and the transfer of some genera to different tribes or to a tribe of their own is proposed, e.g., in the case of Mimulus, Leucocarpus, Halleria, Russelia, Angelonia, Rehmannia , and Campylanthus . Finally, a possible evolutionary trend of the different types of nuclear inclusions based on their structural complexity and on the occurrence in allied families is offered.     

Assembly and breakdown of Cajal bodies in accessory nuclei of Hymenoptera

In some species of insects, oocytes have vesicular organelles, termed accessory nuclei (ANs). The ANs form by budding off from the nuclear envelope of the oocyte and are filled with translucent matrix containing dense inclusions. One type of these inclusions contains coilin and small nuclear ribonucleoproteins (snRNPs) and is homologous to Cajal bodies. We describe the early events in the morphogenesis of Cajal bodies in the ANs (ANCBs) of the common wasp, Vespula germanica, and show that they contain survival of motor neurons (SMN) protein. We present evidence that in the wasp, ANCBs form by the gradual accumulation of aggregates composed of SMN and small nuclear RNAs. We also show that ANCBs break down and disperse within the ANs as the ANs, which initially surround the oocyte nucleus, localize to the oocyte cortex. The components of dispersed ANCBs are retained within ANs until the end of oogenesis, which suggests that their function may be required at the onset of embryonic development. Because the morphology and behavior of ANs and their Cajal body-like inclusions are conserved in two other hymenopteran species, these features might be characteristic of all hymenopterans.     

Ultrastructure of spermiogenesis and synthesis of enigmatic cytoplasmic inclusions in the spirally shaped spermatozoon of the polychaete Spio setosa (Annelida: Spionidae)

The sperm of Spio setosa (Polychaeta, Spionidae) are known to be very unusual in form; here, spermiogenesis and the structure of the spermatozoon in this species are described by transmission electron microscopy. While spermiogenesis is similar to that described for many other polychaetes, two notable exceptions to this process include the synthesis of abundant ring‐shaped and tubular, membrane‐bounded cytoplasmic inclusions in the midpiece, and the differentiation of a spirally shaped sperm head. Spermatids develop as free‐floating tetrads in the male's coelom. A microtubular manchette does not develop during chromatin condensation and nuclear elongation, and the spiral acrosome forms as a single Golgi‐derived vesicle that migrates anteriorly to become housed in a deep anterior nuclear fossa. Early in spermiogenesis, numerous Golgi‐derived, membrane‐bounded cytoplasmic inclusions appear in the cytoplasm; these ultimately occupy the sperm midpiece only. The mature spermatozoon in the male has a 15‐μm‐long head consisting of a nucleus coiled like a spring and a spiral acrosome with differentiated substructure, the posterior two thirds of which sits in an anterior nuclear fossa. The midpiece is wider than the rest of the spermatozoon and contains 9–10 spherical mitochondria surrounding the two centrioles, as well as numerous membrane‐bounded conoid and tubular cytoplasmic inclusions. The axoneme has a 9 + 2 arrangement of microtubules. By contrast, stored sperm in the female's seminal receptacles have lost the midpiece inclusions but contain an abundance of glycogen. The function of the midpiece inclusions remains unresolved, and the significance of their absence in stored sperm within the seminal receptacle and the appearance of midpiece glycogen stores remains unclear and requires additional investigation.     

First record of Mesozoic scroll coprolites: classification,characteristics, elemental composition and probable producers

Multiple, small, cylindrical scroll coprolites having rounded and tapering ends and pertaining to a new ichnotaxon have been recovered from the Upper Triassic Tiki Formation of India. This is the first record of scroll coprolites from the Mesozoic. Based on cross‐sectional geometry, external surface textures, and internal morphology, these coprolites are subdivided into three morphotypes. The coprolites contain several kinds of undigested food material in the form of ganoid fish scales, teeth, lower jaw and skeletal remains of various osteichthyans, chondrichthyans, archosauriforms and indeterminate reptiles. These inclusions are embedded in the groundmass separated by thin mucosal layers. The groundmass contains abundant gas vesicles, and secondarily‐filled shrinkage cracks. EDS analysis shows that the overall composition of the coprolites reflects Ca, P, C and O, suggesting calcium phosphate mineralogy, though other elements such as Fe, Mn, Al, Si are present in lesser proportions. Based on their similarity with the scrolled faeces of extant euryhaline hammerhead sharks, it is deduced that these coprolites were produced by euryhaline hybodontid sharks. At least two hybodontid taxa, Lonchidion and Pristrisodus, show high prevalence in the Tiki vertebrate fauna, suggesting that these were the possible producers. As the coprolite inclusions contain remains of other aquatic animals, these carnivorous hybodonts constituted the dominant predators of the Tiki aquatic ecosystem.     

The surfactant system of the lung in the Mexican axolotl Ambystoma mexicanum

The broncho-alveolar space of axolotl contains numerous osmiophilic structures, which have been classified morphologically into 3 types of inclusions. Type I inclusions exhibited lattices of square to rectangular grid patterns with membranous elements 6 nm thick. This lattice was crossed by a 2 nm dense line. Type II inclusions were composed of 7 nm dark lines and 15.5 nm light lines in an alternating repeating pattern. Furthermore, the light lines showed an intrapenoid line of 2 nm. Type III inclusions consisted of concentric whorls of lamellae similar in form to spider webs. Only type III inclusions were identified in grand alveolar cells.     

Structure in the amphibian germinal vesicle

The germinal vesicle (GV) of Xenopus laevis is an enormous nucleus that contains 18 giant lampbrush chromosomes and thousands of inclusions. The inclusions are primarily of three types: approximately 1500 extrachromosomal nucleoli, 50-100 Cajal bodies, and several thousand B-snurposomes, which correspond to speckles or interchromatin granule clusters in other nuclei. The large size and abundance of the GV organelles, as well as the ease with which they can be studied both in vivo and in vitro, make the GV an ideal object for analysis of nuclear structure and function.     

FINE STRUCTURE OF RAT INTRAFUSAL MUSCLE FIBERS : The Equatorial Region

An ultrastructural study has been undertaken on the equatorial (sensory) region of the rat muscle spindle. Two kinds of intrafusal muscle fibers, a nuclear bag fiber and a nuclear chain fiber, have been identified in this region on the basis of fiber diameter, nuclear disposition, and M-band appearance. The large-diameter nuclear bag fiber contains an aggregation of tightly packed vesicular nuclei, while the small-diameter nuclear chain fiber contains a single row of elongated, well-separated nuclei. Both muscle fibers contain an attenuated peripheral cylinder of myofilaments surrounding a central core of sarcoplasm. Elements of the sarcotubular system, dilatations of the sarcoplasmic reticulum, and the presence of other sarcoplasmic organelles and inclusions are considerably more abundant in the nuclear chain fiber than in the nuclear bag fiber. Leptomeric organelles and membrane-bounded sarcoplasmic granules are present in both intrafusal fiber types and may be situated between the myofibrils or in intimate association with the sarcolemma. The functional significance of some of these structural findings is discussed.     

Unusual structural and chemical characteristics of the Dufour gland in the ant Meranoplus diversus

Abstract The Dufour gland of both queens and workers of Meranoplus diversus is very large. The reservoir is lined with flat epithelial cells, which usually contain smooth endoplasmic reticulum but also show the unusual accumulation of electron‐dense inclusions. Chemically, the gland contains an unusual mixture of long‐chain fatty acids and their derivatives, including hydrocarbons, ketones and amines with long‐chain substituents, hydroquinone, and sugars. Some of the substances identified are already known as insect defensive compounds against other arthropods.     

Feeding behavior,epidermal structure and mucus cytochemistry of the scleractinian Mycetophyllia reesi,a coral without tentacles

The scleractinian Mycetophyllia reesi lacks even the vestiges of tentacles, but quickly captures particulate food by mucus entanglement. Mesenterial filaments emerge through the oral opening, collect the mucus-embedded particulates, and withdraw to the gastrovascular system within 15 min. Mucocytes dominate the outer epidermis with about 3,000 cells/mm(2) and are capable of apocrine discharge en masse. Mucocytes are spumous, typically with web-like inclusions, which for the most part lack electron opacity with ordinary staining, and are only weakly PAS positive. In contrast, the mucus reacts strongly to diamine and other reagents that suggest an appreciable acidic mucopolysaccharide component. The strongest staining reaction occurs in the presence of high iron diamine, suggesting with other tests that the mucus contains significant quantities of sulfated polysaccharides. Cells with cilia anchored by spiriliform microvilli flank the mucocytes and possess small, spumous inclusions that contain acidic, sulfated, and neutral polysaccharides that do not appear to discharge during feeding. These support cells are closely intertwined with narrow, sinuous, secretory cells containing an electron-opaque cytoplasm of unknown composition that is discharged along with mucus during feeding. The outer epidermis also contains scattered cnidae, rather than the clusters or batteries typical of tentacles. The overwhelming abundance of mucocytes is consistent with their importance in feeding. Likewise, the small number of epidermal cnidae suggest they play a minor role in acquiring food. An inner epidermal layer associated with the mesoglea contains epitheliomuscular cells, nerve cells and pigment cells. The two epidermal layers form an essentially pseudostratified, architecturally simple epithelium.     

Characterisation of lipofuscin-like lysosomal inclusion bodies from human placenta

A structural hallmark of lysosomes is heterogeneity of their contents. We describe a method for isolation of particulate materials from human placental lysosomes. After a methionine methyl ester-induced disruption of lysosomes and two density gradient centrifugations we obtained a homogeneous membrane fraction and another one enriched in particulate inclusions. The latter exhibited a yellow-brown coloration and contained bodies lacking a delimiting membrane, which were characterised by a granular pattern and high electron density. The lipofuscin-like inclusion materials were rich in tripeptidyl peptidase I, beta-glucuronidase, acid ceramidase and apolipoprotein D and contained proteins originating from diverse subcellular localisations. Here we show that human term placenta contains lipofuscin-like lysosomal inclusions, a phenomenon usually associated with senescence in postmitotic cells. These findings imply that a simple pelleting of a lysosomal lysate is not appropriate for the isolation of lysosomal membranes, as the inclusions tend to be sedimented with the membranes.     

Ultrastructure of the Lamellated Cytoplasmic Inclusions in Schwann Cells of the Marine Nematode,Deontostoma californicum

Electron microscopy of the photoreceptors in the marine nematode, Deontostoma californicum, revealed numerous lamellated inclusions in the Schwann cells ensheathing the lateral cephalic nerves. Immediately after the axons from the modified bipolar neurons of the photoreceptors enter the lateral nerves, these spherical-to-oval lamellated bodies are observed in the surrounding Schwann cell cytoplasm. These previously undescribed Schwann cell inclusions, approximately 500 nm long and 320 nm in diameter, are lamellated and characterized by the presence of an electron-dense stalk-like process, 80-280 nm long. The lamellated inclusions are bound by a single limiting membrane, 6-7 nm thick, which shows occasional interruptions. The internal structure of the inclusions is characterized by the presence of electron-dense lamellae or bands, 11-16 nm thick, which assume various complex patterns ranging from arrays of parallel linear densities to a reticulate appearance. In addition, the nematode Schwann cell cytoplasm contains the usual organelles, gliosome- and lysosome-like inclusions. Their relationship with lipofuscin pigments is briefly discussed.     

Contribution of the 65-kilodalton protein encoded by the cloned gene cry19A to the mosquitocidal activity of Bacillus thuringiensis subsp. jegathesan.

Two new crystal protein genes, cry19A and orf2, isolated from Bacillus thuringiensis subsp. jegathesan were cloned and characterized. The cry19A gene encodes a 74.7-kDa protein, and the orf2 gene encodes a 60-kDa protein. Cry19A contains the five conserved blocks present in most B. thuringiensis delta-endotoxins. The ORF2 amino acid sequence is similar to that of the carboxy terminus of Cry4 proteins. The cry 19A gene was expressed independently or in combination with orf2 in a crystal-negative B. thuringiensis host. The proteins accumulated as inclusions. Purified inclusions containing either Cry19A alone or Cry19A and ORF2 together were toxic to Anopheles stephensi and Culex pipiens mosquito larvae. They were more toxic to C. pipiens than to A. stephensi. However, inclusions containing Cry19A and ORF2 together were more toxic than inclusions of Cry19A alone but less toxic than the wild-type inclusions of B. thuringiensis subsp. jegathesan.     

Origins and formation of intracellular inclusions associated with two leguminous virus diseases

Summary Two hitherto undescribed viruses were isolated from naturally-infected white clover plants.One induced both cytoplasmic and intranuclear inclusions, and the other caused cytoplasmic inclusions of a new type, which we have called Corner inclusion bodies.All three kinds of inclusion were found in the same cells.Vital observations showed that the two kinds of cytoplasmic inclusion bodies developed principally from the plastids.The intranuclear inclusions were shown to develop from the nucleoli.Cytochemical tests showed that the cytoplasmic inclusions contained protein and ribonucleic acid.On the basis of these findings, the possibility of virus synthesis in plastids and nucleoli is discussed.