The segregation of DNA in epithelial stem cells

It has recently been suggested that stem cells may invariably keep, from one division to the next, the daughter DNA molecules that contain the older of the two parental strands—that is, they may retain a complete set of “immortal strands,” through successive cell divisions (Cairns, 1975). We can test this hypothesis by labeling either the old immortal strands at the time the stem cells are created or the newly synthesized strands during subsequent divisions of the stem cells. In the former case, the stem cells should become permanently labeled; in the latter case, they should eliminate their label on their second division.Experiments of this sort have been conducted with the tongue papilla under steady state conditions and with the regenerating small intestinal crypts. The results clearly show that by far most of the multiplying cells in tongue and intestinal epithelium segregate their DNA “randomly” at mitosis. Nevertheless, the results, though far from conclusive, suggest that there are a small number of cells (1–5 in the stem cell region of each crypt and one at the base of each column of cells in the tongue) that selectively segregate their old and new DNA strands in the expected way. Thus in the immortal strand labeling experiments, there are a few labeled cells that retain their label for up to 4 weeks; conversely, in the new strand labeling experiments, a few cells appear to rid themselves of label after intervals equivalent to approximately two cell cycles.     

Another View of the Ultrastructure of Cucurbita Phloem

The primary phloem of young internodes of Cucurbita maxima wasstudied with the electron microscope. Phloem parenchyma cellsare highly vacuolated and contain nuclei, endoplasmic reticulum,ribosomes, mitochondria, chloro-plasts, and occasional dictyosomes.As compared with parenchyma cells, the most distinctive featuresof companion cells are their extremely dense cytoplasm, lowdegree of vacuolation, lack of chloroplasts, and numerous sieve-elementconnexions. Companion cells contain plastids with few internalmembranes. At maturity the enucleate sieve element is linedby a plasmalemma, one or more cistema-like layers of endoplasmicreticulum, and a membrane which apparently delimits the parietallayer of cytoplasm from a large central cavity. In OsO_4–-andglutaraldehyde-fixed elements, the central cavity is traversedby numerous strands, which run from cell to cell through thepores of sieve plates and lateral sieve areas, and which arederived ontogenetically from the slime bodies of immature cells.Numerous normal-appearing mitochondria are present in the parietallayer of cytoplasm. The pores of sieve plates and lateral sieveareas are lined with cytoplasm. The ultrastructural detailsof young sieve elements differ little from those of other youngnucleate cells. During sieve-element development, the sieveelement increases in vacuolation. At the same time, slime bodiesdevelop in the cytoplasm. With glutaraldehyde fixation, thesebodies often exhibit a double-layered limiting membrane. Asthe sieve element continues to differentiate, the slime bodiesincrease in size and the parietal layer of cytoplasm becomesvery narrow. Presently, the slime bodies begin to disperse andtheir contents fuse. This phenomenon occurs in the parietallayer of cytoplasm, while the latter is still delimited fromthe large central vacuole by a distinct tonoplast. The initiationof slime-body dispersal more or less coincides with perforationof the pore sites, and many pores are traversed by slime earlyin their development. Before slime-body dispersal, all dictyosomesand associated vesicles disappear from the cytoplasm. Eventually,the tonoplast diappears and the slime becomes distributed throughoutthe central cavity in the form of strands. Nuclei and ribosomesdisappear before breakdown of the tonoplast. Sieve elementsare connected with companion cells and parenchyma cells by plasmodesmata.     

Observations on Companion Cells and Specialized Phloem Parenchyma Cells of Nelumbo nucifera Gaertn

The phloem of very young petioles of Nelumbo nucifera Gaertn.(Nelumbium speciosum Willd.) was studied with the light microscope.The elongated, mature sieve elements contain slime, plugs, strands,and numerous plastids. Some sieve elements remain nucleatedfor a brief period even after the sieve plates are well developed.The companion cells numbering 8–14 undergo disintegrationbefore the elongation of the ontogenetically related sieve elementis completed. They are uninucleate to begin with but later becomebinucleate and finally degenerate and obliterate. The variousstages in their ontogeny and disintegration are described. Ofthe very few specialized phloem parenchyma cells present, someare associated with sieve elements. They have slime body-likestructures, and plastid-like bodies which group together andeventually disintegrate.     

Pollen of the Butomaceae and Alismataceae

When the microspores of B. umbellatus are released from the tetrad the developing wall consists only of muri and pilae (bacula). A basal layer is subsequently formed on unitmembrane-like strands. The latter appear to be formed de novo in association with the microspore cell membrane. As a basal layer develops an outer consolidated and an inner stranded layer are differentiated. These are considered ontogenetically equivalent and no terminological distinction appears justified. The basal layer and sexine are not differentiated by basic fuchsin or electron stains. Thus, the stranded (or nonhomogeneous) and amorphous-granular (or homogeneous) types of exine fine structure are histochemically indistinguishable in this species. Concomitant cytoplasmic activity, particularly in the peripheral region of the cell, may be related to substrate or enzyme synthesis, transport, and deposition. Several “unusual” vesicle types are tentatively related to other phases of morphogenetic activity in the protoplast.     

Ontogeny of the vascular bundles and contiguous tissues in the maize leaf blade

The patterns of initiation and early development of the minor and major veins in the flat portion of the leaf blade of maize (Zea mays L.) follow similar patterns. The veins and their associated bundle sheath cells commonly arise from cell assemblages derived from a single cell lineage, or longitudinal file of cells, rather than from two “half vein units” derived from different cell lineages. In addition, apparently, none of the vascular cells derived from the procambium is directly related ontogenetically to a bundle sheath cell. In veins derived from larger cell assemblages, the lateral bundle sheath cells are more closely related ontogenetically to the mesophyll cells, which are derived from the ground meristem, than to the vascular cells, which are derived from procambium. The bundle sheath cells, accordingly, are interpreted as being ground meristem in origin.     

Fine Structure of the Mesothelia and Extracellular Materials in the Coelomic Fluid of the Fin Boxes,Myocoels and Sclerocoels of a Lancelet,Branchiostoma floridae (Cephalochordata = Acrania)

Three ontogenetically related coeloms of a lancelet are described by transmission electron microscopy. The fin box coeloms are lined dorsally and laterally by smooth myomesothelial cells of uncertain function. In contrast, there are no myofilaments in the mesothelial cells of the ventral parts of the fin boxes. Similarly, myofilaments are absent from the mesothelia lining all parts of the sclerocoels and the lateral parts of the myocoels (the medial side of the myocoel is a myomesothelium comprising the striated muscles of the body wall). Lancelet coeloms differ from those of other deuterostomes in containing several kinds of formed extracellular materials. All three kinds of coeloms contain distinctive spherules with ramifying processes; dense strands are limited to the myocoels and sclerocoels; and a finely granular secretion is found only at the coelomic surface of the mesothelium lining the sclerocoels. These extracellular materials, which appear to originate from exocytosis of secretory granules from the mesothelial cells, may function biomechanically and for energy storage. The discussion includes a consideration of the so-called fin rays of lancelets and concludes that none of these structures is homologous with the fin rays of fish.     

Morphogenesis in a community of filamentous cyanobacteria

Reversible differentiation was experimentally discovered in a community of modern filamentous cyanobacteria Oscillatoria terebriformis. Splitting of the initially uniform community into differentiated parts (strands, multiradiate aggregates, networks, etc.) occurs only for the duration of a function facilitating the activity of this community as an integral unit. The structures are formed as a result of regrouping of the filaments, without their specialization. A morphologically regulatory system (polygonal network) was found to develop under the impact of extreme factors. The levels of structural organization of filamentous cyanobacteria and multicellular eukaryotes were compared (individual cells in a filament—cell organelles; filaments—individual cells; community—organism), and the similarities and differences in morphogenesis of these groups were analyzed using the data on the embryonic regulation in multicellular eukaryotes. Spatial information in morphogenesis was shown to result not from direct realization of an inherited program but is created by the elements of integral organisms (cells and filaments) in the course of development.     

Changes in Hechtian strands in cold-hardened cells measured by optical microsurgery

Optical microsurgical techniques were employed to investigate the mechanical properties of Hechtian strands in tobacco (Nicotiana tabacum) and Ginkgo biloba callus cells. Using optical tweezers, a 1. 5-microm diameter microsphere coated with concanavalin A was inserted though an ablated hole in the cell wall of a plasmolyzed cell and attached to a Hechtian strand. By displacing the adhered microsphere from equilibrium using the optical trapping force, the tensions of individual strands were determined. Measurements were made using both normal and cold-hardened cells, and in both cases, tensions were on the order of 10(-12) N. Significant differences were found in the binding strengths of cold-hardened and normal cultured cells. An increased number density of strands in cold-hardened G. biloba compared with normal cultured cells was also observed. Although no Hechtian strands were detected in any Arabidopsis callus cells, strands were present in leaf epidermal cells. Finally, the movement of attached microspheres was monitored along the outside of a strand while cycling the osmotic pressure.     

The Oral Gland Cells of Oikopleura dioica (Tunicata Appendicularia)

Light and electron microscopic studies showed that the oral gland cells have two quite different zones. Medially, the basal zone is in contact with body fluids and the endostyle. Its strongly pyroninophile cytoplasm contains the extremely digitated nucleus and numerous small mitochondria. Laterally, the apical zone contacts the epidermis and it may also send a process between epidermal cells and deliver cell fragments into the primordium of the new house. This cell zone contains numerous membranes. It is concluded that the oral gland cells are light producing glands and that the membrane-rich cell fragments which are incorporated into the house wall are the source of the bioluminiscence which has been reported from empty houses. The ontogenetically related subchordal cells have a similar structure and it is possible that also these cells are light producers.     

植物微管特效解聚剂APM对紫露草雄蕊毛细胞胞质环流的影响

采用体外渗透和显微注射的方法。将植物微管特效解聚剂甲基氨草磷(APM)引入紫露草雄蕊毛细胞后,发现原来沿着胞质束运动的胞质颗粒运动速度渐慢,进而胞质束消失,颗粒运动停止。显微注射后,还发现APM可通过胞间通道由被注射的细胞向两侧细胞扩散,从而也导致两侧细胞胞质束消失,颗粒运动停止。APM对胞质环流的抑制作用是可逆的。结果表明微管可能是胞质束的重要组份之一,植物胞质环流与微管的聚合与解聚状态有密切关系。     

Maintenance of genetic information in stem cells

The available data on DNA cosegregation in some stem cells are reviewed. Cairns was the first to assume cosegregation of template DNA strands for adult stem cells; i.e., all maternal DNA strands are preserved in one daughter cell, which remains a stem cell, while the newly synthesized DNA strands, which may contain errors, appear in the daughter cell that is committed to differentiation and passes to the transitory compartment of the cell population. The role of asymmetric mitosis in DNA cosegregation and maintenance of genetic information in stem cells is discussed.     

ACTIN NETWORK AS A NORMAL COMPONENT OF THE CYTOSKELETON IN MANY VASCULAR PLANT CELLS

F-actin was localized in cells of 17 species of vascular plants, using the F-actin-specific probe Rhodamine-phalloidin. F-actin strands formed a three-dimensional network in these cells, and orientation of the strands varied to some extent depending on the cell shape. The strands often appeared to terminate near or at the plasma membrane. The observations presented, taken together with those from relevant literature, suggest that actin is a normal component of most, if not all, plant cells.     

Freeze-fracture replica studies of tight junctions in normal human bronchial epithelium

Using freeze-fracture techniques, tight junctional networks were observed in the human normal bronchial epithelium. They were morphologically classified into three types: type I was a loosely interconnected, most complicated network consisting of 7-11 roughly parallel wavy strands and situated between ciliated cells; type II was a randomly anastomosing, simple network made up of 2-4 strands and present between goblet cells; type III was an irregularly anastomosing network composed of 4-7 strands and located between a ciliated cell and a goblet cell. Type III junctions, when a goblet cell was strongly bulged, were located on the swollen ridge, the upper surface of which was separated by a deep groove from the bulged apical surface, around the lateral surface of the cell at the level of the luminal surface. The possible relation between the orientation of strands of these networks and extra- or intracellular stress was discussed.     

Maintenance of genetic information in stem cells

The available data on DNA cosegregation in some stem cells are reviewed. Cairns was the first to assume cosegregation of template DNA strands for adult stem cells; i.e., all maternal DNA strands are preserved in one daughter cell, which remains a stem cell, while the newly synthesized DNA strands, which may contain errors, appear in the daughter cell that is committed to differentiation and passes to the transitory compartment of the cell population. The role of asymmetric mitosis in DNA cosegregation and maintenance of genetic information in stem cells is discussed.     

Qualitative and quantitative freeze-fracture studies on olfactory and nasal respiratory epithelial surfaces of frog,ox, rat,and dog

Summary A comparison of the tight-junctions of various cell types in the nasal epithelia of frog, ox, rat and dog shows that Bowman's gland cells have lowest number of strands (4–8), whereas olfactory receptor and supporting, and ciliated respiratory cells show no conspicuous differences and have 6–11 strands. Tight-junctional strand numbers show slight species-dependent variations. In regions where three cells join (observed for receptor and respiratory cells), fracture faces show two parallel strands which fuse at certain points. These strands run perpendicularly to the rest of the tight-junctional belt, which also shows an increased number of strands (13–16) in this region.Tight-junctions of mammalian olfactory dendritic endings usually show strands composed of particles, whereas those of the other three epithelial cell types consist of continuous or discontinuous bars. Tight-junctions of dendritic endings of the frog also conform to the latter type. Differences in strand density are only slight and range from 16–27 strands/m. Small angular gap-junctions were observed only within the tight-junctions of supporting cells in the rat.     

Adhesion, orientation, and movement of cells cultured on ultrathin fibronectin fibers

Summary This study examined the behavior of rat tendon fibroblasts, baby hamster kidney fibroblasts, macrophage-like P388D1 cells, and neurons from rat dorsal root ganglia, cultured on fibronectin strands 0.2–5 μm in diameter. We investigated cell spreading, orientation, formation of focal contacts, the speed of cell movement, and the speed of neurite outgrowth in cells cultured on fibronectin strands, glass covered with fibronectin, and plain, nontreated glass. Fibronectin strands significantly promoted cell spreading and caused a marked alignment of all kinds of cells to the direction of the fiber. The fibers caused the alignment of actin filaments in fibroblasts and focal contacts in fibroblasts and macrophages and increased polymerization of F-actin in cells. Fibronectin fibers also increased the speed and persistence of cell movement and the rate of neurite outgrowth. Macrophages grown on fibronectin fibers produced numerous actin-rich microspikes and adopted a polarized, migratory phenotype. These findings indicate that fibronectin strands, resembling natural components of the extracellular matrix, are more effective in activating various types of cells than two-dimensional, fibronectin-covered substrata. The results also confirm the suitability of the three-dimensionally oriented fibronectin form for use in clinical practice.     

Skin-derived human adult stem cells surprisingly share many features with human pancreatic stem cells

Multiple tissue niches in the human body are now recognised to harbour stem cells. Here, we have asked how different adult stem cell populations, isolated from two ontogenetically distinct human organs (skin, pancreas), actually are with respect to a panel of standard markers/characteristics. Here we show that an easily accessible adult human tissue such as skin may serve as a convenient source of adult stem cell-like populations that share markers with stem cells derived from an internal, exocrine organ. Surprisingly, both, human pancreas- and skin-derived stem/progenitor cells demonstrate differentiation patterns across lineage boundaries into cell types of ectoderm (e.g. PGP 9.5+ and GFAP+), mesoderm (e.g. alpha-SMA+) and entoderm (e.g. amylase+ and albumin+). This intriguing differentiation capability warrants systemic follow-up, since it raises the theoretical possibility that an adult human skin-derived progenitor cell population could be envisioned for possible application in cell replacement therapies.     

The in vivo development of the hamster periodontium and implications for the chin's evolution

The anthropological problem related to the biology and evolution of the human chin was approached by examining the hypothesis that the body of the mandible consists of two bony moieties: (1) alveolar bone and (2) basal bone. The alveolar bone was assumed to be a component of the dentition, ontogenetically distinct from the basal bone on which it is superposed. Each bone moiety exhibits differences in the amount, direction and timing of growth — factors which are presumed to be partially responsible for the origin and evolution of the chin. To provide additional information on this problem, histologic sections from 45 postnatal hamster mandibles were examined with the light microscope. The results revealed that the hamster alveolar bone tissue develops from a hyalinelike precursor which is ontogenetically distinct from the processes of intramembranous bone formation. The maturation of alveolar bone appears to be associated with and highly correlated to the progressive development of the periodontal ligament and the root, a process which suggests epithelio-mesenchymal interactions analogous to those observed during crown formation. The probable relationships between these observations on alveolar bone development and chin formation are discussed.     

DEVELOPMENT OF MUCILAGINOUS SURFACES IN EUGLENOIDS. I. STALK MORPHOLOGY OF COLACIUM MUCRONATUM1

The envelope and stalk of Colacium mucronatum Bourr. & Chad, were examined in living cells with light microscopy and in fixed preparations with scanning electron microscopy using critically point dried (CPD) and freeze dried (FD) preparations. The envelope of palmelloid cells is formed over the entire cell surface by many individual strands attached at right angles to areas of articulation of the pellicular strips. Strands were observed to anastomose on the posterior tip of otherwise naked cells. Stalks of living cells in India ink preparations had an optically dark inner core with a lighter outer sheath. In FD stalks a definite inner core was not evident, whereas CPD stalks had an outer surface composed of thick strands which may be the collapsed and aggregated strands of the FD stalks. In both there was also an amorphous matrix. The stalk forms from the aggregation of many strands from the anterior cell tip back to a point encompassing the cell surface anterior to a cross section of the tip 9 μm diam. The outer surface of the stalk comes from the pellicular surface joining that area and the core from the cell tip in the area of the canal opening. Any possible participation of the inner canal surface in stalk formation could not be determined because of the great density of the mucilage at the cell-tip/stalk junction.     

Linkage of extracellular plasminogen activator to the fibroblast cytoskeleton: colocalization of cell surface urokinase with vinculin

Several cell types display binding sites for [125I]urokinase (Vassalli, J.-D., D. Baccino, D. Belin. 1985. J. Cell Biol. 100:86-92) which in certain cases are occupied with endogenous urokinase. These sites appear to focus urokinase at cell surfaces and hence may participate in tissue matrix destruction and cell invasion. Recently Pollanen et al. (1987) demonstrated that the cell surface urokinase of human fibroblasts and fibrosarcoma cells is deposited underneath the cells in strands, apparently at sites of cell-to-substratum contact. Here, using immunofluorescence double labeling, we show that the urokinase strands present on human foreskin fibroblasts are colocalized with strands of vinculin, an intracellular actin-binding protein that is deposited at cell-to-substratum focal adhesion sites. Thus, this indicates linkage of the plasminogen/plasmin system both to sites of cell adhesion and to the cytoskeleton. The urokinase strands on HT 1080 fibrosarcoma cells are more numerous and have shapes that are more tortuous than those on normal fibroblasts. In intact HT 1080 cells, colocalized vinculin strands are obscured by an intense background of soluble vinculin but are apparent on isolated ventral plasma membranes. Certain properties of the urokinase strands suggest that they are related to the [125I]urokinase-binding sites that have been described by several groups: (a) incubating fibroblasts with dexamethasone for 48 h or at pH 3 at 5 degrees C for 10 min greatly decreases the number and intensity of the urokinase strands; (b) strands reappear when glucocorticoid- treated cells are incubated with exogenous 54-kD (but not 35-kD) urokinase, and this process is inhibited by a previously described 16- amino acid peptide that blocks [125I]urokinase binding to the cells.