PubMed Health⌕ Search

Biomedical subjects

S D Banerjee

Publications and source records attributed to S D Banerjee.

14 recordsLinked to original sources
PubMed2000

Molecular characterization of a novel intracellular hyaluronan-binding protein.

Hyaluronan has well defined functions in extracellular matrices and at the surface of cells. However, several studies have now shown that significant pools of hyaluronan are also present intracellularly, but its function therein is unknown. One avenue of investigation that may assist in defining the function of intracellular hyaluronan is to identify intracellular hyaluronan-binding proteins. In previous studies we identified CDC37, a cell cycle regulatory protein, using a monoclonal antibody that recognizes a novel group of hyaluronan-binding proteins. In this study, we have identified a second hyaluronan-binding protein with this antibody and characterized its properties. This protein, which we have termed IHABP4, was also found to be an intracellular and a specific hyaluronan-binding protein, containing several hyaluronan-binding motifs: (R/K)[X(7)](R/K) (where R/K denotes arginine or lysine and X denotes non-acidic amino acids). Furthermore, we have determined the gene organization of IHABP4 and cloned cDNAs for the chick, mouse, and human homologs. Comparison of the deduced chick, mouse, and human protein sequences showed that the hyaluronan-binding motifs, (R/K)[X(7)](R/K), in these sequences are conserved; both chick and mouse IHABP4 were shown directly to bind hyaluronan. Biochemical fractionation and immunofluorescent localization of epitope-tagged IHABP4 indicated that it is mainly present in the cytoplasm. These data support the possibility that intracellular hyaluronan and its binding proteins may play important roles in cell behavior.

Amino Acid Sequence↗
PubMed1995

A novel glycosaminoglycan-binding protein is the vertebrate homologue of the cell cycle control protein, Cdc37.

Using a monoclonal antibody, IVd4, that recognizes a novel group of hyaluronan-binding proteins, we have immunoscreened a cDNA library constructed from embryonic chick heart muscle mRNA. One of the cDNAs isolated from the library encodes a 29.3-kDa protein homologous to Cdc37, an essential cell cycle regulatory factor previously characterized genetically in yeast and Drosophila; this is the first vertebrate CDC37 gene to be cloned to date. We also present evidence for the existence of a second chick isoform that is identical to the 29.3-kDa protein over the first 175 amino acids but is entirely different at the carboxyl terminus and lacks the IVd4 epitope. The avian Cdc37 binds hyaluronan, chondroitin sulfate and heparin in vitro, and both isoforms contain glycosaminoglycan-binding motifs previously described in several hyaluronan-binding proteins. These findings suggest a role for glycosaminoglycans in cell division control.

Amino Acid Sequence↗
PubMed1992

The role of hyaluronan-binding protein in assembly of pericellular matrices.

Hyaluronan-dependent pericellular matrices or "coats" are expressed by a variety of cell types in culture and modulation of their expression may be important in regulation of cell interactions in vivo during development. Monoclonal antibody IVd4, which recognizes hyaluronan-binding protein with the properties of a hyaluronan receptor, was shown to block formation of these coats by a variety of cells. Using rat fibrosarcoma cells, it was found that the antibody not only blocked initial formation of the coats but also caused their loss when added subsequent to formation. The loss of preformed coats in the presence of antibody occurred at 4 degrees and 37 degrees, implying that the function of hyaluronan-binding protein in coat formation is not in mediating metabolic processes. The antibody also had no significant effect on hyaluronan production by the fibrosarcoma cells. In addition, hyaluronan hexasaccharide, a competitive inhibitor of the interaction between polymeric hyaluronan and its cell surface receptor, was found to inhibit coat formation. Thus it is concluded that a hyaluronan-binding protein with the properties of a hyaluronan receptor is required for pericellular matrix formation.

Animals↗
PubMed1992

Hyaluronan-binding protein in endothelial cell morphogenesis.

Previous studies from several laboratories have provided evidence that interaction of hyaluronan (HA) with the surface of endothelial cells may be involved in endothelial cell behavior. We have recently characterized a mAb, mAb IVd4, that recognizes and neutralizes HA-binding protein (HABP) from a wide variety of cell types from several different species (Banerjee, S. D., and B. P. Toole. 1991. Dev. Biol. 146:186-197). In this study we have found that mAb IVd4 inhibits migration of endothelial cells from a confluent monolayer after "wounding" of the monolayer. HA hexasaccharide, a fragment of HA with the same disaccharide composition as polymeric HA, also inhibits migration. In addition, both reagents inhibit morphogenesis of capillary-like tubules formed in gels consisting of type I collagen and basement membrane components. Immunocytology revealed that the antigen recognized by mAb IVd4 becomes localized to the cell membrane of migrating cells, including many of their lamellipodia. Treatment with high concentrations of HA hexamer causes loss of immunoreactivity from these structures. We conclude that HABP recognized by mAb IVd4 is involved in endothelial cell migration and tubule formation.

Animals↗
PubMed1992

Effects of microgravity or simulated launch on testicular function in rats.

Testes from flight rats on COSMOS 2044 and simulated-launch, vivarium, or caudal-elevation control rats (5/group) were analyzed by subjective and quantitative methods. On the basis of observations of fixed tissue, it was evident that some rats had testicular abnormalities unassociated with treatment and probably existing when they were assigned randomly to the four treatment groups. Considering rats without preexisting abnormalities, diameter of seminiferous tubules and numbers of germ cells per tubule cross section were lower (P less than 0.05) in flight than in simulated-launch or vivarium rats. However, ratios of germ cells to each other or to Sertoli cells and number of homogenization-resistant spermatids did not differ from values for simulated-launch or vivarium controls. Expression of testis-specific gene products was not greatly altered by flight. Furthermore, there was no evidence for production of stress-inducible transcripts of the hsp70 or hsp90 genes. Concentration of receptors for rat luteinizing hormone in testicular tissue and surface density of smooth endoplasmic reticulum in Leydig cells were similar in flight and simulated-launch rats. However, concentrations of testosterone in testicular tissue or peripheral blood plasma were reduced (P less than 0.05) in flight rats to less than 20% of values for simulated-launch or vivarium controls. Thus spermatogenesis was essentially normal in flight rats, but production of testosterone was severely depressed. Exposure to microgravity for greater than 2 wk might result in additional changes. Sequelae of reduced androgen production associated with microgravity on turnover of muscle and bone should be considered.

Animals↗
PubMed1991

Monoclonal antibody to chick embryo hyaluronan-binding protein: changes in distribution of binding protein during early brain development.

A monoclonal antibody, MAb IVd4, that recognizes hyaluronan-binding protein (HABP) from chick embryo brain has been produced and characterized. By immunoblotting, MAb IVd4 was shown to recognize three proteins in chick embryo brain of molecular weight 93, 90, and 69 kDa; this interaction was inhibited by addition of hyaluronan hexasaccharides. Overlay of transblots with [3H]hyaluronan showed binding to proteins of similar molecular weight. MAb IVd4 blocked binding of [3H]hyaluronan to brain HABP and to simian virus-transformed 3T3 cells, indicating a possible relationship with the 85-kDa hyaluronan receptor of these cells. The distribution of HABP during early brain development was analyzed by immunohistochemistry. Immunoreactivity was uniform in newly formed neuroectoderm but became more concentrated in the roof of the brain during the second day of embryonic development. As the neuroectoderm becomes layered, the HABP was increasingly restricted to the forming plexiform layer, an area enriched in neural cell processes. Immunoreactivity was greatly enhanced by pretreatment of tissue with hyaluronidase, presumably due to removal of hyaluronan bound to the HABP, and was abolished on treatment with hyaluronan hexasaccharide, presumably due to inhibition of HABP-antibody interaction. These results suggest that a hyaluronan receptor is involved in early cellular events in brain development.

Animals↗
PubMed1984

Remodelling of the basement membrane: morphogenesis and maturation.

We have analysed the reciprocal interactions between mouse embryo submandibular epithelium and mesenchyme which result in branching morphogenesis of the epithelium. The interactions modify the composition and metabolism of the basal and reticular laminae which comprise the basement membrane lying between these tissues. The mesenchyme remodels the basement membrane by depositing a type I collagen-rich matrix on the basal lamina and by producing a neutral hyaluronidase, which degrades hyaluronate and chondroitin sulphate, components of this basal lamina. By analogy with mouse mammary epithelial cells, the submandibular epithelial cells have a heparan sulphate-rich proteoglycan on their cell surfaces which is anchored to the cells. The extracellular domain of this integral membrane proteoglycan binds to interstitial collagen. Interfering with the collagen-proteoglycan interaction appears to reduce the morphological stability of the cells. Together with other processes, including epithelial cell proliferation, this remodelling leads to branching epithelial morphogenesis. Basement membrane remodelling may be a general process for regulating cell behaviour during development and is one of the mechanisms of morphogenetic tissue interaction. Remodelling may also cause maturation of basement membranes from a dynamic state of high turnover in the embryo to their persistence and stability in the adult organism.

Animals↗
PubMed1977

Basal lamina of embryonic salivary epithelia. Production by the epithelium and role in maintaining lobular morphology.

The role of the basal lamina in maintaining the normal morphology of mouse embryo submandibular epithelia was assessed by examining its production as well as the cellular and organ culture changes associated with its removal and replacement. The lamina was removed from epithelia isolated free of mesenchyme by brief treatment with testicular hyaluronidase in the absence of calcium. The treatment causes rounding-up of the cells, loss of cellular cohesion, appearance of microvilli, and changes in the organization of cytoskeletal structures. The lamina is not removed and the cellular alterations do not occur in the absence of hyaluronidase in calcium-free medium or when both enzyme and calcium are present, possibly because digestion of chondroitin sulfate, a component of the lamina, is inhibited by calcium. Within 2 h after treatment, in the absence of mesenchyme or biological substrata, the epithelia deposits a new lamina, which is identical by several criteria to the preexisting lamina, and reverses the cellular alterations. Epithelia treated with hyaluronidase lose lobular morphology during culture with mesenchyme. Delaying culture with mesenchyme, to allow restoration of the lamina and of normal cellular architecture, prevents the loss of lobular morphology. The results indicate that the basal lamina imposes morphologic stability on the epithelium, while the mesenchyme apparently affects processes involved in changes in morphology, possibly by selective degradation of the basal lamina.

Animals↗
PubMed1977

Basal lamina of embryonic salivary epithelia. Nature of glycosaminoglycan and organization of extracellular materials.

The ultrastructural organization and the composition of newly synthesized glycosaminoglycan (GAG) in the epithelial basal lamina of mouse embryo submandibular glands were assessed. The labeled GAG accumulating in the lamina is distinct from that in its tissue of origin, the epithelium, or from that in the surrounding mesenchyme. In the lamina, hyaluronic acid accounts for approximately 50% of the labeled GAG, chondroitin-4-sulfate is twice the chondroitin-6-sulfate, and there is a low proportion of chondroitin. This composition is constant regardless of whether the lamina is labeled by whole glands or, in the absence of mesenchyme, by isolated epithelia retaining a lamina and by isolated epithelia generating a lamina de novo. The results andicate that the labeled GAG are bona fide components of the lamina, and suggest that laminar GAG is deposited in units of constant composition. Ultrastructural observations following ruthenium red staining or tannic acid fixation extablish that the lamina is a highly ordered specialization of the basal cell surface. Discrete structures in macroperiodic arrays apparently attached to the plasmalemma are visualized. This organization is seen in intact glands and in the laminae produced by epithelia in the absence of mesenchyme or biological substrate. The data are interpreted as indicating that the basal lamina contains supramolecular complexes of hyaluronic acid and proteoglycan which are organized into an extracellular scaffolding which imposes structural form on the epithelium.

Animals↗
PubMed1972

Cytochalasin B: lack of effect on mucopolysaccharide synthesis and selective alterations in precursor uptake.

Synthesis and secretion of mucopolysaccharide in mouse 3T3 fibroblasts and in embryonic submandibular glands are unaffected by amounts of cytochalasin B that alter the morphology of these cells and tissues. The drug markedly and reversibly inhibits incorporation of [(3)H]glucosamine into mucopolysaccharide by preventing cellular uptake of the precursor, but does not affect incorporation of radiosulfate. Cytochalasin does not alter DNA, RNA, or protein synthesis, but stimulates the uptake of orotic acid and markedly inhibits the uptake of glucose. These selective effects on the transport of small molecules suggest that the primary action of the drug may be on cell membranes. Since processes unrelated to microfilament disruption may be altered by cytochalasin, great caution must be exercised in interpreting studies with the drug.

Animals↗
PubMed1972

Acid mucopolysaccharide (glycosaminoglycan) at the epithelial-mesenchymal interface of mouse embryo salivary glands.

Acid mucopolysaccharide (glycosaminoglycan) has been demostrated at the epithelial-mesenchymal interface of mouse embryo submandibular glands by (a) specific staining for polymeric sulfate with Alcian blue 8 GX at various magnesium concentrations, (b) specific staining for polymeric uronic acid by selective oxidation of these residues to Schiff-reactive compounds, (c) electron microscope localization of ruthenium red staining, (d) radioautographic localization of glucosamine-(3)H and (35)SO(4), and (e) by susceptibility of the glucosamine radioactivity at the interface to digestion with protease-free hyaluronidase. Moreover, material labeled with glucosamine-(3)H and (35)SO(4) and with chemical characteristics identical with those of acid mucopolysaccharide were isolated from the glands. Acid mucopolysaccharide is distributed over the entire epithelial surface. The amount of acid mucopolysaccharide, as revealed by the staining procedures, is nearly equivalent at all sites. In contrast, the rate of accumulation of glucosamine-labeled mucopolysaccharide is greater at the surface of the distal ends of the growing and branching lobules. This distribution of newly synthesized acid mucopolysaccharide at the sites of incipient cleft formation suggests that surface-associated acid mucopolysaccharide is involved in the morphogenetic process. A mechanism of branching morphogenesis is proposed which accounts for the distribution of collagen fibers and total and newly synthesized acid mucopolysaccharide at the epithelial surface.

Ammonia↗
PubMed1972

Dependence of salivary epithelial morphology and branching morphogenesis upon acid mucopolysaccharide-protein (proteoglycan) at the epithelial surface.

The morphogenetic role of the acid mucopolysaccharide (glycosaminoglycan) at the epithelial surface of mouse embryo submandibular glands has been studied by comparing the in vitro morphogenesis of epithelia from which the mucopolysaccharide was removed with that of those that retained the mucopolysaccharide. Epithelia isolated free of mesenchyme by procedures which retain the bulk of surface mucopolysaccharide maintain their lobular shape and undergo uninterrupted branching morphogenesis in culture in direct combination with fresh mesenchyme. Under identical culture conditions, epithelia from which surface mucopolysaccharide was removed lose their lobules and become spherical masses of tissue. During continued culture, the spherical epithelia produce outgrowths from which branching morphogenesis resumes. The morphogenetically active mucopolysaccharide is localized within the basal lamina of the epithelial basement membrane and appears to be bound to protein. During culture in combination with mesenchyme, epithelia undergoing uninterrupted morphogenesis show maximal accumulation of newly synthesized surface mucopolysaccharide at the distal ends of the lobules, the sites of incipient branching. In contrast, the material accumulates nearly equivalently over the surface of the spherical epithelia, with the exception that there is greater accumulation of the material at the surfaces of the budding outgrowths, the sites where morphogenesis will resume. Rapidly proliferating cells are localized within the lobules of epithelia undergoing uninterrupted morphogenesis, but are distributed uniformly in the cortex of the spherical epithelia, except for the outgrowths which show a greater localization of proliferating cells. It is concluded that normal salivary epithelial morphology and branching morphegenesis require the presence of acid mucopolysaccharide-protein within the epithelial basal lamina.

Ammonia↗