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Isolation of a cell surface receptor protein for laminin from murine fibrosarcoma cells.

We used affinity chromatography to isolate a specific laminin-binding protein from murine fibrosarcoma cells. These cells bind exogenous laminin to their surface with high affinity (Kd = 2 X 10(-9)M for laminin) with approximately 5 X 10(4) sites per cell. Laminin affinity chromatography of [35S]methionine-labeled cell extracts produced two distinct proteins. One was identified as Type IV (basement membrane) collagen based on its migration pattern on SDS gels and bacterial collagenase sensitivity. The other protein, which migrates as a single band or closely spaced doublet on reduced SDS gels, has a reduced molecular weight of 69,000. Using a nitrocellulose filter disk assay, we found that the latter protein specifically bound 125I-laminin with the same high affinity (Kd = 2 X 10(-9)M for laminin) as did intact fibrosarcoma cells. By iodinating intact cells, we demonstrated that this laminin-binding protein is on the cell surface. We conclude that this protein with reduced molecular weight of 69,000 is a subunit or component of a larger cell surface receptor protein for laminin in this fibrosarcoma model. This laminin receptor may mediate the interaction of the cell with its extracellular matrix.

Animals

T-cadherin expression alternates with migrating neural crest cells in the trunk of the avian embryo.

Trunk neural crest cells and motor axons move in a segmental fashion through the rostral (anterior) half of each somitic sclerotome, avoiding the caudal (posterior) half. This metameric migration pattern is thought to be caused by molecular differences between the rostral and caudal portions of the somite. Here, we describe the distribution of T-cadherin (truncated-cadherin) during trunk neural crest cell migration. T-cadherin, a novel member of the cadherin family of cell adhesion molecules was selectively expressed in the caudal half of each sclerotome at all times examined. T-cadherin immunostaining appeared graded along the rostrocaudal axis, with increasing levels of reactivity in the caudal halves of progressively more mature (rostral) somites. The earliest T-cadherin expression was detected in a small population of cells in the caudal portion of the somite three segments rostral to last-formed somite. This initial T-cadherin expression was observed concomitant with the invasion of the first neural crest cells into the rostral portion of the same somite in stage 16 embryos. When neural crest cells were ablated surgically prior to their emigration from the neural tube, the pattern of T-cadherin immunoreactivity was unchanged compared to unoperated embryos, suggesting that the metameric T-cadherin distribution occurs independent of neural crest cell signals. This expression pattern is consistent with the possibility that T-cadherin plays a role in influencing the pattern of neural crest cell migration and in maintaining somite polarity.

Animals

Antigen-induced changes in lymphocyte circulatory patterns.

The effect of a splenic or lymph node anti-sheep red blood cell response on lymphocyte migration patterns in mice was studied. It was found that trapping of lymphocytes in these stimulated organs was indiscriminate and was followed by a period of restricted cell entry or localization; furthermore, reduced cell localization in the spleen, during the splenic response, was accompanied by a reduction in the number of cells localizing in unstimulated brachial and axillary lymph nodes. These results were taken to indicate that major changes occur in lymphocyte circulation during strong splenic immune responses.

Animals

Inhibitory effect of cetirizine 2HCl on eosinophil migration in vivo.

The effect of a potent antihistamine, cetirizine, was studied on allergic patients and normal subjects by means of an in-vivo 'skin window' technique. All subjects showed significant inhibition of skin-test responses to grass pollen, compound 48/80, histamine and methacholine, after administration of a single dose (10 mg) of cetirizine. Compared to placebo, cetirizine significantly decreased the eosinophils attraction at skin sites challenged with grass pollen and compound 48/80. In allergic patients no change in eosinophil migration pattern was noted with histamine and methacholine skin-tested sites. In normal subjects, compound 48/80 and histamine did not induce eosinophil accumulation and cetirizine did not modify cellular patterns as compared to placebo. These results suggest that cetirizine acts on eosinophil migration by inhibiting the release of mast cell mediators or inhibiting the eosinophilotactic mediators themselves.

Adult

Cells and immunoglobulins in lymph.

Studies of the free-floating lymphocytes and of the immunoglobulins in lymph collected over long periods of time from ducts draining individual tissues of the body as well as from the thoracic duct of the fetus in utero have been reviewed. The findings show that stimuli within the internal milieu act on different classes of lymphocytes to alter their migration pattern, morphology, metabolic activity, and range of immunological potentialities. As the lymphoid cells migrate between the blood, tissue fluid, and lymph, a continual process of reassortment occurs leading to the establishment of heterogeneous lymphoid cell populations in different regions of the lymphatic apparatus. It seems that the biological activities of these cells are not decided only in terms of a thymus or a bone-marrow origin. The immunoglobulins, like other proteins in lymph, are mainly derived by filtration from the circulating plasma. Some of the immunoglobulins and specific antibodies are synthesized, however, by lymphoid cells and secreted directly into the lymph.

Antibody-Producing Cells

Timing and patterns of astrocyte migration from xenogeneic transplants of the cortex and corpus callosum.

The timing, pattern, and pathway of astrocyte migration were investigated in vivo by transplantation of CD-1 mouse cerebral cortex (E13-14) or corpus callosum (P2-3) into neonatal rat cortex. A monoclonal antibody specific for a mouse astrocyte surface antigen (M2) was used to identify the location of the grafts and the migrated donor astrocytes. Within the host cortex, astrocytes from cortical grafts began migration at post-transplantation day (PTD) 7. Over the next 4 days, the most distant displaced donor cells were found progressively further away from the grafts, migrating at a rate of about 220 microns/day. After PTD 11, the migration rate for the farthest displaced donor cells slowed to 25 microns/day, and the cells appeared to stop at about PTD 16 at a distance of 1,100 microns from the edge of the graft. Astrocytes had a faster migration speed in the white matter and covered a longer distance (5 mm) than those in the gray matter, extending on occasion into the contralateral hemisphere. The patterns of astrocyte migration differed depending on local cues around the transplant. Donor astrocytes that had been implanted into the host cortex migrated toward the host cortical surface, sometimes in several radial lines. Astrocytes from grafts, especially callosal grafts, placed in the subcortical white matter migrated along the host fiber tracts. Many astrocytes transplanted into the hippocampus formed laminar patterns close to the hippocampal neuronal layers. These results suggest that the direction, pattern, and speed of astrocyte migration are influenced by local substrates in the host brain.

Animals

Elderly Hispanic migration in the United States.

The specific aim of this study was to extend the analysis of migration patterns to Hispanics (Mexican, Cuban, Puerto Rican, and Other Hispanic) over the age of 60. The 1980 Census public use sample provided the data for this study, which was structured by four research expectations derived from features of the Hispanic population and from migration concepts. First, because of population growth, migration from abroad was expected to be heavy, and about half came from abroad. Second, because of the pattern of Hispanic population concentration and the different settlement patterns of the four subgroups, a relatively small migration system with distinct subsystems for the four nationality groups was expected and found. Florida is the major ethnomagnet. Third, traditional family values, as reflected in dependent living arrangements, were expected to be more prevalent among migrants from abroad, and they were. Finally, returning to one's state of birth was highest among Mexican interstate migrants, as expected.

Aged

Patterns of epithelial migration in the unaffected ear in patients with a history of unilateral cholesteatoma.

Epithelial migration in a centrifugal manner is an established phenomenon in the normal human tympanic membrane. This pattern of migration is symmetrical in both ears of any one individual. We present a prospective study on the pattern of tympanic ink dot migration on the normal drum, in patients with a history of cholesteatoma in one ear. It was demonstrated that patients who develop cholesteatoma have a normal migratory pattern and rate in the unaffected ear. The conclusion from this study is that defective migration is not the initiating factor in the development of acquired cholesteatoma.

Cell Movement

Homozygosity, effective number of alleles, and interdeme differentiation in subdivided populations.

The amount and pattern of genetic variability in a geographically structured population at equilibrium under the joint action of migration, mutation, and random genetic drift is studied. The monoecious, diploid population is subdivided into panmictic colonies that exchange migrants. Self-fertilization does not occur; generations are discrete and nonoverlapping; the analysis is restricted to a single locus in the absence of selection; every allele mutates to new alleles at the same rate. It is shown that if the number of demes is finite and migration does not alter the deme sizes, then population subdivision produces interdeme differentiation and the mean homozygosity and the effective number of alleles exceed their panmictic values. A simple relation between the mean probability of identity and the mean homozygosity is established. The results apply to a dioecious population if the migration pattern and mutation rate are sex independent.

Alleles

Proliferation and migration of rat incisor mesenchymal cells.

Sixty female Wistar rats were injected with tritiated thymidine and killed at intervals between 1 hr and 72 hr after injection. Autoradiographs of paraffin and Vestopal W embedded sagittal sections of the maxillary incisors were prepared. The distribution of labeled cells after 1 hr revealed the basal pulp cells and the preodontoblasts as proliferative cells. The growth rate of the incisor and the migration of the basal pulp cells were established from the position of labeled cells at 72 hr observation time. The number and localization of labeled and unlabeled mitoses were registered in order to determine the duration of the generation cycle of the proliferative cells. The identical cell generation time of preodontoblasts and basal pulp cells and the migration pattern of the latter indicate that the preodontoblasts are renewed by preodontoblast multiplication and not by migration of basal pulp cells.

Animals

Enzymatic heterogeneity of seminomas.

Heterogeneity of placental-like alkaline phosphatase (PLAP-like enzyme) in seminoma was studied. PLAP-like enzyme from seminoma tissues was separated into three areas with different proportions between tumors, while PLAP and PLAP-like enzymes in normal testes were separated into two areas on the basis of hydrophobicity. By use of lectin affinity chromatography, PLAP-like enzyme in seminoma revealed extra sugar chains compared to PLAP, indicating heterogeneity of the carbohydrate moiety. However, the glycosylation patterns were found to be essentially similar between seminoma and normal testis. On isoelectric focusing, differences in migration patterns of PLAP-like enzyme were revealed between seminoma and normal testis as well as between PLAP-like enzyme and PLAP. The differences in charge were mainly due to differences in sialylation of the molecules. The complex pattern on isoelectric focusing was not altered by neuraminidase treatment, indicating a considerable charge heterogeneity within the population of PLAP-like enzyme molecules from seminoma.

Alkaline Phosphatase

Characterization of seminoma-derived placental-like alkaline phosphatase.

Characteristics of placental-like alkaline phosphatase (PLAP-like enzyme) in seminoma was studied. By use of lectin affinity chromatography, PLAP-like enzyme in seminoma revealed extra sugar chains compared to placental alkaline phosphatase (PLAP), indicating heterogeneity of the carbohydrate moiety. However, the glycosylation patterns were found to be essentially similar between seminoma and normal testis. On isoelectric focusing, differences in migration patterns were revealed between seminoma-derived and normal testis-derived PLAP-like enzyme as well as between PLAP-like enzyme and PLAP. The differences in charge were mainly due to differences in sialylation of the molecules. The complex pattern of PLAP-like enzyme from seminoma on isoelectric focusing was not altered by neuraminidase treatment, indicating a considerable charge heterogeneity within the population of the enzyme molecules from the tumor.

Alkaline Phosphatase

The different migratory characteristics of lymphocyte populations from a whole spleen transplant.

Spleens from AS x BN donor rats labelled in vivo by multiple doses of [3H]thymidine were transplanted into syngeneic recipients by anastomosis to the abdominal great vessels. The recipients were killed 1-5 days after receiving the whole spleen transplants and the numbers and location of the [3H]thymidine-labelled cells which had migrated from the labelled donor spleen traced by means of autoradiographs of sections, imprints and smears of various recipient lymphoid tissues. These results were compared with the migration pattern of labelled dissociated spleen cell suspensions injected intravenously. The latter consists almost entirely of small lymphocytes which migrate to T or B areas of recipient spleen, lymph nodes and Peyer's patches. The labelled whole spleens also contained cells which migrated to the T and B areas of recipient lymphoid tissues, but in addition contained many lymphoid cells which migrated to the red pulp of the recipient spleen and to the lamina propria of the gut. These experiments showed, therefore, that the spleen contains mobile elements which have not been detected by transfer of spleen cell suspensions.

Animals

Analysis of genetic variation of epizootic hemorrhagic disease virus and bluetongue virus field isolates by coelectrophoresis of their double-stranded RNA.

Thirty-two bovine field isolates of bluetongue virus (BTV), 6 field isolates of epizootic hemorrhagic disease virus (EHDV) from deer, 4 BTV prototype serotypes (10, 11, 13, and 17), and 2 EHDV prototype serotypes (1 and 2) were coelectrophoresed, using polyacrylamide gels. Field isolates were obtained from various regions of the United States. Analysis of polyacrylamide gels and scattered plots generated for comparison of migration patterns for different isolates within each serotype of BTV revealed wide variation among the individual segments. The BTV serotypes 10 and 11 had more variation, compared with BTV serotypes 13 and 17, especially for migration of genome segment 5. A definitive correlation was not seen between the double-stranded RNA migration profiles on polyacrylamide gel electrophoresis, geographic origin, herd of origin, or year of collection. One BTV field isolate contained more than 1 electropherotype, with 2 bands at the segment-7 position, and it was further characterized as BTV serotype 11. Segments 2 and 5 of EHDV isolates were more variable in their migration than were the other gene segments. Generally, migration profiles for EHDV double-stranded RNA were more variable, compared with those of BTV isolates. Although a correlation was found between migration profiles and serotype of 2 isolates of EHDV, a study of additional EHDV isolates is required before the diversity of electrophoretic patterns of EHDV can be determined.

Animals

Generation of avian cells resembling osteoclasts from mononuclear phagocytes.

Several lines of indirect evidence suggest that a monocyte family precursor gives rise to the osteoclast, although this hypothesis is controversial. Starting with a uniform population of nonspecific esterase positive, tartrate-sensitive, acid phosphatase-producing, mannose receptor-bearing mononuclear cells, prepared from dispersed marrow of calcium-deprived laying hens by cell density separation and selective cellular adherence, we generated multinucleated cells in vitro. When cultured with devitalized bone, these cells show, by electron microscopy, the characteristic osteoclast morphology in that they are mitochondria-rich, multinucleated, and, most importantly, develop characteristic ruffled membranes at the matrix attachment site. Moreover, as documented by scanning electron microscopy, these cells pit bone slices in a manner identical to freshly isolated osteoclasts. In addition, isoenzymes of acid phosphatase from generated osteoclasts, separated by 7.5% polyacrylamide gel electrophoresis at pH 4, are identical to those of mature osteoclasts in migration pattern and tartrate resistance, although the precursor cells from which the osteoclasts are generated produce an entirely different isoenzyme, which is tartrate-sensitive and migrates less rapidly at pH 4. The fused cells also exhibit a cAMP response to prostaglandin E2. Therefore, osteoclast-like cells can be derived by in vitro culture of a marrow-derived monocyte cell population.

Animals

In vivo migration and tissue localization of highly purified lymphokine-activated killer cells (A-LAK cells) in tumor-bearing rats.

Our laboratory has previously reported that the adoptive transfer of highly purified lymphokine-activated killer cells (adherent-LAK, A-LAK) into Fischer 344 (F344) rats bearing established lung or liver micrometastases effectively reduced the resultant tumor growth more than 90%, leading to significant increases in animal survival (Cancer Res. 49, 1441, 1989). To begin to investigate the mechanism(s) by which A-LAK cells mediate this anti-tumor effect, we studied their migration patterns in F344 rats bearing experimentally induced lung and liver metastases as well as subcutaneous tumors. A-LAK cells which were phenotypically 95 to 100% natural killer cells/large granular lymphocytes were labeled with either 51Chromium or fluorescein diacetate (so as to be visualized microscopically). Intravenous injection of such labeled A-LAK cells did not show significant differences in their tissue distribution patterns in tumor-bearing versus normal rats, even when high levels of exogenous recombinant interleukin-2 (rIL-2) was administered. A-LAK cells first migrated to the lungs and then subsequently migrated to the liver and spleen as early as 2 to 6 hr following iv injection. The kinetics of exit of A-LAK cells from the pulmonary capillary beds was not significantly different in rats bearing 3-day micrometastases or 14-day macrometastases compared to normal rats. Moreover, the presence of metastases in the liver did not alter the extent or kinetics of entry of A-LAK cells into the liver even in the presence of exogenously administered rIL-2. Finally, in rats bearing subcutaneous tumors, no evidence could be obtained that A-LAK cells were selectively localized to the tumor site. Tissue sections of livers from metastases-bearing animals injected with fluorescein diacetate labeled A-LAK cells did not demonstrate significant numbers of A-LAK cells infiltrating tumor nests with or without the administration of exogenous IL-2. These data suggest that A-LAK cells may mediate tumor regression in vivo by direct and indirect mechanisms, possibly through the secretion of cytokines and/or the recruitment of secondary effector cells.

Adenocarcinoma

Invited review: guidance cue patterns and cell migration in multicellular organisms.

In multicellular organisms, guidance cues are either diffusible molecules or cellular or extracellular surfaces that are found in reproducible locations and that orient migrating cells and cell processes. The pattern of the guidance cues usually determines the complex in vivo migration routes of motile cells and cell processes. Within organisms, guidance cues are found to be organized in two general patterns: a) broad gradients--such as diffuse chemotactic gradients; b) discrete routes (substrate pathways)--such as chemotactic gradients confined to long channels, and such as the axon surface which represents a long specific highway for migrating Schwann cells.

Amphibians

[Transplantation of embryonic cerebella into adult rat cerebella].

Cerebellar primordia of rat embryos (embryonic days of 14-20) were transplanted into mature cerebella of normal adult rats (Fischer 344). After 1-12 months, animals were sacrificed, and the development of grafts and the interaction between host and graft were examined by light and electron microscopy. Grafts from E14 rat cerebella showed the highest survival rate (85%), which decreased with increasing donor age. In some experiments, the donor tissue developed and differentiated to form folia with the trilaminar organization of the cerebellar cortex. Synaptic connections between neuronal elements in the graft showed basically the normal pattern. Thus, mossy terminals formed synaptic contacts with dendrites of granule cells, and axons of basket cells made synaptic contacts with somata of Purkinje cells. Many spines of Purkinje dendrites were contacted with parallel fibers, while others were surrounded by processes of astroglia. Migration of donor Purkinje and granule cells in the host molecular layer was frequently observed. Some donor granule cells, implanted in the superficial aspect of the host molecular layer, migrated down along processes of Bergmann glia, which resembled the migration pattern of the normal development. Other granule cells attached to neuroglial processes which ensheathed capillary walls. While the migration of granule cells was found to be glial-dependent, the glial role in the migration of Purkinje cells was not clear. Purkinje cells probably migrated into the host molecular layer under favorable conditions where the loss of host Purkinje cells had been occurred to some extent and consequently followed by the increase of free axon endings of parallel fibers and interneurons of the host.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals