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Biomedical subjects

D B Wilson

Publications and source records attributed to D B Wilson.

At least 343 records · Page 19Linked to original sources

Histological defects in the cerebellum of adult lurcher (Lc) mice.

The adult cerebellum of the heterozygous lurcher mouse (Lc/+) shows severe defects which affect particularly the Purkinje cells and granule cells. The cytoarchitecture of the cerebellar cortex is disturbed, and the granular and molecular layers are poorly defined. The population of Purkinje cells is depleted, except for a few large neurons scattered in the granular layer and white matter. Although a discrete Purkinje cell layer does not exist, pyridine-silver preparations reveal a row of arborizing basket cell axons and climbing fiber terminals which suggests the site previously occupied by Purkinje cell bodies. Granule cells are also affected and are distributed throughout the molecular layer, granular layer, and white matter.

Animals↗

Properties of the entry and exit reactions of the beta-methyl galactoside transport system in Escherichia coli.

The Km, Vmax, and Ki of the entry reaction were determined for three substrates of the beta-methyl galactoside transport system: D-galactose, D-glycerol-beta-D-galactoside, and beta-methyl-D-galactoside. Although the data for D-galactose and D-glycerol-beta-D-galactoside followed simple Michaelis-Menten kinetics, the results for beta-methyl-D-galactoside deviated from Michaelis-Menten kinetics in that the Ki for beta-methyl-D-galactoside inhibition of both of the other two substrates was 10-fold greater than the Km for beta-methyl-D-galactoside entry. Furthermore, two partial mgl- strains retain 56% of the parental level of the beta-methyl-D-galactoside entry reaction, but only 12% of the parental level of transport of the other two substrates. The exit reaction of beta-methyl-D-galactoside was shown to be first order. It was stimulated sixfold when the cells were provided with an energy source. This stimulation required adenosine 5'-triphosphate or a related compound. The exit reaction was not altered by mutations in any of the three cistrons which inactivate the beta-methyl-D-galactoside entry reaction, was not increased by growth in the presence of inducers of the entry reaction, and was not repressed by growth on glucose. The striking differences between the entry and exit reactions suggest that they either use different carriers or that none of the three cistrons which are currently known to code for components of the beta-methyl galactoside transport system code for its membrane carrier.

Adenosine Triphosphate↗

SPECIFIC positive and negative selection of rat lymphocytes reactive to strong histocompatibility antigens: activation with alloantigens in vitro and in vivo.

This study compares the functional properties of rat thoracic duct lymphocytes (TDL) after stimulation with strong alloantigens of the major histocompatibility complex (MHC) either in vitro in preparative mixed lymphocyte interactions (MLI) or in vivo in systemic graft-vs-host (GVH) reactions. Comparisons were made of PHA responses and reactivity to the specific priming haplotypes or to third party haplotypes in analytical MLI and in GVH reactions either before or after the activated populations were "parked" in syngenetic T cell-deprived (B) rats. These comparisons can be summarized as follows: 1) TDL populations primed in bulk MLI cultures (MLI-TDL) slowed some evidence of specific positive selection when tested immediately; MLI responses to specific alloantigens were both relatively large and accelerated in tempo, whereas responses to third party alloantigens were diminished but also accelerated in tempo. Specific GVH responses were more marked than in third party recipients but they were also decreased relative to normal, and displayed an abberant dose/response slope. MLI-TDL populations tested after they had been stored in syngeneic B rats showed clear evidence of stable-specific positive selection; specific MLI and GVH responses were enriched relative to third party responses and also in comparison to normal, unselected TDL populations. This finding indicates that GVH and MLI reactivity are probably both functional capacities of the same lymphocyte subpopulation since positive selection by one function (MLI) also enriched for a second (GVH). 2) Parental strain TDL activated in vivo in the systemic GVH reaction in irradiated F1 animals and recovered from the thoracic duct 3 to 4 days later (late GVH-TDL) consisted mainly of blast cells, however, in contrast to MLI-TDL these populations showed no evidence of positive selection when tested before or after parking in B rats. MLI responses to specific alloantigens were minimal, and greatly reduced in magnitude compared to normal. GVH responses to specific haplotypes could be detected, but these were not enriched compared to normal, despite the content in the late GVH-TDL populations of a significant proportion of blast cells presumably activated by host alloantigens. 3) Early collections (less than 40 hr) of parental strain GVH-TDL collected from F1 recipients contained no blast cells and showed impressive degrees of negative selection; they were markedly depleted of both GVH and MLI activity to specific alloantigens but displayed normal reactivity to third party alloantigens. Moreover, specific negative selection was persistent in these populations parked for several weeks in B rats, and indication that a specific subpopulation of reactive cells had been physically eliminated. 4) PHA responses of both MLI- and GVH-activated TDL populations tested either before or after parking in B rats were approximately normal on a per T cell basis...

Animals↗

Specific selection of cytotoxic effector cells: the generation of cytotoxic T cells in rat thoracic duct lymphocyte populations positively or negatively selected for reactivity to specific strong histocompatibility alloantigens.

These studies consider the generation of cytotoxic T lymphocytes (CTL) from precursors (CTLP) present in rat thoracic duct lymphocytes after stimulation with strong alloantigens. Also, they explore the relationship between CTLP and "initiator" (I) lymphocytes responsible for specific GVH and MLI reactions. Positively selected TDL populations prepared in bulk MLI cultures show enriched GVH and MLI reactivity for the selecting major histocompatibility complex (MHC) haplotype, but no cytotoxic activity, raising the possibility that I and CTLP may belong to different subpopulations, and the latter failed to differentiate or to survive under these culture conditions. Restimulation of these cells in Marbrook culture vessels with the original priming haplotype under conditions suitable for generating killer cells in vitro resulted in greatly increased specific CTL activity with accelerated kinetics soon after priming and normal kinetics later. These findings indicate that "memory" killer cells can be generated in a previously stimulated lymphocyte population that had no overt cytotoxic activity. Restimulation with third party haplotypes failed to give CTL activity either to specific or to third party targets. Negatively selected TDL populations prepared by "filtration" through x-irradiated F1 rats, depleted of specific GVH and MLI responses, were also depleted of the ability to generate CTL in Marbrook cultures stimulated with the selecting haplotype. Stimulation with third party haplotypes, or with both third party and specific haplotypes together, gave CTL effective only against the third party target.

Animals↗

Proliferation gradients in the inner ear of the monkey (Macaca mulatta) embryo.

Radioautographic studies were conducted on early developmental stages of the inner ear in monkey (Macaca mulatta) embryos ranging in age from stage 11 to stage 20 (25 to 39 days' gestation) and labeled for one hour with [3H] thymidine. The most active period of proliferation occurred at stage 13, at which time all regions of the otocyst, including the endolymphatic duct, were heavily labeled. In subsequent stages the dorsal portion of the endolymphatic duct failed to incorporate label, whereas proliferation continued in the ventral portion of the duct near its communication with the medial wall of the otocyst. The duct thus appeared to grow upward as a result of cell division from below. Mitotic activity continued in the remainder of the otocyst although there was a progressive temporal decrease in the labeling index. Spatial gradients also occurred, with dorsal and lateral regions less active than ventral and medial ones. As the otocyst differentiated, the cells in various areas became attenuated. These flattened cells were still capable of undergoing cell division, as indicated by the presence of labeled nuclei in all regions of the utricle, saccule, semicircular ducts, and cochlear duct.

Animals↗

Collaboration of allogeneic T and B lymphocytes in the primary antibody response to sheep erythrocytes in vitro.

This study provides a direct quantitative comparison of the helper effects of allogeneic and syngeneic rat T cells in the production of direct SRBC plaque-forming cell (PFC) responses by B cells in culture. In syngeneic T-B combinations, log-log plots of the number of PFC generated after 5.5 days in culture vs. the number of T cells employed as helpers showed a linear response between 10(4) and 2.5 times 10(5) T cells added. Allogeneic T-B combinations, in which the T cells possess the capacity for reactivity to major alloantigens of the B-cell donor, showed a different dose/response relationship in which PFC responses were decreased at high T/B ratios and augmented at low T/B rations. In this system responses were detected with as few as 10(3) allogeneic T cells. Use of negatively selected allogeneic T populations, specifically depleted of mixed lymphocyte interaction (MLI) and graft-vs-host reactivity for B-cell alloantigens, as helpers gave dose/response curves quantitatively identical to responses with syngeneic T-B combinations and also with F1 T-cell parental B-cell combinations. These data indicate that rat T and B cells need not share a major histocompatibility complex haplotype in order to collaborate effectively in a primary direct PFC response to SRBC in culture. In addition, the PFC response required the combinaed presence of T and B cells as well as antigen in the cultures, a finding consistent with the two signal model of B-cell activation. Finally, the dose/response data obtained suggest the possibility that although SRBC antigen is required in the cultures helper activity with low numbers of normal allogeneic T cells may not depend on T cells having specificity for this antigen.

Animals↗

Tryptophan and cystein residues of the acetylcholine receptors of Torpedo species. Relationship to binding of cholinergic ligands.

Several methods were used to analyze for tryptophan in the acetylcholine (ACh) receptors purified from the electric organs of the electric rays, Torpedo californica and Torpedo marmorata. The best value of tryptophan was 2.4 mol %. When excited at 290 nm, both receptors fluoresced with a maximum at 336, but there was no change in the fluorescence emission spectra upon binding of carbamylcholine, d-tubocurarine, ACh, or decamethonium. The free SH content of the Torpedo receptors varied in different preparations, and was highest in that purified from fresh T. californica using deaerated solutions and dialysis under nitrogen, and lowest in that prepared from the aged lyophilized membranes of T. marmorata. The maximum free SH content was 20 nmol/mg of protein or 0.22 mol %, equal to at most 18% of the total cysteic acid residues. Reaction of either 33% or of all the SH residues with p-chloromercuribenzoate reduced maximum ACh binding to the pure receptor prepared from fresh T. californica by only 23%.

Acetylcholine↗

Proliferation in vivo of T lymphocytes reactive to histocompatibility alloantigens: a correction.

Rat lymphocytes in mixed cultures can reutilize tritiated thymidine from labeled granulocytes. Shortly after thymidine injections in vivo, major effects on the frequency of labeled lymphocyte mitoses in peripheral blood cultures are introduced by 10-20% polymorph contamination, even though transfer of label via supernates is not demonstrable. Cold thymidine in the cultures prevents reutilization, and has permitted reevaluation of several previous conclusions concerning the life history of lymphocytes reactive to major histocompatibility alloantigens (HARC). Rather than being predominantly recently divided cells, HARC do not appear to have an age distribution, in blood or lymph, significantly different from the general recirculating lymphocyte population. However, the ability of immunization across strong allogeneic differences to increase markedly the proportion of young HARC among the specifically responsive population has been confirmed.

Animals↗

Surface defects in ventricular cells of brains of mouse embryos homozygous for the Loop-tail gene: scanning electron microscopic study.

Luminal surfaces in the mesencephalon and rhombencephalon in normal mouse embryos and those homozygous for Lopp-tail were studied by means of scanning electron microscopy. Ventricular cells in the ventrolateral regions of normal day-10 and -11 brains showed single apical cilia and microvilli, whereas those in ventromedial regions showed a dense network of microvilli and bulbous projections which tended to obscure the apical cilia and cellular outlines. Similar regional differences occurred in the Loop-tail brains, although there was a marked decrease in the number and density of microvilli and bulbous projections. At days 12-14 of gestation the latter brains also showed a flattening of cell surfaces, shallow depressions, and craterlike ruptures in the plasma membranes.

Animals↗

Acetylcholinesterase activity in the cerebellum of the lurcher (Lc) mutant mouse.

The activity and distribution of the enzyme acetylcholinesterase in the cerebellum of adult lurcher (Lc) mutant mice and of their normal littermates was investigated using biochemical assay and light microscopic histochemistry. The biochemical assay demonstrated an approximate two-fold increase of enzyme activity in the lurcher cerebellum compared to the values obtained for the normal controls. Acetylcholinesterase activity in the cerebellum of the normal adult mouse was predominantly evident in the granular layer, corresponding to the location of the glomeruli. In contrast, the lurcher cerebellum exhibited enzyme activity in both molecular and granular layers. In the molecular layer the staining appeared to be associated with ectopic granule cells. In both normal and lurcher mice, the Golgi cells, subcortical white matter and deep nuclei also showed varying degrees of staining for acetylcholinesterase.

Acetylcholinesterase↗

Specific positive selection of lymphocytes reactive to strong histocompatibility antigens.

Selected populations of thymus-derived (T) rat lymphocytes having specific immunological reactivity to chosen histocompatibility (H) alloantigens are found among the cellular products of the mixed lymphocyte interaction (MLI). Such specific selection seems to depend on (a) the antigen-induced proliferation of specific H antigen reactive cells (HARC), and (b) the disappearance of nonreactive cells from the cultures. When the surviving cells from this lymphocyte-antigen interaction are transferred into thymectomized, X-irradiated, marrow-reconstituted syngeneic recipients (B rats) which lack detectable T-lymphocyte functions, the lymphocyte populations subsequently recovered from the hosts possess the capacity to react in the MLI and in the graft-vs.-host (GVH) reaction, and the reactions have specificity for the original priming alloantigens. In addition, these findings identify the cell that reacts in the MLI with the GVH reactive cell.

Animals↗

Source of energy for the Escherichia coli galactose transport systems induced by galactose.

The beta-methyl-galactoside- and galactose-specific transport systems of Escherichia coli were shown by experiments involving inhibitors and the use of an adenosine triphosphatase mutant strain to utilize adenosine 5'-triphosphate or a related compound to drive active transport. These systems were shown to be unable to use the activated-membrane state. The galactose-specific transport system was shown to behave most like a member of the binding-protein class of transport systems by its response to osmotic shock and vesicle formation. These results extended to two sugar transport systems: the correlation between the source of energy and class of transport system found by Berger (1973) for amino acid transport systems. That is, binding-protein systems utilized adenosine 5'-triphosphate whereas membrane-bound systems utilized the activated-membrane state to drive active transport.

Adenosine Triphosphatases↗