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Biomedical subjects

E M Johnson

Publications and source records attributed to E M Johnson.

At least 325 records · Page 18Linked to original sources
PubMed1983

Retrograde axonal transport of intravenously administered 125I-nerve growth factor in rats with streptozotocin-induced diabetes.

We have previously demonstrated a reproducible axonopathy, involving but not confined to sympathetic neurons, which involves selectively the mesenteric nerves of chronically diabetic rats. We describe a series of experiments aimed at correlating changes in the retrograde transport of 125I-nerve growth factor (125I-NGF) with development of the lesions. 125I-NGF was injected systemically and the time course of accumulation compared in superior cervical ganglion (SCG) and superior mesenteric ganglion (SMG). 125I-NGF accumulated in the SCG with a sharp peak at 12 h, whereas in the SMG, accumulation reached a plateau, remaining at relatively constant amounts for 8 h before decreasing. There was a marked decrease in the accumulation of 125I-NGF (averaging 44% in 11 separate experiments) in the SMG at early times (approximately 12 h). These decreases were seen in animals diabetic for times ranging from 2 days to 10 mo. In contrast, no consistent decrease was observed in the SCG. The time course of 125I-NGF accumulation was unaltered in the SCG of diabetic rats but was clearly retarded in the SMG of diabetic animals. Morphometric examination of the SMG of diabetic animals and controls showed little or no atrophy or neuron loss due to diabetes. We conclude that decreases in the retrograde transport of NGF occur selectively in those ganglia (SMG) serving the alimentary tract in which mesenteric nerves develop distal axonopathy, but not in ganglia whose nerves do not ultimately develop such lesions (SCG). The decreases in retrograde transport precede the development of morphologic lesions in this system. These results are consistent with the suggestion that impairment of axonal transport may play a role in the development of axonopathy.

Animals↗
PubMed1983

Retrograde transport of nerve growth factor in lesioned goldfish retinal ganglion cells.

Previous experiments have shown that nerve growth factor (NGF) enhances regeneration of goldfish optic nerve after local application of NGF at the site of the lesion. However, the site and mechanism of action of NGF are not yet known. One possibility is that NGF is taken up at the site of the lesion and retrogradely transported to the cell bodies of the retinal ganglion cells and thereby exerts its trophic effects. The present work was carried out to assess the role of retrograde transport of NGF in this enhanced regeneration of goldfish retinal ganglion cells. In intact retinal ganglion cells of the goldfish, 125I-labeled NGF was found not to be retrogradely transported from the optic tectum to the retina, suggesting that retinal ganglion cells do not possess specific NGF receptors. However, if [125I]NGF was injected at the site of an optic nerve lesion at the time of lesion, [125I]NGF was retrogradely transported from the site of a lesion of the optic nerve to the cell body of retinal ganglion cells. The accumulated radioactivity was shown to be intact NGF by SDS-PAGE. The ability of NGF to decrease the time required for recovery of visual function was observed only when NGF was administered at the time of the injury. Likewise, no transport of [125I]NGF was observed when it was injected at the crush site 16 hr or longer after crush. Thus, there is a temporal correlation between the ability of intact [125I]NGF to be retrogradely transported from a lesion site to the retina and the regenerative effect of NGF. Autoradiography showed that the [125I]NGF accumulated only in retinal ganglion cells. The transport of NGF in the lesioned goldfish visual system was not specific for NGF in that other proteins (cytochrome c, bovine serum albumin) were transported equally well. Likewise, transport of [125I]NGF was not prevented by concomitant administration of excess unlabeled NGF. The retrograde transport of [125I]NGF therefore was not selective and did not appear to be mediated by specific NGF receptors in this system. This nonspecific transport of [125I]NGF did not occur in the axotomized spinal motor neurons in the neonatal or adult rat or in the newt. However, receptor-mediated transport is seen in lesioned sensory neurons in both species.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1982

Effect of exposure to anti-NGF on sensory neurons of adult rats and guinea pigs.

Nerve growth factor (NGF) deprivation was produced in adult rats and guinea pigs by immunization against mouse NGF. Exposure of adult animals to anti-NGF had no effect on sensory ganglion neuronal number or size-frequency histograms. However, the substance P content of sensory ganglia, spinal cord and hind paw skin decreased to as great an extent as was seen in animals exposed to anti-NGF in utero. We conclude that NGF has two separate effects on sensory neurons: adult sensory neurons require NGF for normal function whereas developing neurons require NGF for both survival and transmitter expression.

Animals↗
PubMed1982

Immunosuppressive agents prevent guanethidine-induced destruction of rat sympathetic neurons.

Chronic administration of guanethidine to rats causes destruction of peripheral sympathetic neurons. Neuronal destruction, characterized morphologically by small cell infiltration and the reduction in the number of neurons within sympathetic ganglia, and biochemically by a marked reduction in tyrosine hydroxylase activity, occurred reproducibly by day 7 of treatment following 5 daily injections of 50 mg/kg guanethidine sulfate. Several observations in the literature suggested that guanethidine-induced destruction may occur by an immunologically mediated mechanism. Experiments were therefore designed to test the effects of immunosuppressive agents on guanethidine sympathectomy. A single exposure to either gamma-irradiation or cyclophosphamide, administered 8 h prior to the initiation of guanethidine treatment, protected against guanethidine-induced destruction in a dose-related manner and was virtually complete with either 900 rads of irradiation or with 100 or 150 mg/kg of cyclophosphamide. Cyclophosphamide afforded complete protection only if administered immediately prior to guanethidine treatment suggesting that it was acting during the proliferative phase of an immune response rather than non-specifically. Pretreatment with either irradiation or cyclophosphamide had no effect on the sympathectomy produced by treatment with either 6-hydroxydopamine or antibodies to nerve growth factor, nor did it prevent the accumulation of guanethidine within the sympathetic ganglia. Concurrent treatment with either azathioprine or dexamethazone also provided partial protection against guanethidine sympathectomy. These results strongly suggest that the destruction of sympathetic neurons induced by guanethidine occurs by an immunologically mediated mechanism.

Animals↗
PubMed1982

Effects of autoimmune NGF deprivation in the adult rabbit and offspring.

An experimental autoimmune approach to the production of nerve growth factor deprivation, which we have previously described in the rat and guinea pig, has been applied to the rabbit. This species was chosen for study because of several potential advantages. The rabbit produces large litters and has a relatively short gestation period. More importantly, rabbits generate high titers of antibody against mouse NGF and large amounts of maternal antibody are passively transferred to the developing rabbit fetus compared to most other species, particularly the rat. The sympathetic nervous system of adult rabbit immunized against mouse NGF underwent degeneration with up to an 85% decrease in neuronal numbers in the superior cervical ganglion after 10 months of immunization, thus providing further evidence that NGF is required for the survival of mature sympathetic neurons. Despite the fact that newborn rabbits born to anti-NGF producing mothers had much higher titers of anti-NGF than did rats, the effects on the developing sympathetic and sensory nervous systems were not found to be any greater than in rats. Reductions in norepinephrine levels in the heart and spleen of adult rabbits born to anti-NGF producing mothers were greater than in small intestine. Prenatal exposure to maternal anti-NGF caused reductions (up to 70%) in the number of neurons in the dorsal root ganglia. Substance-P immunoreactivity was reduced in the substantia gelatinosa of the spinal cord of rabbit exposed to maternal anti-NGF. These changes, however, were not greater than seen in the rat. We conclude that although the rabbits offers some advantage in the study of the effects of NGF deprivation in the adult animal, it appears less well suited than the rat or guinea pig to the study of the effects of NGF deprivation on development.

Animals↗
PubMed1982

Time and dose dependencies of effects of nerve growth factor on sympathetic and sensory neurons in neonatal rats.

Nerve growth factor (NGF) plays a role in the development of several components of the sympathetic and sensory nervous systems. The objectives of this study were to examine the time and dose dependencies of some of the well known effects of NGF on sympathetic ganglia and to examine qualitatively and quantitatively the recently described effects on sensory ganglia of neonatal rats. Single doses of NGF as low as 0.1 mg/kg produce increases in tyrosine hydroxylase (TOH) activity in superior cervical ganglia (SCG), and doses of 3 mg/kg produce maximal effects. Larger doses and longer treatments are required to see increases in protein content of the SCG. Larger doses are also required to affect TOH activity in the adrenal gland. Increases in TOH activity in SCG can be observed within 18 h of injection. Chronic NGF treatment for three weeks produces no change in blood pressure or heart rate in neonatal rats. chronic administration of NGF (1 or 3 mg/kg/day) results in dose-related increases in the protein content of dorsal root ganglia (DRG). The increase in protein content of the DRG was associated with an increase in the diameter of smaller neurons (those less than 30 micron in diameter), but NGF caused no change in the number of neurons.

Adrenal Glands↗
PubMed1982

The Hydra attenuata system for detection of teratogenic hazards.

By a uniformly applied protocol, adult hydra are exposed to a test substance over a broad range of concentrations, and the minimal toxic concentration is determined to within one-tenth log. In a second experiment, dissociated hydra cells are manipulated into a configuration wherein, if undisturbed, they will achieve the developmental events characteristic of any embryo and undergo total whole-body regeneration. During its 4-day ontogenesis this artificial "embryo" is exposed to the test substance by the same protocol as the adult and the minimal developmentally toxic concentration is determined to within one-tenth log. The ratio of the adult (A) to the developmentally (D) toxic concentration is calculated. A small A/D ratio indicates that the substance disrupts development only at or near the concentration also toxic to the adult (a developmentally nonhazardous substance or coeffective teratogen). A large A/D ratio indicates that a substance disrupts developmental events at a small fraction of the exposure toxic to adults (a developmental hazard). The system is directly predictable of a putative teratogen's hazard potential (A/D ratio) in standard laboratory animals and man. It provides an objective and reliable means to prioritize otherwise untested substances according to the need for further study of their developmental toxicity.

Animals↗
PubMed1982

Intralitter variation of neonatal rat lung morphology following prenatal maternal hypervitaminosis A.

Prenatal exposure to excess vitamin A (160,000 USP units/day) from days 15 through 19 of gestation results in altered lung morphology, characterized by thickened septal walls and/or large areas of atelectasis and an associated high neonatal mortality. Marked variability in both morphological and physiological expression from this prenatal insult is commonly seen between litters and littermates, making analyses difficult. The present study documents morphological intralitter variation observed in vitamin A-exposed 2-day-old rats as compared to controls. Representative midhilar coronal histological sections of each lung were examined by two methods and the results compared. The first method, although less sensitive, demonstrated that five out of eight experimental rat lungs had a significantly greater percentage of tissue to airway space as compared with undisturbed controls (greater than 43%). The second method, using several morphological criteria and a grid system to score parenchyma into classifications based on the degree of morphological variation and projected functional capability, clearly found significantly increased percentages of poor or nonfunctional lung tissue (P greater than 0.01) in seven out of eight pups exposed to excess vitamin A. This method of ranking the severity of adverse effects on tissue morphology allowed identification of drug-affected newborn and provided a means to quantify the alterations. Such a means for identifying affected individuals from littermates is essential to research methodologies for detecting substances which, when administered during fetal life, produce decrements of postnatal function but no gross structural abnormalities.

Animals↗
PubMed1982

Genetic and molecular studies of the regulation of atypical citrate utilization and variable Vi antigen expression in enteric bacteria.

1. The atypical citrate-utilizing ability to two strains of E. coli has been shown to be plasmid-encoded. Strain V414 carries a 130 Mdal conjugative Cit+ plasmid that also specifies Tcr and Cmr. Strain V517 carries 9 different plasmid species but only the 36 Mdal species is correlated with Cit+ ability. These plasmids are different from previously reported Cit+ plasmids of E. coli and Salmonella, which express thermosensitive conjugal transfer systems. 2. A 9 kb Pstl fragment, carrying the Cit+ genes of pWR60, has been cloned into the pBR325 plasmid. 3. Metabolic studies indicate that intact citrate is not incorporated directly into whole cells. Rather, atypical citrate utilization by these E. coli strains appears to involve partial metabolism of citrate at the cell surface before or during uptake. 4. The expression of atypical Cit+ ability by the parental pWR60 plasmid or by the recombinant pWR61 plasmid appears reversible and may involve an expression switch mechanism (i.e., insertion sequence element). 5. Two widely separated genetic loci, viaA and viaB, are necessary for Vi antigen synthesis in Salmonella and Citrobacter. In some strains of C. freundii, Vi antigen expression is reversible, a phenomenon which can be visualized by a colonial morphology transition between Vi-expressing and -nonexpressing forms. 6. The C. freundii viaB locus appears to encode the Vi antigen as well as the genetic "switch" mechanism controlling reversible Vi antigen expression. The viaA locus, which is found in several different bacterial species, may encode some common property (e.g., cell surface structure or enzymatic activity) that is needed for Vi antigen expression. 7. S. typhi and E. coli K12 hybrid strains which carry the C. freundii viaB locus have been constructed. These hybrid strains express reversible Vi antigen expression, even in the absence of general recombination (i.e., functional recA gene product). 8. The C. freundii viaB locus was transposed via Mu-mediated events to an F'lac plasmid in the E. coli K12 hybrid strain WR2376. F' plasmids carrying the viaB locus should serve as a highly enriched source of viaB DNA for physical examination of the switch mechanism. 9. Genetic manipulations such as those described herein can be used to study virtually any plasmid, viral, or chromosomally-encoded property. The resultant better understanding of biochemical pathways and of genetic regulatory control systems, and the isolation of desired gene sequences should provide ample information and materials for improving chemical processes and constructing vaccines against various organisms.

Antigens, Bacterial↗
PubMed1982

Evaluation of two Salmonella typhimurium hybrids as challenge organisms in a system for the assay of typhoid vaccines.

A mouse-virulent Salmonella typhimurium hybrid (H42), which expresses the Salmonella typhi Vi antigen in addition to S. typhi O antigens 9 and 12, and a mouse-virulent S. typhimurium hybrid (H1), which expresses only the 9 and 12 antigens of S. typhi, were compared in their behavior as challenge organisms in a system developed to assay the protective capabilities of typhoid vaccines. Swiss-Webster white mice, vaccinated intraperitoneally with live Escherichia coli hybrids expressing the S. typhi O antigens 9 and 12, were significantly protected against death from intraperitoneal challenge with each of the S. typhimurium hybrid strains. Vaccination with an E. coli hybrid expressing the S. typhi Vi antigen in addition to O antigens 9 and 12 was seen to confer no advantage in protection against either S. typhimurium hybrid challenge organism over that obtained by vaccination with an E. coli hybrid expressing only the O antigens of S. typhi. However, a notable difference in the behavior of the two S. typhimurium hybrids was seen in mice vaccinated with the parent of the E. coli hybrid vaccinating strains, E. coli F464, which expresses no surface antigens common to either of these S. typhimurium hybrid challenge organisms. A nonspecific (with respect to the vaccinating strain) protective effect, believed to be associated with Vi antigen expression by the challenge organism, was seen against the challenge with S. typhimurium hybrid H42 after F464 vaccination, whereas no protection was conferred by F464 vaccination against the challenge with Vi-nonexpressing S. typhimurium hybrid H1. Inasmuch as neither S. typhimurium hybrid discriminates between the expression or nonexpression of the Vi antigen in a vaccinating strain, it is concluded that the Vi-nonexpressing S. typhimurium hybrid H1, which more clearly indicates the vaccine-specific protective role of the S. typhi O antigens and does not exhibit the nonspecific protection response of hybrid H42, is the better choice as challenge organism for this vaccine assay system.

Animals↗
PubMed1981

Introduction of late gestational teratogenesis in rat lung by hypervitaminosis A.

Long-Evans black-hooded rats treated via stomach tube with 160,000 USP units of retinyl acetate (vitamin A) in 0.5 ml Mazola corn oil on gestational days 15--19 deliver normal-sized litters with significantly decreased viability. Vitamin A is known to effect the differentiation and to stimulate the growth of epithelial cells. Additionally, lung epithelia undergo marked morphologic and physiologic changes late in gestation. Thus the effects of hypervitaminosis A on developing lung constitute an excellent system for the study of teratogenesis late in gestation. Non-hilar, right lower-lobe sections of lungs from the vehicle control and experimental groups, compared via quantitative light microscopy, revealed no significant difference in gross overall histologic appearance on any given day, either in the total number of airways present in the volume of lung sectioned or in the percent area of any individual airway occupied by cells or by lumen. The only significant difference was in the number of cells per square micrometer in that region of an airway occupied by cells. Additionally, there was a significant difference between the control and experimental mitotic indices on gestational days 18 and 19. Thus in the experimental group the number of cells lining the developing airways increases, while the absolute thickness of this cellular layer remains constant. Transmission electron microscopy (TEM) fails to reveal any morphologic differences between control and treated type II pneumatocytes. The increased number of respiratory cells in what otherwise appears to be normal lung may create a diffusion-perfusion imbalance or other difficulties contributing to the heightened neonatal mortality resulting from teratogen exposure late in gestation.

Animals↗
PubMed1981

Protein tightly bound near the termini of the Physarum extrachromosomal rDNA palindrome.

The genes coding for ribosomal RNa in plasmodia of Physarum polycephalum are arranged palindromically on extrachromosomal rDNA molecules of 61 kb (kilobasepairs). Incubation of mildly extracted rDNA with the 125I Bolton-Hunter reagent results in incorporation of label not removed by SDS, CsCl, or various organic solvents. Labeled protein is preferentially associated with terminal rDNA restriction fragments, as detected after gel electrophoresis of the DNA. Antibody reaction with dinitrophenylated protein-rDNA complexes allows visualization of protein located from 1 to 2 kb from the termini, in a region containing multiple inverted repeat sequences and single-strand gaps. DNase I treatment of either rDNA or rDNA termini releases primarily two labeled protein bands of 5,000 and 13,000 daltons as well as less prominent bands of higher molecular weight. We discuss mechanisms for involvement of terminal protein in replication of 3' ends and chromosomal integration of the rDNA.

Base Sequence↗
PubMed1981

Transported enzymes in sciatic nerve and sensory ganglia of rats exposed to maternal antibodies against nerve growth factor.

The accumulations by axoplasmic transport of selected enzyme activities proximal and distal to a ligature placed on the sciatic nerve were monitored in rats exposed in utero to maternal antibodies to nerve growth factor (NGF) and in control rats. Littermates of the animals exposed to anti-NGF were shown elsewhere to have had a 70% reduction in the number of sensory neurons in dorsal root ganglia and a 90% reduction in number of neurons in superior cervical (sympathetic) ganglion. The accumulation of F(-)-sensitive acid phosphatase activity was depressed 75% both proximal and distal to the tie. Accumulation of F(-)-resistant acid phosphatase activity was depressed nearly 50% proximal to the tie. Distal accumulation of this activity did not occur in either group of rats. Accumulation of acetylcholinesterase activity was depressed 30%. Distal accumulation of the activities of beta-glucuronidase and hexokinase was depressed 50%. In the lumbar dorsal root ganglia, dry weight was reduced 40%, and the activities of peroxide-sensitive, F(-)-resistant acid phosphatase and of the mitochondrial enzymes hexokinase, glutamic dehydrogenase, glutamic-oxalacetic transaminase, and NAD-dependent isocitric dehydrogenase were all reduced a little more, 45--50% per ganglion. However, the activities of the lysosomal enzymes, F(-)-sensitive acid phosphatase and beta-glucuronidase, of the peroxide-resistant, F(-)-resistant acid phosphatase, and of the mitochondrial enzyme glutaminase were all reduced about 60% per ganglion. The results of these measurements were interpreted to suggest that much, and perhaps all, of the F(-)-sensitive acid phosphatase activity in motion in peripheral nerve in rat is confined to sensory axons.

Acid Phosphatase↗