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Biomedical subjects

J S King

Publications and source records attributed to J S King.

At least 19 recordsLinked to original sources

Ontogenesis of cerebellar afferents identified by cholecystokinin-like immunoreactivity.

The present account provides a developmental timetable for the maturation of cholecystokinin (CCK)-positive fibers in the cerebellar cortex and cerebellar nuclei of the opossum. CCK-positive fibers are in the cerebellar peduncle by postnatal day (PD) 1, however they wait until PD 7 to penetrate the cerebellar anlage. Between PD 7 and PD 20 the fibers wait again in the medullary core of the cerebellum. After PD 20, there are 2 distinct patterns of CCK localization within the overlying cortical layers. The first pattern develops between PD 20-26 when CCK puncta are present in restricted foci within the Purkinje cell layer of the anterior lobe vermis. They distribute in 4 parasagittal bands, 2 on either side of the midline, that extend from the primary fissure rostrally into the anterior lobe of the cerebellum. By PD 33 two additional parasagittal bands are present in the posterior lobe vermis. The vast majority of these CCK puncta are transient in nature as all but a few disappear by PD 84. Those that remain progress through a series of developmental stages characteristic of climbing fiber ontogeny. These climbing fibers persist in lobules V, VII and VIII of the adult cerebellum. Further, there is a transient expression of CCK-immunoreactivity within inferior olivary neurons. These observations support the interpretation that the transient population of CCK-IR puncta are immature climbing fiber axons derived from the inferior olive. The second pattern of CCK localization is evident between PD 30-33, the time when granule cells first can be recognized in a histologically distinct internal granule cell layer (IGL). Between PD 30 and PD 68 there is a differential pattern of distribution of CCK-IR profiles within the lobules of the cerebellum. Initially, CCK-IR axons are only present in the anterior vermis where they are aligned in register with the bands of CCK puncta in the Purkinje cell layer. CCK-IR puncta are not present in the posterior lobe vermis or hemispheres until later stages of development. Further, a sagittal organization is not evident in either of these latter 2 areas. Initially, CCK-IR profiles in the IGL cannot be identified as mossy fibers based on their terminal morphology. When they first enter the IGL they appear as punctate elements. Over time they become increasingly more complex in shape and between PD 68-84 develop morphological characteristics of adult mossy fiber rosettes. The cerebellar nuclei can be distinguished histologically by PD 18, but CCK-IR fibers are not evident among these neurons until PD 36 which corresponds to about the time they can be visualized in the IGL. In addition, CCK-IR cell bodies first appear in the cerebellar nuclei between PD 26-30; these are present in the adult.(ABSTRACT TRUNCATED AT 400 WORDS)

Aging

Differential modulation of Purkinje cell activity by enkephalin and corticotropin releasing factor.

Several peptides have been localized within afferents to the opossum's cerebellum, including cholecystokinin (15), enkephalin (16, 17) and corticotropin releasing factor (7, 9). Each of these peptides has a heterogeneous laminar and lobular distribution. Two peptide, enkephalin (ENK) and corticotropin releasing factor (CRF) are present in specific populations of climbing fibers and mossy fibers, which have an overlapping distribution in several areas of the cerebellum, in particular the lateral aspect of vermal lobules VII and VIII. Within this location ENK and CRF are co-localized in individual climbing fibers and mossy fibers (7). In the present study, the physiological effects of these peptides on Purkinje cell activity were analyzed. The data indicate that ENK and CRF have opposite effects on Purkinje cell activity. ENK suppresses spontaneous activity as well as that induced by application of glutamate and aspartate, as described previously (5). In contrast, CRF enhances both spontaneous and amino acid-induced unit activity. Further, when applied simultaneously, CRF blocks the suppressive effect induced by ENK. Previous studies have shown that climbing fibers, as well as the mossy fiber-parallel fiber pathway, are excitatory to Purkinje cells (11). However, immunohistochemical data have shown that these afferents are heterogeneous with respect to their chemical content (7-9, 15-17, 25). As found in the current and previous studies (3, 5) peptides in climbing and mossy fibers modulate the responsiveness of Purkinje cells to primary excitatory neurotransmitters in selected areas of the cerebellar cortex. However, the firing rate of individual Purkinje cells is differentially altered depending on which neurochemical messenger(s) are released.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials

A rapid, accurate, immunohistochemical method to label proliferating cells in the digestive tract. A comparison with tritiated thymidine.

A rapid immunohistochemical method using 5'-bromo-2'-deoxyuridine (BrdU), a thymidine analog, for labeling proliferating epithelial cells was modified and tested for accuracy against standard tritiated thymidine autoradiography (3H-TdR) in rat esophagus, stomach, duodenum, and colon. Either BrdU or 3H-thymidine or both compounds (simultaneously) were injected IP. Histological sections of these tissues were immunostained with monoclonal anti-BrdU antibody, linked to horseradish peroxidase by standard avidin-biotin techniques and stained with diaminobenzidine, or sections were dipped for autoradiography, or both techniques were applied to the same tissue section. Results showed that (a) BrdU labeled the same number of proliferating cells in all organs as 3H-TdR; (b) BrdU colabeled with 3H-thymidine; (c) immunostaining was complete in 3-4 days but standard 3H-TdR took 2 weeks; and (d) qualitative analysis took 50% less time with BrdU than with standard 3H-TdR.

Animals

A mathematical model of interference for use in constructing linkage maps from tetrad data.

In determining genetic map distances it is necessary to infer crossover frequencies from the ratios of recombinant and parental progeny. To do this accurately, in intervals where multiple crossovers may occur, a mathematical model of chiasma interference must be assumed when mapping in organisms displaying such interference. In Saccharomyces cerevisiae the model most frequently used is that of R.W. Barratt. An alternative to this model is presented. This new model is implemented using a microcomputer and standard numerical methods. It is demonstrated to fit ranked tetrad data from Saccharomyces more closely than the Barratt model and thus generates more accurate estimates of map distances when used with two-point data. A computer program implementing the model has been developed for use in calculating map distances from tetrad data in Saccharomyces.

Chromosome Mapping

Distribution and brainstem origin of cholecystokinin-like immunoreactivity in the opossum cerebellum.

In order to determine the distribution of the peptide cholecystokinin (CCK) within the cerebellum and medullary precerebellar nuclei of the adult opossum, sections of these brain regions were processed for peroxidase-antiperoxidase immunohistochemistry. Within the inferior and superior cerebellar peduncles, fine-beaded fibers are evident and a beaded plexus of fibers is present in all the cerebellar nuclei. In the overlying cerebellar cortex, CCK-positive mossy fiber rosettes are present in all lobules, where their morphology varies from simple enlargements to more complex rosettes. However, their distribution varies particularly in vermal lobules II, III, VII, and IX where they are organized in parasagittal bands. Climbing fibers that are positive for CCK are present in very restricted areas of vermal lobules IV, VII, and VIII. After colchicine pretreatment, CCK-positive cell bodies are seen in restricted regions of the posterior interposed and fastigial nuclei as well as within several precerebellar nuclei known to give rise to mossy fibers. Such nuclei include the lateral cuneate nucleus, the nucleus prepositis hypoglossi, the nucleus reticularis lateralis, the nucleus raphe obscurus, the paramedian reticular nucleus, the nucleus reticularis gigantocellularis, and the medial vestibular nucleus. To localize the brainstem origin(s) of the CCK fibers in the cerebellum, a double-label paradigm employing a retrograde tracer and CCK immunohistochemistry was used. These experiments indicate that CCK mossy fibers originate primarily within the lateral cuneate nucleus, the perihypoglossal complex, and the lateral reticular nucleus. Some also originate within the medial vestibular nucleus and the nucleus reticularis gigantocellularis. In addition, double-labeled cell bodies are present within the caudal medial accessory inferior olive, the likely source of the CCK-positive climbing fibers. These data indicate that specific populations of climbing fibers and mossy fibers may utilize CCK to alter the firing rate of their target neurons.

Animals

Random-breakage mapping, a rapid method for physically locating an internal sequence with respect to the ends of a DNA molecule.

We describe a method for determining the position of a cloned internal sequence with respect to the ends of a DNA molecule. The molecules are randomly broken at low frequency and the fragments are subjected to electrophoresis. Southern hybridization using the cloned DNA as a probe identifies only those fragments containing the sequence. The size distribution of these fragments is such that two threshold changes in intensity of signal are seen in the smear pattern below the unbroken molecules. The positions of the changes represent the distances from the sequence to each molecular end. The intensity changes arise because the natural ends of the molecules influence the fragment distribution obtained. From once-broken molecules, no fragments can arise that contain a given sequence and are shorter than the distance between that sequence and the nearest molecular end. We tested the method by using x-rays to induce breakage in yeast DNA. Genes of independently known position were mapped within whole chromosomes or Not I restriction fragments using Southern blots from gels of irradiated molecules. We present equations to predict fragment distribution as a function of break-frequency and position of the probed sequence.

Base Sequence

Coexistence of corticotropin releasing factor and enkephalin in cerebellar afferent systems.

In this study, we tested the hypothesis that corticotropin-releasing factor (CRF) and enkephalin (ENK) coexist within restricted brainstem-cerebellar circuits. Antisera for CRF and ENK were applied simultaneously or to serial sections of adult opossum brain and were processed for fluorescence or peroxidase, antiperoxidase immunohistochemistry, respectively. CRF and ENK were colocalized in 1) climbing fibers within the flocculus and in bilateral foci in the cerebellar vermis; 2) fibers with morphological characteristics of simple mossy fiber terminals or climbing fiber glomeruli that were located within the granule cell layer of the vermis underlying the foci of CRF/ENK-IR climbing fibers; 3) mossy fiber terminals within the flocculus; 4) neuronal perikarya in subnucleus A and C of the medial accessory olive and in the dorsal cap of Kooy, nuclei known to project as climbing fibers; and 5) cell bodies in nucleus prepositus, the subtrigeminal reticular nucleus, and the reticular formation, likely origins of CRF/ENK colocalized mossy fibers. The demonstration that single climbing and mossy fibers contain two peptides extends previous studies that have described chemical heterogeneity within cerebellar afferent pathways. Furthermore, these results support suggestions that this heterogeneity may provide a substrate for differential regulation of signal transduction by chemically coded cerebellar afferents.

Animals

A polymerization model of chiasma interference and corresponding computer simulation.

A model of chiasma interference is proposed and simulated on a computer. The model uses random events and a polymerization reaction to regulate meiotic recombination between and along chromosomes. A computer simulation of the model generates distributions of crossovers per chromosome arm, position of events along the chromosome arm, distance between crossovers in two-event tetrads, and coincidence as a function of distance. Outputs from the simulation are compared to data from Saccharomyces cerevisiae and the X chromosome of Drosophila melanogaster. The simulation demonstrates that the proposed model can produce the regulation of recombination observed in both genetic and cytological experiments. While the model was quantitatively compared to data from only Drosophila and Saccharomyces, the regulation observed in these species is qualitatively similar to the regulation of recombination observed in other organisms.

Animals

Effect of vacuum fluctuation during milking on the development of intramammary infection from teat duct colonization by Staphylococcus aureus.

Twelve of twenty-one udder quarters with colonized teat ducts became infected within 13 d when milked with a machine producing large cyclic and irregular vacuum fluctuations in the teatcup; of 23 quarters milked under the same conditions, but with metal shields fitted inside the liners to protect the teat apex (Thiel, 1974), only 2 quarters became infected in the same period. The introduction of a post-milking teat disinfectant teat dip resulted in the elimination of orifice colonization from 15 teats dipped in Na hypochlorite solution (40 g/l available chlorine) and from all but 2 of 15 teats dipped in an iodophor solution (5 g/l available iodine).

Animals

The synaptic terminations of certain midbrain-olivary fibers in the opossum.

The nuclear origin and distribution of midbrain-olivary fibers has been described in a previous study utilizing axonal transport techniques (Linauts and Martin, '78a). The present report extends their results to the electron microscopic level and details the postsynaptic distribution of such fibers. Lesions within the ventral periaqueductal grey and adjacent tegmentum, the red nucleus or the nucleus subparafascicularis result in electron dense axon terminals within the olive at survival times of 48, 72 and 96 hours. At 72 hours, many degenerating presynaptic profiles shrink, become irregular in shape and are totally or partially surrounded by glial processes. The principal olivary nucleus contains the majority of these profiles. However, the subparafascicular terminals are more abundant in the rostral and intermediate parts of the medial accessory nucleus and the rubral terminals are concentrated within the dorsal lamella of the principal nucleus. The nuclear location of the degenerating terminals was determined by examination of 1 micrometer plastic sections cut in the transverse plane from each block face prior to thin sectioning. Degenerating terminals were counted in three cases, one from each of the three lesion sites described above. When taken together these cases show that just over 50% of the degenerating terminals are presynaptic to spiny appendages and are located within the synaptic clusters (glomeruli) described previously (King, '76). The percentage of degenerating terminals in the glomeruli increases to 70% when the lesion is in the ventral periaqueductal grey and adjacent tegmentum. The remaining degenerating terminals contact dendritic shafts outside the astrocytic boundaries of the synaptic clusters. The synpatic vesicle populations within the degenerating terminals vary with the location of the lesion. Lesions in the ventral periaqueductal grey and the adjacent tegmentum result in the degeneration of terminals with either clear spherical vesicles or endings with both clear spherical vesicles and a variable number of large dense core vesicles. In contrast, the primary degenerative changes that occur after destruction of the red nucleus or the nucleus subparafascicularis are in terminals with clear spherical vesicles. When the synaptic complex was present in the plane of section, regardless of the site of the lesion, the degenerating terminals could be classified as Gray's type I. Thus, we have demonstrated that afferents from the mesencephalon terminate within synpatic clusters located in the principal and medial accessory (part A) subnuclei of the inferior olive. Although the mesencephalic afferents have multiple origins (Linauts and Martin, '78a), many of their synaptic terminals contact spiny appendages within the synaptic clusters. This postsynaptic site also receives cerebellar terminals (King et al., '76). The origin of presynaptic profiles within the synaptic clusters that contain clear pleomorphlic vesicles is yet to be determined.

Animals

The bactericidal activity of a teat dip containing chlorhexidine and cetrimide.

Using an in vivo test on teat skin the disinfectant activity of a teat dip containing chlorhexidine and cetrimide was compared with two iodophor solutions, one containing the recommended concentration of 0.5 per cent available iodine and the other a 10-fold dilution of this (0.05 per cent iodine). The test organisms used were Staphylococcus aureus and Escherichia coli and for both species the 0.5 per cent iodophor was significantly more bactericidal than either the diluted iodophor or the chlorhexidine/cetrimide teat dip (P less than 0.01). In the test against S aureus, chlorhexidine/cetrimide and the 0.05 per cent iodophor showed similar bactericidal activity, but the iodophor was significantly more bactericidal against E coli (P less than 0.01). It is argued that due to its low bactericidal activity this formulation of chlorhexidine/cetrimide is likely to be inferior to 0.5 per cent iodophor solution as a disinfectant teat dip.

Animals

Disinfection properties of some bovine teat dips.

The efficacy of 18 disinfectant teat dips was tested on teats artificially contaminated with a milk suspension of Staphylococcus aureus. A solution of Na hypochlorite with 40 g/l available chlorine was significantly more bactericidal than one containing 1 g/1 available chlorine and than most other disinfectants tested. The method was not able to distinguish differences in efficacy between solution containing 40g/1 and 10g/1 available chlorine nor between these and some of the iodophors containing 5 g/1 available iodine. The additon of 190-416 g/1 (15-33% v/v) glycerol significantly reduced the bactericidal properties of 3 iodophors (5 g/1 available iodine), but soluble lanolin at approximately 20 g/1 did not appear to lower the efficiency of NaOC1 (45 g/1 available chlorine) or of an iodophor (5 g/1 available iodine).

Animals