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Biomedical subjects

J Broman

Publications and source records attributed to J Broman.

31 records · Page 2Linked to original sources
PubMed1990

Ascending afferents to the lateral cervical nucleus are enriched in glutamate-like immunoreactivity: a combined anterograde transport-immunogold study in the cat.

To investigate whether glutamate (Glu) may be a transmitter in terminals of ascending afferents to the lateral cervical nucleus (LCN), these terminals were identified by anterograde transport of wheatgerm agglutinin-horseradish peroxidase from the spinal cord, and their content of Glu-like immunoreactivity (Glu-LI) was assessed at the ultrastructural level by the immunogold technique. The gold particle density over the peroxidase-positive terminals of the spinocervical tract (SCT) was significantly higher (by a factor of 2.44) than over a reference terminal population containing flattened or pleomorphic vesicles. Further, LCN neurons were densely labeled by the Glu antiserum, although the gold particle density over neuronal cell bodies was not as high as in the SCT terminals. Previous investigations have shown enrichment of Glu-LI in putative glutamatergic terminals in other parts of the CNS. Hence, the present observations indicate that Glu may be a transmitter in the synapses between SCT terminals and LCN neurons. The cell body labeling in the LCN is more difficult to interpret because of possible interference of metabolic pools of glutamate.

Afferent Pathways↗
PubMed1990

Serotonergic innervation of the lateral cervical nucleus: an immunohistochemical study in cats and monkeys (Aotus trivirgatus).

A serotonergic input to the lateral cervical nucleus of cats and monkeys (Aotus trivirgatus) was demonstrated with immunohistochemical methods. In both species, the lateral cervical nucleus was found to contain a network of serotonin-immunoreactive fibers. However, the density of labeled fibers was greater in the monkeys than in the cats. Most labeled fibers were thin and had irregularly spaced varicosities. Electron microscopic examination showed that labeled varicosities were in apposition with dendrites, neuronal somata and unlabeled terminals, but synapses were rare. The results demonstrate that the lateral cervical nucleus receives a serotonergic innervation, as is the case with other somatosensory relay structures such as the spinal dorsal horn and the dorsal column nuclei. The presence of a serotonergic innervation suggests that the transmission of somatosensory information through the lateral cervical nucleus is modulated by a descending pathway. However, its effect on the response properties of neurons in the lateral cervical nucleus is unknown.

Animals↗
PubMed1989

Substance P-like immunoreactivity in the lateral cervical nucleus of the owl monkey (Aotus trivirgatus): a comparison with the cat and rat.

The location of substance P (SP) in the lateral cervical nucleus (LCN) of monkeys (Aotus trivirgatus), cats, and rats was investigated with immunohistochemical methods. Light microscopic analysis showed that SP-positive fibers and terminals are evenly distributed throughout the LCN of the monkey and rat, whereas the SP labeling in the LCN of the cat is concentrated in the medial part of the nucleus, with only very sparse labeling in the lateral part. Electron microscopic examination of the monkey LCN revealed the presence of SP-like immunoreactivity within terminal boutons and unmyelinated axons. The SP-positive boutons are in synaptic contact with dendrites and, occasionally, cell bodies; they contain densely packed, clear, round synaptic vesicles, as well as dense-core vesicles. The distribution of SP-like immunoreactivity in the LCN of monkeys, cats, and rats is similar to that of nociceptive-responsive neurons demonstrated in electrophysiological experiments. The possible role of the SP-containing fibers in the transmission of nociceptive information through the LCN is discussed.

Animals↗
PubMed1989

GABA-immunoreactive neurons and terminals in the lateral cervical nucleus of the cynomolgus monkey.

An antiserum against the inhibitory transmitter substance gamma-aminobutyric acid (GABA) was used to investigate the distribution of GABAergic nerve terminals and cell bodies in the lateral cervical nucleus (LCN) of the cynomolgus monkey. Light microscopic immunohistochemistry demonstrated GABA-immunoreactive puncta, suggestive of nerve terminals, scattered throughout the LCN. The terminal-like profiles are often present along the somata of unlabeled neurons, but most are located in the neuropil. GABA-immunoreactive neurons are present in the LCN, but constitute a very small number of the LCN neurons. Electron microscopy showed that the GABA-positive neurons are small with a relatively large nucleus. They are contacted by few somatic boutons. Numerous GABA-immunoreactive terminals containing densely packed round to oval synaptic vesicles were also found. Most GABA-positive terminals make synaptic contact with dendrites, but synapses with cell bodies are also present. Synaptic contacts between labeled and unlabeled terminals were not observed. Some GABA-positive terminals make contact with GABA-positive neurons. The present findings suggest that GABA is a major inhibitory transmitter substance in the LCN of the monkey. However, in comparison with other somatosensory relay nuclei, there are few GABA-immunoreactive neurons in the LCN. This may imply that the GABA-positive neurons branch extensively in the LCN or that an extrinsic source of GABAergic input exists.

Animals↗
PubMed1989

Subunit structure of the galactose and N-acetyl-D-galactosamine-inhibitable adherence lectin of Entamoeba histolytica.

The galactose and N-acetyl-D-galactosamine-inhibitable adherence lectin of Entamoeba histolytica is a cell surface protein which mediates parasite adherence to human colonic mucus, colonic epithelial cells, and other target cells. The amebic lectin was purified in 100-micrograms quantities from detergent-solubilized trophozoites by monoclonal antibody affinity chromatography. The adherence lectin was purified 500-fold as judged by radioimmunoassay. The nonreduced lectin had a molecular mass of 260 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an isoelectric point of pH 6.2. The amebic lectin reduced with beta-mercaptoethanol consisted of 170- and 35-kDa subunits. Both subunits could be labeled on the cell surface with 125I, and both were metabolically labeled with [3H]glucosamine. The amino termini of the subunits had unique amino acid sequences, and polyclonal antisera to the heavy subunit did not cross-react with the light subunit. The yield of phenylthiohydantoin derivatives from the second and third positions in the sequence of the heavy and light subunits gave a molar ratio of one 170- to one 35-kDa subunit. Antibodies directed to the heavy subunit inhibited amebic adherence to Chinese hamster ovary cells by 100%, suggesting that the heavy subunit is predominantly responsible for mediating amebic adherence.

Acetylgalactosamine↗
PubMed1989

Antigenic stability and immunodominance of the Gal/GalNAc adherence lectin of Entamoeba histolytica.

Immunoprecipitation of Entamoeba histolytica proteins was performed with the sera of patients recovered from amebic liver abscess and colitis. The patients' amebic infection had been acquired in diverse areas of the world. The amebic galactose and N-acetyl-D-galactosamine-inhibitable adherence lectin was the major amebic antigen immunoprecipitated. The adherence lectin was recognized by all of the patients' sera tested regardless of the site (liver abscess vs. colitis) or geographic region that the amebic infection had occurred.

Animals↗
PubMed1988

GABA-immunoreactive neurons and terminals in the lateral cervical nucleus of the cat.

Previous findings have indicated the presence of local circuit neurons in the lateral cervical nucleus (LCN). An immunohistochemical study with gamma-aminobutyric acid (GABA) antiserum was therefore performed both to investigate whether GABA-immunoreactive neurons are present in the LCN, and if so, to compare their characteristics with those previously assigned to probable internuncial neurons in the nucleus. The fine structure and synaptology of GABA-positive boutons in the LCN were also studied. Transversely cut sections from the upper cervical spinal cord of three cats were processed for GABA immunohistochemistry with the free-floating PAP technique. On light microscopic examination immunoreactive neurons were observed within the ventromedial half of the LCN. Their total number was estimated to be 42.5 +/- 11.7 in the entire LCN on one side of the cervical spinal cord, but this may have been an underestimation, as the penetration by the antisera was limited. The labeled neurons were small and had a relatively large nucleus and a low bouton covering ratio. In their number, localization, and ultrastructural appearance the GABA-positive neurons closely resembled the population of neurons previously suggested to be local circuit neurons. Immunoreactive bouton-sized puncta were scattered throughout the LCN. Ultrastructural examination showed labeled terminals with a mean sectional area of 0.85 micron 2 and a relatively high density of synaptic vesicles. The vast majority of GABA-positive terminals were in contact with dendrites and only a minority had synaptic contact with cell bodies. No axoaxonal synapses were observed. The GABA-positive boutons probably derive at least partly from the observed GABA-positive neurons, but there is also a possibility of extrinsic GABAergic input.

Animals↗
PubMed1988

Light and electron microscopic immunohistochemical demonstration of GABA-immunoreactive astrocytes in the brain stem of the rat.

An antiserum against gamma-aminobutyric acid (GABA) was used in an immunohistochemical investigation of the nature of GABA-immunoreactive profiles in various regions of the brain stem of the rat. In accordance with findings in previous biochemical studies, but at discrepancy with recent immunohistochemical results, GABA-like immunoreactivity was demonstrated not only in neurons but also in glial cells. Electron microscopy revealed that the GABA-positive glial cells were astrocytes. Cells identified as oligodendrocytes were unlabelled as were pericytes and endothelial cells. In the labelled astrocytes, immunohistochemical reaction product was seen throughout the nucleus and cytoplasm, and in thin sheet-like processes surrounding neuronal elements and in end-feet lining the basal membrane of capillaries. These observations support the concept that astrocytes play a role in the metabolism of GABA. It is suggested that the failure to demonstrate the presence of GABA in glial cells in previous immunohistochemical studies may have been due to some factor in the tissue preparation. However, it is also possible that labelled glial cells may have been mistaken for small labelled neurons.

Animals↗
PubMed1987

Postnatal development of the feline lateral cervical nucleus: I. A quantitative light and electron microscopic study.

With the aim of obtaining some basic information for future developmental studies, the lateral cervical nucleus (LCN) was investigated in 32 kittens of different ages by electron microscopic and stereologic methods. Corresponding light microscopic measurements of neuronal and nuclear profiles and of the total LCN volume were also performed. The total LCN volume increased sixfold between the ages of 12 hours and 120 days, the most rapid increase occurring during the first month. The neuronal size was fairly constant up to the age of 9 days, whereafter it showed greater variation. The mean profile area increased rapidly during the second week and then more slowly. The relative volume of boutons increased significantly between birth and the age of 34 days and then decreased slightly up to 120 days postnatally. The total bouton volume showed a steady increase, which was most pronounced between the ages of 9 and 34 days. The relative dendritic volume decreased during the 120 days of observation, whereas the total volume of dendrites increased up to the age of 92 days and then decreased. The total volume of glial cells increased during the 120-day observation period, as did both the relative and total volumes of myelinated axons. The changes in the relative volumes of mitochondria in boutons and dendrites were very similar, with increases that were most marked between the ages of 9 and 34 days and between 92 and 120 days.

Animals↗
PubMed1987

Recognition of the galactose- or N-acetylgalactosamine-binding lectin of Entamoeba histolytica by human immune sera.

Cure of amebic liver abscess is associated with resistance to recurrent invasive amebiasis and the development of a humoral and cell-mediated immune response. We determined whether human immune sera contain blocking antibody for the 170-kilodalton (kDa) galactose or N-acetylgalactosamine (Gal/GalNAc)-binding lectin of Entamoeba histolytica. By Western blot (immunoblot) of whole amebae subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, all eight immune sera studied here prominently recognized a 170-kDa amebic protein. Western blot of the purified Gal/GalNAc lectin with pooled human immune sera (PHIS) confirmed that the 170-kDa band was the adherence lectin. Immunoprecipitation of [35S]methionine-metabolically-labeled amebae with the antilectin monoclonal antibody H8-5 and with PHIS demonstrated that the 170-kDa lectin was the major antigen recognized by PHIS. The in vitro adherence of E. histolytica trophozoites to CHO cells at 4 degrees C was inhibited by prior exposure of amebae to greater than or equal to 1.0% PHIS. The humoral response to the Gal/GalNAc-binding lectin of the parasite may contribute to the development of protective immunity against invasive amebiasis.

Adhesiveness↗