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1.
Prenat Diagn ; 22(1): 1-4, 2002 Jan.
Article in English | MEDLINE | ID: mdl-11810640

ABSTRACT

The aim of this study was to evaluate the detection of fetal structural abnormalities by the 11-14 week scan. 2853 pregnant women were submitted to a routine ultrasound scan between the 11th and 14th week and the fetal skull, brain, spine, abdominal wall, limbs, stomach and bladder were examined. Following the scans the patients were examined in the second or third trimester of pregnancy. An isolated increased nuchal translucency was not considered an abnormality. However, these patients had an early echocardiography assessment. Fetal structural abnormalities were classified as major or minor and of early or late onset. A total of 130 (4.6%) defects were identified and 29 (22.3%) of these were diagnosed at the 11-14 week scan, including nine cardiac defects associated with increased nuchal translucency. The antenatal ultrasound detection rate was 71.5%, and 31.2% were detected in the first-trimester assessment. 78.8% of the major defects were diagnosed by the prenatal scan and 37.8% by the 11-14 week scan. Fetal structural abnormalities at the 11-14 week scan were detected in approximately 22.3% of the cases, therefore, a second-trimester anomaly scan is important in routine antenatal care to increase the prenatal detection of fetal defects.


Subject(s)
Congenital Abnormalities/diagnostic imaging , Gestational Age , Ultrasonography, Prenatal , Adult , Central Nervous System/abnormalities , Chromosome Aberrations , Craniofacial Abnormalities/diagnostic imaging , Digestive System Abnormalities , False Negative Reactions , Female , Heart Defects, Congenital/diagnostic imaging , Humans , Limb Deformities, Congenital/diagnostic imaging , Pregnancy , Spine/abnormalities , Urinary Tract/abnormalities
2.
J Neurobiol ; 48(3): 204-14, 2001 Sep 05.
Article in English | MEDLINE | ID: mdl-11466707

ABSTRACT

We prepared living slice preparations of the peripheral retina of adult goldfish to examine electrical membrane properties of progenitor cells at the retinal margin. Cells were voltage-clamped near resting potential and then stepped to either hyperpolarizing or depolarizing test potentials using whole-cell voltage-clamp recordings. Electrophysiologically examined cells were morphologically identified by injecting both Lucifer Yellow (LY) and biocytin. All progenitor cells examined (n = 37) showed a large amount of passively flowing currents of either sign under suppression of the nonjunctional currents flowing through K(+) and Ca(2+) channels in the cell membrane. They did not exhibit any voltage-gated Na(+) currents. Cells identified by LY fills were typically slender. As the difference between the test potential and the resting potential increased, 13 out of 37 cells exhibited symmetrically voltage- and time-dependent current decline on either sign at the resting potential. The symmetric current profile suggests that the current may be driven and modulated by the junctional potential difference between the clamping cell and its neighbors. The remaining 24 cells did not exhibit voltage dependency. A gap junction channel blocker, halothane, suppressed the currents. A decrease in extracellular pH reduced coupling currents and its increase enhanced them. Dopamine, cAMP, and retinoic acid did not influence coupling currents. Injection of biocytin into single progenitor cells revealed strong tracer coupling, which was restricted in the marginal region. Immature ganglion cells closely located to the retinal margin exhibited voltage-gated Na(+) currents. They did not reveal apparent tracer coupling. These results demonstrate that the marginal progenitor cells couple with each other via gap junctions, and communicate biochemical molecules, which may subserve or interfere with cellular differentiation.


Subject(s)
Gap Junctions/physiology , Goldfish/physiology , Retina/cytology , Retina/physiology , Stem Cells/physiology , Animals , Cellular Senescence/physiology , Electrophysiology , Fluorescent Dyes , In Vitro Techniques , Isoquinolines , Lysine/analogs & derivatives , Staining and Labeling , Uncoupling Agents/pharmacology
3.
J Comp Neurol ; 419(3): 297-305, 2000 Apr 10.
Article in English | MEDLINE | ID: mdl-10723006

ABSTRACT

Multipotent progenitor cells at the retinal margin of adult goldfish give rise to all cell types in the rest of the retina. We took advantage of this spatial arrangement of progenitor and mature cells in slices of peripheral retina, to investigate the appearance and maturation of voltage-activated Na(+) current. We divided the peripheral retina into three broad regions (marginal, intermediate, and mature) on the basis of their morphological development. Whole-cell patch-clamp recordings were performed in ruptured-patch mode, so that cells from which currents were recorded could be identified by Lucifer Yellow fills. No voltage-activated Na(+) current was detected in the slender, peripherally located marginal cells. Voltage-activated Na(+) currents were detected in rounded cells found alongside or near marginal cells, facing the vitreal side of the retina. Some of these "intermediate cells" had a long axon-like process which ran along the vitreal surface. Intermediate cells adjacent to the marginal region tended to have smaller Na(+) currents than intermediate cells closer to the mature region. On average, the maximum Na(+) current amplitude recorded from intermediate cells was roughly 6-fold smaller than that of mature ganglion cells. In addition, the activation threshold of the Na(+) current in intermediate cells was nearly 14 mV more positive than that of mature ganglion cells. The results indicate that voltage-activated Na(+) current, as a possible marker of retinal ganglion cells, begins to develop well before these cells migrate to their adult position within the retina.


Subject(s)
Goldfish/physiology , Retina/cytology , Retinal Ganglion Cells/physiology , Stem Cells/physiology , Animals , Cell Differentiation/physiology , Cellular Senescence/physiology , Electric Conductivity , In Vitro Techniques , Patch-Clamp Techniques , Retinal Ganglion Cells/cytology , Sodium/physiology , Stem Cells/cytology
4.
J Neurobiol ; 42(2): 258-69, 2000 Feb 05.
Article in English | MEDLINE | ID: mdl-10640332

ABSTRACT

Gap junctional coupling between progenitor cells of regenerating retina in the adult newt was examined by a slice-patch technique. Retinal slices at the early regeneration stage comprised one to two layers of cells with mitotic activity, progenitor cells. These cells were initially voltage-clamped at a holding potential of -80 mV, near their resting potentials, and stepped to either hyperpolarizing or depolarizing test potentials under suppression of voltage-gated membrane currents. About half the cells showed passively flowing currents that reversed polarity around their resting potentials. The currents often exhibited a voltage- and time-dependent decline. As the difference between the test potential and resting potential increased, the time until the current decreased to the steady-state level became shorter and the amount of steady-state current decreased. Thus, the overall current profile was almost symmetrical about the current at the resting potential. Input resistance estimated from the initial peak of the currents was significantly smaller than that expected in isolated progenitor cells. In a high-K(+) solution, which decreased the resting potential to around 0 mV, the symmetrical current profile was also obtained, but only when the membrane potential was held at 0 mV before the voltage steps. These observations suggest that the current was driven and modulated by the junctional potential difference between the clamping cell and its neighbors. In addition, we examined effects of uncoupling agents on the currents. A gap junction channel blocker, halothane, suppressed the currents almost completely, indicating that the currents are predominantly gap junctional currents. Furthermore, injection of biocytin into the current-recorded cells revealed tracer coupling. These results demonstrate that progenitor cells of regenerating retina couple with each other via gap junctions, and suggest the presence of their cytoplasmic communication during early retinal regeneration.


Subject(s)
Gap Junctions/physiology , Nerve Regeneration/physiology , Retina/physiology , Salamandridae/physiology , Stem Cells/physiology , Anesthetics, Inhalation/pharmacology , Animals , Calcium/metabolism , Cells, Cultured , Dopamine/pharmacology , Fluorescent Dyes/administration & dosage , Gap Junctions/drug effects , Halothane/pharmacology , In Vitro Techniques , Intracellular Fluid/metabolism , Isoquinolines/administration & dosage , Lysine/administration & dosage , Lysine/analogs & derivatives , Membrane Potentials/drug effects , Membrane Potentials/physiology , Microinjections , Mitosis , Patch-Clamp Techniques , Potassium/metabolism , Retina/cytology , Retina/growth & development , Stem Cells/cytology
5.
Mod Rheumatol ; 10(4): 225-9, 2000 Dec.
Article in English | MEDLINE | ID: mdl-24383634

ABSTRACT

Abstract Expression of the proto-oncogene c-fos was investigated by in situ hybridization and a study of the immunohistochemistry of the interfacial membranes surrounding the femoral component of failed cementless total hip arthroplasty (THA). The values of the proto-oncogene c-fos gene and protein were high in these interfacial membranes. High expression of the c-fos gene and protein was observed in the macrophage-like cells and fibroblast-like cells. However, there was little expression in multinucleated giant cells. We demonstrated the significance of the proto-oncogene c-fos in interfacial membranes - the first report of c-fos in total joint replacement failure.

6.
J Dermatol ; 26(9): 611-4, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10535258

ABSTRACT

We report the case of a 47-year-old woman who began to experience stiffness and Raynaud's phenomenon of her fingers and toes as well as easy fatigability approximately one year before initial evaluation. Histopathologic examination revealed dense fibrosis and perivascular lymphoid cell infiltration in the dermis. The patient's diagnosis was systemic sclerosis (SSc). Esophageal sclerosis was not revealed, but an early type IIb carcinoma of the gastric antrum was noted by endoscopic examination. Other recent reports have discussed the association of SSc with various malignant carcinomas. We also reviewed the literature for associations between SSc and gastric cancer in Japanese patients.


Subject(s)
Carcinoma, Signet Ring Cell/complications , Scleroderma, Systemic/complications , Stomach Neoplasms/complications , Biopsy, Needle , Carcinoma, Signet Ring Cell/pathology , Carcinoma, Signet Ring Cell/surgery , Female , Follow-Up Studies , Gastrectomy , Gastroscopy , Humans , Middle Aged , Scleroderma, Systemic/pathology , Stomach Neoplasms/pathology , Stomach Neoplasms/surgery , Treatment Outcome
7.
Jpn J Antibiot ; 51(5): 305-18, 1998 May.
Article in Japanese | MEDLINE | ID: mdl-9693964

ABSTRACT

Tazobactam/Piperacillin (TAZ/PIPC) is a newly developed intravenous antibiotics, in which TAZ, a new potent inhibitor of beta-lactamases, is combined with PIPC, a well-established beta-lactam antibiotics, at the ratio of 1:4. In this study, we clinically evaluated efficacy of the drug in 14 pediatric patients with various infections, and pharmacokinetic study was applied to 3 patients. Range of age was from 1-month to 15 1/4-year. Patients consisted of 9 cases of pneumonia, 3 urinary tract infection, 1 acute otitis media, and 1 left sacroiliitis with sepsis. Standard dose of TAZ/PIPC was 50 mg/kg/dose and administered 2-4 times per day with intravenous injection or drip infusion. Two cases of pneumonia were excluded because of non-bacterial infection. Nine causative pathogens including 3 Gram-positive cocci and 6 Gram-negative bacilli were detected in 7 patients, of which 5 Gram-negative strains produced bete-lactamase. All of cases showed 100% of efficacy rate and bacteriological eradication rate. It was noted that beta-lactamase-producing E. coli and B. catarrhalis were eradicated efficiently by TAZ/PIPC, which should be resistant to PIPC alone according to MIC data. Non-serious diarrhea and discomfort of back with nausea were observed in one each patients as side effects. Both of side effects were transient, and improved with anti-diarrheic agent or cessation of the drug, respectively. As abnormal laboratory test results, moderate increases of the eosinophils and platelets counts as well as moderate elevation of the transaminases were observed in 2 separate patients. Pharmacokinetics study showed that Cmax, T1/2, and AUC were similar to the data reported in adult patients. Urinary recovery rate in the first 6 hours also resemble the data from adult patients. Based on above results, TAZ/PIPC is a useful agents pediatric infections by beta-lactamase producing strains also.


Subject(s)
Bacterial Infections/drug therapy , Drug Therapy, Combination/administration & dosage , Adolescent , Bacteria/drug effects , Bacteria/isolation & purification , Bacterial Infections/microbiology , Child , Child, Preschool , Drug Resistance, Microbial , Drug Therapy, Combination/pharmacokinetics , Drug Therapy, Combination/pharmacology , Female , Humans , Infant , Infant, Newborn , Infusions, Intravenous , Injections, Intravenous , Male , Penicillanic Acid/administration & dosage , Penicillanic Acid/analogs & derivatives , Penicillanic Acid/pharmacokinetics , Penicillanic Acid/pharmacology , Piperacillin/administration & dosage , Piperacillin/pharmacokinetics , Piperacillin/pharmacology , Piperacillin, Tazobactam Drug Combination
8.
Brain Res ; 785(1): 171-7, 1998 Feb 23.
Article in English | MEDLINE | ID: mdl-9526076

ABSTRACT

The appearance of endogenous glutamate during retinal regeneration in the newt was examined by immunohistochemistry. Glutamate-like immunoreactivity (Glu-LI) first appeared in prospective ganglion cells along the vitreal margin of retinas that were about six cells thick, in prospective photoreceptors immediately before segregation of retinal plexiform layers and then in prospective bipolar cells immediately after the initial appearance of thin plexiform layers. In retinas nearing complete regeneration, Müller cells showed immunoreactivity. The appearance of glutamatergic phenotypes during retinal regeneration seemed to follow the order of cell differentiation [T. Saito, Y. Kaneko, F. Maruo, M. Niino, Y. Sakaki, Study of the regenerating newt retina by electrophysiology and immunohistochemistry (bipolar- and cone-specific antigen localization), J. Exp. Zool. 270 (1994) 491-500]. However, changes in the amount of endogenous glutamate during retinal regeneration were more complex. On the one hand, Glu-LI at the prospective ganglion cell layer temporarily increased during the initial period of segregation of the inner plexiform layer. On the other hand, immunoreactivity in the photoreceptor layer declined during segregation of the outer plexiform layer. The transient expression of immunoreactivity may represent a function of glutamate in events such as cell survival or neurite extension during retinal regeneration.


Subject(s)
Glutamic Acid/metabolism , Nerve Regeneration/physiology , Photoreceptor Cells/physiology , Retina/physiology , Retinal Ganglion Cells/physiology , Animals , Immunohistochemistry , Photoreceptor Cells/cytology , Retina/cytology , Retinal Ganglion Cells/cytology , Salamandridae
9.
Gastroenterology ; 113(6): 1828-35, 1997 Dec.
Article in English | MEDLINE | ID: mdl-9394722

ABSTRACT

BACKGROUND & AIMS: Antineutrophil cytoplasmic antibodies (ANCAs) have been detected in approximately 60% of sera from patients with ulcerative colitis. The aim of this study was to examine the presence of ANCAs and distribution of ANCA-producing B cells in the lymphoid tissue of T-cell receptor alpha-deficient (TCR-alpha-/-) mice that develop chronic colitis resembling human ulcerative colitis. METHODS: Sera from 87 TCR-alpha-/- mice were tested for the presence of ANCAs by enzyme-linked immunosorbent assay against a human neutrophil extract and selected antigens and by immunofluorescence study using human neutrophils. Enzyme-linked immunospot assay was used for detecting ANCA-producing cells from spleen, mesenteric lymph node (MLN), or colon. RESULTS: Approximately 70% of sera from TCR-alpha-/- mice showed reactivity against human neutrophil extracts by enzyme-linked immunosorbent assay. Sixty percent of ANCA-positive sera showed a perinuclear reaction pattern. By enzyme-linked immunospot, ANCA-producing cells were detected in MLN and colon and less often in the spleen of TCR-alpha-/- mice with chronic colitis. The predominant immunoglobulin isotype of these autoantibodies was immunoglobulin A in the colon but not in the MLN and spleen. CONCLUSIONS: TCR-alpha-/- mice produce ANCAs. The ANCA (immunoglobulin A isotype)-producing B cells exist primarily in the diseased colonic mucosa of TCR-alpha-/- mice.


Subject(s)
Antibodies, Antineutrophil Cytoplasmic/analysis , Colitis/immunology , Colitis/metabolism , Receptors, Antigen, T-Cell, alpha-beta/deficiency , Animals , Antibodies, Antineutrophil Cytoplasmic/immunology , Blood/immunology , Cell Extracts/immunology , Chronic Disease , Colitis/pathology , Colon/immunology , Colon/metabolism , Colon/pathology , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody Technique, Indirect , Histones/immunology , Humans , Immunoglobulin A/metabolism , Lymph Nodes/immunology , Mesentery , Mice , Mice, Inbred C57BL , Mice, Mutant Strains , Neutrophils/immunology
10.
Brain Res Dev Brain Res ; 98(2): 204-10, 1997 Feb 20.
Article in English | MEDLINE | ID: mdl-9051262

ABSTRACT

Appearance and maturation of the GABA (gamma-aminobutyric acid) system during newt retinal regeneration were studied by electrophysiological, immunohistochemical, and biochemical techniques. (1) Responses to GABA appeared in neurons dissociated from regenerating retinae before the segregation of the plexiform layers; whereas (2) GABA immunoreactivity appeared at sites of the presumptive horizontal cell and amacrine cell layers at the beginning of the segregation of these layers. During subsequent regeneration, GABA-immunoreactive cells at the amacrine cell layer increased in number and extended lateral processes, forming a GABA-immunoreactive inner plexiform layer. Also GABA immunoreactivity increased in the region of the outer plexiform layer, but not their somata which showed decreased GABA immunoreactivity. (3) GABA synthesis in the retina increased significantly at the beginning of the segregation of the plexiform layers. These results suggest that the increase of GABA synthesis during retinal regeneration correlates well with the development of GABA-immunoreactive cells and that functional GABA receptors appear earlier than increased GABA synthesis.


Subject(s)
Nerve Regeneration/physiology , Receptors, GABA/analysis , Retina/physiology , gamma-Aminobutyric Acid/analysis , Animals , Chromatography, High Pressure Liquid , Immunohistochemistry , Patch-Clamp Techniques , Reference Values , Salamandridae
11.
J Exp Med ; 184(2): 707-15, 1996 Aug 01.
Article in English | MEDLINE | ID: mdl-8760824

ABSTRACT

T cell receptor-alpha mutant mice (TCR-alpha-/-), created by gene targeting of the TCR-alpha gene in embryonic stem cells, spontaneously develop inflammatory bowel disease (IBD) resembling human ulcerative colitis. Since gut-associated lymphoid tissue is likely to play an important role in the development of chronic intestinal inflammation, we examined the changes in the appendix lymphoid follicle (ALF) and Peyer's patches (PP) in these mice. We found the structure of the ALF to be remarkably similar to that of the PP in the small intestine; in both instances, lymphoid follicles covered by surface epithelium (dome-formation) were found. The amount of proliferation in the lymphoid follicles of the appendix estimated by in vivo incorporation of 5-bromo-2'deoxyuridine was more than two times that of PP in TCR-alpha-/- mice. ELISPOT assay showed an increase of IgA, IgG1, and IgG2a, but not IgM-secreting B cells in ALF of TCR-alpha-/- mice compared to TCR-alpha+/- control mice. Furthermore, TCR-alpha-/- mice revealed an increase of autoantibody-producing B cells against the cytoskeletal protein tropomyosin in ALF as compared to PP. When TCR-alpha-/- mice underwent appendectomy at a young age (3-5 wk), the number of mesenteric lymph nodes cells at 6-7 mo were markedly less than in the sham-operated TCR-alpha-/- mice. Furthermore, appendectomy at 1 mo of age suppressed the development of IBD, with only 3.3% of these mice developing IBD in the 6-7-mo period of observation. In contrast, approximately 80% of controls, including the sham-operated TCR-alpha-/- mice, developed IBD during this period. These results suggest that ALF, rather than PP, is the priming site of cells involved in the disease process and plays an important role in the development of IBD in TCR-alpha-/- mice.


Subject(s)
Appendix/physiopathology , Inflammatory Bowel Diseases/immunology , Receptors, Antigen, T-Cell, alpha-beta/immunology , Animals , Appendectomy , Autoantibodies/biosynthesis , B-Lymphocytes/immunology , Cell Division , Immunity, Mucosal , Immunophenotyping , Intestinal Mucosa/cytology , Leukocyte Common Antigens/analysis , Mice , Mice, Knockout , Peyer's Patches/cytology , Peyer's Patches/immunology , Receptors, IgE/analysis , T-Lymphocytes/immunology
12.
J Exp Med ; 183(3): 847-56, 1996 Mar 01.
Article in English | MEDLINE | ID: mdl-8642289

ABSTRACT

Spontaneous inflammatory bowel disease (IBD) resembling human ulcerative colitis develops in mice mutant for the T cell receptor alpha gene (TCR-alpha-/-). TCR-alpha-/- mice lack TCR-alpha/beta+ cells but contain TCR-gamma/delta+ cells and a small population of a unique CD4+, TCR-alpha-/beta+(low) cells. Since all the immunoglobulin (Ig) classes are present in these mice, help to B cells must be provided by cells other than TCR-alpha/beta+ cells. In the present study, we found serum levels of IgG1 and IgG2 to be markedly increased in TCR-alpha-/- mice with IBD as compared to TCR-alpha-/- mice without IBD or TCR-alpha+/- controls. An increase in IgG1-, IgG2a- and IgA- but not IgM-secreting mesenteric lymph node (MLN) B cells was detected in TCR-alpha-/- mutant mice. There was also a marked increase in MLN B cells secreting autoantibody (IgG) to tropomyosin, a cytoskeletal protein. Examination of the hyperplastic MLN showed a marked increase in the number of B, TCR-delta+, and CD4+ TCR-alpha-/beta+ cells, similar to the cell population observed at the site of colonic inflammation. Analysis of spontaneous cytokine production by MLN cells using an enzyme-linked immunospot assay, immunohistochemistry, and reverse transcription/polymerase chain reaction showed a decrease of interleukin 2 (IL-2) but a marked increase of IL-4 and interferon gamma (IFN-gamma) production in TCR-alpha-/- mice with IBD as compared to TCR-alpha-/- mice without IBD and TCR alpha+/- control mice. Both TCR-alpha-/beta+ and TCR-delta+ cells were found to be capable of producing IL-4; IFN-gamma was produced mostly by non-T cells, many of which were shown to be CD3- NK 1.1+ cells. We propose that the cytokine imbalance present in these mice results in expansion of B cells, production and switching of autoantibodies to IgG2 subclass, and development of IBD. It is possible that the unusual CD4+ TCR-alpha-/beta+ population and expanded TCR-gamma/delta+ population present in TCR-alpha-/- mice plays a central role in this abnormal immune response.


Subject(s)
Autoantibodies/biosynthesis , Cytokines/biosynthesis , Inflammatory Bowel Diseases/genetics , Inflammatory Bowel Diseases/immunology , Receptors, Antigen, T-Cell, alpha-beta/genetics , T-Lymphocytes/immunology , Animals , Autoimmune Diseases/genetics , B-Lymphocytes/immunology , Cells, Cultured , Colitis, Ulcerative/immunology , Cytokines/analysis , DNA Primers , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Humans , Immunoglobulin Isotypes/analysis , Immunoglobulin Isotypes/biosynthesis , Immunophenotyping , Intestinal Mucosa/immunology , Mice , Mice, Mutant Strains , Polymerase Chain Reaction , T-Lymphocyte Subsets/immunology
13.
Clin Diagn Lab Immunol ; 2(3): 322-4, 1995 May.
Article in English | MEDLINE | ID: mdl-7664179

ABSTRACT

Cytokine (interleukin 6 [IL-6] and tumor necrosis factor alpha [TNF-alpha]) activity in nasopharyngeal secretions of 21 infants and children (19 days to 16 months old) infected with primary respiratory syncytial virus was determined by an enzyme-linked immunosorbent assay. IL-6 and TNF-alpha were detectable in 100% (21 of 21) and 67% (14 of 21) of cases during the course of infection, respectively. Generally, TNF-alpha activity was high in the acute phase and declined thereafter, sometimes to undetectable levels. IL-6 activity was also highest in the acute phase and declined thereafter in infants younger than 5 months, while in patients older than 5 months, it-increased during the course of the disease to peak in the early convalescent phase. These observations suggest that inflammatory cytokines are produced in vivo in infants and children in response to primary respiratory syncytial virus infection and may be involved in disease pathogenesis. However, the mechanism of induction of cytokines may be different for infants and children in different age groups.


Subject(s)
Interleukin-6/metabolism , Nasopharynx/virology , Respiratory Syncytial Virus Infections/immunology , Tumor Necrosis Factor-alpha/metabolism , Antibody Specificity , Cytokines/immunology , Cytokines/metabolism , Female , Humans , Immunoglobulin A/blood , Immunoglobulin A/immunology , Infant , Infant, Newborn , Kinetics , Male , Nasopharynx/immunology , Nasopharynx/metabolism , Time Factors
14.
Jpn J Physiol ; 45(5): 849-67, 1995.
Article in English | MEDLINE | ID: mdl-8713181

ABSTRACT

Whole-cell currents activated by L-glutamate, kainate (KA), alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA), N-methyl-D-aspartate (NMDA), gamma-aminobutyric acid (GABA), and glycine were recorded from spiking cells enzymatically dissociated from adult newt retina. Most spiking cells responded to both AMPA and KA. KA tended to be a more potent agonist for them than AMPA. The reversal potential of both AMPA and KA responses was about -7 mV; this value was similar to that (about -5 mV) of glutamate response. AMPA- and KA-induced currents may be carried by monovalent cations, such as Na+, because their reversal potentials were sensitive to external Na+. About half of the spiking cells examined responded to mixtures of NMDA and glycine. Extracellular Mg2+ blocked completely the response to NMDA plus glycine in spiking cells held at negative membrane potentials, but not at positive membrane potentials. Almost all spiking cells responded to the inhibitory amino acids GABA and glycine. Dose-dependent desensitization was observed in both GABA and glycine responses. Currents activated by GABA and glycine were carried by Cl-. Bicuculline and strychnine strongly suppressed the responses to GABA and glycine, respectively, suggesting the existence of GABAA receptors and conventional glycine receptors in the spiking cells.


Subject(s)
Amino Acids/pharmacology , Receptors, Amino Acid/drug effects , Retina/drug effects , Animals , Dose-Response Relationship, Drug , Glutamic Acid/pharmacology , Kainic Acid/pharmacology , N-Methylaspartate/pharmacology , Patch-Clamp Techniques , Salamandridae , alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid/pharmacology
15.
Jpn J Physiol ; 45(5): 869-87, 1995.
Article in English | MEDLINE | ID: mdl-8713182

ABSTRACT

Responsiveness to amino acids in solitary spiking cells dissociated from regenerating newt retinae at different stages were studied by whole-cell patch-clamp methods in comparison with that in the normal retina. Cells from regenerating retinae of 1-2 cell thickness ("early" stage) lacked the receptors for kainate (KA), alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA), N-methyl-D-aspartate (NMDA), gamma-aminobutyric acid (GABA), and glycine. AMPA/KA, NMDA, GABAA, and glycine receptors, each of which were electrophysiologically and pharmacologically similar to those in the normal retina, appeared in spiking cells before regenerating retina had segregated into two synaptic layers. During subsequent regeneration stage, the time course of appearance of amino acid receptors seems to be different between excitatory and inhibitory ones. Cells which responded to GABA and glycine gradually increased in number to normal levels, and the current amplitude induced by these amino acids also increased monotonically. The number of cells responding to the excitatory amino acids appeared to be maximal at a period of synaptic segregation. During subsequent regeneration, however, cells responded to AMPA and NMDA decreased in number and returned to the number of cells in the normal retina, while cells which responded to KA were maintained at the same number as those in the normal retina. The amplitudes of the current induced by excitatory amino acids also became maximal at a period of synaptic segregation. During subsequent regeneration, AMPA- and KA-induced currents declined to the normal level, while NMDA-induced currents were maintained.


Subject(s)
Amino Acids/pharmacology , Regeneration/physiology , Retina/physiology , Action Potentials/physiology , Animals , Kainic Acid/pharmacology , N-Methylaspartate/pharmacology , Patch-Clamp Techniques , Receptors, Amino Acid/physiology , Salamandridae , Strychnine/pharmacology , alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid/pharmacology
16.
Neuroreport ; 5(4): 489-92, 1994 Jan 12.
Article in English | MEDLINE | ID: mdl-8003681

ABSTRACT

In vertebrate retinae, APB (2-amino-4-phosphonobutyrate) selectively activates glutamate receptors of ON-bipolar cells. Thus, APB has been used as a pharmacological tool for studying the functional retinal circuitry and the inputs to higher-order pathways of the visual centres. Direct action of APB on ganglion cells has not previously been demonstrated. We now report, using whole-cell patch-clamp recordings, that APB has a direct inhibitory effect on ganglion cells isolated from newt retina. APB-induced currents were dependent upon the intracellular Cl- concentration. Their reversal potential values agreed with theoretical values of Cl- equilibrium potential. Strychnine, a glycine receptor blocker, inhibited the APB response. These results may suggest that APB activates the Cl- channels of ganglion cells through glycine receptors.


Subject(s)
Aminobutyrates/pharmacology , Chlorides/physiology , Retinal Ganglion Cells/drug effects , Animals , Cell Separation , Electric Conductivity , GABA Antagonists , In Vitro Techniques , Receptors, Glycine/antagonists & inhibitors , Salamandridae
17.
Neuroreport ; 4(6): 775-8, 1993 Jun.
Article in English | MEDLINE | ID: mdl-7688592

ABSTRACT

Neuroblasts dissociated from early regenerating newt retina (18-20 days after surgery) survived in culture and extended neurites. They were classified into three groups on the basis of neurite outgrowth patterns; monopolar, bipolar, and multipolar varieties. Freshly dissociated neuroblasts (3-15 h in culture) were characterized by slow-rising action potentials with long duration, mediated by inward Na+ currents. After a few days in culture, these cells produced much briefer action potentials mediated by the characteristic Na+, Ca2+ and K+ currents of ganglion cells dissociated from normal retina. The results suggest that neuroblasts in early regenerating retina differentiate in vitro into spiking cells, possibly ganglion cells, characterized by their morphology and voltage-dependent ionic currents.


Subject(s)
Ion Channel Gating/physiology , Ion Channels/metabolism , Neurons/metabolism , Retina/metabolism , Action Potentials/physiology , Animals , Cells, Cultured , Nerve Regeneration/physiology , Neurites/metabolism , Retina/cytology , Retinal Ganglion Cells/metabolism , Salamandridae
18.
J Comp Physiol A ; 170(6): 773-86, 1992 Jul.
Article in English | MEDLINE | ID: mdl-1359128

ABSTRACT

1. Transmitters of motoneurons in the stomatogastric ganglion (STG) of Squilla were identified by analyzing the excitatory neuromuscular properties of muscles in the posterior cardiac plate (pcp) and pyloric regions. 2. Bath and iontophoretic applications of glutamate produce depolarizations in these muscles. The pharmacological experiments and desensitization of the junctional receptors elucidate the glutamatergic nature of the excitatory junctional potentials (EJPs) evoked in the constrictor and dilator muscles. The reversal potentials for the excitatory junctional current (EJC) and for the glutamate-induced current are almost the same. 3. Some types of dilator muscle show sensitivity to both glutamate and acetylcholine (ACh) exogenously applied. The pharmacological evidence and desensitization of the junctional receptors indicate the glutamatergic nature of neuromuscular junctions in these dually sensitive muscles. The reversal potentials for the EJC and for the ACh-induced current are not identical. 4. Glutamate is a candidate as an excitatory neuro-transmitter at the neuromuscular junctions which the STG motoneurons named PCP, PY, PD, LA and VC make with the identified muscles. Kainic and quisqualic acids which act on glutamate receptors are potent excitants of these muscles. Extrajunctional receptors to ACh are present in two types of the muscle innervated by LA and VC. 5. Neurotransmitters used by the STG motoneurons of stomatopods are compared to those of decapods.


Subject(s)
Decapoda/physiology , Ganglia/physiology , Glutamates/physiology , Motor Neurons/physiology , Acetylcholine/physiology , Animals , Electrophysiology , Ganglia/ultrastructure , Iontophoresis , Membrane Potentials/physiology , Neuromuscular Junction/physiology , Neurotransmitter Agents/physiology
19.
J Comp Physiol A ; 169(6): 737-50, 1991 Dec.
Article in English | MEDLINE | ID: mdl-1795238

ABSTRACT

The pyloric constrictor muscles of the stomach in Squilla can generate spikes by synaptic activation via the motor nerve from the stomatogastric ganglion. Spikes are followed by slow depolarizing afterpotentials (DAPs) which lead to sustained depolarization during a burst of spikes. 1. The frequency of rhythmic bursts induced by continuous depolarization is membrane voltage-dependent. A brief depolarizing or hyperpolarizing pulse can trigger or terminate bursts, respectively, in a threshold-dependent manner. 2. The conductance increases during the DAP response. The amplitude of DAP decreases by imposed depolarization, whereas it increases by hyperpolarization. DAPs from successive spikes sum to produce a sustained depolarizing potential capable of firing a burst. 3. The spike and DAP are reduced in amplitude by decreasing [Ca]o, enhanced by Sr2+ or Ba2+ substituted for Ca2+, and blocked by Co2+ or Mn2+. DAPs are selectively blocked by Ni2+, and the spike is followed by a hyperpolarizing afterpotential. 4. The spike and DAP are prolonged by intracellular injection of the Ca2+ chelator EGTA. A hyperpolarizing afterpotential is abolished by EGTA and enhanced by increasing [Ca]o. The DAP is diminished in Na(+)-free saline and reduced by tetrodotoxin. 5. It is concluded that the muscle fiber is endowed with endogenous oscillatory properties and that the oscillatory membrane events result from changes of a voltage- and time-dependent conductance to Ca2+ and Na+ and a Ca2+ activated conductance to K+.


Subject(s)
Decapoda/physiology , Muscles/physiology , Animals , Egtazic Acid/pharmacology , Electrophysiology , Female , Ions , Male , Muscle Contraction , Neuromuscular Junction/physiology , Oscillometry , Stomach , Tetrodotoxin/pharmacology
20.
Behav Neural Biol ; 54(2): 184-90, 1990 Sep.
Article in English | MEDLINE | ID: mdl-2241761

ABSTRACT

Female gerbils (Meriones unguiculatus) of three distinct coat colors (agouti, black, and sandy or pink-eyed dilution) were tested in a Y-maze whose arms led to compartments containing unfamiliar male gerbils of varying coat colors. The stimulus animals were separated from the females by a Plexiglas door. The trials lasted for 2 min and each female was exposed to the following four combinations: two males of the same coloring as the female; one male of the same color and another of a different color from the female; both males of different color from the female. The number of crossings to the left and right arms was relayed by photocells to an IBM PC computer. The results indicate that agouti females preferred visiting the arm occupied by agouti males while those of the other coat colors showed no preference for the "wild-type" males. Instead, sandy and black female gerbils preferred to be in proximity with those of non-wild types.


Subject(s)
Gerbillinae/physiology , Hair Color/physiology , Sexual Behavior, Animal/physiology , Species Specificity , Animals , Arousal/physiology , Choice Behavior/physiology , Discrimination Learning/physiology , Female , Male , Orientation/physiology
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