Biomedical subjects

K Kamino

Publications and source records attributed to K Kamino.

At least 19 recordsLinked to original sources

PubMed2004

WKY/Ztm-ter: a new rat inbred strain on the WKY/Ztm genetic background with congenital teratomas.

We describe a new rat model for teratomas (WKY/Ztm-ter) which arose through a spontaneous mutation in the inbred WKY/Ztm rat strain. When the tumours of the gonads became clinically apparent, affected males were 14 to 224 days of age, whereas the females only developed tumours between days 21 and 63. Tumour incidence is not gender-dependent. However, almost all females develop bilateral tumours, while 50% of the males show unilateral tumours. Histologically, all examined tumours (n = 65) represent partially undifferentiated teratocarcinomas.

Animals↗

PubMed2004

Orthotopic transplantation of rat ovaries as a tool for strain rescue.

Spontaneous mutations in inbred strains provide valuable resources for new disease models. Unfortunately, these mutations may affect reproduction, which require considerable efforts in breeding management. We transplanted ovaries of such mutant rat strains orthotopically into ovariectomized premature coisogenic recipients. A reproductive cycle was established in each of the recipients within 5 to 6 weeks after transplantation. Moreover, one-third became pregnant and had litters an average of 3 months after transplantation. These experiments demonstrate that orthotopic transplantation of ovaries can be used in the management of subfertile rat colonies.

Animals↗

PubMed2001

Bacterial response to siderophore and quorum-sensing chemical signals in the seawater microbial community.

BACKGROUND: Oceans are iron-deficient and nutrient-poor environments. These conditions impart limitations on our understanding of and our ability to identify microorganisms from the marine environment. However, less of knowledge on the influence of siderophores and N-acyl homoserinelactone as interspecies communication signals on the bacterial diversity of seawater has been understood. RESULTS: In the presence of 0.1 nM of the commercial siderophore desferroixamine and the known quorum-sensing chemical signals, synthetic N-(3-oxo)-hexanoylhomoserine lactone (0.1 nM) or N-octanoylhomoserine lactone (0.1 nM), the total numbers of bacteria in S9905 seawater increased nearly three-fold, and nearly eight-fold in S0011 seawater as determined by DAPI staining and counting, and increased three-fold by counting colony forming units in S9905 seawater after 7 days of incubation. Similar bacterial changes in bacterial abundance were observed when high concentration of desferroixamine (1 microM) and each of homoserine lactone compounds (1 microM) were presented in seawater samples. The number of cultivable bacterial species observed was also found to increase from 3 (without addition) to 8 (with additions) including three unknown species which were identified by phylogenetic analysis of 16S rDNA sequences. The growth of unknown species was found to be related to their siderophore production with response to the addition of desferroixamine and N-acyl homoserine lactones under iron-limited conditions. CONCLUSION: Artificial addition of siderophores and HSLs may be a possible method to aid in the identification and isolation of marine bacterial species which are thought to be unknown.

4-Butyrolactone↗

PubMed2001

Novel barnacle underwater adhesive protein is a charged amino acid-rich protein constituted by a Cys-rich repetitive sequence.

Barnacle cement is an underwater adhesive that is used for permanent settlement, and is an insoluble protein complex. A method for rendering soluble the cement of Megabalanus rosa has been developed, and three major proteins have been identified in a previous study. To survey the M. rosa cement proteins in a lower molecular mass range, the cement proteins were separated by reversed-phase HPLC and a previously unidentified protein named 20 kDa M. rosa cement protein (Mrcp-20k) was found. Mrcp-20k cDNA was cloned to reveal its primary structure. This cDNA was 902 bp long and encoded a 202 amino acid-long open reading frame, including 19 amino acids of the signal sequence. The molecular mass in the disulphide form was calculated to be 20357 Da and the isoelectric point of the mature polypeptide was 4.72. Mrcp-20k was characterized by an abundance of Cys residues and charged amino acids. The most common amino acid was Cys (17.5%), with Asp (11.5%), Glu (10.4%) and His (10.4%) following in order of magnitude. The alignment of the Cys residues indicated the primary structure of this protein to consist of six degenerated repeats, each about 30 residues long. Mrcp-20k has no intermolecular disulphide bonds and no free thiol groups of Cys in the insoluble cement complex. Abundant Cys is thought to play a role in maintaining the topology of charged amino acids on the molecular surface by intramolecular disulphide-bond formation. The possible function of abundant charged amino acids, including the interaction with a variety of surface metals on the substratum, is discussed.

Adhesives↗

PubMed2001

Effects of anisomycin on LTP in the hippocampal CA1: long-term analysis using optical recording.

Long-term potentiation (LTP) in the hippocampal CA1 region and in the dentate gyrus consists of different stages: early LTP lasting minutes or several hours, and late LTP lasting longer than 4 h. It has been suggested that the late phase of LTP is dependent on protein synthesis. However, the experimental results of the effects of protein synthesis inhibitors are still confusing. We applied optical recording techniques to rat hippocampal slices, and re-evaluated the effects of a protein synthesis inhibitor, anisomycin, on LTP. Using a voltage-sensitive oxonol dye, NK3630 (RH482), LTP in the CA1 region could be monitored optically for a long-term period (7-8 h). In the presence of anisomycin, the potentiation of the EPSP (excitatory postsynaptic potential) lasted about 2-3 h, followed by a gradual decline in the signal amplitude. Statistically, significant effects of anisomycin were observed 6 h after LTP induction for 100 Hz tetanus and 8 h after LTP induction for 400 Hz tetanus. These results suggest that the early phase of LTP is independent of protein synthesis, while the late phase of potentiation (> 3-5 h) depends on protein synthesis.

Animals↗

PubMed2001

Optical responses to micro-application of GABA agonists in the embryonic chick brain stem.

We investigated optical responses induced by micro-application of muscimol and baclofen in the embryonic chick brain stem. Muscimol evoked biphasic optical signals which were similar to those induced by GABA. The first component was a dye-dependent absorption change that reflected membrane depolarization, and the second component was an intrinsic optical change coupled with changes in the membrane potential. On the other hand, baclofen did not elicit any optical change. The optical responses induced by muscimol persisted in the presence of picrotoxin and 2-hydroxysaclofen, suggesting that they contain a component which is not mediated by classical GABA receptors.

Animals↗

PubMed2001

Optical approaches to embryonic development of neural functions in the brainstem.

The ontogenetic approach to physiological events is a useful strategy for understanding the functional organization/architecture of the vertebrate brainstem. However, conventional electrophysiological techniques are difficult or impossible to employ in the early embryonic central nervous system. Optical techniques using voltage-sensitive dyes have made it possible to monitor neural activities from multiple regions of living systems, and have proven to be a useful tool for analyzing the embryogenetic expression of brainstem neural function. This review describes recent progress in optical studies made on embryonic chick and rat brainstems. Several technical issues concerning optical recording from the embryonic brainstem preparations are discussed, and characteristics of the optical signals evoked by cranial nerve stimulation or occurring spontaneously are described. Special attention is paid to the chronological analyses of embryogenetic expression of brainstem function and to the spatial patterning of the functional organization/architecture of the brainstem nuclei. In addition, optical analyses of glutamate, GABA, and glycine receptor functions during embryogenesis are described in detail for the chick nucleus tractus solitarius. This review also discusses intrinsic optical signals associated with neuronal depolarization. Some emphases are also placed on the physiological properties of embryonic brainstem neurons, which may be of interest from the viewpoint of developmental neurobiology.

Animals↗

PubMed2001

Spreading depolarization waves triggered by vagal stimulation in the embryonic chick brain: optical evidence for intercellular communication in the developing central nervous system.

Throughout experiments on multiple-site voltage-sensitive dye recordings of neural activity in embryonic chick brain preparations, we have found a novel type of depolarization waves which spread widely from the brainstem to the whole brain region at a rapid rate (mm/s). This depolarization wave was triggered by glutamate-mediated postsynaptic potentials and was especially correlated to N-methyl-D-aspartate receptor function. Evidence that the spreading depolarization wave is eliminated by octanol or 18beta-glycyrrhetinic acid suggests that the depolarization wave depends on functions of gap junctions. The profile obtained with Ca(2+)-imaging experiments also suggests that the propagation of the depolarization wave is accompanied by a calcium wave. These results provide new evidence for intercellular functional communication between neural cells in the vertebrate central nervous system during embryonic development.

Animals↗

PubMed2001

Spectrum and age-related incidence of spontaneous tumours in a colony of Han:AURA hamsters.

One-hundred-and-forty-four male and 184 female untreated Syrian golden hamsters (strain Han:AURA) were kept for life under standard laboratory conditions. They were examined with regard to spontaneously occurring tumours in relation to their survival periods. The mean survival rate of the males was 106 +/- 26 weeks and that of the females 97 +/- 20 weeks. Tumours were found in 71% of males and 67% of females. Adenomas and carcinomas of the adrenal glands were the most frequently observed tumours in both sexes (male: 66%; female: 38%) and in the early stages of life. Malignant lymphoma (8%), adenomas and carcinomas of pancreatic islet-cells (8%) and papillomatous benign and malignant squamous cell tumours of the forestomach (7%) showed relatively high incidences in males, whilst in females, leiomyoma (10%) and endometrial adenocarcinoma (7%) of the uterus and adenomas and carcinomas in the pars distalis of the pituitary gland (9%) occurred frequently.

Animals↗

PubMed2001

Multiple-site optical recording reveals embryonic organization of synaptic networks in the chick spinal cord.

We examined embryonic expression of postsynaptic potentials in stages 26-31 (E5 to E7) chick spinal cord slices. Slow optical signals related to the postsynaptic potentials which were evoked by electrical stimulation of afferent fibers were identified in the dorsal grey matter and the ventral motoneuronal area. In cervical spinal cord (C13) preparations, the dorsal slow signal appeared from stage 28 (E6), whilst the ventral slow signal was recognized from stage 29. At stages 26 and 27 (E5), no slow signal was observed in either the dorsal or ventral regions. On the other hand, in lumbosacral spinal cord (LS5) preparations, the dorsal, as well as ventral, slow signals appeared from stage 29; at stage 28 no slow signal was detected in the dorsal or ventral regions. These results suggest that there are differences in the ontogenetic expression of synaptic functions between the dorsal and ventral regions, and between the cervical and lumbosacral spinal cords. In embryos older than stage 29, removal of Mg2+ from the bathing solution markedly enhanced the amplitude and incidence of the ventral slow signal. In addition, in C13 preparations at stage 28, removal of Mg2+ elicited small slow signals in the ventral region in which no synaptic response was evoked in normal Ringer's solution. The slow signals induced in the Mg2+-free solution were blocked by 2-amino-5-phosphonovaleric acid (APV), showing that they are attributable to N-methyl- D-aspartate (NMDA) receptors. These results suggest that functional synaptic connections via polysynaptic pathways are already generated on motoneurons, but are suppressed by a Mg2+ block on the NMDA receptors at developmental stages when synaptic transmission from the primary afferents to the dorsal interneurons is initially expressed in the dorsal region.

Animals↗

PubMed2001

Optical imaging of spreading depolarization waves triggered by spinal nerve stimulation in the chick embryo: possible mechanisms for large-scale coactivation of the central nervous system.

Using a multiple-site optical recording technique with a voltage-sensitive dye, we found that widely spreading depolarization waves were evoked by dorsal root stimulation in embryonic chick spinal cords. Spatiotemporal maps of the depolarization waves showed that the signals were mainly distributed in the ventral half of the slice, with the highest activity in the ventrolateral area. The propagation velocity of the waves was estimated to be in the order of mm/s. Depolarization waves were evoked in the ventral root-cut preparation, but not in the dorsal root-cut preparation, suggesting that the wave was triggered by synaptic inputs from the primary afferents, and that activation of the motoneurons was not essential for wave generation. In intact spinal cord-brain preparations, the depolarization wave propagated rostrally and caudally for a distance of several spinal segments in normal Ringer's solution. In a Mg(2+)-free solution, the amplitude and extent of the signals were markedly enhanced, and the depolarization wave triggered in the cervical spinal cord propagated to the brainstem and the cerebellum. The depolarization wave demonstrated here had many similarities with the vagus nerve-evoked depolarization wave reported previously. The results suggest that functional cell-to-cell communication systems mediated by the depolarization wave are widely generated in the embryonic central nervous system, and could play a role in large-scale coactivation of the neurons in the spinal cord and brain.

Animals↗

PubMed2001

Age-related incidence of spontaneous non-neoplastic lesions in a colony of Han:AURA hamsters.

In toxicologic testing or experimental studies using animals, an adequate knowledge of spontaneously occurring lesions is required. 144 male and 184 female untreated Syrian golden hamsters (strain Han:AURA) were kept for life under standard laboratory conditions and an investigation of non-neoplastic lesions in relationship to the lifespan was performed. The average lifespan of the males was 106 weeks and that of the female hamsters 97 weeks. While cartilage degeneration of the sternum and fatty degeneration of the femoral bone marrow occurred already in the first half of life with high incidence, the majority of lesions were observed in the second half. The most frequent non-neoplastic changes in various organs were fatty change, calcification, cystic change, hyperplasia and amyloidosis. Such spontaneous lesions were discussed in connection with the same alterations which can also be induced by chemical or hormonal agents.

Aging↗

PubMed2001

Herpesvirus saimiri Tip gene causes T-cell lymphomas in transgenic mice.

New World primates develop T-cell lymphomas on infection with Herpesvirus saimiri. To investigate the oncogenic potential of the Tip gene of Herpesvirus saimiri strain C488, we tried to establish transgenic mice that should express Tip under control of a constitutive promoter. Although transgene-positive embryos were found, lines could not be established. However, using a system in which the transgene has to be activated by a Cre recombinase-mediated deletion, we were able to obtain several Tip transgenic lines. At high expression levels, the mice developed T-cell lymphomas. Thus, Tip can induce lymphomas and is therefore very likely responsible for the oncogenicity of Herpesvirus saimiri.

Animals↗

PubMed2001

Effect of exogenous siderophores on iron uptake activity of marine bacteria under iron-limited conditions.

More than 60% of species examined from a total of 421 strains of heterotrophic marine bacteria which were isolated from marine sponges and seawater were observed to have no detectable siderophore production even when Fe(III) was present in the culture medium at a concentration of 1.0 pM. The growth of one such non-siderophore-producing strain, alpha proteobacterium V0210, was stimulated under iron-limited conditions with the addition of an isolated exogenous siderophore, N,N'-bis (2,3-dihydroxybenzoyl)-O-serylserine from a Vibrio sp. Growth was also stimulated by the addition of three exogenous siderophore extracts from siderophore-producing bacteria. Radioisotope studies using (59)Fe showed that the iron uptake ability of V0210 increased only with the addition of exogenous siderophores. Biosynthesis of a hydroxamate siderophore by V0210 was shown by paper electrophoresis and chemical assays for the detection of hydroxamates and catechols. An 85-kDa iron-regulated outer membrane protein was induced only under iron-limited conditions in the presence of exogenous siderophores. This is the first report of bacterial iron uptake through an induced siderophore in response to exogenous siderophores. Our results suggest that siderophores are necessary signaling compounds for growth and for iron uptake by some non-siderophore-producing marine bacteria under iron-limited conditions.

Animals↗

PubMed2001

Developmental changes in trial-to-trial variations in whisker barrel responses studied using intrinsic optical imaging: comparison between normal and de-whiskered rats.

We used an intrinsic optical imaging technique to examine postnatal developmental changes in the rat barrel response to a single whisker movement. We compared the optical response patterns between control and de-whiskered rats, from which whiskers were removed except for the D1 whisker just after birth. Barrel responses were evoked by D1-whisker movement stimulation, and the intrinsic optical signals were detected from the somatosensory cortex through the dura mater. In the control rats, the area of the barrel response decreased gradually as postnatal development proceeded from 2 to 7 wk, until reaching the adult pattern. On the other hand, in the de-whiskered rats, the barrel response area did not change during development and showed a larger size than in the control rats. We also compared the trial-to-trial variations in the barrel responses between the two groups. In the control rats, trial-to-trial variations in the optical responses were observed under the same conditions of whisker stimulation, and the extent of the variations decreased with postnatal development up to 7 wk. In the de-whiskered rats, trial-to-trial variations were also observed, but the extent was larger and unchanged during development. In both groups, the positions of the response area were the same with respect to the bregma. These results suggest that the decrease in the area and variations in the optical responses are caused by interactions of the corresponding whisker barrel with neighboring barrels and that these interactions are necessary for the developmental stabilization of the intracortical horizontal connections, which are widespread and have high plasticity in early postnatal periods.

Animals↗

PubMed2001

Optical mapping approaches to cardiac electrophysiological functions.

Recently, optical methods for monitoring membrane potential with fast voltage-sensitive dyes have been introduced as a powerful tool for studying cardiac electrical functions. These methods offer two principal advantages over more conventional electrophysiological techniques. One is that optical recordings may be made from very small cells that are inaccessible to microelectrode impalement, and the other is that multiple sites/regions of a preparation can be monitored simultaneously to provide spatially resolved mapping of electrical activity. The former has made it possible to record spontaneous electrical activities in early embryonic precontractile hearts, and the latter has been applied for mapping of the propagation patterns of electrical activities in the cardiac tissue. In this article, optical studies of the electrophysiological function of the vertebrate heart are reviewed.

Animals↗

PubMed2000

Optical detection of embryogenetic expression of vagal excitability in the rat brain stem.

We traced and identified the ontogenetic expression of neural excitability related to the vagus nerve in the embryonic rat brain stem. Multiple-site optical recordings of neural activities revealed two response areas in the E12 rat brain stem: one corresponding to the dorsal motor nucleus of the vagus nerve, and the other reflecting the activities of sensory nerve fibers. In embryos younger than E11, no optical response was identified, suggesting that excitability of the motoneurons and/or sensory nerve fibers is first generated no later than E12. A contour line map of the neural responses suggested that, in contrast to older embryos, the functional organization of the vagal nucleus is not orderly at the time of the initial expression of neural excitability.

Animals↗

PubMed2000

Srf(-/-) ES cells display non-cell-autonomous impairment in mesodermal differentiation.

The serum response factor (SRF) transcription factor is essential for murine embryogenesis. SRF+(-/-) embryos stop developing at the onset of gastrulation, lacking detectable mesoderm. This developmental defect may reflect cell-autonomous impairment of SRF(-/-) embryonic cells in mesoderm formation. Alternatively, it may be caused by a non-cell-autonomous defect superimposed upon inappropriate provision of mesoderm-inducing signals to primitive ectodermal cells. We demonstrate that the ability of SRF(-/-) embryonic stem (ES) cells to differentiate in vitro into mesodermal cells is indeed impaired. However, this impairment can be modulated by external, cell-independent factors. Retinoic acid, but not dimethylsulfoxide, permitted activation of the mesodermal marker gene T(Bra), which was also activated when SRF was expressed in SRF(-/-) ES cells. Embryoid bodies from SRF(-/-) ES cell aggregates also activated mesodermal marker genes, but displayed unusual morphologies and impairment in cavitation. Finally, in nude mice, Srf(-/-) ES cells readily differentiated into mesodermal cells of SRF(-/-) genotype, including cartilage, bone or muscle cells. We demonstrate that SRF contributes to mesodermal gene expression of ES cells and that SRF(-/-) ES cells display a non-cell-autonomous defect in differentiation towards mesoderm.

Animals↗