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Biomedical subjects

S Inoué

Publications and source records attributed to S Inoué.

At least 37 records · Page 2Linked to original sources
PubMed2000

Amino acid release from the rat oral pontine reticular nucleus across the sleep-wakefulness cycle.

To elucidate a functional involvement of amino acids in the regulation of vigilance states, we investigated time-course changes of glycine (Gly), glutamate (Glu), and glutamine (Gln) in the extracellular fluid of the oral pontine reticular nucleus (PnO) in freely behaving rats by an in vivo microdialysis technique. The average concentration of Gly in dialysates collected during rapid eye movement sleep (REMS) was significantly higher than that collected during wakefulness (W) and non rapid eye movement sleep (NREMS) under perfusion with normal artificial cerebrospinal fluid (aCSF). These state-dependent changes in Gly, however, failed to occur when the PnO was perfused with Ca2+-free aCSF. In contrast, the average concentration of neither Glu nor Gln exhibited state dependency under the perfusion with normal or Ca2+-free aCSF; however, the concentrations of all three amino acids showed no consistent time-sequential changes specifically in the first three minutes from the beginning of each vigilance state. Although it is suggested that in the PnO the average concentration of Gly was higher during REMS than during NREMS and W by a Ca2+-dependent mechanism, however, time-course changes in the amino acids are complicated and other mechanisms may also be influential.

Algorithms↗
PubMed1999

Architectural dynamics and gene replacement of coronin suggest its role in cytokinesis.

Coronin is a ubiquitous actin-binding protein representing a member of proteins portraying a WD-repeat sequence, including the beta-subunits of trimeric G-proteins. Coronin has been suggested to participate in multiple, actin-based physiological activities such as cell movement and cell division. Although the slow growth of coronin deletion mutants has been attributed to a defect in the fluid-phase uptake of nutrients, the exact role of coronin in cytoskeletal organization has not been elucidated. In this study, we examined a role of coronin in cytokinesis by analyzing the effect of coronin deletion on the actin cytoskeleton and its dynamic distribution using a green fluorescent protein (GFP)-coronin fusion protein. We show that GFP-coronin works similarly to natural coronin in vivo and in vitro. In live cells, GFP-coronin was found to accumulate into the cleavage furrow during cytokinesis. The fluorescence pattern suggests its association to the contractile ring throughout cytokinesis. Interestingly, a substantial amount of coronin was also bound to F-actin at the prospective posterior cortex of the daughter cells. We also show that the coronin null cells reveal irregularities in organization of actin and myosin II and divide by a process identical to the traction-mediated cytofission reported in myosin II mutants. Overall, this study suggests that coronin is essential for organizing the normal actin cytoskeleton and plays a significant role in cell division.

Actins↗
PubMed1999

Effects of prolactin on sleep in cyclic rats.

We previously demonstrated that pregnancy-associated sleep enhancement is correlated with the daily surges of prolactin (PRL). However, in spite of a surge of PRL in the proestrous night, a reduction of nocturnal sleep occurs in phase with proestrus. Therefore, to clarify the physiological role of PRL in sleep regulation during the estrous cycle, time-course changes in sleep were analyzed in bromocriptine (CB-154)-treated and/or PRL-supplemented female rats. Sleep patterns characteristic of proestrus-to-estrus were not affected by the CB-154 treatment. In contrast, nocturnal rapid eye movement sleep (REMS) was significantly increased after the PRL supplementation. The CB-154 treatment diminished the REMS-enhancing effect of PRL. Thus, the results suggest that the endogenous PRL is not crucial for the regulation of sleep during the estrous cycle, while exogenous PRL can enhance REMS.

Animals↗
PubMed1999

Colony-stimulating factors in rapid eye movement sleep and non-rapid eye movement sleep regulation.

Although several cytokines are known to be somnogenic, no study has been conducted to examine whether colony-stimulating factors (CSF) affect sleep. Therefore, we studied the effects of granulocyte-macrophage CSF (GM-CSF) and macrophage CSF (M-CSF) on sleep in rats and their possible mechanism of action. At the dose of 10 pmol, GM-CSF or M-CSF significantly increased both non-rapid eye movement and rapid eye movement (REM) sleep or REM sleep only when infused intracerebroventricularly during the dark period. When injected locally in the hypothalamus, GM-CSF and M-CSF increased nitric oxide (NO) production. Thus, NOergic neural signals in the hypothalamus may take part in the somnogenic action of CSF.

Animals↗
PubMed1999

Windows to dynamic fine structures, then and now.

How can we learn about dynamic fine structures that are far too small to be resolved with the light microscope without destroying the active living cell? Examples spanning the last half century show how polarized light microscopy can-and should-continue to provide an attractive window for such studies. Long before microtubules were found with electron microscopy, or their assembly properties were biochemically characterized in isolated cell-free systems, the dynamic fine structure of the mitotic spindle and assembly properties of its microtubules were revealed in living cells by polarized light microscopy. More recently, the polarizing microscope was improved, by invention of the new Pol-Scope, so that quantitative measurements of bire-fringence retardation and axes could be made rapidly for all image pixels independent of their birefringence axis orientation. In addition, the centrifuge polarizing microscope, just developed, allows us to follow the dynamic ordering of fine structures in living cells as they become stratified or restructured by centrifugal acceleration of up to ten thousand times gravity. The significance of these technological advances is discussed-Inoué, S. Windows to dynamic fine structures, then and now.

Cells↗
PubMed1998

Methylcobalamin amplifies melatonin-induced circadian phase shifts by facilitation of melatonin synthesis in the rat pineal gland.

Effects of methylcobalamin (methyl-B12), a putative drug for treating human circadian rhythm disorders, on the melatonin-induced circadian phase shifts were examined in the rat. An intraperitoneal injection of 1-100 microg/kg melatonin 2-h before the activity onset time (CT 10) induced phase advances of free-running activity rhythms in a dose-dependent manner (ED50=1.3 microg/kg). Injection of methyl-B12 (500 microg/kg) prior to melatonin (1 microg/kg) injection induced larger phase advances than saline preinjected controls, while the injection of methyl-B12 in combination with saline did not induce a phase advance. These results indicate amplification of melatonin-induced phase advances by methyl-B12. Pinealectomy abolished the phase alternating effect of methyl-B12, suggesting a site of action within the pineal gland. In fact, methyl-B12 significantly increased the content of melatonin in the pineal collected 2-h after activity onset (CT 14). In contrast, no difference in melatonin content was found at CT 10, indicating that the effect of methyl-B12 may be gated after the activity onset time when endogenous melatonin synthesis is known to increase. These results suggest that methyl-B12 amplifies melatonin-induced phase advances via an increase in melatonin synthesis during the early subjective night at a point downstream from the clock regulation.

Animals↗
PubMed1998

An animal model for pregnancy-associated sleep disorder.

We studied basic sleep changes in pregnant rats in order to understand how pregnancy alters sleep. In the rat, pregnancy increased nocturnal nonREM sleep across the entire period but increased REM sleep only in the early period. By the end of pregnancy, diurnal sleep was decreased, showing that pregnancy in rats causes biphasic sleep changes as it does in humans. Termination of pregnancy returned the enhanced sleep to baseline as in the estrous cycle. Therefore, significant changes in the pattern of sleep occurred during pregnancy in rats, suggesting that the animal model may contribute to understanding the mechanism of sleep disorders related to human pregnancy.

Animals↗
PubMed1997

Central administration of vitamin B12 aggravates cataplexy in canine narcolepsy.

Experimental evidence in canine narcolepsy suggests that central cholinergic systems are critically involved in the regulation of cataplexy, an abnormal manifestation of REM sleep atonia. In the current study, we found that intracerebroventricular perfusion of methyl-B12, (10(-5)-10(-2) M), significantly aggravated cataplexy and enhanced REM sleep in narcoleptic dogs. Choline, a direct precursor of acetylcholine, was also found to aggravate cataplexy, while cyano-B12, a vitamin B12 analog without methyl donating abilities, had no effect on cataplexy. Since both methyl-B12 and choline are reported to enhance acetylcholine synthesis, enhancement of the biosynthesis of acetylcholine may be involved in the effects observed in canine narcolepsy. Our results suggest that central administration of methyl-B12 has the potential to modulate both normal and pathological REM sleep.

Analysis of Variance↗
PubMed1997

Circadian rhythms in vitamin B12 content of the rat brain.

The whole brain content of vitamin B12 (VB12) in the rat was assayed by the chemiluminescent immunoassay at 4 h intervals in 12:12 h light-dark cycles (LD) and five different circadian times (CTs) under constant dim illumination (dim LL). In LD-entrained rats, the content of VB12 exhibited a day-night rhythm with a peak 2 h after the dark onset time and a trough 2 h before the light onset time. In freerunning rats under dim LL, the content of VB12 also exhibited a circadian variation with a peak ca. 2 h after the activity onset time (CT14) and a trough ca. 12 h after the activity onset time (CT 0). These findings clearly indicate that the brain VB12 content decreased during the active phase and increased during the resting phase regardless of the lighting schedule. On the other hand, the drinking behavior, as an index of the intake activity, was observed less frequently in the light phase of LD cycles and the resting phase of dim LL. Since most of the digested VB12 is known to be continuously stored in the liver, it is demonstrated that the rhythm of the brain content of VB12 may be caused by brain consumption of VB12 during the active phase and the transportation from the peripheral storage during the resting phase, independent of intake activities.

Animals↗
PubMed1997

Amoeboid movement anchored by eupodia, new actin-rich knobby feet in Dictyostelium.

To date, protrusion of pseudopodia has been considered to be primarily responsible for translocation of free-living amoebae and leukocytes of higher organisms. Although there is little question that the pseudopodium plays an important role, little attention has been given to the cortical structures that are responsible for cell-substratum anchorage in amoeboid movement. Here, we report on a new knobby foot-like structure in amoebae of a cellullar slime mold, Dictyostelium discoideum. These feet, each about 1 micron in diameter, appear transiently in multiple units at the base of certain pseudopodia where the amoeba contacts a partially deformable substrate. The feet were discovered, and their spatial and temporal behavior relative to pseudopodial anchorage and invasive locomotion were observed, by examining Dictyostelium amoebae using a DIC video microscope providing an 0.3 micron depth of field. Key evidence for the anchoring role of the knobby feet was obtained by investigating amoebae, flattened in a specially devised observation chamber, and attracted by chemotaxis towards 3',5' cyclic-adenosine monophosphate (cAMP). The cAMP was released by highly localized, pulsed UV-microbeam irradiation of caged cAMP. We show by indirect immunofluorescence that the knobby feet contain a high concentration of filamentous (F-) actin, myoB (a member of Dictyostelium myosin-I family), and alpha-actinin (an actin-binding protein). Interestingly, myoB exhibits a circular disposition around each foot. Neither myosin-II (conventional myosin) nor the 269 kD protein, which has been recently identified as a talin homologue of Dictyostelium [Kreitmeier et al., 1995: J. Cell Biol. 129:179-188], are concentrated at the feet. We propose that the knobby feet provide anchorage to the substratum needed by lamellipodia to exert projectile forces for invading narrow spaces or otherwise for a flattened amoeba to secure itself to the deformable substratum. Some forms of adhesion plaques in higher organisms such as "podosomes" or "invadopodia" may perform functions similar to the knobby feet, but appear to differ in life time, cytoskeletal organization and composition. We have named the knobby foot "eupodium."

Animals↗
PubMed1997

The role of microtubule assembly dynamics in mitotic force generation and functional organization of living cells.

This article summarizes the author's presentation at the Baylor Medical School Symposium on the Biophysics of Microtubules, held April 12 to 14, 1996, in Houston, Texas. It presents a brief historical sketch and discusses the role that assembly/disassembly of microtubules is likely to be playing in force generation for chromosome movement and related organellar positioning in living cells. The article starts out with how polarized light microscopy of living cells had laid the foundation for this concept in the 1950s and 1960s, but was then eclipsed for some 2 decades following the discovery of force generation by microtubule sliding powered by an ATP-hydrolyzing motor protein, dynein. The intriguing recent discoveries: that microtubules undergo dynamic instability; that they both assemble and disassemble right at the kinetochore where they are attached to the chromosome; and that assembling and disassembling microtubules can of themselves push and pull reasonable loads in model experiments, even in the absence of hydrolyzable nucleotides, have refocused serious attention on the probable role played by assembly/disassembly of microtubules. This mode of force generation may well be intricately coupled, and interact, with force-generating and/or dynamic attachment roles played by "motor" proteins, especially at the kinetochore.

Animals↗
PubMed1997

Vitamin B12 enhances GABA content but reduces glutamate content in the rat suprachiasmatic nucleus.

The present study attempted to elucidate effects of vitamin B12 (VB12) on the rat suprachiasmatic nucleus (SCN), which is the locus of the circadian clock in mammals, by evaluating the contents of its major neurotransmitters, gamma-aminobutyric acid (GABA) and glutamate (Glu). First, contents of these amino acids were analyzed from tissue samples prepared at three different circadian times (CTs) in free-running rats under constant dim illumination. The content of GABA was highest in the middle of the resting phase (CT 6), intermediate early in the active phase (CT 14), and lowest late in the active phase (CT 20), whereas Glu content indicated inverse circadian variations. Subsequently, effects of intracerebroventricularly infused VB12 were assayed at CT 20. Compared with the saline-infused control, the infusion of VB12 (30 nmol/30 microliters) at CT 12-15 significantly increased GABA content but decreased Glu content. The quantitative balance of these amino acids after VB12 infusion was similar to that at CT 6 in noninfused rats. These results suggest that infusion of VB12 to active rats shifts the ratio of SCN neurotransmitters toward that observed in resting rats.

Animals↗