Biomedical subjects

K Kanda

Publications and source records attributed to K Kanda.

At least 145 records · Page 8Linked to original sources

PubMed1989

Alpha-actinins, calspectin (brain spectrin or fodrin), and actin participate in adhesion and movement of growth cones.

We have used biochemical and immunocytochemical techniques to investigate the possible involvement of membrane cytoskeletal elements such as alpha-actinin, calspectin (brain spectrin or fodrin), and actin in growth cone activities. During NGF-induced differentiation of PC12 cells, alpha-actinin increased in association with neurite outgrowth and was predominantly distributed throughout the entire growth cone and the distal portion of neurites. Filopodial movements were sensitive to Ca2+ flux. Two types of alpha-actinin, with Ca2(+)-sensitive and -insensitive actin binding abilities, were identified in the differentiated cells. Ca2(+)-sensitive alpha-actinin and actin filaments were concentrated in filopodia. The Ca2(+)-insensitive protein was distributed from the body of the growth cone to the distal portion of neurites, corresponding to the substratum-adhesive sites. The location of calspectin in growth cones was similar to that of the Ca2(+)-insensitive alpha-actinin. These results are consistent with the hypothesis that Ca2(+)-sensitive alpha-actinin and actin filaments are involved in Ca2(+)-dependent filopodial movement and Ca2(+)-insensitive alpha-actinin and calspectin are associated with adhesion of growth cones.

Actinin↗

PubMed1989

The cytoskeletal architecture of the presynaptic terminal and molecular structure of synapsin 1.

We have examined the cytoskeletal architecture and its relationship with synaptic vesicles in synapses by quick-freeze deep-etch electron microscopy (QF.DE). The main cytoskeletal elements in the presynaptic terminals (neuromuscular junction, electric organ, and cerebellar cortex) were actin filaments and microtubules. The actin filaments formed a network and frequently were associated closely with the presynaptic plasma membranes and active zones. Short, linking strands approximately 30 nm long were found between actin and synaptic vesicles, between microtubules and synaptic vesicles. Fine strands (30-60 nm) were also found between synaptic vesicles. Frequently spherical structures existed in the middle of the strands between synaptic vesicles. Another kind of strand (approximately 100 nm long, thinner than the actin filaments) between synaptic vesicles and plasma membranes was also observed. We have examined the molecular structure of synapsin 1 and its relationship with actin filaments, microtubules, and synaptic vesicles in vitro using the low angle rotary shadowing technique and QF.DE. The synapsin 1, approximately 47 nm long, was composed of a head (approximately 14 nm diam) and a tail (approximately 33 nm long), having a tadpole-like appearance. The high resolution provided by QF.DE revealed that a single synapsin 1 cross-linked actin filaments and linked actin filaments with synaptic vesicles, forming approximately 30-nm short strands. The head was on the actin and the tail was attached to the synaptic vesicle or actin filament. Microtubules were also cross-linked by a single synapsin 1, which also connected a microtubule to synaptic vesicles, forming approximately 30 nm strands. The spherical head was on the microtubules and the tail was attached to the synaptic vesicles or to microtubules. Synaptic vesicles incubated with synapsin 1 were linked with each other via fine short fibrils and frequently we identified spherical structures from which two or three fibril radiated and cross-linked synaptic vesicles. We have examined the localization of synapsin 1 using ultracryomicrotomy and colloidal gold-immunocytochemistry of anti-synapsin 1 IgG. Synapsin 1 was exclusively localized in the regions occupied by synaptic vesicles. Statistical analyses indicated that synapsin 1 is located mostly at least approximately 30 nm away from the presynaptic membrane. These data derived via three different approaches suggest that synapsin 1 could be a main element of short linkages between actin filaments and synaptic vesicles, and between microtubules and synaptic vesicles, and between synaptic vesicles in the nerve terminals.(ABSTRACT TRUNCATED AT 400 WORDS)

Actins↗

PubMed1989

A novel phage genome integrated into a plasmid in Bacillus thuringiensis strain AF101.

Bacillus thuringiensis strain AF101 possesses a single plasmid (pAF101) with a molecular size of 42 MDa (69 kb). During plasmid curing experiments in strain AF101, we found that a phage (J7W-1) was induced by ethidium bromide treatment. Moreover, the phage genome (48 kb) hybridized only with pAF101 on a Southern blot of the DNA of a cleared lysate prepared from strain AF101. Comparison of the restriction patterns of pAF101 and J7W-1 phage DNA revealed that pAF101 contains not only the entire phage DNA but also a plasmid-specific DNA region. These results indicate that the J7W-1 genome has been stably integrated into pAF101 in strain AF101. Integration of the J7W-1 genome into a plasmid was also observed after phage infection of the type strain of B. thuringiensis subsp. israelensis.

Bacillus thuringiensis↗

PubMed1989

Potentiation of atrial natriuretic peptide-stimulated cyclic guanosine monophosphate formation by glucocorticoids in cultured rat renal cells.

1. The effect of steroid hormones on atrial natriuretic peptide (ANP)-stimulated cyclic guanosine monophosphate (cyclic GMP) formation was studied in cultured rat renal cells. 2. ANP increased cyclic GMP formation in a dose-dependent manner, while cyclic AMP was not changed by ANP. 3. Steroid hormones did not affect basal cyclic GMP levels in cultured rat renal cells. 4. Dexamethasone at 10(-8) M increased ANP (human and rat ANP)-stimulated cyclic GMP dose-dependently in cultured rat renal cells. Cortisol, corticosterone and aldosterone at a concentration of 10(-7) M also potentiated ANP-stimulated cyclic GMP formation, although triiodothyronine, oestradiol and testosterone were ineffective. Potentiation of ANP action by these steroids seems to parallel glucocorticoid activity. 5. Dexamethasone did not affect cyclic GMP formation stimulated by sodium nitroprusside which stimulates soluble guanylate cyclase in the cytosol. Therefore, the potentiating action of dexamethasone may be mediated through the action on particulate guanylate cyclase at the plasma membrane. 6. It is suggested that the diuretic action of glucocorticoids may, at least in part, be mediated through the potentiating effect of glucocorticoids on cyclic GMP response to ANP.

Animals↗

PubMed1989

Changes in properties of the medial gastrocnemius motor units in aging rats.

1. The properties of motor units were investigated in the medial gastrocnemius (MG) of old rats [27.5 +/- 1.6 (SD) mo old, n = 18]. Individual motor units were functionally isolated by ventral root fiber splitting and grading stimulus intensity. The muscle-unit portion of the motor unit was identified by the glycogen depletion method. The physiological properties of 77 motor units in 6 animals and the histological results of 7 slow-twitch (type S) muscle units were compared with data from motor units in the same muscle of middle-aged rats (12.8 +/- 1.6 mo old, n = 33). 2. The motor units were classified into four types of categories [FF (fast-twitch motor units with a fatigue index less than or equal to 0.5), FI (fast-twitch motor units with a fatigue index greater than 0.5 but less than 0.75), FR (fast-twitch motor units with a fatigue index greater than or equal to 0.75), S (slow-twitch motor units with a fatigue index greater than 0.75)] using the same criteria (i.e., presence or absence of the "sag" property and fatigability) used for middle-aged rats. No significant difference in the relative distributions of these unit types was detected, although the MG muscle in old rats exhibited a relatively high proportion of type S units and fewer type FR units. 3. The mean tetanic tensions for type FF + FI and FR units were significantly smaller than those in the middle-aged rats. On the other hand, type S motor units produced more tension than in the middle-aged rats. 4. The conduction velocity of motor axons was considerably slower in any unit type of old motor units, and the most marked change was found in type FR units. 5. The general morphological features of the old rat MG were fiber-type grouping, disseminated atrophic or angulated fibers, a decrease in the total number of muscle fibers, and an increase in the number of type I muscle fibers. The major distribution patterns of fibers of different types were the same as those in the middle-aged MG. 6. Seven type S units that produced large tetanic tension were depleted of glycogen in the muscle-unit portions. These units had a large innervation ratio compared with those in the middle-aged rats, whereas the mean cross-sectional area of muscle fibers and the calculated specific tension remained unaltered.(ABSTRACT TRUNCATED AT 400 WORDS)

Aging↗

PubMed1989

[Afferent projections to the uvula in the cat. I. Climbing fiber projections].

The inferior olive afferents to the cerebellar uvula were studied by means of retrograde axonal transport of horseradish peroxidase (HRP) in the cat. Following large and small injections of HRP into the uvula, the distributions of labeled cells in the inferior olive were investigated. As to the climbing fiber projection, it is revealed that there are six longitudinal parasagittal zone extending throughout the dorsal and ventral uvula. That is, the caudal aspect of the nucleus beta projects to a most medially oriented zone (caudal beta zone), the rostral aspect of the nucleus beta projects to a zone (rostral beta zone) oriented lateral to the caudal beta zone, the caudal aspect of the medial accessory olive (MAO) projects to a zone oriented lateral to the rostral beta zone, the dorsomedial cell column projects to a zone (dorsomedial cell column zone) oriented in the intermediate part of the uvula, the ventral lamella of the principal olive (vlPO) projects to a more lateral zone (vlPO zone), and the rostral aspect of the MAO projects to the most lateral zone (rostral MAO zone).

Afferent Pathways↗

PubMed1989

[Afferent projections to the uvula in the cat. II. Mossy fiber projections].

The mossy fiber projections to the uvula of the cerebellum were studied by means of retrograde axonal transport of horseradish peroxidase (HRP) in the cat. Following large and small injections into the uvula, distribution of the labeled cells in the brainstem nuclei was investigated. The results showed different afferent projections between the dorsal and ventral uvula. Major sources projecting to the dorsal uvula were the peduncular, paramedian, and lateral nuclei of the pontine nuclei. Labeled cells found in the pontine nuclei amounted to 81.6% of the total number of labeled cells in cat 1. On the other hand, major sources projecting to the ventral uvula were the caudal aspect of the medial and inferior vestibular nuclei, the x- and f-groups of the vestibular nuclei, the dorsal and central aspect of the superior vestibular nucleus, the rostral dorsomedial aspect of the paramedian nucleus of the pontine nuclei, the caudal aspect of the prepositus hypoglossal nucleus, and the infratrigeminal nucleus. Labeled cells in the vestibular nuclei amounted to 72.1% of the total number of the labeled cells in the cat 40. It was revealed that the lateral aspect of the ventral uvula receives inputs from the pontine nuclei, whereas the medial part of the ventral uvula receives inputs from the vestibular nuclei. Mediolateral differences were not found in the dorsal uvula. These mossy fiber zones were mediolaterally wide, and the dorsal uvula was different from the ventral uvula with regard to mossy fiber projection.

Afferent Pathways↗

PubMed1989

Localization of 4.1 related proteins in cerebellar neurons.

Localization of 4.1 related proteins in neurons was studied with immunofluorescence microscopy and with immunoelectron microscopy on ultrathin cryosections. In rat cerebellum, 4.1 immunoreactive proteins were demonstrated in Purkinje cell bodies, dendrites and other neurons in the cerebellar cortex. Some glial cells showed staining, but no labeling was found in myelinated axons of the white matter and of the glomeruli in the granule cell layer. At the ultrastructural level, the 4.1 related proteins were localized mainly in the cytoplasmic matrix, while some labeling was found underneath the plasma membrane. To determine whether 4.1 related proteins in neuronal cytoplasm exist as part of the cytoskeleton or not, PC12 cells cultured in the presence of nerve growth factor were stained with the anti-4.1 antibody. Since cytoplasmic staining was retained after detergent treatment, the 4.1 related proteins seem to exist as a component of the neural cell cytoskeleton. Localization of 4.1 related proteins during the postnatal development of the cerebellum was also studied. In Purkinje cells, localization of 4.1 related proteins changed according to the stages of the postnatal development. The present data suggest that 4.1 related proteins in neurons localized mainly in the cytoplasm and may play some role in organizing cytoskeletal networks in the cytomatrix. Their distribution is developmentally regulated in some neurons, possibly in relationship to their maturation in the cytoskeleton.

Animals↗

PubMed1988

Localization of pp60c-src in growth cone of PC12 cell.

By immunocytochemical and biochemical techniques, we observed the localization and expression of pp60c-src in nerve growth factor (NGF)-treated PC12 cells. Immunostaining of pp60c-src is detected in the neuronal soma and the tips of neurites (growth cones). Immunofluorescence in the neurites is less significant. High-resolution microscopy reveals that the location of pp60c-src in growth cone is in good agreement with the adhesive site of growth cone to the substratum. The pp60c-src kinase activity and the pp60c-src protein level increase 3.1- to 3.5-fold and 2.0-fold during differentiation of PC12 cells, respectively. The pp60c-src levels in the neurite fraction are also higher than those in the neuronal soma fraction. These results support the immunocytochemical finding that pp60c-src is localized in growth cones of differentiated PC12 cells. Furthermore, we discuss the possible role of pp60c-src in growth cone.

Adrenal Gland Neoplasms↗

PubMed1988

Molecular orbital study on the metabolic pathway through the diol epoxide form of carcinogenic benzene in comparison with benzo[a]pyrene.

The stabilization energy for the hydronium-ion-catalyzed hydrolysis of benzene diol epoxide (BDE) in the configuration of anti- or syn-form has been estimated by using the semi-empirical molecular orbital calculations with the CNDO/2 method. The values for the formation of carbonium ion from BDE are compared with those from benzo[a]pyrene, and it is suggested that the anti-form BDE belongs to a relatively strong reactive group with benzo[a]pyrene and benz[a]anthracene. The reactivity of BDE to the cation is completely different from that of polycyclic aromatic hydrocarbon diol epoxides (PAHDEs) from the viewpoint of the electronic structure; the cation from the anti-form BDE has a three center-four electron bond, whereas cations from PAHDEs do not have such a bond and the aromaticity still remains.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗

PubMed1988

Target neurons of floccular middle zone inhibition in medial vestibular nucleus.

Unitary activities of 288 neurons were recorded extracellularly in the medial vestibular nucleus (MV) in anesthetized cats. In 19 neurons, located in the rostral part of the MV adjacent to the stria acustica, floccular middle zone stimulation resulted in cessation of spontaneous discharges. Systematic microstimulation in the brainstem during recording of 16 of 19 target neurons of floccular middle zone inhibition revealed that the target neurons projected to the ipsilateral abducens nucleus (ABN), and not to the contralateral ABN nor the oculomotor nucleus. The conjugate ipsilateral horizontal eye movement elicited by middle zone stimulation may be mediated by this pathway to motoneurons and internuclear neurons in the ipsilateral ABN. In additional experiments, the MV neurons responding antidromically to ipsilateral ABN stimulation and orthodromically to ipsilateral 8 nerve stimulation were recorded extracellularly. In only 7 of 36 recorded neurons, middle zone stimulation depressed the orthodromic and spontaneous activities. Many neurons were free of floccular inhibition. As to the route of floccular inhibitory control over the vestibulo-ocular reflex (VOR) during visual-vestibular stimulation, we propose that the interaction of target and VOR relay neurons takes place at the ipsilateral ABN and modulates the VOR, in addition to well known Ito's proposal that the interaction of the floccular output and the VOR takes place at secondary vestibular neurons and modulates the VOR.

Animals↗

PubMed1988

Medial gastrocnemius motor nucleus in the rat: age-related changes in the number and size of motoneurons.

The age-related alterations in the number and size of alpha- and gamma-motoneurons were studied in the medial gastrocnemius (MG) motor nuclei in rats at four ages: young (5 months), middle aged (10-13 months), old (26 months), and very old (31 months). Small volumes (0.1-0.5 microliter) of 40% horseradish peroxidase (HRP) solution were injected into the cut MG nerve bilaterally by using glass micropipettes and a pressure injection system. The number, position, and soma size (average soma diameter) of MG motoneurons were determined by using photographic maps of each TMB-stained section. The total number of myelinated axons was counted in seven MG nerves from the same animals. The average soma diameters in each MG nucleus were distributed bimodally; cells with average diameter greater than 21.0-24.0 micron were presumed to be alpha-motoneurons and those with smaller diameters were presumed to be gamma. The mean number of presumed alpha-motoneurons was significantly less in the old and very old groups as compared with the young and middle-aged. In contrast, the number of presumed gamma-motoneurons was the same across age groups. The mean average soma diameter of both alpha- and gamma-motoneurons was smaller in the old animals. The apparent decrease in the total number of labeled motoneurons in old animals was also reflected in a decrease in myelinated axon counts. We conclude that there is a significant decrease in the absolute numbers of motoneurons in rats aged 26 months and older, with most of the decrease occurring among the larger alpha-motoneurons.

Aging↗

PubMed1988

Caldesmon: a common actin-linked regulatory protein in the smooth muscle and nonmuscle contractile system.

Caldesmon was originally purified from gizzard smooth muscle as a major calmodulin-binding protein which also interacts with actin filaments. It has an alternative binding ability to either calmodulin or actin filaments depending upon the concentration of Ca2+ ("flip-flop binding"). Two forms of caldesmon (Mr's in the range of 120-150 kDa and 70-80 kDa) have been demonstrated in a wide variety of smooth muscles and nonmuscle cells. Immunohistochemical studies suggest that caldesmon is colocalized with actin filaments in vivo. Considering its abundance, the Ca2+-dependent flip-flop binding ability to either calmodulin or actin filaments, and its intracellular localization, caldesmon is expected to be involved in contractile events. Recent results from our laboratory have led to the conclusion that caldesmon regulates the smooth muscle and nonmuscle actin-myosin interaction and the smooth muscle actin-high Mr actin-binding protein (ABP or filamin) interactin in a flip-flop manner. It might function in cell motility by regulating the contractile system.

Animals↗

PubMed1988

Tumor promoter induces reorganization of actin filaments and calspectin (fodrin or nonerythroid spectrin) in 3T3 cells.

We have used immunofluorescence, differential-interference-contrast, and interference-reflection microscopy to examine the translocation of actin filaments and calspectin (fodrin or nonerythroid spectrin) in 3T3 cells induced by phorbol 12-myristate 13-acetate (PMA). The two cytoskeletal proteins were observed to localize in dot structures that corresponded to the cell-substratum contact sites (focal contact) of the cytoplasmic surface of the plasma membrane. The induction of these cytoskeletal changes was specific for tumor promoters. High-resolution microscopy revealed that calspectin was intensely concentrated in ring-like structures surrounding actin dots. It was also located within the areas of actin dots, but to a lesser extent. Trifluoperazine and other phenothiazine derivatives inhibited the formation of those dot structures that appeared after the addition of PMA. Some serine protease inhibitors were also demonstrated to influence cytoskeletal changes by PMA. Our results provide evidence that calspectin is closely associated with actin filaments in dot structures induced by PMA. Possible mechanisms for these cytoskeletal changes produced by PMA are discussed.

Actins↗

PubMed1988

Effect of ovarian steroids on cyclic adenosine 3':5'-monophosphate production stimulated by arginine vasopressin in rat renal monolayer cultured cells.

Renal resistance to antidiuretic hormone (ADH) has been speculated to be a mechanism of transient nephrogenic diabetes insipidus occurring during late pregnancy. In order to study possible involvement of ovarian steroids in this mechanism, their effect on cyclic adenosine 3':5'-monophosphate (cAMP) response to arginine vasopressin (AVP) was examined utilizing rat and human renal medullary cells in monolayer culture. In both rat and human cells, estradiol significantly reduced cAMP response to AVP; estradiol at 1.84 x 10(-8) M, 1.84 x 10(-7) M and 1.84 x 10(-6) M decreased cAMP production stimulated by 10(-8) M AVP to 78 +/- 5%, 67 +/- 2% (P less than 0.05) and 52 +/- 1% (P less than 0.001) of the control in rat renal cells, respectively, and in human renal cells the effect of estradiol was comparable to that in rat cells. In rat renal cells, progesterone also reduced cAMP response to AVP dose-dependently; progesterone at 1.59 x 10(-7) M, 1.59 x 10(-6) M and 1.59 x 10(-5) M decreased cAMP production stimulated by 10(-8) M AVP to 87 +/- 1%, 72 +/- 5% (P less than 0.001) and 37 +/- 5% (P less than 0.001) of the control, respectively. On the other hand, corticosterone and dexamethasone at concentrations ranging from 10(-8) M to 10(-5) M and aldosterone at concentrations ranging from 10(-9) M to 10(-5) M did not alter cAMP response to AVP significantly. The suppressive effect of estradiol increased with time until six hours and thereafter it reached a plateau.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1987

Role of Mg2+ in the ribozyme system.

Recently, several kinds of catalytic activities have been identified from L-19 IVS RNA or 'ribozyme'. In the catalytic process of ribozyme, Mg2+ is required. We investigated the role of Mg2+ by molecular orbital calculation. Based on the results of the calculation, a model for the self-splicing reaction was constructed.

Animals↗

PubMed1987

Choline acetyltransferase and acetylcholinesterase activities in muscles of aged mice.

The activities of choline acetyltransferase (CAT) and acetylcholinesterase (AChE) were assayed in intact diaphragm, extensor digitorum longus (EDL), and soleus muscles or their homogenates of young (2-6 months) and aged (24-34 months) mice. CAT activity (per mg of protein) was significantly higher in diaphragm and soleus of old mice in comparison with the young but the age change in EDL was negligible. On the other hand, AChE activity (per mg of protein) was significantly higher in EDL of old mice but in diaphragm and soleus muscles the enzyme activity did not show any significant change statistically. The diaphragm muscle was divided into two fractions, one being neuromuscular (NM) fraction and the other the remainder of the muscle (M fraction). No appreciable change in the ratio of the enzyme activities of NM fraction to the one of M fraction was obtained between the young and aged preparations. Thus, it seems likely that there is an age-related change in CAT and AChE activities which might be affected by the degree to which muscle activity is maintained.

Acetylcholinesterase↗

PubMed1987

Expression of high and low molecular weight caldesmons during phenotypic modulation of smooth muscle cells.

We investigated the expression of two molecular weight forms of caldesmon in a wide range of tissues and cells. The distribution of high molecular weight caldesmon (h-caldesmon, Mr 120,000-150,000) was restricted to smooth muscles where it was found in large quantity. The low molecular weight protein (l-caldesmon, Mr 70,000-80,000) was widely distributed in nonmuscle tissues and cells. Therefore, the expression of h-caldesmon might be much more specific to smooth muscles. We then examined the expressional changes of two caldesmons during phenotypic modulation of smooth muscle cells (SMCs). In developing gizzards, the expression of caldesmons switched from the l- to the h-form. Contrarily, the expression turned from h- to l-caldesmon in association with dedifferentiation of aortic SMCs in primary culture. In agreement with these observations, the levels of those mRNAs that direct the synthesis of both caldesmons were apparently in proportion to the quantities of protein, as determined by use of an in vitro translation system. In addition, h-caldesmon in smooth muscle-like BC3H1 cells increased in its amount with a concomitant reduction of l-caldesmon following serum-depleted and contact-inhibited cytodifferentiation. These results suggest that the expressional changes of two caldesmons are closely correlated with the phenotypic modulation of SMCs.

Animals↗