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1.
Rei NAKANO Kazuya EDAMURA Tomohiro NAKAYAMA Kenji TESHIMA Kazushi ASANO Takanori NARITA Ken OKABAYASHI Hiroshi SUGIYA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(1):27-35
We investigated the in
vitro differentiation of canine bone marrow stromal cells (BMSCs) into voltage-
and glutamate-responsive neuron-like cells. BMSCs were obtained from the bone marrow of
healthy beagle dogs. Canine BMSCs were incubated with the basal medium for neurons
containing recombinant human basic fibroblast growth factor (bFGF; 100
ng/ml). The viability of the bFGF-treated cells was
assessed by a trypan blue exclusion assay, and the morphology was monitored. Real-time
RT-PCR was performed to evaluate mRNA expression of neuronal, neural stem cell and glial
markers. Western blotting and immunocytochemical analysis for the neuronal markers were
performed to evaluate the protein expression and localization. The Ca2+
mobilization of the cells was evaluated using the Ca2+ indicator Fluo3 to
monitor Ca2+ influx. To investigate the mechanism of bFGF-induced neuronal
differentiation, the fibroblast growth factor receptor inhibitor, the phosphoinositide
3-kinase inhibitor or the Akt inhibitor was tested. The bFGF treatment resulted in the
maintenance of the viability of canine BMSCs for 10 days, in the expression of neuronal
marker mRNAs and proteins and in the manifestation of neuron-like morphology. Furthermore,
in the bFGF-treated BMSCs, a high concentration of KCl and L-glutamate induced an increase
in intracellular Ca2+ levels. Each inhibitor significantly attenuated the
bFGF-induced increase in neuronal marker mRNA expression. These results suggest that bFGF
contributes to the differentiation of canine BMSCs into voltage- and glutamate-responsive
neuron-like cells and may lead to the development of new cell-based treatments for
neuronal diseases. 相似文献
2.
Mizuki MAEDA Sayaka KITAUCHI Tetsushi HIRANO Yoshinori IKENAKA Misaki NISHI Asuka SHODA Midori MURATA Youhei MANTANI Yoshiaki TABUCHI Toshifumi YOKOYAMA Nobuhiko HOSHI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(3):542
Recently, it has been reported that neonicotinoid pesticides (NNs) are transferred from mother to child and are assumed to affect the next generation, but the behavioral effects of NN exposure at different developmental stages have not been investigated. We exposed mice to no-observed-adverse-effect level (NOAEL) doses of clothianidin (CLO) during the fetal and lactational period, and then evaluated the neurobehavioral effects in juvenile and adult mice. Significant increases in anxiety-like behavior and locomotor activity were observed in juveniles and adults, respectively, and neuronal activity and neurogenesis in the hippocampal dentate gyrus were affected in both stages. These results suggest that fetal and lactational exposure to CLO may inhibit neurogenesis and cause different behavioral abnormalities at different developmental stages. 相似文献
3.
Sayaka KITAUCHI Mizuki MAEDA Tetsushi HIRANO Yoshinori IKENAKA Misaki NISHI Asuka SHODA Midori MURATA Youhei MANTANI Toshifumi YOKOYAMA Yoshiaki TABUCHI Nobuhiko HOSHI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(4):746
Recently, developmental exposure to clothianidin (CLO) has been shown to cause reproductive toxicity in male mice, but the effects in female mice remain to be clarified. Pregnant C57BL/6N mice were given a no-observed-adverse-effect-level (NOAEL) dose of CLO until weaning. We then examined ovaries of 3- or 10-week-old female offspring. In the CLO-administered group, morphological changes, a decrease in the immunoreactivity of the antioxidant enzyme glutathione peroxidase 4 (GPx4), and activation of genes in the steroid hormone biosynthesis pathway were observed in 3-week-old mice, and decreases of GPx4 immunoreactivity, 17OH-progesterone and corticosterone levels were observed in 10-week-old mice, along with high rates of infanticide and severe neglect, providing new evidence that developmental exposure to CLO affects juvenile and adult mice differently. 相似文献
4.
Yan Sai Junfeng Chen Feng Ye Yuanpeng Zhao Zhongmin Zou Jia Cao Zhaojun Dong 《Journal of toxicologic pathology》2013,26(2):149-157
Rotenone is an inhibitor of mitochondrial complex I that produces a model of Parkinson’s
disease (PD), in which neurons undergo dopamine release dysfunction and other features. In
neurons, exocytosis is one of the processes associated with dopamine release and is
dependent on Ca2+ dynamic changes of the cell. In the present study, we have
investigated the exocytosis of dopamine and the involvement of Ca2+ in dopamine
release in PC12 cells administrated with rotenone. Results demonstrated that rotenone led
to an elevation of intracellular Ca2+ through Ca2+ influx by opening
of the voltage-gated Ca2+ channel and influenced the soluble N-ethylmaleimide
attachment protein receptor (SNARE) proteins expression (including syntaxin,
vesicle-associated membrane protein 2 (VAMP2) and synaptosome-associated
protein 25 (SNAP-25)); pretreatment with a blocker of L-type voltage-activated
Ca2+ channels (nifedipine) decreased the intracellular dopamine levels and
ROS formation, increased the cell viability and enhanced the neurite outgrowth and
exocytosis of synaptic vesicles. These results indicated that the involvement of
intracellular Ca2+ was one of the factors resulting in suppression of dopamine
release suppression in PC12 cells intoxicated with rotenone, which was associated with the
rotenone-induced dopamine neurotoxicity. 相似文献
5.
6.
Jumpei YASUDA Muneyoshi OKADA Hideyuki YAMAWAKI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(10):1289-1291
Endostatin, a fragment of collagen XVIII, is known as an endogenous angiogenesis
inhibitor, and its serum concentration increases in various cardiovascular diseases.
T-type Ca2+ channel, low voltage-activated Ca2+ channel, is not
expressed in adult ventricular myocytes. Re-expression of T-type Ca2+ channels
in cardiac myocytes is thought to be involved in the development of cardiac hypertrophy.
We examined the effects of endostatin on T-type Ca2+ channel current by
whole-cell patch clamp technique in freshly isolated adult guinea pig ventricular
myocytes, which exceptionally express T-type Ca2+ channels. Although endostatin
300 ng/ml had no effect on L-type Ca2+
current, it significantly inhibited T-type Ca2+ current. These data indicate
that endostatin can be an endogenous inhibitor of T-type Ca2+ channels in the
cardiac myocytes. 相似文献
7.
Ehab MOSSAAD Masahito ASADA Daichi NAKATANI Noboru INOUE Naoaki YOKOYAMA Osamu KANEKO Shin-ichiro KAWAZU 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(1):53-58
Bovine babesiosis is a livestock disease known to cause economic losses in
endemic areas. The apicomplexan parasite Babesia bovis is able to invade
and destroy the host’s erythrocytes leading to the serious pathologies of the disease,
such as anemia and hemoglobinuria. Understanding the egress mechanisms of this parasite is
therefore a key step to develop new therapeutic strategies. In this study, the possible
involvement of Ca2+ in the egress of B. bovis merozoites from
infected erythrocytes was investigated. Egress was artificially induced in
vitro using calcium ionophore A23187 and thapsigargin to increase
Ca2+ concentration in the cytosol of the parasite cells. The increased
intracellular Ca2+ concentration following these treatments was confirmed using
live cell Ca2+ imaging with confocal laser scanning microscopy. Based on our
findings, we suggest a Ca2+ signalling pathway in the egress of B.
bovis merozoites. 相似文献
8.
Yumi Umeda Tatsuya Kasai Misae Saito Hitomi Kondo Tadao Toya Shigetoshi Aiso Hirokazu Okuda Tomoshi Nishizawa Shoji Fukushima 《Journal of toxicologic pathology》2013,26(2):131-140
To evaluate pulmonary toxicity of multi-walled carbon nanotubes (MWCNTs), F344 rats of
both sexes were exposed by inhalation to 0.2, 1 or 5 mg/m3 MWCNT aerosol for 6
h/day, 5 days/week for 2 weeks using a whole-body exposure system. At the end of the
2-week exposure period, one-half of the rats were necropsied, and at the end of an
additional 4-week postexposure period, the remaining rats were necropsied. MWCNTs were
deposited in the lungs of all MWCNT-exposed groups and mostly remained in the lungs
throughout the 4-week postexposure period. Granulomatous changes in the lung were found in
the rats exposed to 5 mg/m3 MWCNTs, and these changes were slightly aggravated
at the end of the 4-week postexposure period. In the bronchoalveolar lavage fluid (BALF),
the numbers of neutrophils, percentages of bi- and multinucleated alveolar macrophages,
levels of ALP activity and concentrations of total protein and albumin were elevated in
the rats exposed to 1 and 5 mg/m3 MWCNTs. At the end of the 4-week postexposure
period, the values of the BALF parameters tended to remain elevated. In addition, goblet
cell hyperplasias in the nasal cavity and nasopharynx were observed in the rats exposed to
1 and 5 mg/m3 MWCNTs, but these lesions had largely regressed by the end of the
postexposure period. Based on the histopathological and inflammatory changes, the
no-observed-adverse-effect level (NOAEL) for inhalation of MWCNTs for 2 weeks was 0.2
mg/m3. 相似文献
9.
Yoshio MINAMI Seiko YAMANO Minako KAWAI Atsushi HIRAGA Hirofumi MIYATA 《Journal of Equine Science》2009,20(3):33-40
To find a new parameter indicating muscle fitness in Thoroughbred horses, we examined
time-dependent recovery of glycogen content and sarcoplasmic reticulum (SR)
Ca2+-ATPase activity of skeletal muscle after intensive treadmill running.
Two repeated 50-sec running sessions (13 m/sec) were performed on a flat treadmill
(approximately 90%VO2max). Muscle samples of the middle gluteal muscle were
taken before exercise (pre) and 1 min, 20 min, 60 min, and 24 hr after exercise. Muscle
fiber type composition was determined in the pre muscle samples by immunohistochemical
staining with monoclonal antibody to myosin heavy chain. SR Ca2+-ATPase
activity of the muscle and glycogen content of each muscle fiber type were determined with
biochemical analysis and quantitative histochemical staining, respectively. As compared to
the pre value, the glycogen content of each muscle fiber type was reduced by 15–27% at 1
min, 20 min, and 60 min after the exercise and recovered to the pre value at 24 hr after
exercise test. These results indicate that 24 hr is enough time to recover glycogen
content after short-term intensive exercise. The mean value of the SR
Ca2+-ATPase activity showed a slight decrease (not significant) immediately
after exercise, and complete recovery at 60 min after exercise. There were no significant
relationship between the changes in glycogen content of each muscle fiber type and SR
Ca2+-ATPase. Although further studies are needed, SR Ca2+-ATPase
is not a useful parameter to detect muscle fitness, at least in Thoroughbred horses. 相似文献
10.
Takeharu KANEDA Erika SAITO Hidenori KANDA Norimoto URAKAWA Kazumasa SHIMIZU 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(10):1269-1274
The effects of papaverine on carbachol (CCh) -and high K+- induced contraction
in the bovine abomasum were investigated. Papaverine inhibited CCh (1 µM)
-and KCl (65 mM) -induced contractions in a concentration-dependent manner. Forskolin or
sodium nitroprusside inhibited CCh-induced contractions in a concentration-dependent
manner in association with increases in the cAMP or cGMP contents, whereas papaverine
increased cGMP contents only at 30 µM. Changes in the extracellular
Ca2+ from 1.5 mM to 7.5 mM reduced verapamil-induced relaxation in high
K+-depolarized muscles, but papaverine-induced relaxation did not change.
Futhermore, papaverine (30 µM) and NaCN (300 µM)
decreased the creatine phosphate contents. These results suggest that the relaxing effects
of papaverine on the bovine abomasum are mainly due to the inhibition of aerobic energy
metabolism. 相似文献
11.
12.
Dugar DELGERMURUN Shigeo ITO Toshio OHTA Soichiro YAMAGUCHI Ken-ichi OTSUGURO 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2016,78(1):71-76
Chemoreceptor cells aggregating in clusters in the chicken thoracic aorta
contain 5-hydroxytryptamine (5-HT) and have voltage-dependent ion channels and nicotinic
acetylcholine receptors, which are characteristics typically associated with neurons. The
aim of the present study was to investigate the effects of 5-HT uptake inhibitors,
fluvoxamine, fluoxetine and clomipramine (CLM), and amphetamine derivatives,
p-chloroamphetamine (PCA) and methamphetamine (MET), on endogenous 5-HT
outflow from the isolated chick thoracic aorta in vitro. 5-HT was
measured by using a HPLC system with electrochemical detection. The amphetamine
derivatives and 5-HT uptake inhibitors caused concentration-dependent increases in
endogenous 5-HT outflow. PCA was about ten times more effective in eliciting 5-HT outflow
than MET. The 5-HT uptake inhibitors examined had similar potency for 5-HT outflow. PCA
and CLM increased 5-HT outflow in a temperature-dependent manner. The outflow of 5-HT
induced by PCA or 5-HT uptake inhibitors was independent of extracellular Ca2+
concentration. The 5-HT outflow induced by CLM, but not that by PCA, was dependent on the
extracellular NaCl concentration. These results suggest that the 5-HT uptake system of
5-HT-containing chemoreceptor cells in the chicken thoracic aorta has characteristics
similar to those of 5-HT-containing neurons in the mammalian central nervous system
(CNS). 相似文献
13.
Yingxiao Fu Jianhong Gu Yi Wang Yan Yuan Xuezhong Liu Jianchun Bian Zong-Ping Liu 《Journal of veterinary science (Suw?n-si, Korea)》2015,16(2):151-156
The purpose of this study was to determine whether the Ca2+ signaling pathway is involved in the ability of osteoprotegerin (OPG) to inhibit osteoclast differentiation and maturation. RAW264.7 cells were incubated with macrophage colony-stimulating factor (M-CSF) + receptor activator of nuclear factor-κB ligand (RANKL) to stimulate osteoclastogenesis and then treated with different concentrations of OPG, an inhibitor of osteoclast differentiation. The intracellular Ca2+ concentration [Ca2+]i and phosphorylation of Ca2+/calmodulin-dependent protein kinase II (CaMKII) in the different treatment groups were measured by flow cytometry and Western blotting, respectively. The results confirmed that M-CSF + RANKL significantly increased [Ca2+]i and CaMKII phosphorylation in osteoclasts (p < 0.01), and that these effects were subsequently decreased by OPG treatment. Exposure to specific inhibitors of the Ca2+ signaling pathway revealed that these changes varied between the different OPG treatment groups. Findings from the present study indicated that the Ca2+ signaling pathway is involved in both the regulation of osteoclastogenesis as well as inhibition of osteoclast differentiation and activation by OPG. 相似文献
14.
Shusei Mizushima Tomohiro Sasanami Tamao Ono Norio Kansaku Asato Kuroiwa 《The Journal of Poultry Science》2022,59(2):175
We previously reported that egg activation in Japanese quail is driven by two distinct types of intracellular Ca2+ ([Ca2+]i): transient elevations in [Ca2+]i induced by phospholipase Czeta 1 (PLCZ1) and long-lasting spiral-like Ca2+ oscillations by citrate synthase (CS) and aconitate hydratase 2 (ACO2). Although the blockade of inositol 1,4,5-trisphosphate receptors (ITPRs) before microinjections of PLCZ1, CS, and ACO2 cRNAs only prevented transient increases in [Ca2+]i, a microinjection of an agonist of ryanodine receptors (RYRs) induced spiral-like Ca2+ oscillations, indicating the involvement of both ITPRs and RYRs in these events. In this study, we investigated the isoforms of ITPRs and RYRs responsible for the expression of the two types of [Ca2+]i increases. RT-PCR and western blot analyses revealed that ITPR1, ITPR3, and RYR3 were expressed in ovulated eggs. These proteins were degraded 3 h after the microinjection of PLCZ1, CS, and ACO2 cRNAs, which is the time at which egg activation was complete. However, degradation of ITPR1 and ITPR3, but not RYR3, was initiated 30 min after a single injection of PLCZ1 cRNA, corresponding to the time of the initial Ca2+ wave termination. In contrast, RYR3 degradation was observed 3 h after the microinjection of CS and ACO2 cRNAs. These results indicate that ITPRs and RYR3 differentially mediate in creases in [Ca2+]i during egg activation in Japanese quail, and that downregulation of ITPRs and RYR3-mediated events terminate the initial Ca2+ wave and spiral-like Ca2+ oscillations, respectively. 相似文献
15.
The water and electrolyte balance of Bulinus (Bulinus) tropicus, a freshwater pulmonate, was determined when subjected to different sublethal concentrations of copper sulphate. It was found that the lethal dose (LD50), which is the dose lethal to 50% of the snails, is 1,0 ppm in water of pH 8,5. Increasing dosages disturb the ionic and water balance. Haemolymph concentrations of Na + , Ca2+ and Cl- decrease markedly. The rates of ion influx and efflux for Ca2+ , Na + , Cl- and 3H20, measured With their corresponding radioisotopes, were drastically changed. The results indicate that the physiological effects of copper are not limited to particular organs or tissues but probably occur throughout the snail body. The possible effect of copper sulphate on the in- tegumental exchange mechanisms for Ca2+, HCO3-, Na+ , K+ and Cl- are discussed. 相似文献
16.
Antolín Morillo Rodríguez Cristina Ortega Ferrusola Beatriz Macías García Jose A. Tapia Fernando J. Peña 《Journal of Equine Veterinary Science》2013
We studied the effect of 1,2-bis-(o-aminophenoxy)-ethane-N,N,N′,N′-tetraacetic acid, tetraacetoxymethyl ester (BAPTA-AM) on the outcome of cryopreservation of stallion spermatozoa and whether reextension of thawed sperm in a more physiological and Ca2+-containing medium might improve the characteristics of thawed stallion spermatozoa. Individual ejaculates from six stallions were collected and split into three subsamples. The first two samples were supplemented with the membrane-permeable Ca2+ chelator BAPTA-AM at final concentrations of 5 and 10 μM, respectively, while the third subsample served as control. After 4 weeks of storage, samples were thawed in a water bath at 37°C and evaluated using flow cytometry and computer-assisted sperm analysis (CASA). In a second experiment, in order to determine whether restoring Ca2+ could improve sperm quality after cryopreservation, thawed semen was washed by centrifugation and resuspended in Tyrode's complete medium. BAPTA-AM supplementation did not modify the outcome of cryopreservation; however, changing the spermatozoa from INRA 96 to Tyrode's complete medium resulted in significant improvements in the percentages of live sperm and total motility post thaw. 相似文献
17.
《动物营养(英文)》2021,7(4):1115-1123
This study aimed to explore the mechanism underlying arginine-promoted myogenesis of myoblasts. C2C12 cells were cultured with a medium containing 0.1, 0.4, 0.8, or 1.2 mmol/L arginine, respectively. Cell proliferation, viability, differentiation indexes, cytoplasmic Ca2+ concentration, and relative mRNA expression levels of myogenic regulatory factors (MRF) and key Ca2+ channels were measured in the absence or presence of 2 chemical inhibitors, dantrolene (DAN, 10 μmol/L) and nisoldipine (NIS, 10 μmol/L), respectively. Results demonstrated that arginine promoted myogenic differentiation and myotube formation. Compared with the control (0.4 mmol/L arginine), 1.2 mmol/L arginine upregulated the relative mRNA expression levels of myogenin (MyoG) and Myomaker at d 2 during myogenic induction (P < 0.05). Cytoplasmic Ca2+ concentrations were significantly elevated by arginine supplementation at d 2 and 4 (P < 0.05). Relative mRNA expression levels of Ca2+ channels including the type 1 ryanodine receptor (RyR1) and voltage-gated Ca2+ channel (Cav1.1) were upregulated by 1.2 mmol/L arginine during 2-d myogenic induction (P < 0.01). However, arginine-promoted myogenic potential of myoblasts was remarkably compromised by DAN and NIS, respectively (P < 0.05). These findings evidenced that the supplementation of arginine promoted myogenic differentiation and myotube formation through increasing cytoplasmic Ca2+ concentration from both extracellular and sarcoplasmic reticulum Ca2+. 相似文献
18.
Su Li Liang Qian Jun Zhao Xiang Chen Li Ya Ping Jin Yi Peng Wang Xiao Hua Su Wei Jun Guan Yue Hui Ma 《Journal of veterinary science (Suw?n-si, Korea)》2011,12(2):133-142
Mammalian oocyte maturation and early embryo development processes are Ca2+-dependent. In this study, we used confocal microscopy to investigate the distribution pattern of Ca2+ and its dynamic changes in the processes of bovine oocytes maturation, in vitro fertilization (IVF), parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) embryo development. During the germinal vesicle (GV) and GV breakdown stage, Ca2+ was distributed in the cortical ooplasm and throughout the oocytes from the MI to MII stage. In IVF embryos, Ca2+ was distributed in the cortical ooplasm before the formation of the pronucleus. In 4-8 cell embryos and morulas, Ca2+ was present throughout the blastomere. In PA embryos, Ca2+ was distributed throughout the blastomere at 48 h, similar to in the 4-cell and 8-cell phase and the morula. At 6 h after activation, there was almost no distribution of Ca2+ in the SCNT embryos. However, Ca2+ was distributed in the donor nucleus at 10 h and it was distributed throughout the blastomere in the 2-8 cell embryos. In this study, Ca2+ showed significant fluctuations with regularity of IVF and SCNT groups, but PA did not. Systematic investigation of the Ca2+ location and distribution changes during oocyte maturation and early embryo development processes should facilitate a better understanding of the mechanisms involved in oocyte maturation, reconstructed embryo activation and development, ultimately improving the reconstructed embryo development rate. 相似文献
19.
We analyzed the histopathological changes and the number of motor neurons (MNs) in the lumbar spinal cord of Cu/Zn superoxide dismutase transgenic (SOD1G93ATg) mice, which are frequently used as a disease model of amyotrophic lateral sclerosis (ALS). In SOD1G93ATg mice, hyaline inclusions and foamy vacuoles in the neuronal cell body were observed at 7 weeks of age before neurologic symptoms, and large vacuoles, spheroid formation, and nerve cell aggregation became prominent after 13 weeks of age. The number of healthy MNs was 28.7 to 37.1 cells/animal in wild-type mice and 9.3 to 13.6 cells/animal in transgenic (Tg) mice. Furthermore, the number of MNs, including degenerative neurons, in Tg mice was 27.3–36.1 cells/animal at 18 weeks of age and 17.8–19.6 cells/animal at 21 weeks of age. The present results provide useful information for the development of drugs in ALS treatment. 相似文献
20.
A. Burmańczuk T. Grabowski M. Osypiuk B. Polska C. Kowalski 《Journal of veterinary pharmacology and therapeutics》2017,40(5):552-560
The aim of this study was to perform a comparative analysis of the characteristics of cloxacillin (CLO) (MRL of withdrawal in bovine milk is 30 ng/g) after a single intramammary (IMM) dose in the dry period (DP) and lactation (LP), and to establish a high‐performance liquid chromatography (HPLC) analytical method for CLO detection in milk. The research was conducted on a group of 10 cows in DP and 10 in LP. A single dose of 600 mg of CLO was administrated by the IMM route for a single quarter in DP and 500 mg for a single quarter in LP. CLO concentration was analyzed by HPLC. CLO was monitored at a wavelength of 206 nm. Pharmacokinetic calculations were performed using Phoenix® WinNonlin® 6.4 software. The calibration curve was linear over the range of 13.03–28 019.00 ng/g with the coefficient of determination R2 > 0.999. CLO withdrawal in both the LP and DP group had a biphasic nature. The total CLO elimination in the DP and LP group was reached after 36 and 6.5 days, respectively. A quantitative and confirmatory method for the determination of CLO in fresh milk has been established. We have confirmed that the withdrawal of CLO in the DP group is not a linear process and has a stepwise character. 相似文献