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1.
J Comp Pathol ; 136(2-3): 145-55, 2007.
Article in English | MEDLINE | ID: mdl-17416234

ABSTRACT

A new type of inherited chondrodysplasia is described in Japanese Brown cattle, but the basic defects of the epiphyseal growth plate (EGP) in the limb long bones, and proliferation and differentiation of the chondrocytes in the EGP, are not yet understood. In the present study, the EGPs of the limb long bones in eight cases of chondrodysplasia and four normal (control) cattle were examined histologically and immunohistochemically. In the control cattle, proliferative chondrocytes (PCs) and hypertrophic chondrocytes (HCs) were arranged in columns parallel to the long axis of the bone, and HCs were situated on the metaphyseal side of the EGP. In all the affected cattle, many chondrocytes with a hypertrophic appearance were detected in the inner areas of the central portion of the EGP. The PC columns were short and arranged irregularly. Bone tissue and small blood vessels were found frequently in these areas. Six affected cattle showed complete EGP-closure. Backscattered electron (BSE) imaging showed that the calcified cartilage matrix was restricted to the lower region of the hypertrophic zone (HZ) of the EGP in the control cattle, while the calcified cartilage matrix and bone tissue were scattered in the inner areas of the EGP in all the chondrodysplastic cattle. Immunohistochemistry revealed type X collagen in the HCs and cartilage matrix of the HZ in the control cattle. In all the affected cattle, type X collagen was detected in apparently hypertrophic chondrocytes in the inner areas of the EGP. Type II collagen was detected in the entire EGP in all the affected cattle, as in the controls. BrdU (5-bromo-2'-deoxyuridine), injected intravenously 1h before euthanasia was detected in many PCs in the EGP in the control cattle; none, however, was detected in the central portion of the EGP in any affected animal. These observations indicate that differentiation into HCs and calcification of cartilage matrix occur in the inner areas of the central portion of the EGP in chondrodysplasia of Japanese Brown cattle. Differentiation into the HCs at this abnormal site may be caused by the inadequate proliferation and disorganization of the PCs. Premature EGP-closure, observed commonly in chondrodysplasia of Japanese Brown cattle, was thought to be caused by replacement of the calcified cartilage in the inner areas of the EGP by bone tissue.


Subject(s)
Cattle Diseases/pathology , Growth Plate/pathology , Osteochondrodysplasias/veterinary , Animals , Biomarkers/metabolism , Cattle , Cattle Diseases/metabolism , Cell Differentiation , Cell Proliferation , Chondrocytes/pathology , Collagen Type II/metabolism , Collagen Type II/ultrastructure , Collagen Type X/metabolism , Collagen Type X/ultrastructure , Female , Growth Plate/metabolism , Immunoenzyme Techniques/methods , Immunoenzyme Techniques/veterinary , Male , Osteochondrodysplasias/metabolism , Osteochondrodysplasias/pathology
2.
Ann N Y Acad Sci ; 1074: 90-6, 2006 Aug.
Article in English | MEDLINE | ID: mdl-17105906

ABSTRACT

Dihydropyrimidinase-related protein 2 (DRP-2 or DPYSL-2)mediates the intracellular response to collapsin, a repulsive extracellular guidance cue or axonal outgrowth. DRP-2 is also referred to as collapsin response mediator protein 2 (CRMP-2). We have previously demonstrated that the DRP-2 gene is associated with susceptibility to schizophrenia, but not to bipolar disorders. In addition, a genetic association was observed with paranoid-type schizophrenia, but not with hebephrenic-type schizophrenia. It has been well documented that repeated abuse of methamphetamine (METH) for a long period frequently produces psychotic symptoms, such as auditory hallucinations and delusions that are hardly distinguishable from those of paranoid-type schizophrenia. Therefore, we hypothesized that a certain genetic variant of the DRP-2 gene may affect individual vulnerability to the development of METH-induced psychosis. We examined the genetic association by a case-control method. The polymorphism *2236T>C in the 3' untranslated region of the DRP-2 gene, which has been shown to be a negative genetic risk factor for paranoid-type schizophrenia, was analyzed in 198 patients with METH psychosis and 221 corresponding healthy controls in a Japanese population. No significant association of the DRP-2 gene with METH psychosis was found. Neither did we find an association with the clinical phenotype of METH psychosis, such as the age of first consumption of METH, latency to development of psychosis after METH abuse, prognosis of psychosis after detoxification from METH use, complication of spontaneous relapse of psychosis without reconsumption of the drug, or multisubstance abuse status. These findings indicate that a genetic variant of the DRP-2 gene may not affect the risk of METH psychosis or any clinical phenotype of the disorder.


Subject(s)
Amphetamine-Related Disorders/genetics , Dopamine Agents/pharmacology , Gene Frequency , Intercellular Signaling Peptides and Proteins/genetics , Methamphetamine/pharmacology , Nerve Tissue Proteins/genetics , Polymorphism, Single Nucleotide , Adult , Amphetamine-Related Disorders/etiology , Case-Control Studies , Female , Humans , Male , Middle Aged
3.
J Hered ; 95(1): 81-4, 2004.
Article in English | MEDLINE | ID: mdl-14757734

ABSTRACT

Hymenopteran insects (sawflies, ants, bees, and wasps) have an unusual genetic system called haplodiploidy, where parthenogenetically produced haploid eggs become males, and fertilized, diploid eggs become females. Several hypotheses have been proposed to explain the mechanism of such sex determination, including control at a single polymorphic locus. From experiments of mother-son mating and using a genetic marker, we show that a single multiallele locus controls sex determination in the turnip sawfly (Athalia rosae). We estimated the number of alleles at this single locus in a field population by analyzing the rate of diploid males in the field and the rate of diploid males by random crossing in the laboratory. Only one diploid male was discovered in 1306 diploid larvae collected in the field. However, the number of alleles calculated by random crossing in the laboratory was 45-50. We suggest that the effective population size may be much larger than that from the areas where we collected larvae, and that there are mechanisms for avoiding inbreeding, including protogyny, dispersion, and sperm displacement by second-mated males.


Subject(s)
Alleles , Animals, Wild/genetics , Hymenoptera/genetics , Sex Determination Processes , Animals , Crosses, Genetic , Diploidy , Female , Male
4.
Cells Tissues Organs ; 175(1): 43-50, 2003.
Article in English | MEDLINE | ID: mdl-14605494

ABSTRACT

Laminar bone or primary plexiform tissue, not Haversian bone, shows an alternative concentric pattern of laminar-bone units or plates around the bone marrow periphery of long bones, although the laminar bone is gradually replaced by osteons during the growth period. One laminar-bone unit is constructed with a hypercalcified line in the center, woven bone on both sides of the line, and lamellar bone with laminated appositional lines. Such a laminar bone showing a homogeneous calcification has been reported in young calves and some young large animals, but it has not been reported in foals although a previous report proposed that the bone structure was distinguishable from plexiform tissue. In this study, we compared young calves with foals by backscattered electron imaging mainly of transverse ground sections of mid-diaphysis. Foals had many hypercalcified lines arranged concentrically around the bone marrow periphery, which were similar to those of young calves. However, rows of cylindrical osteon-like structures with Haversian canal-like canals running along the long-bone axis were arranged between the concentric hypercalcified lines. Each Haversian canal-like structure was enclosed with laminated appositional rings of lamellar bone deposited on the woven bone. In the developing period, the bone units containing the concentric hypercalcified lines were basically equal to the laminar-bone units. The osteon-like structures or 'pseudo-osteons' were gradually replaced by 'true osteons' during the growth period. The blood vessels in the Haversian canal-like canals of foals ran along the long-bone axis, whereas the blood vessels in the concentrically prolonged bone cavities of young calves ran transversely to obliquely against the long-bone axis. Thus, the long-bone cortex of foals showing an alternative concentric pattern of a row of the osteon-like structures arranged between the hypercalcified lines will be histologically classified into a variety of laminar bone caused by the different arrangement of blood vessels. Such a laminar bone may have a biomechanical structure against physical stress, especially the modified laminar bone of foals with osteon-like structures, when compared with the typical concentric laminar bone of young calves and also Haversian bone possessing variously calcified numerous osteons caused by bone remodeling.


Subject(s)
Cattle/anatomy & histology , Horses/anatomy & histology , Tibia/cytology , Tibia/growth & development , Animals , Animals, Newborn , Biological Evolution , Bone Development , Decalcification Technique , Haversian System/cytology , Species Specificity , Tibia/blood supply
5.
J Electron Microsc (Tokyo) ; 50(1): 57-64, 2001.
Article in English | MEDLINE | ID: mdl-11291961

ABSTRACT

By using a dentifrice or toothpaste for sensitive teeth, the brushing-induced effects on dentinal tubule occlusion and abrasion of human sound dentin were investigated with a scanning electron microscope and a scanning laser microscope. The dentifrice contained diatomaceous earth and silica as abrasives and strontium chloride hexahydrate as an active ingredient. Thirty dentin pieces of human premolar teeth with an average of 20% occluded dentinal tubules were attached to resin plates and exposed to the oral cavities of five adult subjects for 2, 4, and 8 weeks. Brushing with and without dentifrice was performed 1 min per day, respectively. Brushing with the dentifrice gradually decreased the mean average of occluded tubules from about 91 to 77% during 2 to 8 weeks, although there were no significant differences among the individual values. However, the mean abrasive loss of the dentin surfaces brushed with dentifrice significantly increased from about 52 to 143 microm in depth. The brushed surfaces of the dentin showed a rough topography with numerous toothbrush scratches but no organic pellicle was found. On the other hand, brushing without dentifrice caused about 99% of the dentinal tubules to occlude in 2 and 4 weeks and 100% in 8 weeks. The brushed dentin surfaces at 8 weeks were entirely covered with organic pellicle containing fine mineral granules derived from saliva, and the abrasive loss was about 1.4 microm in mean depth. Such results indicate that brushing with abrasive dentifrices for sensitive teeth remarkably erodes dentin, and suggest that the brushing should cause the dentinal tubules to open again for a certain period of time.


Subject(s)
Dentifrices/adverse effects , Dentin/pathology , Tooth Abrasion , Adolescent , Adult , Child , Dentin/ultrastructure , Female , Humans , Male , Microscopy, Confocal , Microscopy, Electron, Scanning
6.
J Electron Microsc (Tokyo) ; 49(1): 195-202, 2000.
Article in English | MEDLINE | ID: mdl-10791437

ABSTRACT

Using backscattered electron (BSE) imaging and scanning electron microscopy, subsequent to a combined treatment of sodium hypochlorite (NaOCl) and ethylenediamine tetra-acetic acid (EDTA) or only with EDTA etching, we observed some structures of non-collagenous calcified matrixes with the aim of revealing the correlation of deposition between calcification degree and organic amount. In human tooth enamel, the NaOCl-EDTA method eroded more intensively the hypocalcified prisms of enamel tufts containing a relatively large amount of EDTA-insoluble organic matter than the hypercalcified normal prismatic enamel containing a small amount of the organic matter. Afibrillar cementum, one of the non-collagenous calcified tissues similar to the enamel, has been reported to consist of organic-rich and poor incremental lamellae. The BSE imaging showed an alternation pattern of hypocalcification and hypercalcification. The hypocalcified lamellae were retained by EDTA etching, while the hypercalcified lamellae showed a resistance against the NaOCl-EDTA method. In the non-collagenous calcareous concretions of human pineal body, organic-rich and poor, and hyper- and hypocalcified incremental lamellae have been reported. The deposition pattern of calcification degree and organic amount was similar to that in afibrillar cementum, and the hypercalcified lamellae showed a resistance against the NaOCl-EDTA method. In conclusion, the high and the lower calcified regions of non-collagenous calcified matrixes contained smaller and larger amounts of EDTA-insoluble organic matter respectively. Moreover, scanning electron microscopy subsequent to the NaOCl-EDTA method corresponding to the BSE imaging clearly showed fine calcified structures compared with the BSE imaging.


Subject(s)
Dental Cementum/ultrastructure , Dental Enamel/ultrastructure , Edetic Acid/pharmacology , Microscopy, Electron, Scanning/methods , Pineal Gland/ultrastructure , Sodium Hypochlorite/pharmacology , Calcification, Physiologic , Humans
7.
J Vet Med Sci ; 62(3): 293-9, 2000 Mar.
Article in English | MEDLINE | ID: mdl-10770602

ABSTRACT

The histological disorders related to the focal disappearance of the epiphyseal growth plate were examined histochemically in the proximal tibia of rats administered a high dose of vitamin A. Animals were given 100,000 IU/100 g body weight/day of vitamin A for 5 days from 4 weeks after birth (VA rats) or given deionized water as control and sacrificed on Day 12 and 19 of the experiment. Tibiae were examined by immunohistochemistry for type I, II and X collagens, lectin-histochemistry for Helix pomatia and backscattered electron imaging. On Day 12, the abnormally developed calcified cartilage matrix was detected within the epiphyseal growth plate in VA rats. The uncalcified cartilage matrix contained type I collagen but lacked type II collagen. In addition, the eroded regions accompanied with numerous osteoclasts and osteoblasts were detected in the epiphyseal growth plate. On day 19, eroded regions penetrated the epiphyseal growth plate to result in its focal disappearances with the eroded surfaces entirely covered with bone tissue in VA rats. These findings suggested that the cartilage matrix of the epiphyseal growth plate was abnormally calcified and showed the phenotypes like bone matrix. The eroded regions of the epiphyseal growth plate seemed to be caused by the invasion of osteoclasts into the altered cartilage matrix and might develop to the focal disappearances by the modeling or remodeling due to action of osteoclasts and osteoblasts.


Subject(s)
Growth Plate/drug effects , Vitamin A/pharmacology , Animals , Cattle , Dose-Response Relationship, Drug , Helix, Snails , Immunohistochemistry , Rats , Rats, Wistar , Tibia/cytology , Tibia/growth & development , Vitamin A/administration & dosage
8.
J Biol Chem ; 275(1): 605-12, 2000 Jan 07.
Article in English | MEDLINE | ID: mdl-10617657

ABSTRACT

We examined the effect of 2-arachidonoylglycerol, an endogenous cannabinoid receptor ligand, on the intracellular free Ca(2+) concentrations in HL-60 cells that express the cannabinoid CB2 receptor. We found that 2-arachidonoylglycerol induces a rapid transient increase in intracellular free Ca(2+) concentrations in HL-60 cells. The response was affected by neither cyclooxygenase inhibitors nor lipoxygenase inhibitors, suggesting that arachidonic acid metabolites are not involved. Consistent with this notion, free arachidonic acid was devoid of any agonistic activity. Importantly, the Ca(2+) transient induced by 2-arachidonoylglycerol was blocked by pretreatment of the cells with SR144528, a CB2 receptor-specific antagonist, but not with SR141716A, a CB1 receptor-specific antagonist, indicating the involvement of the CB2 receptor but not the CB1 receptor in this cellular response. G(i) or G(o) is also assumed to be involved, because pertussis toxin treatment of the cells abolished the response. We further examined the structure-activity relationship. We found that 2-arachidonoylglycerol is the most potent compound among a number of naturally occurring cannabimimetic molecules. Interestingly, anandamide and N-palmitoylethanolamine, other putative endogenous ligands, were found to be a weak partial agonist and an inactive ligand, respectively. These results strongly suggest that the CB2 receptor is originally a 2-arachidonoylglycerol receptor, and 2-arachidonoylglycerol is the intrinsic natural ligand for the CB2 receptor that is abundant in the immune system.


Subject(s)
Arachidonic Acids/metabolism , Cannabinoids , Glycerides/metabolism , Palmitic Acids/metabolism , Receptors, Drug/agonists , Amides , Calcium Signaling , Camphanes/pharmacology , Cyclohexanols/pharmacology , Cyclooxygenase Inhibitors/pharmacology , Drug Interactions , Endocannabinoids , Ethanolamines , HL-60 Cells , Humans , Ligands , Lipoxygenase Inhibitors/pharmacology , Molecular Mimicry , Pertussis Toxin , Piperidines/pharmacology , Polyunsaturated Alkamides , Pyrazoles/pharmacology , RNA, Messenger/analysis , Receptors, Cannabinoid , Receptors, Drug/antagonists & inhibitors , Receptors, Drug/classification , Receptors, Drug/genetics , Rimonabant , Structure-Activity Relationship , Virulence Factors, Bordetella/pharmacology
9.
J Electron Microsc (Tokyo) ; 48(2): 167-72, 1999.
Article in English | MEDLINE | ID: mdl-10356789

ABSTRACT

Automatic toothbrushing with a commercial dentifrice containing hydroxyapatite (HAP) crystals was performed on the tangential polished surfaces of sound human enamel, mainly consisting of biological apatite similar to HAP, for 10 min in vitro. The X-ray diffraction peaks of HAP, brushite (DCPD), and monetite (DCP) crystals were detected from the dentifrice. After brushing, the enamel surfaces were observed with a scanning electron and a confocal scanning laser microscope. The brushing caused larger abrasive loss and more remarkable roughness of the enamel surfaces following the broad traces of brush bristles and the exposure of prism structures than brushing with a dentifrice containing only DCPD, which we previously reported. We claim that the fine granular-shaped HAP crystals of the dentifrice indicated as an active ingredient for preventing enamel caries possess stronger abrasivity of sound enamel than the DCPD and DCP as abrasives on account of their Mohs hardness values rather than sizes and shapes. The HAP crystals of dentifrices may not occlude the small defects of early caries enamel, but erode them more strongly as an abrasive than the other abrasives.


Subject(s)
Dental Enamel/ultrastructure , Dentifrices/chemistry , Durapatite , Tooth Abrasion/etiology , Toothbrushing/adverse effects , Adolescent , Calcium Phosphates , Child , Humans , Microscopy, Confocal , Microscopy, Electron, Scanning , X-Ray Diffraction
10.
J Vet Med Sci ; 61(3): 233-8, 1999 Mar.
Article in English | MEDLINE | ID: mdl-10331194

ABSTRACT

The initial disorders of the epiphyseal growth plate cartilage were immunohistochemically examined in the proximal tibia of rats administered a high dose of vitamin A. Male Wistar rats were given 100,000 IU/100 g body weight/day of vitamin A for administration periods of 1 to 5 days (Day 1 to 5) from 4 weeks after birth or were given deionized water and used as control. They were sacrificed after 5-bromo-2'-deoxyuridine (BrdU) injection on Day 1 to Day 5 to remove the tibiae. The tibiae were processed for immunohistochemical examinations using antibodies against type I, II, X collagens and BrdU. BrdU-incorporated chondrocytes and type X collagen-negative area were reduced since Day 2 and type X collagen-positive area was reduced since Day 4. The cartilage matrix partially lost type II collagen and deposited type I collagen in the epiphyseal growth plate near the periosteum on Day 5. These findings suggest that a high dose of vitamin A initially disturbed the differentiation from resting to proliferating chondrocytes, subsequently inhibited the differentiation from proliferating to hypertrophic chondrocytes, caused the chondrocytes to deviate from the process of normal differentiation, and finally resulted in the deformation of the epiphyseal growth plate.


Subject(s)
Cartilage Diseases/chemically induced , Growth Plate/drug effects , Vitamin A/administration & dosage , Vitamin A/toxicity , Animals , Cell Differentiation/drug effects , Cell Division/drug effects , Chondrocytes/cytology , Chondrocytes/drug effects , Immunohistochemistry , Male , Rats , Rats, Wistar
11.
J Vet Med Sci ; 61(2): 101-6, 1999 Feb.
Article in English | MEDLINE | ID: mdl-10081745

ABSTRACT

The transverse sections of radius diaphysis in an 11-year-old giant Holstein cow with dermal dysplasia of a collagen disorder-related skin fragility (Cow 1), probably based on increasing turnover of the dermal collagen as reported previously, were morphologically and physico-chemically investigated. Cow 1 had about one and a half times as much as the body weight of normal Holstein cows, aged 5 to 6.5 years with stabilized growth. The bone samples were compared with those of a 12-year-old Holstein cow as controls (Cow 2). It has been reported that the long-bone diaphysis of young calves and some herbivorous dinosaurs are occupied with laminar bone showing a concentric appositional formation, and that such a laminar bone is characteristically seen during the growing period of some farm animals and large dogs that show very rapid growth rates. Cow 1 had a smaller number of osteons than Cow 2 in the outer-half layer of the diaphysis, and showed an intermediate type between Cow 2 and a 1-year-old Holstein ox in the entire layers, although their bone volumes were similar among them. There were no significant differences in Ca and P concentrations and the Vickers microhardness values between the bone matrix of Cow 1 and Cow 2. The bone-collagen fibrils of Cow 1 showed uneven diameters and a disordered arrangement. Thus, there may be some relation in collagen formation between the bone matrix of Cow 1 and the dermis. From the remaining volume of laminar bone, Cow 1, aged 11 years, had probably shown growth until quite recently, so that we consider that Cow 1 became a giant animal, in the same way as some herbivorous dinosaurs.


Subject(s)
Cattle Diseases/metabolism , Haversian System/metabolism , Radius/metabolism , Skin Diseases/veterinary , Animals , Bone and Bones/ultrastructure , Cattle , Collagen/ultrastructure , Diaphyses/metabolism , Diaphyses/pathology , Female , Male , Microscopy, Electron, Scanning/veterinary , Radius/pathology , Skin Diseases/complications , Skin Diseases/metabolism
12.
J Biol Chem ; 274(5): 2794-801, 1999 Jan 29.
Article in English | MEDLINE | ID: mdl-9915812

ABSTRACT

An endogenous cannabimimetic molecule, 2-arachidonoylglycerol, induces a rapid, transient increase in intracellular free Ca2+ concentrations in NG108-15 cells through a cannabinoid CB1 receptor-dependent mechanism. We examined the activities of 24 relevant compounds (2-arachidonoylglycerol, its structural analogues, and several synthetic cannabinoids). We found that 2-arachidonoylglycerol is the most potent compound examined so far: its activity was detectable from as low as 0.3 nM, and the maximal response induced by 2-arachidonoylglycerol exceeded the responses induced by others. Activities of HU-210 and CP55940, potent cannabinoid receptor agonists, were also detectable from as low as 0.3 nM, whereas the maximal responses induced by these compounds were low compared with 2-arachidonoylglycerol. Anandamide was also found to act as a partial agonist in this assay system. We confirmed that free arachidonic acid failed to elicit a response. Furthermore, we found that a metabolically stable ether-linked analogue of 2-arachidonoylglycerol possesses appreciable agonistic activity, although its activity was apparently lower than that of 2-arachidonoylglycerol. We also confirmed that pretreating cells with various cannabinoid receptor agonists nullified the response induced by 2-arachidonoylglycerol, whereas pretreating cells with other neurotransmitters or neuromodulators did not affect the response. These results strongly suggested that the cannabinoid CB1 receptor is originally a 2-arachidonoylglycerol receptor, and 2-arachidonoylglycerol is the intrinsic physiological ligand for the cannabinoid CB1 receptor.


Subject(s)
Arachidonic Acids/chemistry , Cannabinoids/metabolism , Receptors, Drug/metabolism , Animals , Arachidonic Acids/metabolism , Benzoxazines , Cannabinoids/antagonists & inhibitors , Cell Line , Cyclohexanols/pharmacology , Dronabinol/analogs & derivatives , Dronabinol/pharmacology , Endocannabinoids , Models, Chemical , Morpholines/pharmacology , Naphthalenes/pharmacology , Piperidines/pharmacology , Polyunsaturated Alkamides , Pyrazoles/pharmacology , Rats , Receptors, Cannabinoid , Receptors, Drug/agonists , Rimonabant , Structure-Activity Relationship
13.
J Vet Med Sci ; 60(7): 815-21, 1998 Jul.
Article in English | MEDLINE | ID: mdl-9713809

ABSTRACT

Epiphyseal growth plates of proximal tibiae in rats with high doses of vitamin A (V-A) were observed. Of 4 groups, each consisting of 5 rats, three groups were given V-A at doses, IU/100 g by body weight/day, of 50,000, 100,000, and 150,000, respectively. The other group rats were given no V-A (control). Rats were administered V-A for the 5 days from 4 weeks after birth and sacrificed at 12 weeks after birth. Three rats of the 150,000 IU group died during the period of observation. The decalcified sections were stained with hematoxylin-eosin or toluidine blue. In the ground sections, microradiography, backscattered electron imaging, and energy-dispersive X-ray microanalysis were performed. These observations suggest that the local disappearance of epiphyseal growth plates under high doses of V-A goes in the order of the increased doses through the process of (1) calcified cartilage areae appearing in the resting cell zone, (2) some of the calcified areae extending in the growth plate towards the diaphysial side, (3) bone tissue replacing the calcified areae, and (4) the local disappearing of the growth plate. Such a local disappearance may be formed in the stressed proximal regions of tibiae.


Subject(s)
Growth Plate/pathology , Hypervitaminosis A/pathology , Animals , Animals, Newborn , Calcification, Physiologic , Growth Plate/cytology , Growth Plate/diagnostic imaging , Hypervitaminosis A/diagnostic imaging , Male , Radiography , Rats , Rats, Wistar , Tibia/cytology , Tibia/diagnostic imaging , Tibia/pathology , Time Factors , Vitamin A/administration & dosage , Vitamin A/toxicity
14.
FEBS Lett ; 429(2): 152-6, 1998 Jun 12.
Article in English | MEDLINE | ID: mdl-9650580

ABSTRACT

The molecular species compositions of monoacylglycerols obtained from various rat tissues were examined by reverse-phase high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) analyses. We confirmed that 2-arachidonoylglycerol, an endogenous cannabinoid receptor agonist, is one of the most abundant molecular species of monoacylglycerols in the brain. Substantial amounts of 2-arachidonoylglycerol were also found in the liver, spleen, lung and kidney, but the levels were considerably lower than that in the brain. We found that a small amount of 2-arachidonoylglycerol was generated in a brain homogenate during incubation in the absence of Ca2+. Importantly, the generation of 2-arachidonoylglycerol was markedly augmented in the presence of Ca2+, suggesting that Ca2+ plays a key role in regulation of the generation of 2-arachidonoylglycerol in this tissue.


Subject(s)
Arachidonic Acids , Brain/metabolism , Calcium/metabolism , Glycerides/metabolism , Receptors, Drug/agonists , Animals , Cannabinoids/metabolism , Chromatography, High Pressure Liquid , Endocannabinoids , Gas Chromatography-Mass Spectrometry , Male , Rats , Rats, Wistar , Receptors, Cannabinoid , Tissue Distribution
15.
Arch Biochem Biophys ; 354(2): 303-10, 1998 Jun 15.
Article in English | MEDLINE | ID: mdl-9637740

ABSTRACT

Changes in the levels of various molecular species of N-acylethanolamine in CdCl2-administered rat testis were examined. We found that the levels of various N-acylethanolamines including anandamide (N-arachidonoylethanolamine), an endogenous cannabinoid receptor ligand, were dramatically increased in CdCl2-admin-istered rat testis. Such changes were particularlyprominent for saturated and monoenoic species such as N-palmitoyl species (39-fold at 9 h) and N-stearoyl species (21-fold at 9 h), compared with unsaturated fatty acid-containing species such as anandamide (5-fold at 9 h). Noticeably, increased levels were observed of not only N-acylethanolamines but also several species of N-acylphosphatidylethanolamine, potential precursors for N-acylethanolamines. We confirmed that the rat testis microsomal fraction contains phosphodiesterase activity catalyzing the release of N-acylethanolamine from N-acylphosphatidylethanolamine and transacylase activity catalyzing the formation of N-acylphosphatidylethanolamine from phosphatidylethanolamine and phosphatidylcholine. These enzyme activities were not dramatically different in the microsomal fraction obtained from CdCl2-administered rat testis compared with that in the case of control rat testis, at least when estimated in cell-free assay systems, suggesting that the accessibility of the substrates to the enzymes may be increased in CdCl2-administered rat testis to generate a large amount of N-acylethanolamine. Possible pathophysiological implications of the augmented generation of N-acylethanolamine including anandamide in CdCl2-administered rat testis were discussed.


Subject(s)
Arachidonic Acids/metabolism , Ethanolamines/metabolism , Orchitis/metabolism , Testis/metabolism , Animals , Cadmium Chloride , Disease Models, Animal , Endocannabinoids , Male , Polyunsaturated Alkamides , Rats , Rats, Wistar
16.
J Electron Microsc (Tokyo) ; 47(1): 57-65, 1998.
Article in English | MEDLINE | ID: mdl-9602527

ABSTRACT

Pulp stones of human teeth usually contain biological apatite. In the dental pulp of a cow, aged 12 years, we observed spherulitic pulp stones, up to approximately 200 microns in diameter, consisting of radially arranged crystals with several kinds of light microscopy and scanning electron microscopy including the secondary electron and the backscattered electron signals. The crystals extending fan-like towards the periphery showed a rectangular shape, and they had subunits of elongated crystallites and fine slits of NaOCl-dissolved substances, probably including organic matter and amorphous mineral. By using energy-dispersive X-ray microanalysis, large amounts of Ca and P, and trace elements of Na, Mg, Cl, and Fe were detected from the stones. The calcium phosphate crystals were identified as biological brushite by shape, Ca/ P molar ratio, and electron diffraction pattern. Core regions of the stones containing granular structures, probably erythrocytes, were always present in blood vessels, and rectangular crystals frequently penetrated the vessel walls. Under the decalcified sections, the nidi were completely or incompletely dissolved, but the peripheries showed a radial arrangement of erythrocytes. Thus, the nidi may be thrombus or necrotic blood cells including erythrocytes. Such calcospherulites in the dental pulp of a cow were similar to the spherulitic pulp stones in human deciduous teeth reported previously, however, their nidi were present in different regions.


Subject(s)
Calcium Phosphates , Cattle Diseases/pathology , Dental Pulp Calcification/veterinary , Animals , Cattle , Crystallization , Dental Pulp/blood supply , Dental Pulp/ultrastructure , Dental Pulp Calcification/pathology , Dentin/ultrastructure , Erythrocytes/ultrastructure , Incisor/ultrastructure , Microradiography , Microscopy, Electron , Microscopy, Electron, Scanning , Microscopy, Polarization
17.
Biochem Biophys Res Commun ; 243(3): 838-43, 1998 Feb 24.
Article in English | MEDLINE | ID: mdl-9501013

ABSTRACT

Human vascular endothelial cells were found to generate and release 2-arachidonoylglycerol, an endogenous cannabinoid receptor ligand, upon stimulation with thrombin or A23187. We confirmed that vascular smooth muscle cells as well as endothelial cells possess cannabinoid CB1 receptor mRNA. 2-Arachidonoylglycerol, generated in vascular tissues, may play an important role in modulating vascular tone through acting on the cannabinoid CB1 receptor expressed on vascular smooth muscle cells, endothelial cells as well as peripheral nerve terminals.


Subject(s)
Arachidonic Acids , Endothelium, Vascular/chemistry , Glycerides/analysis , Muscle, Smooth, Vascular/chemistry , RNA, Messenger/analysis , Receptors, Drug/genetics , Aorta , Arachidonic Acid/metabolism , Calcimycin/pharmacology , Cells, Cultured , Diglycerides/metabolism , Endocannabinoids , Endothelium, Vascular/drug effects , Endothelium, Vascular/metabolism , Glycerides/metabolism , Glycerides/pharmacology , Humans , Kinetics , Muscle, Smooth, Vascular/drug effects , Muscle, Smooth, Vascular/metabolism , Receptors, Cannabinoid , Receptors, Drug/drug effects , Receptors, Drug/physiology , Thrombin/pharmacology , Tritium , Umbilical Veins
18.
J Biochem ; 122(4): 890-5, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9399597

ABSTRACT

The effects of delta9-tetrahydrocannabinol and 2-arachidonoylglycerol on the intracellular free Ca2+ concentration ([Ca2+]i) in NG108-15 cells were examined in detail. We found that delta9-tetrahydrocannabinol induces a rapid, modest increase in [Ca2+]i. The response was detectable with 3 nM delta9-tetrahydrocannabinol. We also found that very low concentrations of 2-arachidonoylglycerol elicit a rapid, more prominent increase in [Ca2+]i. Such a response was observed not only in NG108-15 cells but also in N18TG2 cells. The response induced by 2-arachidonoylglycerol in either NG108-15 cells or N18TG2 cells was abolished by pretreatment of the cells with a cannabinoid CB1 receptor specific antagonist, SR141716A, suggesting that 2-arachidonoylglycerol interacts with the CB1 receptor to induce the response. The results of an experiment involving a phospholipase C inhibitor suggested that phospholipase C is involved in the rapid increase in [Ca2+]i induced by 2-arachidonoylglycerol. We also found that 1(3)-arachidonoylglycerol exhibits similar activity to that of 2-arachidonoylglycerol, although its activity at low concentrations was somewhat weak compared with that of 2-arachidonoylglycerol. We further confirmed that several structural analogues of 2-arachidonoylglycerol were less active compared with 2-arachidonoylglycerol. These results suggest that the structure of 2-arachidonoylglycerol is strictly recognized by the CB1 receptor, which raises the possibility that the CB1 receptor is originally a 2-arachidonoylglycerol receptor.


Subject(s)
Calcium/metabolism , Diglycerides/metabolism , Receptors, Drug/metabolism , Cell Line , Protein Conformation , Receptors, Cannabinoid , Receptors, Drug/chemistry
19.
Biochem Biophys Res Commun ; 233(1): 207-10, 1997 Apr 07.
Article in English | MEDLINE | ID: mdl-9144424

ABSTRACT

2-Arachidonoylglycerol was found to inhibit the depolarization-induced increase in [Ca2+]i in NG108-15 cells differentiated with prostaglandin E1 and theophylline in a dose-dependent manner. Such an effect appears to be rather specific to polyunsaturated fatty acid-containing monoacylglycerols such as 2-arachidonoylglycerol. Neither 2-palmitoylglycerol nor free arachidonic acid exhibited appreciable inhibitory activity. These observations raise the possibility that 2-arachidonoylglycerol attenuates the increase in [Ca2+]i, thereby modulating several neural functions in this type of cell.


Subject(s)
Arachidonic Acids , Calcium/metabolism , Glioma/metabolism , Glycerides/pharmacology , Neuroblastoma/metabolism , Neurotransmitter Agents/pharmacology , Endocannabinoids , Glioma/pathology , Hybrid Cells , Neuroblastoma/pathology , Tumor Cells, Cultured
20.
Biochem Biophys Res Commun ; 229(1): 58-64, 1996 Dec 04.
Article in English | MEDLINE | ID: mdl-8954083

ABSTRACT

Low concentrations of 2-arachidonoylglycerol were found to induce rapid, transient elevation of intracellular free Ca2+ in NG108-15 cells (EC50 was 150 nM). Free arachidonic acid, 2-palmitoylglycerol, 2-oleoylglycerol, 2-linoleoylglycerol and 2-docosahexaenoylglycerol were inactive. Anandamide acted as a partial agonist. Importantly, desensitization was observed upon sequential challenge with 2-arachidonoylglycerol. Furthermore, cross-desensitization was observed between 2-arachidonoylglycerol and WIN 55212-2, a cannabinoid receptor agonist. Pretreatment of the cells with SR141716A, a cannabinoid receptor antagonist, abolished the activities of both 2-arachidonoylglycerol and WIN 55212-2. These results strongly suggest that 2-arachidonoylglycerol and WIN 55212-2 bind to a common cannabinoid receptor to elicit cellular responses and that 2-arachidonoylglycerol has some physiological role in nervous tissues.


Subject(s)
Calcium/metabolism , Glycerides/pharmacology , Neurons/drug effects , Receptors, Drug/metabolism , Arachidonic Acids/pharmacology , Benzoxazines , Cannabinoids/metabolism , Dose-Response Relationship, Drug , Drug Interactions , Endocannabinoids , Glioma , Glycerides/chemistry , Hybrid Cells , Ligands , Morpholines/pharmacology , Naphthalenes/pharmacology , Neuroblastoma , Piperidines/pharmacology , Platelet Activating Factor/pharmacology , Polyunsaturated Alkamides , Pyrazoles/pharmacology , Receptors, Cannabinoid , Receptors, Drug/agonists , Receptors, Drug/antagonists & inhibitors , Rimonabant
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