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Solid phase microextraction(SPME)in combination with high-resolution mass spectrometry was employed for the determination of metabolomic profile of mouse melanoma growth within in vitro 2D,in vitro 3D,and in vivo models.Such multi-model approach had never been investigated before.Due to the low-invasiveness of SPME,it was possible to perform time-course analysis,which allowed building time profile of biochemical reactions in the studied material.Such approach does not require the multiplication of samples as subsequent analyses are performed from the very same cell culture or from the same individual.SPME already reduces the number of animals required for experiment;therefore,it is with good concordance with the 3Rs rule(replacement,reduction,and refinement).Among tested models,the largest number of compounds was found within the in vitro 2D cell culture model,while in vivo and in vitro 3D models had the lowest number of detected compounds.These results may be connected with a higher metabolic rate,as well as lower integrity of the in vitro 2D model compared to the in vitro 3D model resulting in a lower number of compounds released into medium in the latter model.In terms of in vitro-in vivo extrapolation,the in vitro 2D model performed more similar to in vivo model compared to in vitro 3D model;however,it might have been due to the fact that only compounds secreted to medium were investigated.Thus,in further experiments to obtain full metabolome infor-mation,the intraspheroidal assessment or spheroid dissociation would be necessary.
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Objective To establish a C57BL/6 mouse model of intestinal infection induced by S.Typhimurium.Methods In order to improve the infectious sensitivity of S.Typhimurium, C57BL/6 mice were intragastrically given 5% (w/v) NaHCO3.Then mice were challenged with S.Typhimurium.The health condition, survival and body weight of mice were observed from day 0 to day 7 after the bacterial infection.The pathological changes were also examined.Results the mice challenged with S.Typhimurium showed decreased body weight and typical clinical signs, including in appetence, piloerection and low survival rate.Macroscopic dissection revealed that intestinal hyperemia and swelling were founded in the mice challenged with S.Typhimurium.Histopathology showed intestinal epithelial and mucosal damages.Conclusions We have successfully established a C57BL/6 mouse model of S.Typhimurium infection.This model may be of crucial significance for studying the biological functions of associated immunological molecules or cytokines in the process of inflammatory bowel disease induced by S.Typhimurium.
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OBJECTIVE To explore the expression of Bcl-2, on mRNA and protein levels, in the different age of C57BL/6 mice cochleae and the expression localization in the cochleae.METHODS Using ABR to test the hearing level in C57BL/6 mice. Surface preparation of cochlear basil membrane is used to observe the morphology and amout of the outer and inner hair cells in different age of C57BL/6 mice. Fluorescent quantitative real time PCR, immunofluorescence histochemical method and western blot are used to detect the expression of Bcl-2 on the mRNA and protein levels in the C57BL/6 mice cochlea of different age groups ('young group', 'elderly group').RESULTS ABR results showed that the hearing threshold of 'older' C57BL/6 mice is much higher than that in the 'young' mice, and surface preparation of cochlear basil membrane showed the hair cell localized in the cochlear basil turn of 'old' mice arranged in a disorder station and part of hair cells were lost. Also, the spiral ganglion cells arranged sparsely and messily. Fluorescence quantitative real-time PCR results suggest the expression of Bcl-2 on/at the mRNA level of the 'old' mice cochleae decreases significantly, compared to that in the 'young' mice. The results of Immunofluorescence and Western blot suggest the expression of Bcl-2 on/at the protein levels of the 'old' mice cochlea decreased, compared to that in the 'young' mice. Also, the Bcl-2 is located in the cytoplasm, and the expression of Bcl-2 in the inner hair cells seems higher than that in the outer hair cells. CONCLUSION The expression of Bcl-2 significantly deceased in the 'old' C57BL/6 mice cochleae, both on mRNA and protein level, which may be related to the hearing loss and loss of hair cells.
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Objective To improve the gene targeting efficiency with C57BL/6 embryonic stem ( ES) cells.Meth-ods Three different genetically modified C57BL/6 ES cell lines, named TLX3, Ai3K and SL, were microinjected into ICR, B6( Cg)-Tyrc-2J and BALB/c mouse blastocysts, respectively.The efficiency was statistically evaluated according to three aspects:blastocyst collection, chimera production and germline transmission.Results None of the three ES cell lines was germline transmitted with B6(Cg)-Tyrc-2J mice as blastocyst donors, while it was achieved with both BALB/c and ICR mouse blastocysts.Compared in the aspect of blastocysts collection, ICR mouse was much better than BALB/c mouse (P<0.05), and the chimera production efficiency of ICR mouse was comparable to that of BALB/c mouse (P =0.115). As to the germline transmission efficiency, that of BALB/c mice is significantly higher than that of the ICR mice ( P<0.01).Conclusions The germline transmission efficiency of BALB/c mouse is highest among these three mouse strains. However, it has the disadvantages of blastocyst collection, developmental delay and zona pellucida fragility, compared with ICR mouse.Therefore, ICR mouse is also a good candidate as blastocyst donor for embryonic stem cell microinjection.
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Sargassum fusiforme has traditionally been widely consumed in Asia as a food, and it has gained much attention due to its high nutritional, pharmaceutical, and industrial value. This study aimed to examine the promotional effects of ethanol extract (ET) and fraction obtained from ethyl acetate (FR) of S. fusiforme on hair growth in C57BL/6 mice and HaCaT cells. Five-week-old mice were used to compare hair regrowth during application of ET and FR for 21 days. Hair regrowth was evaluated by macroscopic observation and verified by hematoxylin-eosin tissue staining. Levels of mRNA expression of factors relevant to the hair growth cycle such as keratinocyte growth factor (KGF), vascular endothelial growth factor (VEGF), and transforming growth factor-beta1 (TGF-beta1) were examined by quantitative polymerase chain reaction (qPCR). Our results showed that ET and FR successfully promoted hair regrowth in shaved C57BL/6 mice at a dose >20 mg/kg. Moreover, ET and FR were effective in stimulating expression of KGF and VEGF mRNAs in a dose-dependent manner, whereas TGF-beta1 was not activated. These results indicate that ET and FR of S. fusiforme effectively promoted hair growth and gene expression relevant to hair growth cycles in both in vitro and in vivo models.
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Animals , Mice , Alopecia , Asia , Ethanol , Fibroblast Growth Factor 7 , Gene Expression , Hair , Polymerase Chain Reaction , RNA, Messenger , Sargassum , Transforming Growth Factor beta1 , Vascular Endothelial Growth Factor AABSTRACT
Background C57BL/6(B6) is a kind of routine mouse specie used in experimental autoimmune uveitis (EAU) research.Previous studies showed that the pathogenesis of uveitis related to inflammatory cytokines secreted by different helper T(Th) cells.However,the interaction of different Th cells in EAU is unclear.Objective This study was to investigate the dynamic changes of inflammatory factors in the spleen and serum after immunization in EAU mice.Methods Forty-four SPF B6 mice were immunized by injection of interphotoreceptor retinoid-binding protein (IRBP) and complete Freund adjuvant (CFA) emulsion via caudal vein and footpad.Indirect ophthalmoscope was used to examine the eyes 3 times per week and the inflammatory response was scored based on Thurau's criteria.In the thirty day after injection,20 model eyes were extracted and the sections of eye tissue were prepared for histopathological examination.The spleens of model mice were enucleated before injection and 2,5,10,15,20,25,30 days after injection,and reverse transcriptase PCR (RT-PCR) was used to detect the contents of interleukin-17 (IL-17) mRNA,interferon-γ (IFN-γ) mRNA,tumor necrosis factor-α (TNF-α) mRNA and IL-10 mRNA,and the contents of IL-17,IFN-γ,TNF-α and IL-10 in model serum were assayed by ELISA in 24 model mice.The experimental protocol and use of the animals were approved by Ethic Committee for Care and Use of Laboratory Animals of Shandong University of Traditional Chinese Medicine.Results Mild inflammatory response was seen in 12 days under the indirect ophthalmoscope with the scores of 0.5.The inflammatory scores peaked in 13-15 days with the scores of 1.0 and alleviated after that with the inflammatory scores of 0.5 in 30 days after injection.The histopathological score was consistent with the clinical score in the models on the 30 days.The serum IL-17 content of model mice was (0.98±0.05) ng/L before injection and increased to (51.85 ±2.42) ng/L on the fifth day,and decreased to (4.01±0.06)ng/L on the fifteen day.But,the serum IL-17 level increased to (25.00±0.94)ng/L again on the 25th day,and then lowed to (6.01 ±0.21)ng/L 30 days after injection,showing a significant elevation in comparison with that of before injection (P=0.000).The serum IFN-γ content of the model mice was (1.02±0.09)ng/L before injection and increased to (50.54±0.48) ng/L on the fifth day,and (73.21±0.12) ng/L on the tenth day,and then it declined gradually until (5.15±0.18)ng/L in the 30th day,which was still higher than that of before injection (P=0.000).After injection of IRBP+CFA,the serum TNF-α level upregulated from the second day to fifth day with the peak values (134.25±0.59)ng/L,and declined to valley on 15th day.A repeat elevation of serum TNF-α level was found on the 20th day with the values (60.54±0.62)ng/L and followed by decrease till the 30th day,which was higher than that of before injection (P=0.660).Serum IL-10 was detectable in the tenth day and peaked on the 15th day.Then a slight decrease was seen till the 30th day,compared with before injection(P =0.000).The contents of IL-10 mRNA,IL-17 mRNA,TNF-α mRNA,IFN-γmRNA in mice spleens followed the same pattern with serum levels of their proteins.Conclusions IL-17,IFN-γ,TNF-α and IL-10 are key inflammatory factors of Th1,Th2 and Th17,they present with specific changes during EAU,it confirming that IFN-γ probably play a pathogenic role in EAU,IL-17 and TNF-α levels probably associated with the chronic and recurrent procedure of uveitis,IL-10 plays an inhibit role in EAU.
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Background Pupillary light reflex has been widely used in the diagnosis and evaluation of visual system and nervous system diseases.However,in animal experiments,functional evaluation of the visual system and nervous system needs more advanced technology and are affected by many factors.Objective This study was to explore the use of the dynamic pupillometer in evaluating pupillary light reflex and to discuss the influence of brightness of stimulate on relevant curve parameters in C57BL/6 mouse.Methods Ten healthy SPF male C57BL/6 mice were collected in this experiment.White light of five luminance levels (2,8,32,128,256 cd/m2) was used to stimulate the mice following a 2-hour dark adaptation.The stimulation was given at the 60-second intervals,for a duration of 100 ms at every stimulation.An infrared camera and video capture card were used to capture digital images during the measuring process in a scotopic environment,at a speed of 60 frames per second.Measuring outcome was saved in the*.AVI format.A software that was developed by our group was used to determine pupil diameter and output pupillary light reflex curve offline.Pupil initial diameter (R1),constriction amplitude (CA),constriction velocity (CV),latency (T1),time for maximum velocity (T2),time for maximum constriction (T3),time for maximun con-striction to 10.1％ R1 re-dilation (RT)and re-dilation velocity (RV)were assessed,and the correlations between luminosity and measuring parameters were analyzed using the Spearman rank correlation.The use of animals followed the Regulations for thd Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results R1 values showed no statistically significant difference among the 5 different luminosity groups(F=1.117,P=0.361).A positive linear correlation was found between stimulating luminosity and CA(r=0.508,P＜ 0.01),but negative correlations were seen between stimulating luminosity and CV or RV (r=-0.625,-0.609,P＜0.01).T1 and T2 values in the 5 different luminosity groups were not statistically significant (F =0.202,P =0.936 ; F =1.584,P =0.195).The different levels of stimulating luminosity showed positive linear correlations with T3 and RT values (r =0.791,0.609,P＜ 0.01).Conclusions The dynamic pupillometer can quantitatively measure the pupillary light reflex of C57BL/6 mice.The pupillary light reflex dynamic curve parameters of mouse were affected by stimulus luminosity levels.These outcomes offer a basis for the application of the dynamic pupillometer system for measuring pupillary light reflex in animal models.
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Background Recently,there were many studies on corneal innervations during mammalian development.However,there were fewer studies on discussing corneal innervations before and after mouse eye openings.Objective The present study was to investigate the change in the regulation of corneal nerve fiber length and density before and after mouse eye openings to offer a basis for clinical research in human.Methods Thirty SPF C57BL/6 mice were divided into postnatal 1 day(P1 d),P7 d,P13 d(1 day before eye opening),P14 d(eye halfopened),P17 d(1 day after eye opening)and P23 d(7 day after eye opening)groups,with 5 mice and 10 eyes for each group.Entire corneal stretches were prepared and immunostaining with an anti-neuron-specific β-Ⅲ tubulin antibody was performed to label the corneal nerve fibers.Confocal microscopic pictures from the corneal dorsal-nasal region (DN),dorsal-temporal(DT),ventral-nasal region(VN)and ventral-temporal(VT)were taken using Delta Vision Core.From these pictures,the mouse corneal area,total length and density of nerve fibers in the 4 regions were calculated.The use of the animals complied with Statement of ARVO.Results Corneal areas of P1 d,P7d,P13 d,P14 d,P17 d and P23 d mice were(0.404±0.007),(1.362±0.154),(1.573±0.080),(1.603±0.046),(1.847±0.052),(2.445±0.798)mm2,respectively ; the total lengths of nerve fibers were(3.718±1.044),(19.065±3.350),(23.687±0.907),(27.309±2.477),(31.989±3.976),(41.214±1.573)mm,respectively ; the densities of nerve fibers were(9.592±1.138),(14.506±1.908),(15.088±1.241),(16.772±1.897),(16.821±2.102),(17.660±1.216)mm/mm2,respectively,all showing significant increases with age(F =22.906,P =0.000 ; F =0.424,P =0.000 ; F =2.375,P=0.000).A positive correlation of the increasing corneal areas and increasing lengths of nerve fibers was found(r=0.983,P＜0.01).Nerve fiber densities in the four corneal regions significantly increased with age(DN region:F =0.159,P =0.000 ; DT region:F =2.1 72,P =0.001 ; VN region:F =1.998,P =0.000 ; VT region:F=2.352,P=0.000).From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference(t =0.589,P =0.572); and the corneal nerve fiber densities in the DT region,VN region and VT region decreased by 4.6％,5.5％ and 0.1％,respectively,without significant difference from P14 d to P17 d(t=0.549,P=0.596;t=0.701,P=0.501 ;t=-0.100,P=0.919).Conclusions The development of nerve fibers in the whole cornea or the four corneal regions is influenced by eye opening in mouse to various extents.From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference.From P14 d to P17 d,the corneal nerve fiber densities in the DT region,VN region and VT region decrease by 4.6％,5.5％ and 0.1％,respectively,without significant difference.Afterwards,the growth of nerve fibers increased in pace and the growth rate is recovered.
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PURPOSE: Skin grafting is one of the most commonly used methods in reconstructive plastic surgery field, but complications such as color change, contracture or hypertrophy are common problems. However, pathophysiology of the color change after skin graft is not yet determined and no animal model is established. METHODS: Full thickness skin grafts were performed on the dorsum of C57BL/6 mice. Serial chronological gross inspection for color change and pigmentation were examined. Melanin pigments were traced by Fontana-Masson staining and semi-quantitative analysis was performed. In addition, immunohistochemical staining of S-100, Micropthalmia related Transcription Factor(MITF) and Melan-A antibodies were also performed to observe melanocytes and their changes. RESULTS: After skin graft, color change and pigment spots were observed in the graft. Fontana-Masson staining showed melanin pigments in the epidermal and dermal layers in all mice. Immunohistochemistry staining to S-100, MITF, Melan-A antibodies showed melanocytes at the basal layer of epidermis and dermis. CONCLUSION: In conclusion, we have established an animal model for skin pigmentation after skin graft. We believe this study may be useful in understanding of the behavior of melanocytes after skin graft.
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Animals , Mice , Antibodies , Contracture , Epidermis , Hypertrophy , Immunohistochemistry , MART-1 Antigen , Melanins , Melanocytes , Models, Animal , Pigmentation , Skin , Skin Pigmentation , Skin Transplantation , Surgery, Plastic , TransplantsABSTRACT
Objective: We have reported previously that mice deficient in nuclear erythroid 2 p45-related factor 2 (Nrf2), which regulates the expression of antioxidant and detoxification genes, showed significant susceptibility to airway inflammatory responses when exposed to diesel exhaust particles for eight weeks2. As disruption of Nrf2 promotes immune cells that stimulate Th2-like immunoresponsiveness, Nrf2-deficient mice may be resistant to M. tuberculosis infection. Setting: Nrf2-deficient mice were infected with M. tuberculosis aerially, and the size of their granulomas and cytokine mRNA expression were compared with those of wild-type mice. Results: Significant reduction of granuloma formation and tubercle bacilli in granulomas was noted in the deficient mice 27 weeks after infection, concurrently with higher expression of IL-2 and IL-13 mRNA. Conclusion: It is concluded that Nrf2 inversely regulates M. tuberculosis-induced granuloma development at the late stage.
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Background An ethylnitrosourea(ENU)-induced mutant strain C57BL/6 mouse model has been established by our research group.This model is proved to have the spontaneous phenotype of corneal opacity and the typical pathological process similar to human keratitis.Therefore,this model is expected to be a good animal model in the research of the mechanism,hereditary property,and development of drugs for corneal infectious diseases.Objective The present study is to investigate the biological features of opportunistic pathogens using a mouse Staphylococcus-infected corneal model(C57BL/6 mouse) induced by N-ethyl-N-nitrosourea(ENU),and offers an evidence of stability in this animal model.Methods Ten-week-old male C57BL/6 mice were treated with ENU at 150mg/kg by intraperitoneal injection,and then mated with female mice after 60 days.Corneal opacity mutant mice in the F1 generation were selected to backcross with C57BL/6 mice.The bacteria were isolated from the eyeballs of the mutants and cultivated,purified and identified.Drug sensitivity assay was carried out to screen for effective antibiotics for clinic medical care.Results The staphylococcus-infected corneal mouse model(B6-Co) was established successfully,and the Staphylococcus sciuri strain was separated and purified,and then the sensitive antibiotics were distinguished from resistant ones.The sensitive drugs for Staphylococcus sciuri included azithromycin,clindamycin,chloramphenicol,gentamicin,rifampicin,tetracycline,amikacin,sulfamethoxazole compound sinomin,minocycline,levofloxacin,cephalothin,cefotaxime,and furazolidone;whereas this Staphylococcal strain was resistant to cefoxitin,penicillin,ampicillin,novobiocin.Nitrofurantoin showed an intermediate sensitivity.Conclusion The C57BL/6 mouse model is a spontaneous-derived animal model that is infected by coagulase-negative staphylococci,among which the most abundant strain is Staphylococcus sciuri.
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Objective To compare the condition of illness and pathological characteristics of experimental autoimmune encephalomyelitis (EAE)in C57 BL/6 mouse models induced by myelin oligodendrocyte glycoprotein 35-55 (MOG35-55)at different doses,and provide a reliable animal model for further study of multiple sclerosis(MS).Methods Male SPF-grade C57 BL/6 mice were divided randomly into four groups:normal group and three EAE model groups (MOG35-55 high-dose,middle-dose and low-dose model groups).200,100,50μg MOG35-55/mice were mixed with complete Freund's adjuvant(CFA),respectively,to prepare complete antigen in different concentrations.The mice were anesthetized and injected s.c.over flanks with the complete antigen and injected i.P.with pertussis toxin to establish immunization-induced C57BL/6 mouse-model of EAE.The mice of the normal group were injected with normal saline instead.Since the day of immunization,the incidence,body weight and neurological score of the mice were observed.The mice of different neurological scores in different periods were anesthetized and perfused with saline and followed by 4% paraformaldehyde.The brain and spinal cord of the mice were removed and fixed in the same fixative solution.The brains and spinal cords of the mice were examined by histopathology with hematoxylin-eosin(HE) staining.The mice on the 40th day were sacrificed and perfused with 2% paraformaldehyde and 2% glutaraldehyde, 1 mm~3 pieces of cerebral white matter and intumescentia lumbalis of the spinal cord were taken and ultrathin sections were prepared according to conventional techniques for electron microscopy. Results All the MOG_(35-55) in three different doses induced mouse models of EAE. The disease was with an incidence rate of 100% and a chronic monophasic course. The body weight of the mice in the three groups decreased obviously compared with those in the normal group. The maximum value of neurological score was 1.33,2.25 and 2.50 in the mice of high-, middle-and low-dose groups, respectively. The major histopathological changes observed in the brain and spinal cord of the EAE mice were different degrees of inflammatory cell infiltration around small vessels showing sleeve-like changes, dcmyelination and neuronal karyopyknosis in the acute and remission stages. The main site of the brain inflammation was in white matter around encephalocoele, and also in the DG and CA zones of hippocampus. The spinal cord inflammation was most severe in the lumbosacral region. The above mentioned pathological changes in the low-dose group were more prominent than those in the middle-dose and high-dose groups. The major ultrastructural changes were scattered around encephalocoele, interstitial edema, especially around small blood vessels, and swollen mitochondria with damaged cristae, and some karyopyknosis in vascular endothelial cells. Some tight junctions were blurred. Some dispersed lymphocytes and mononuclear cells were seen in the perivascular space. In lumbar intumescentia of the spinal cord, there were some myelin figures in the white matter myelin sheath. Some of them showed demyelization and structurtal fusion. The cytoplasmic organelles of axons were considerably reduced or even disappeared. The vascular basement membrane showed an increased thickness and focal necrosis in some areas. Conclusion The mouse models of immune-induced EAE are successfully established with MOG_(35-55), especially that induced with MOG in a dose of 50 μg. This mouse model is stable, with a high incidence and low mortality rate, and can be applied for EAE research in the future.
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BACKGOUND: Imiquimod enhances both acquired and innate immune responses by inducing the synthesis of IFN-alpha and other cytokines (TNF-alpha, IL-1, 6, 8, 12, GM-CSF). Imiguimod was investigated in relation to melanoma, a highly resistant cancer in its immunogenic nature, but also an attractive target for immunotherapy. OBJECTIVE: The purpose of this study was to investigate the anti-tumor effect of 5% imiquimod cream in vivo. METHOD: C57BL/6 mice were inoculated intradermally into the left flank with 2X103 murine B16 melanoma cells. Five groups (including vehicle only, imiquimod cream, imiquimod cream with dimethyl sulfoxide (DMSO), vehicle with ultrasound, and imiquimod cream with ultrasound) were treated once daily at the inoculation site of the melanoma cells. The tumor growth was evaluated daily by using a caliper. RESULTS: The difference in tumor growth was seen around day 12 of treatment, and only the group of 5% imiquimod cream with ultrasound showed a decrease in tumor growth when compared to the control group. CONCLUSION: These results indicate that topical application of 5% imiquimod cream on mouse skin melanomas induces inhibition of tumor growth, and imiquimod cream could be used for the combination treatment of malignant melanoma.
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Animals , Mice , Cytokines , Dimethyl Sulfoxide , Immunity, Innate , Immunotherapy , Interleukin-1 , Melanoma , Melanoma, Experimental , Skin , UltrasonographyABSTRACT
The regional distribution and relative frequency of the pancreatic endocrine cells in the C57BL/6 mouse were studied by immunohistochemical method using four types of specific mammalian antisera against insulin, glucagon, somatostatin and human pancreatic polypeptide (PP). The pancreas of mouse could be divided into three portions; pancreatic islets, pancreatic duct and exocrine portions, and pancreatic islets were further subdivided into three regions (central, mantle and peripheral regions) according to their located types of immunoreactive cells and pancreatic duct portions were also subdivided into two regions (epithelial and connective tissue regions). In the pancreatic islet portions, although some cells were also demonstrated in the mantle regions, most of insulin-immunoreactive cells were located in the central regions and they were randomly dispersed in the whole pancreatic islets. Glucagon-immunoreactive cells were detected in the mantle and peripheral regions. Their relative frequencies in the peripheral regions were somewhat numerous than those of the mantle regions. Somatostatin-immunoreactive cells were detected in the mantle and peripheral regions. However, no PP-immunoreactive cells were demonstrated in the pancreatic islets of C57BL/6 mouse. In the pancreatic duct portions, rare glucagon-immunoreactive cells were situated in the epithelial regions. Cell clusters that consisted of glucagon- or somatostatin- immunoreactive cells were found in some case of connective tissue regions of pancreatic ducts. However, insulin- and PP-immunoreactive cells were not detected in the epithelial nor connective tissue regions. In the exocrine portions, all four types of immunoreactive cells except for PP cells were demonstrated in the C57BL/6 mouse. However, no PP-immunoreactive cells were demonstrated. In conclusion, regional distribution of endocrine cells in the pancreas of C57BL/6 mouse was similar to that of mammals, especially other rodents except for topographically different distribution of endocrine cells compared to that of other rodents.
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Animals , Female , Male , Mice , Glucagon/metabolism , Immunohistochemistry , Insulin/metabolism , Islets of Langerhans/cytology , Mice, Inbred C57BL/metabolism , Pancreatic Polypeptide/metabolism , Somatostatin/metabolismABSTRACT
The regional distributions and relative frequencies of some gastrointestinal endocrine cells in the three portions (cecum, colon and rectum) of the large intestinal tract of C57BL/6 mice were examined with immunohistochemical method using 7 types of specific antisera against chromogranin A (CGA), serotonin, somatostatin, human pancreatic polypeptide (HPP), glucagon, gastrin and cholecyctokinin (CCK)-8. In this study, all 3 types of immunoreactive (IR) cells were identified. Most of these IR cells in the large intestinal portion were generally spherical or spindle in shape (open-typed cell) while cells with a round shape (close-typed cell) were found in the intestinal gland. Their relative frequencies varied according to each portion of the large intestinal tract. CGA-IR cells were found throughout the whole large intestinal tract but were most predominant in the colon. Serotonin-IR cells were detected throughout the whole large intestinal tract and showed highest frequency in the colon. Peculiarly, glucagon-IR cells were restricted to the colon with a low frequency. However, no somatostatin-, HPP-, gastrin- and CCK-8-IR cells were found in the large intestinal tract. In conclusion, some peculiar distributional patterns of large intestinal endocrine cells were identified in C57BL/6 mice.
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Animals , Female , Male , Mice , Chromogranin A , Chromogranins/metabolism , Enteroendocrine Cells/metabolism , Gastrins/metabolism , Glucagon/metabolism , Immunohistochemistry/veterinary , Intestine, Large/cytology , Mice, Inbred C57BL , Pancreatic Polypeptide/metabolism , Serotonin/metabolism , Sincalide/metabolism , Somatostatin/metabolismABSTRACT
PURPOSE: Changes in the balance between MMP and TIMP can have a profound effect on the composition in the extracellular matrix (ECM) and affect various cellular functions including adhesion, migration, differentiation of cells, and fibrosis and invasion and metastasis of cancer cells. Radiation therapy is a popular treatment modality for benign and malignant tumor, but the study for radiation effect on MMP and TIMP is scarce. In the current study, we have examined the expression of TIMP in fibrosis-prone (C57BL/6) mice after radiation. METHODS AND MATERIALS: Adult female mice of 10~12 weeks were used. The whole body were irradiated using a Varian CL-4/100 with 2 and 10 Gy. Immunohistochemical staining was performed according to Avidin Biotin complex method and evaluated by observing high power field. For TIMP-1, TIMP-2 antibodies, reactivity was assessed in the parenchymal cell and in the stromal cell. The scale of staining was assessed by combining the quantitative and qualiative intensity of staining. RESULTS: TIMP-1 immunoreactivity did not change in lung. But, in liver, TIMP-1 immunoreactivity was localized in cytoplasm of hepatocyte and Kupffer cell. In kidney, TIMP-1 immunoreactivity was localized in cytoplasm of some tubular cell. Temporal variations were not seen. Dose-response relationship was not seen except kidney. TIMP-2 immunoreactivity in lung was a score (++) at 0 Gy and elevated to a score (+++) at 2 Gy. TIMP-2 immunoreactivity was a score (++) in liver at 0 Gy. TIMP-2 immunoreactivity was localized in cytoplasm of hepatocyte and Kupffer cell as same as patterns of TIMP-1 immunoreactivity. The TIMP-2 immunoreactivity in liver was elevated to (+++) at 2 Gy. Immunoreactivity to TIMP-2 in kidney was a score (+++) at 0 Gy and was not changed at 10 Gy. The score of TIMP-2 immunoreactivity was reduced to (++) at 2 Gy. TIMP-2 immunoreactivity was confined to tubules in kidney. Temporal variation of TIMP-2 immunoreactivity was irregular. Dose-response relationship of TIMP-2 immunoreactivity was not seen. CONCLUSIONS: Differences between intensity of expression of TIMP-1 and TIMP-2 in each organ was present. Expression of TIMP was localized to specific cell in each organ. Irradiation increased TIMP-1 immunoreactivity in the liver and the kidney. Irradiation increased TIMP-2 immunoreactivity in the lung. But, in the liver and the kidney, TIMP-2 expression to radiation was irregular. Temporal variation of TIMP-2 immunoreactivity was irregular. Dose-response relationship of TIMP-2 immunoreactivity was not seen. In the future, we expect that the study of immunohistochemical staining of longer period of postirradiation and quantitative analysis using western blotting and northern blotting could define the role of TIMP in the radiation induced tissue fibrosis.
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Adult , Animals , Female , Humans , Mice , Antibodies , Avidin , Biotin , Blotting, Northern , Blotting, Western , Cytoplasm , Extracellular Matrix , Fibrosis , Hepatocytes , Immunohistochemistry , Kidney , Liver , Lung , Neoplasm Metastasis , Radiation Effects , Stromal Cells , Tissue Inhibitor of Metalloproteinase-1 , Tissue Inhibitor of Metalloproteinase-2ABSTRACT
AIM: To study the effect of effective parts of Acanthopanax senticosus on Parkinson's disease mouse model(from C_(57) BL/6 strain) concerning Gammaamino butyric acid(GABA),nigra-striatum tyrosine hydrozylase(TH) and hippocampal glial fibrillary acidic protein(GFAP). METHODS: In MPTP(1-methyl-4-phenyl-1,2,3,6-tetrahydropytidine)-induced PD mouse model(accumulative dose amounted to 150 mg/kg) pole-climbing time was determined of mice after receiving effective parts of Acanthopanax senticosus for 20 d,and GARA,TH and GFAP expression were assayed simultaneously with the help of immunohistochemical method. RESULTS: Mice's pole-climbing time was measured,GABA and TH made a significant difference between treated group and control group(P