ABSTRACT
<p><b>OBJECTIVE</b>To explore a method for combining Fluoro-Jade B (FJB) staining with immunofluorescent staining in rats with focal cortical infarction.</p><p><b>METHOD</b>Permanent distal middle cerebral artery occlusion (dMCAO) was induced in rats by electrocoagulation. The rat models were randomized into two groups, and frozen sections of the brain tissues from each group were stained with FJB followed by immunofluorescent staining or in the reverse order.</p><p><b>RESULTS</b>FJB staining followed by immunofluorescence staining clearly visualized both FJB-positive and immunofluorescence-positive cells in the frozen sections, but the staining protocol in the reverse sequence failed to clearly show the immunofluorescence-positive cells.</p><p><b>CONCLUSION</b>FJB staining prior to immunofluorescence staining does not affect the staining effect of protein immunofluorescent staining and better visualizes the positive cells.</p>
Subject(s)
Animals , Rats , Brain , Pathology , Fluoresceins , Chemistry , Fluorescent Antibody Technique , Methods , Fluorescent Dyes , Chemistry , Infarction, Middle Cerebral Artery , Staining and Labeling , MethodsABSTRACT
<p><b>OBJECTIVE</b>To analyze the expression of inducible nitric oxide synthase (iNOS) in the testis tissues of Fmr1 (fragile X mental retardation 1) knockout and wild-type male mice in different developmental stages, and provide background information for researches on fragile X syndrome.</p><p><b>METHODS</b>This study included 4, 6, 8 and 10 weeks old Fmr1 knockout and wild-type male mice, 6 in each age group. We identified the genotype of the mice by PCR, and detected and compared the expression of iNOS in the testis tissues of the Fmr1 knockout and wild-type mice by immunohistochemistry.</p><p><b>RESULTS</b>The iNOS expression was weakly positive in the Leydig cells of the 4-week-old mice, moderately positive in the 6-week-old ones, and strongly positive in 8- and 10-week-old ones, significantly weaker in the Fmr1 knockout than in the wild-type ones.</p><p><b>CONCLUSION</b>The expression of iNOS significantly decreases in the testis of Fmr1 knockout mice, suggesting that iNOS may be involved in the pathogenesis of fragile X syndrome.</p>
Subject(s)
Animals , Male , Mice , Fragile X Mental Retardation Protein , Genetics , Fragile X Syndrome , Genetics , Gene Expression Regulation, Developmental , Mice, Knockout , Nitric Oxide Synthase Type II , Metabolism , Testis , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To explore the regulatory effect of fragile X mental retardation protein (FMRP) on the translation of microtubule associated protein 1B (MAP1B).</p><p><b>METHODS</b>The expressions of MAP1B protein and MAP1B mRNA in the brains of 1-week and 6-week old fragile X mental retardation-1 (Fmr1) knockout (KO) mice were investigated by immunohistochemistry, Western blot, and in situ hybridization, with the age-matched wild type mice (WT) as controls.</p><p><b>RESULTS</b>The mean optical density (MOD) of MAP1B was significantly decreased in each brain region in KO6W compared with WT6W, whereas in KO1W, this decrease was only found in the hippocampus and cerebellum. MAP1B in 6-week mice was much less than that in 1-week mice of the same genotype. The results of Western blot and in situ hybridization showed that MAP1B protein and MAP1B mRNA were significantly decreased in the hippocampus of both KO1W and KO6W.</p><p><b>CONCLUSION</b>The decreased MAP1B protein and MAP1B mRNA in the Fmr1 knockout mice indicate that FMRP may positively regulate the expression of MAP1B.</p>
Subject(s)
Animals , Mice , Age Factors , Animals, Newborn , Brain , Metabolism , Fragile X Mental Retardation Protein , Genetics , Gene Expression Regulation, Developmental , Genetics , Mice, Knockout , Microtubule-Associated Proteins , Metabolism , Mutation , Physiology , RNA, MessengerABSTRACT
Objective To explore the possible role of voltage-gated potassium channel-interacting protein 1 (KChIP1) in the pathogenesis of epilepsy. Methods Sprague Dawley female adult rats were treated with pentylenettrazole (PTZ) to develop acute and chronic epilepsy models. The approximate coronal sections of normal and epilepsy rat brain were processed for immunohistochemistry. Double-labeling confocal microscopy was used to determine the coexistence of KChIP1 and gamma-aminobutyric acid (GABA). Results KChIP1 was expressed abundantly throughout adult rat brain. KChIP1 is highly co-localize with GABA transmitter in hippocampus and cerebral cortex. In the acute PTZ-induced convulsive rats, the number of KChIP1-postive cells was significantly increased especially in the regions of CA1 and CA3 (P < 0.05); whereas the chronic PTZ-induced convulsive rats were found no changes. The number of GABA-labeled and co-labeled neurons in the hippocampus appeared to have no significant alteration responding to the epilepsy-genesis treatments. Conclusion KChIP1 might be involved in the PTZ-induced epileptogenesis process as a regulator to neuronal excitability through influencing the properties of potassium channels. KChIP1 is preferentially expressed in GABAergic neurons, but its changes did not couple with GABA in the epileptic models.