Monocarboxylate transporter 4 plays a significant role in the neuroprotective mechanism of ischemic preconditioning in transient cerebral ischemia

Monocarboxylate transporters（MCTs）, which carry monocarboxylates such as lactate across biological membranes, have been associated with cerebral ischemia/reperfusion process. In this study, we studied the effect of ischemic preconditioning（IPC） on MCT4 immunoreactivity after 5 minutes of transient cerebral ischemia in the gerbil. Animals were randomly designated to four groups（sham-operated group, ischemia only group, IPC ＋ sham-operated group and IPC ＋ ischemia group）. A serious loss of neuron was found in the stratum pyramidale of the hippocampal CA1 region（CA1）, not CA2/3, of the ischemia-only group at 5 days post-ischemia; however, in the IPC ＋ ischemia groups, neurons in the stratum pyramidale of the CA1 were well protected. Weak MCT4 immunoreactivity was found in the stratum pyramidale of the CA1 in the sham-operated group. MCT4 immunoreactivity in the stratum pyramidale began to decrease at 2 days post-ischemia and was hardly detected at 5 days post-ischemia; at this time point, MCT4 immunoreactivity was newly expressed in astrocytes. In the IPC ＋ sham-operated group, MCT4 immunoreactivity in the stratum pyramidale of the CA1 was increased compared with the sham-operated group, and, in the IPC ＋ ischemia group, MCT4 immunoreactivity was also increased in the stratum pyramidale compared with the ischemia only group. Briefly, present findings show that IPC apparently protected CA1 pyramidal neurons and increased or maintained MCT4 expression in the stratum pyramidale of the CA1 after transient cerebral ischemia. Our findings suggest that MCT4 appears to play a significant role in the neuroprotective mechanism of IPC in the gerbil with transient cerebral ischemia.

Iodine deficiency up-regulates monocarboxylate transporter 8 expression of mouse thyroid gland

Background Iodine deficiency is a major factor affecting thyroid auto-regulation,the quantity of iodine may greatly influence the synthesis of thyroid hormones （THs）.It has long been believed that TH enters the cell through passive diffusion.Recent studies have suggested that several transporters could facilitate transportation of TH.The monocarboxylate transporter 8 （MCT8） was identified as a very active and specific TH transporter.The purpose of this study was to investigate whether iodine insufficient affected the expression of MCT8 in the thyroid gland.Methods Sixty BALB/c mice were randomly divided into two groups：control group was fed with standard feed （iodine concentration of 300 μg/kg）; while low-iodine （LI） group received iodine-insufficient feed （iodine concentration of 20-40 μg/kg）.After 3 months,10 mice of each group were sacrificed.The remaining 20 mice of each group were kept till 6 months.From the LI group,we randomly selected 15 mice and injected triiodothyronine （T3,100 μg/kg body weight per day） intraperitoneally for 24,48 or 72 hours （5 mice for each time-point）.Then,all the mice were sacrificed.Mouse serum thyroxine （T4）,T3,and thyroid-stimulating hormone （TSH） levels were determined by chemiluminescence immunoassay （CIA）.The protein content or messenger RNA （mRNA） level of thyroid MCT8 was measured by Western blotting analysis or real time RT-PCR respectively.MCT8 subcellular location in thyroid tissues was probed with immunohistochemistry （IHC） assay.Results We found that mouse serum T3 and T4 levels decreased and TSH level increased by the end of the third month.Consistent with these findings,there was significant goiter and hypothyroidism in the LI group.Meanwhile,the MCT8 mRNA increased to 1.36-fold of the level in the control group at the 3rd month.At 6th month,the serum T4 level in LI mice remained at a lower level,and MCT8 mRNA expression continued rising to nearly 1.60-fold compared with the control group.The protein content was also about 3 times higher than that in the control group.IHC results also revealed MCT8 was of higher expression and localized in the cytoplasm of thyroid follicular cells.After providing exogenous T3 to iodine deficient mice,the serum T3 and T4 gradually increased,whereas MCT8 mRNA and protein both started to decrease and returned to the same level as the control group.Conclusion There is a compensatory increase in thyroid MCT8 expression to enhance its capability to transport TH from thyroid to the blood circulation in iodine deficient mice.

Interaction between basigin and monocarboxylate transporter 2 in the mouse testes and spermatozoa

Basigin is a member of the immunoglobulin superfamily and plays various important roles in biological events including spermatogenesis. To examine the basigin molecular variants during spermatogenesis and sperm maturation in the mouse, immunoprecipitated basigin samples from testis and epididymal spermatozoa were analyzed by liquid chromatography-mass spectrometry/mass spectrometry （LC-MS/MS）. The results demonstrated that basigin molecules from the testis and spermatozoa were separable into two major bands and that the differences in the molecular sizes were possibly because of an endoproteolytic cleavage. Since basigin is known to be a chaperone for the monocarboxylate transporter 1 （MCT1）, the localization of basigin, MCT1 and MCT2 was examined during postnatal testicular development. Immunohistochemical studies showed different expression patterns of MCT1 and MCT2. MCT1 was localized on the surface of spermatogonia, spermatocytes, and spermatids. In contrast, MCT2 appeared on the principal piece of spermatozoa in the testis, where basigin was also observed. In mature epididymal spermatozoa, MCT2 was located on the midpiece, where basigin co-localized with MCT2 but not with MCT1. Furthermore, MCT2 was immunoprecipitated with basigin in mouse testes and sperm. These results suggest that basigin has a functional role as a binding partner with MCT2 in testicular and epididymal spermatozoa.

Loss of monocarboxylate transporter 1 aggravates white matter injury after experimental subarachnoid hemorrhage in rats

Monocarboxylic acid transporter 1(MCT1)maintains axonal function by transferring lactic acid from oligodendrocytes to axons.Subarachnoid hemorrhage(SAH)induces white matter injury,but the involvement of MCT1 is unclear.In this study,the SAH model of adult male Sprague-Dawley rats was used to explore the role of MCT1 in white matter injury after SAH.At 48 h after SAH,oligodendrocyte MCT1 was significantly reduced,and the exogenous overexpression of MCT1 significantly improved white matter integrity and long-term cognitive function.Motor training after SAH significantly increased the number of ITPR2+SOX10+oligodendrocytes and upregulated the level of MCT1,which was positively correlated with the behavioral ability of rats.In addition,miR-29b and miR-124 levels were significantly increased in SAH rats compared with non-SAH rats.Further intervention experiments showed that miR-29b and miR-124 could negatively regulate the level of MCT1.This study confirmed that the loss of MCT1 may be one of the mechanisms of white matter damage after SAH and may be caused by the negative regulation of miR-29b and miR-124.MCT1 may be involved in the neurological improvement of rehabilitation training after SAH.

Cloning and Characterization of karmoisin Homologue Gene(Nlka) in Two Brown Planthopper Strains with Different Eye Colors

The brown planthopper （BPH）, Nilaparvata lugens, is a destructive insect pest of rice throughout Asia. Different from brown-eye color wild type, BPH also has red-eye color mutation phenotype. As a visible genetic marker, the red-eye mutant in BPH is a valuable material. To reveal the eye color mutation mechanism, a karmoisin homologue gene （named as Nlka） was cloned from BPH. And karmoisin is always deemd as a xanthommatin-related gene in other insects, encoding phenoxazinone synthetase （PHS）. Nlka is consisted of 7 exons and encodes a protein with 502 amino acids （NIKA）. NIKA showed high amino acid identities with its insect homologues （48.8%-51.8%）. Nlka transcripts can be detected at all the developmental stages and in all tissues tested, including egg, nymph, adult, body wall, ovary, fat body, midgut and Malpighian tubule. However, no constant In/Del or non-synonymous mutation was observed between the mutant and the wild type strains. Quantitative real-time PCR experiment also showed that Nlka transcript level had no significant differences between them. These results indicated that Nlka is not the target gene causing the red-eye color mutation phenotype of BPH. Through the second structure and motif analysis, the present study also showed that all the proteins deduced from the karmoisin genes in insects may be members of monocarboxylate transporters （MCTs） rather than PHSs.

Association between Alzheimer's disease pathogenesis and early demyelination and oligodendrocyte dysfunction

The APPSwe/PSEN1 dE9（APP/PS1） transgenic mouse model is an Alzheimer＇s disease mouse model exhibiting symptoms of dementia, and is commonly used to explore pathological changes in the development of Alzheimer＇s disease. Previous clinical autopsy and imaging studies suggest that Alzheimer＇s disease patients have white matter and oligodendrocyte damage, but the underlying mechanisms of these have not been revealed. Therefore, the present study used APP/PS1 mice to assess cognitive change, myelin loss, and corresponding changes in oligodendrocytes, and to explore the underlying mechanisms. Morris water maze tests were performed to evaluate cognitive change in APP/PS1 mice and normal C57 BL/6 mice aged 3 and 6 months. Luxol fast blue staining of the corpus callosum and quantitative reverse transcription-polymerase chain reaction（q RT-PCR） for myelin basic protein（MBP） mRNA were carried out to quantify myelin damage. Immunohistochemistry staining for NG2 and qRT-PCR for monocarboxylic acid transporter 1（MCT1） mRNA were conducted to assess corresponding changes in oligodendrocytes. Our results demonstrate that compared with C57 BL/6 mice, there was a downregulation of MBP mRNA in APP/PS1 mice aged 3 months. This became more obvious in APP/PS1 mice aged 6 months accompanied by other abnormalities such as prolonged escape latency in the Morris water maze test, shrinkage of the corpus callosum, upregulation of NG2-immunoreactive cells, and downregulation of MCT1 mRNA. These findings indicate that the involvement of early demyelination at 3 months and the oligodendrocyte dysfunction at 6 months in APP/PS1 mice are in association with Alzheimer＇s disease pathogenesis.

Role of rno-miR-124-3p in regulating MCT1 expression in rat brain after permanent focal cerebral ischemia

This study aimed to assess the role of microRNAs(miRNAs)in regulating monocarboxylate transporter-1(MCT1)expression in rat brain after permanent focal cerebral ischemia to identify a new target for early treatment of cerebral ischemia.Focal cerebral ischemia was induced by permanent middle cerebral artery occlusion(pMCAO)in rats.Morphology and protein expression levels of MCT1 were assessed by immunofluorescence and Western blotting.Using bioinformatics and double luciferase reporter assays,rno-miR-124-3p was selected as a direct target for rat MCT1.Expression of rno-miR-124-3p after pMCAO was detected.Then,rats were treated with rno-miR-124-3p agomir via lateral ventricle injection,and after 6 h or 24 h ischemia,rno-miR-124-3p expression and gene and protein expression of MCT-1 were detected by qRT-PCR and Western blotting.Brain infarction was identified by 2,3,5-triphenyltetrazolium chloride(TTC)staining.Results showed that pMCAO induced brain infarction and increased the expression of MCT1.The levels of rno-miR-124-3p after pMCAO were in contrast to those of MCT1 protein in ischemic region,while declined after 3,6 and 12 h of pMCAO in ischemic penumbra.After administration of rno-miR-124-3p agomir,MCT1 mRNA and protein levels were increased after 6 h of pMCAO,while decreased after 24 h of pMCAO.Meanwhile,rno-miR-124-3p levels increased after both times.TTC staining showed treatment with rno-miR-124-3p agomir reduced brain infarction.The role of rno-miR-124-3p in regulating MCT1 was as a positive regulator after 6 h of pMCAO,while a negative regulator after 24 h of pMCAO,however,both activities had protective effects against cerebral ischemia.

Urothelial bladder cancer progression: lessons learned from the bench

Urothelial bladder carcinoma(UBC)is an intricate malignancy with a variable natural history and clinical behavior.Despite developments in diagnosis/prognosis refi nement and treatment modalities,the recurrence rate is high,and progression from non-muscle to muscle invasive UBC commonly leads to metastasis.Moreover,patients with muscle-invasive or extra-vesical disease often fail the standard chemotherapy treatment,and overall survival rates are poor.Thus,UBC remains a challenge in the oncology fi eld,representing an ideal candidate for research on biomarkers that could identify patients at increased risk of recurrence,progression,and chemo-refractoriness.However,progress toward personalized medicine has been hampered by the unique genetic complexity of UBC.Recent genome-wide expression and sequencing studies have brought new insights into its molecular features,pathogenesis and clinical diversity,revealing a landscape where classical pathology is intersected by the novel and heterogeneous molecular groups.Hence,it seems plausible to postulate that only an integrated signature of prognostic/predictive biomarkers inherent in different cancer hallmarks will reach clinical validation.In this review,we have summarized ours and others’research into novel putative biomarkers of progression and chemoresistance that encompass several hallmarks of cancer:tumor neovascularization,invasion and metastasis,and energy metabolism reprogramming of the tumor microenvironment.