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In JoVE (1)
Other Publications (13)
- British Journal of Pharmacology
- Brain Research
- Acta Biochimica Et Biophysica Sinica
- Neuroscience Letters
- European Journal of Cardio-thoracic Surgery : Official Journal of the European Association for Cardio-thoracic Surgery
- Cellular & Molecular Biology Letters
- Hippocampus
- European Journal of Pharmacology
- Acta Biochimica Et Biophysica Sinica
- Psychopharmacology
- European Journal of Pharmacology
- Molecular Neurobiology
- Basic & Clinical Pharmacology & Toxicology
Articles by Zhiyuan Ma in JoVE
JoVE General
Tracking Dynamics of Muscle Engraftment in Small Animals by In Vivo Fluorescent Imaging
1Department of Anesthesia, Brigham and Woman's Hospital, 2Department of Radiology, Brigham and Woman's Hospital
We describe an in vivo fluorescence imaging protocol to monitor muscle regeneration by GFP-labeled myoblasts after transplantation into skeletal muscles of both healthy and dystrophic mice. This protocol can be adapted to study muscle regeneration by transplantation of other types of cells and in other muscular conditions as well.
Other articles by Zhiyuan Ma on PubMed
Effect of Fentanyl on 5-HT Efflux Involves Both Opioid and 5-HT1A Receptors
1. Fentanyl is a micro -opioid analgesic that might disinhibit 5-HT neurons and thus increase 5-HT efflux. However, fentanyl also binds to 5-HT(1A) receptors, and if it activates 5-HT(1A) somatodendritic autoreceptors, the resultant inhibition might offset opioid-mediated increases in 5-HT efflux. To test this hypothesis, we used microdialysis to study effects of fentanyl on extracellular 5-HT in the dorsal raphe nucleus (DRN) of unanesthetized rats. 2. Systemic administration of fentanyl (0.01-0.2 mg kg(-1), s.c.) increased 5-HT efflux in the DRN. An intermediate dose of fentanyl (0.05 mg kg(-1)) produced the maximum increase in 5-HT to approximately 180% of baseline levels in the DRN. Naltrexone (10 mg kg(-1), s.c.) blocked the increase in response to systemic fentanyl (0.05 mg kg(-1)). 3. In contrast, during infusion into the DRN, fentanyl (10-1000 micro M) induced a dose-dependent decrease in 5-HT. Naltrexone and nor-binaltorphimine failed to block the decrease suggesting that micro - and kappa-opioid receptors did not mediate this effect. 4. Systemic (-)-pindolol (8 mg kg(-1), s.c.) or infusion of WAY-100635 (100 micro M) into the DRN blocked the decrease, and instead 5-HT increased in response to local infusion of fentanyl (100 micro M). WAY-100635 (0.3 mg kg(-1), s.c.) also potentiated the effect of systemic fentanyl (0.2 mg kg(-1), s.c.). (-)-Pindolol and WAY-100635 block 5HT(1A) receptors, indicating that inhibition of 5-HT neuronal activity resulting from fentanyl binding to somatodendritic autoreceptors attenuated opioid-mediated increases in 5-HT efflux. 5. These results provide novel evidence that besides stimulating micro -opioid receptors, fentanyl is a 5-HT(1A) receptor agonist. Possibly, this contributes to lethality of fentanyl overdose.
Evidence of Reuptake Inhibition Responsible for Mecamylamine-evoked Increases in Extracellular Serotonin
The present report using microdialysis approach investigates the neurochemical mechanism of mecamylamine in the regulation of extracellular serotonin in the dorsal raphe nucleus of freely behaving rats. These results suggest that mecamylamine may block serotonin reuptake, the effect consistent with its efficacy of antidepressant.
Suppression of Type 1 Insulin-like Growth Factor Receptor Expression by Small Interfering RNA Inhibits A549 Human Lung Cancer Cell Invasion in Vitro and Metastasis in Xenograft Nude Mice
Cancer invasion and metastasis, involving a variety of pathological processes and cytophysiological changes, contribute to the high mortality of lung cancer. The type 1 insulin-like growth factor receptor (IGF-1R), associated with cancer progression and invasion, is a potential anti-invasion and anti-metastasis target in lung cancer. To inhibit the invasive properties of lung cancer cells, we successfully down-regulated IGF-1R gene expression in A549 human lung cancer cells by small interfering RNA (siRNA) technology, and evaluated its effects on invasion-related gene expression, tumor cell in vitro invasion, and metastasis in xenograft nude mice. A549 cells transfected with a plasmid expressing hairpin siRNA for IGF-1R showed a significantly decreased IGF-1R expression at the mRNA level as well as the protein level. In biological assays, transfected A549 cells showed a significant reduction of cell-matrix adhesion, migration and invasion. Consistent with these results, we found that down-regulation of IGR-1R concomitantly accompanied by a large reduction in invasion-related gene expressions, including MMP-2, MMP-9, u-PA, and IGF-1R specific downstream p-Akt. Direct tail vein injections of plasmid expressing hairpin siRNA for IGF-1R significantly inhibited the formation of lung metastases in nude mice. Our results showed the therapeutic potential of siRNA as a method for gene therapy in inhibiting lung cancer invasion and metastasis.
CART Peptides Increase 5-hydroxytryptamine in the Dorsal Raphe and Nucleus Accumbens of Freely Behaving Rats
Cocaine and amphetamine-regulated transcript peptides (CART) are implicated in the antidepressant effect. This may involve in 5-hydroxytryptamine (5-HT) in the CNS. The aim of the present studies was to investigate the effect of CART peptides on extracellular 5-HT in the dorsal raphe nucleus (DRN) and nucleus accumbens (NAcc) using a microdialysis approach in freely behaving rats. Reverse infusion of CART61-102 in the DRN produced a concentration (10-100 microM) -dependent increase in 5-HT in the DRN. Similarly, CART62-76 (10-100 microM) infused into the DRN and NAcc elevated 5-HT in the DRN and NAcc, respectively. Thus, CART increases extracellular 5-HT in both the DRN and NAcc. In addition, infusion of CART62-76 (100 microM) in the DRN produced a significant increase in 5-HT in the NAcc, implying an existence of CART receptors responsible for the depolarization-dependent release. In summary, the results of the present studies suggest that CART peptides may have an antidepressant effect through increases in extracellular 5-HT.
Does Sleeve Lobectomy Concomitant with or Without Pulmonary Artery Reconstruction (double Sleeve) Have Favorable Results for Non-small Cell Lung Cancer Compared with Pneumonectomy? A Meta-analysis
It has been reported that sleeve lobectomy (SL) concomitant with or without pulmonary artery reconstruction (PAR) might be an alternative procedure for pneumonectomy (PN) in non-small cell lung cancer (NSCLC). The aim of this study was to assess whether SL or PN offers a low morbidity and mortality and better long-term survival. We performed a meta-analysis of studies published in English between 1996 and 2006 to comprehensively compare the postoperative mortality, morbidity, locoregional recurrences, and time-to-event outcomes of SL and PN in NSCLC, and reviewed the recent literatures on PAR in the corresponding period as well. Twelve studies met the defined criteria including a total of 2984 subjects, and five studies for PAR. The odds ratio for postoperative mortality (SL vs PN) was 0.65 (95% confidence interval (CI): 0.42-1.01), 1.01 (95% CI: 0.70-1.44) for postoperative complications, and 0.91 (95% CI: 0.45-1.82) for locoregional recurrences. The weighted mean operative mortality for PAR was 3.3%, and 32.4% for complications. The estimated combined hazard ratio for overall survival in 10 studies was 0.70 (95% CI: 0.62-0.79) in favor of SL group. The median overall survival was 60 months for the SL group, 26 months for the PN group, and 30 months for PAR group. Survival difference in patients with pN0 or pN1 at 1 year demonstrated a pooled risk difference (SL vs PN) of 0.03 (95% CI: -0.08-0.13), 0.13 (95% CI: 0.00-0.25) in patients with pN2 at 1 year, 0.21 (95% CI: 0.07-0.36) in patients with pN0 or pN1 at 5 years, and 0.06 (95% CI: -0.10-0.21) in patients with pN2 at 5 years. Our results suggests that SL with or without PAR can be accomplished safely in selected patients without increasing the morbidity and mortality as compared to PN, that SL even with PAR offers better long-term survival than does PN, and that a more radical operation such as PN is not a more appropriate procedure, even in higher stage tumors.
Silencing of the Type 1 Insulin-like Growth Factor Receptor Increases the Sensitivity to Apoptosis and Inhibits Invasion in Human Lung Adenocarcinoma A549 Cells
The type 1 insulin-like growth factor receptor (IGF-1R), which is over-expressed or activated in many human cancers, including lung cancer, mediates cancer cell proliferation and metastasis. Several studies indicate that blocking IGF-1R expression can inhibit tumor cell proliferation and metastasis. In this study, inhibition of the endogenous IGF-1R by recombinant adenoviruses encoding short hairpin RNAs against IGF-1R was found to significantly suppress IGF-1R expression, arrest the cell cycle, enhance the apoptotic response, and inhibit proliferation, adhesion, invasion and migration in A549 cells. Moreover, silencing IGF-1R decreases the expression of invasive-related genes including matrix metalloproteinase-2 (MMP-2), MMP-9, and urokinase-plasminogen activator (u-PA), and the phosphorylation of Akt and ERK1/2. These results suggest that the silencing of IGF-1R has the potential to be an effective cancer gene therapy strategy for human lung cancer.
Hippocampus Modulates the Behaviorally-sensitizing Effects of Nicotine in a Rat Model of Novelty-seeking: Potential Role for Mossy Fibers
Present experiments investigate interactions between a rat model of the novelty-seeking phenotype and psychomotor sensitization to nicotine (NIC) in adolescence, and the potential role of hippocampal mossy fibers in mediating the behaviorally-sensitizing effects of NIC. Outbred rats were phenotype-screened as high-responders (HR; locomotor reactivity to novelty score ranking in the upper third of the population) or low-responders (LR; locomotor reactivity to novelty score ranking in the lower third of the population). In Experiment 1, both phenotypes were trained with four NIC injections (at 3-d intervals on postnatal days 33-44), and lidocaine microinfusion was used to temporarily inactivate the hippocampal hilus at each NIC injection. Systemic saline and microinjection of artificial cerebral spinal fluid (CSF) were used as controls. During NIC training, lidocaine inactivation caused augmented locomotor response to NIC in HRs compared to LRs irrespective of injection days. Following 1 week of abstinence, all animals were challenged with a low dose of NIC. During challenge, previously NIC/CSF trained LRs and HRs were divided into two; one half receiving lidocaine inactivation of the hippocampal hilus and the other half receiving CSF control microinjection. Only HRs showed behavioral sensitization to the challenge dose of NIC, which was enhanced with lidocaine inactivation. In Experiment 2, a single NIC exposure was found sufficient to induce sensitization to the challenge dose of NIC in HRs, and concurrently an enlarged supra-pyramidal mossy fiber (SP-MF) terminal field. The increase in the SP-MF volume in HRs was greater with repeated NIC training. In both single and repeated NIC training cases, a significant positive morphobehavioral correlation was observed between challenge NIC-induced locomotion and the SP-MF terminal field volume. These findings suggest that the HR hippocampal mossy fibers are vulnerable to neuroadaptive alterations induced by NIC, which may be a substrate for the observed behavioral vulnerability to NIC.
Characterization of Serotonin-toxicity Syndrome (toxidrome) Elicited by 5-hydroxy-l-tryptophan in Clorgyline-pretreated Rats
Patients are at high risk of developing serotonin-toxicity syndrome (toxidrome) when they take multiple serotonergic drugs, particularly co-administered with monoamine oxidase inhibitors or 5-hydroxytryptamine (5-HT) reuptake blockers. The toxidrome can vary from mild to severe. The primary goal of the present study was to understand the relationship between behavioral signs and degrees of toxidrome induced by 5-hydroxy-l-tryptophan (5-HTP) in clorgylinized rats. The severity was obtained by scoring behavioral signs including head shakes, penile erection, forepaw treading, hind limb abduction, Straub tail and tremor. It was found that 5-HTP produced a dose-dependent increase in degrees of the toxidrome. Furthermore, correlation between the toxidrome and changes in body-core temperature (delta Tcor) was determined. There was hypothermia in the mild toxidrome (delta Tcor<-1 degrees C), high hyperthermia in the severe toxidrome (delta Tcor>+2 degrees C) and a small change in T(cor) in the moderate toxidrome (-1 degrees C
Knockdown of Insulin-like Growth Factor 1 Receptor Enhances Chemosensitivity to Cisplatin in Human Lung Adenocarcinoma A549 Cells
The effects of RNA interference-mediated insulin-like growth factor 1 receptor (IGF1R) gene silencing in response to cisplatin (DDP) in the lung cancer cell line A549 in vivo and in vitro were investigated using two plasmids expressing short hairpin RNA (shRNA) to IGF1R. A549 cells were transfected with plasmids expressing each shRNA and then treated with DDP. Semi-quantitative reverse transcription-PCR and Western blot analysis were used to detect the expression of IGF1R. MTT assay, flow cytometry and tumor growth assay in athymic nude mice were used to assess the chemosensitivity to DDP following IGF1R knockdown. Our data showed that the transfection of A549 cells with shRNA resulted in specific silencing of IGF1R by 78.9% at the mRNA level and by 89.8% at the protein level. Down-regulation of IGF1R significantly enhanced cell sensitivity to DDP, decreased the IC50 of DDP in A549 cells at 24 h, 48 h and 72 h, and retained 77.5% of A549 cells in the G0/G1 phase. Furthermore, shRNA-mediated silencing of IGF1R in combination with DDP treatment enhanced the suppression of tumor growth in both size and weight by more than 60% and increased apoptosis by more than 75% when compared with the controls in vivo. Suppression of IGF1R gene expression by shRNA enhances the chemosensitivity of A549 cells to DDP both in vitro and in vivo, indicating the therapeutic potential of RNA interference as a method for gene therapy in treating lung cancer.
Effects of a Cannabinoid Receptor (CB) 1 Antagonist AM251 on Behavioral Sensitization to Nicotine in a Rat Model of Novelty-seeking Behavior: Correlation with Hippocampal 5HT
There are marked individual differences in the efficacy of mainstream nicotine cessation agents in preventing relapse. A rat model of novelty-seeking phenotype was reported to have predictive value for psychostimulant taking behavior where locomotor reactivity to novelty is used to rank high (HR, highest 1/3) versus low (LR, lowest 1/3) responsiveness to novelty in outbred rats. We tested the hypothesis that a cannabinoid receptor (CB) 1 antagonist that is in clinical trials for smoking cessation may reverse behaviorally sensitizing effects of nicotine in HRs and repeated nicotine-induced elevations in hippocampal 5HT.
Assessment of 5-hydroxytryptamine Efflux in Rat Brain During a Mild, Moderate and Severe Serotonin-toxicity Syndrome
Serotonin (5-hydroxytryptamine; 5-HT)-toxicity syndrome, an iatrogenic brain disorder induced by excessive efflux of 5-HT, has received much attention because of increasing incidents of serotonergic antidepressants. However, the neural mechanism by which extracellular 5-HT is elevated to a toxic level for the syndrome remains to be determined. The goal of the present study was to test the hypothesis that extracellular 5-HT is composed of two component effluxes responsible for distinct aspects of the syndrome. The first set of experiments was to characterize the syndrome by measuring changes in neuromuscular signs, body-core temperature and mortality rate. Our results indicate that the syndrome severity can be categorized into mild, moderate and severe levels. The second set of experiments was to determine a threshold of extracellular 5-HT for induction of each level of the syndrome. Our results demonstrate that there were an 11-fold increase in the mild syndrome and an over 55-fold increase in the severe syndrome. In the last series of experiments, the excessive increases in 5-HT were pharmacologically separated into primary and secondary component effluxes with the 5-HT2A receptor antagonists cyproheptadine and ketanserin and NMDA receptor antagonist (+)-MK-801. Our results suggest that the primary component efflux was caused by direct drug effects on 5-HT biosynthetic and metabolic pathways and secondary efflux ascribed to indirect drug effect on a positive-feedback circuit involving 5-HT2A and NMDA receptors. In summary, the primary efflux could be an initial cause for the induction of the syndrome while the secondary efflux might involve deterioration of the syndrome.
Post-MPTP Treatment with Granulocyte Colony-stimulating Factor Improves Nigrostriatal Function in the Mouse Model of Parkinson's Disease
The neuroprotective effects of granulocyte colony-stimulating factor (G-CSF) were reported in several neurological disease models, including Parkinson's disease (PD). In the present study, we investigated the therapeutic effect of G-CSF after the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of PD was established. G-CSF was subcutaneously administered into C57BL/6 mice that had undergone systemic MPTP injections. We found that G-CSF treatment markedly increased the number of dopaminergic neurons in the substantia nigra pars compacta (SNpc) of the G-CSF-treated group. Consistent with this finding, we found a significant increase in dopamine release under high K(+) stimulation in the striatum of the G-CSF-treated animals compared to the MPTP-exposed mice. Finally, we observed a persistent recovery of locomotor function in the G-CSF-treated animals. These results suggest the potential therapeutic value of G-CSF in treating PD. However, our bromodeoxyuridine labeling experiment failed to identify any newly generated dopaminergic neurons in SNpc. This might indicate an indirect effect of G-CSF on cell proliferation. The underlying mechanism of G-CSF is under further investigation.
Involvement of 5-HT2A Receptors in the Serotonin (5-HT) Syndrome Caused by Excessive 5-HT Efflux in Rat Brain
Previous studies have demonstrated that serotonin (5-HT) syndromes, particularly for the malignant cases, can be alleviated by ice water mists, cooling blankets and many other external cooling measures. In this study, we tested the hypothesis that external cooling measures reduce the responsivity of 5-HT(2A) receptors to excessive 5-HT efflux, which may be a possible mechanism underlying the treatment of serotonin syndrome. To test this, rat experiments were carried out in the standard and cool ambient temperature (T(amb) ) by administration of the 5-HT precursor 5-hydroxy-L-tryptophan combined with the monoamine oxidase inhibitor clorgyline. The first set of experiments was to assess severity of the syndromes by measuring body temperature responses. Consistent with the hypothesis, we found that the syndrome was malignant at the standard T(amb) of 22°C but alleviated at 12 or 6°C, these results being similar to those in rats pre-treated with the 5-HT(2A) receptor antagonist ketanserin. The second set of experiments was to utilize microdialysis to determine the relationship between the syndrome severity and 5-HT levels at the above-mentioned T(amb) . We found that excessive 5-HT efflux consisted of primary and secondary components through two distinct mechanisms. Furthermore, the secondary component efflux, which can be ascribed to 5-HT(2A) receptor activation, was proportionally reduced at the cool T(amb) of 12 and 6°C. In conclusion, results of this study support the hypothesis that cooling T(amb) reduces the functional activity of 5-HT(2A) receptors, thus alleviating the malignant syndrome.
