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EFFECTS OF GLUTAMATERGIC BLOCKADE ON BEHAVIORAL AND NEURONAL CHANGES IN PARKINSONS DISEASE RAT MODEL Ying-Jui Ho1, Ying-Cih Hung1, Chin-Yen Chou1, Shih-Chun Ho1, Guo-Joe Huang1, Tan-Yu Liao1, Guan-Da Huang1, Shih-Hui Hsu2, Sheng-Huei Wu School of Psychology, 2School of Medical Laboratory and Biotechnology, Chung Shan Medical University (Taichung City, Taiwan, ROC) yjho@csmu.edu.tw In addition to motor dysfunction, up to 20-30% of patients with Parkin sons disease (PD) suffer from dementia also, which is known as Parkinsons disease dementia (PDD). The features of PDD include impairments of working memory and recognition. Hyper-glutamatergic activity has been implicated in the pathophysiology of PD. Although NMDA receptor- and metabotropic glu tamate receptor (mGluR)-mediated excitotoxicity may take part in neuronal de generation in PD, role of these two kinds of receptors in PDD remains unclear.

To address this issue, systemic administration with 2-methyl-6-(phenylethynyl) pyridine (MPEP), a selective antagonist of mGluR5, and MK-801, a noncompet itive antagonist of NMDA receptors, were used to evaluate the role of glutama tergic activity in neurodegeneration and behavioral deficits in 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced rat PD model. One day after MPTP lesioning, the rats received daily treatment with MPEP (2 mg/kg, i.p.) or MK-801 (0.2 mg/kg/day, i.p.) for 14 days. Motor function and working memory were measured by using the bar test and T-maze test, respectively. MPTP caused significant motor deficit, compared to the sham-operated group, on day 1. But this impairment recovered spontaneously on day 7. A decrease in working memory was observed in MPTP-lesioned rats, which was restored by both MPEP and MK-801 treatment. Further, the administration of MPEP and MK 801 prevented the disturbance of object recognition in MPTP-lesioned rats.

Moreover, the administration of MPEP and MK-801 significantly ameliorated MPTP-induced neuroinflammation, dopaminergic degeneration in the nigrostria tal system, and neuronal loss in the hippocampal CA1 area. These results sug gest that suppression of glutamatergic activity may have clinical potential in the treatment of dementia associated with PD.

SIMULTANEOUS FILTRATION AND FOCUSING OF BIOLOGICAL CELLS ON AN INSULATOR-BASED DIELECTROPHORETIC MICRODEVICE Chun-Ping Jen1, Wei-Fu Chen1 and Tamara G. Amstislavskaya National Chung Cheng University (Chia-Yi 621, Taiwan, R.O.C.) Institute of Cytology and Genetics SB RAS (Novosibirsk, Russia) imecpj@ccu.edu.tw Manipulating and separating biological cells of interest using microfluidic and micro total analysis system (TAS) devices has potential applications in clinical diagnostics and medicine. For example, in investigations of the nervous system, glial cells can enwrap neuronal cells and thus shield neurons from a di rect extracellular access. Cellular focusing in microfluidic devices is a prerequi site for medical applications, such as cell sorting, cell counting or flow cytome try. In the present study, an insulator-based dielectrophoretic microdevice is de signed for the simultaneous filtration and focusing of biological cells. The cells are introduced into the microchannel and pre-confined hydrodynamically by the funnel-shaped insulating structures close to the inlet. Dead and viable human carcinoma (HeLa) cells were cultured for the experiment demonstrating hydro dynamic separation herein. There are ten sets of X-patterned insulating struc tures in the microfluidic channel. The main function of the first five sets of insu lating structures is to guide the cells by negative dielectrophoretic responses (vi able HeLa cells) into the center region of the microchannel. The positive dielec trophoretic cells (dead HeLa cells) are attracted to regions with a high electric field gradient generated at the edges of the insulating structures. The remaining five sets of insulating structures are mainly used to focus negative dielectropho retic cells that have escaped from the upstream region. Experiments employing a mixture of dead and viable HeLa cells are conducted to demonstrate the effec tiveness of the proposed design. The results indicate that the performance of both filtration and focusing improves with the increasing strength of the applied electric field and a decreasing inlet sample flow rate, which agrees with the trend predicted by the numerical simulations. The filtration efficiency, which is quantitatively investigated, is up to 88% at an applied voltage of 50 volts peak to-peak (1 kHz) and a sample flow rate of 0.5 L/min. The proposed device can focus viable cells into a single file using a voltage of 35 volts peak-to-peak ( kHz) at a sample flow rate of 1.0 L/min.

This work was supported by grant No. 148 SB RAS, Russia - NSC, Tai wan Research Cooperation.

LONG-TERM ADVERSE EFFECT OF NEONATAL GLUCOCORTICOID TREATMENT ON AMYGDALA Kwok-Tung Lu 1, Meng-Chang Ko1, Yu-Hui Hung1, Yi-Ling Yang Department of Life Science, National Taiwan Normal University, Taipei, Taiwan Department of Biochemical Science and Technology, National Chia-Yi University, Chia-Yi, Taiwan ktlu@ntnu.edu.tw In addition to neonatal adversity, some neonatal medications are suspect ed to produce subtle changes in brain maturation, the impact of which may be come apparent only in the late period of life. Dexamethasone (DEX) is a syn thetic glucocorticoid and is frequently used to lessen the progression of chronic lung disease in extremely low birth weight (ELBW) infants. Even though clini cal trials failed to consistently demonstrate improvement in mortality or length of hospitalization, exposures of up to 42 days were frequently used as a thera peutic agent to lessen the morbidity of chronic lung disease in premature infants.

Recent studies pointed out that the neonatal DEX treatment alters hippocampal synaptic plasticity in juvenile. Little is known about the long-term effect of neo natal DEX treatment on amygdale function. The current study was performed to evaluate the long-term effect of neonatal DEX treatment on amygdale synaptic plasticity. Male adult Wister rats were subjected to receive subcutaneous injec tion (sc) of tapering doses of DEX (0.5 mg/kg, 0.3 mg/kg and 0.1mg/kg) from postnatal day 1 to 3, PN1~PN3. Animals were then subjected for fear potentiated startle (FST) and electrophysiological recording at the age of 6 or weeks. Results showed that neonatal DEX treatment temporary decrease body weight in the young rats, but not affected in the later life when compared with the control group. In addition, neonatal DEX treatment affected the formation of amygdaloid long-term potentiation. Furthermore, neonatal DEX treated animals showed deficit on the acquisition of fear-potentiated startle (FST) which re vealed neonatal DEX treatment has long-term adverse effect on amygdale func tion and resulted in adverse consequence in the later life.

MATERNAL SEPARATION OF DIFFERENT INTERVALS INDUCES SEX DIFFERENCES IN ANXIETY RESPONSES IN POST-WEANING RATS Yuan-Feen Tsai1, Yi-Sian Wu1, Hsiao-Fung Pu2, Mei-Ying Yeh1, Su-Ping Wang1, Mei-Yun Tai Institute of Physiology, College of Medicine, National Taiwan University, Taiwan, ROC Department of Physiology, School of Medicine, National Yang-Ming University, Taiwan, ROC yftsai@ntu.edu.tw Early life experience has prolonged effects on emotionality and endocrine functions at adulthood. Maternal separation has been described to induce differ ent anxiety responses at adult age, but rare studies have investigated the effects of maternal separation in post-weaning rats. In the present study, we used both male and female Long-Evans rats at 22 days of age to reveal the effects of ma ternal separation on anxiety-like behavior, the basal levels of corticosterone and adrenal gland weights. Neonatal pups were randomly assigned to three groups:

control, maternal separation 15 minutes (MS-15), and maternal separation minutes (MS-180). Infant rats were deprived of maternal contact between the 4th and the 21st postnatal days. The elevated plus-maze was used to establish the anxiety-like behavior profile of animals. In female rats, the MS-15 group showed significantly lower anxiety-like behavior, serum corticosterone levels and lighter adrenal glands than control rats. However, the female MS-180 ani mals had significantly lower serum corticosterone levels compared to the female controls, but their anxiety-like performance was not significantly different from that in the controls. In contrast, although there was no significant difference in the anxiety-like behavior between these three male groups, the male MS-15 rats had lighter adrenal glands than those of the male MS-180 rats. Both the male MS-15 and MS-180 groups had lower serum corticosterone levels than the male controls. Together, these results suggest that neonatally maternal separation of different intervals may cause sex differences in anxiety-like behavior in post weaning rats.

STRESS EFFECT OF THYROIDECTOMY ON YHE SECRETION OF GLUCOCORTICOIDS IN MALE RATS Paulus S. Wang1,2,3,4,5, Yung-Hsing Yeh1, Cai-Yun Jian1, Shyi-Wu Wang Dept. Physiology, Sch. Med., National Yang-Ming University, Taipei, Dept. Med. Res. Edu., Taipei Veteran General Hospital, Taipei, Dept. Med. Res. Edu., Taipei City Hospital, Taipei, Grad. Inst. Exerc. Sports Sci., Taipei Physical Edu. Coll., Taipei, Ph.D. Program for Aging, China Med. University, Taichung, Dept. Physiol. Pharmacol., Coll. Med., Chang-Gung University, Taoyuan, Taiwan, Republic of China pswang@ym.edu.tw Both adrenocorticotropin (ACTH) and glucocorticoids are stress hor mones for regulating psychoneuroendocrine functions. It has been shown that the pituitary-adrenal functions are modulated by hypothyroidism. Thyroid hor mones increases the secretion of ACTH and consequently corticosterone pro duction. Hypothyoid men have higher levels of plasma cortisol. However, the detail mechanisms of the effects of thyroid hormones on glucocorticoid produc tion are unclear. In the present studies, the effects of thyroid hormones and thy roidectomy on glucocorticoid secretion were studied. Blood samples were col lected from male rats after receiving intravenous 3,5,3-triiodothyronine (T3) or thyroxine (T4). Zona fasciculata-reticularis (ZFR) cells were treated with ACTH, T3, T4, ACTH plus T3, or ACTH plus T4 at 37C for 2 h. Male rats were thyroidectomized (Tx) or sham Tx for 2 weeks before challenging with ACTH. Blood samples were collected at different time intervals. In another in vitro study, rat ZFR cells were incubated with or without forskolin, 8-Br-cAMP, or ACTH at 37 for 1 h. The concentrations of corticosterone in plasma and medium samples were measured by radioimmunoassay. Administration of T and T4 suppressed the basal and the ACTH-stimulated levels of plasma corti costerone. In ZFR cells, both thyroid hormones inhibited ACTH-stimulated cor ticosterone secretion, but the basal corticosterone was inhibited only with T310-10M or T4 10-8M. Likewise, T3 or T4 inhibited the basal- and ACTH stimulated levels of intracellular cAMP. Thyroidectomy resulted in an increase of the basal level of plasma corticosterone and spontaneous release of corti costerone from rat ZFR cells. The maximal response of plasma corticosterone to ACTH challenge was attenuated by thyroidectomy. Thyroidectomy caused an increased response of corticosterone production in vitro to the administration of forskolin but not 8-Br-cAMP. These results suggest that removal of thyroid hormones is a stress to increase the production of glucocorticoids via a mecha nism at least in part associated with an activation of adenylyl cyclase. Mean while, thyroid hormones counteract ACTH in adrenal steroidogenesis through their inhibition of cAMP production in ZFR cells.

RESVERATRON PROTECTS RATS FROM A-INDUCED NEUROTOXICITY BY THE REDUCTION OF iNOS EXPRESSION AND LIPID PEROXIDATION Yi-Ling Yang1, T.C. Huang2, K. T. Lu2, Y.Y. P. Wo1, Y.J. Wu Department of Biochemical Science and Biotechnology, National Chia-Yi University, Chia Yi, Taiwan, 2Department of Life Science, National Taiwan Normal University, Taipei, Taiwan ylyang@mail.ncyu.edu.tw Alzheimer disease (AD) is an age-dependent neurodegenerative disease characterized by the formation of amyloid (A)-containing senile plaque. The disease could be induced by the administration of Ab peptide, which was also known to upregulate inducible nitric oxide synthase (iNOS) and stimulate neu ronal apoptosis. The present study is aimed to elucidate the cellular effect of resveratrol, a natural phytoestrogen with neuroprotective activities, on A induced hippocampal neuron loss and memory impairment. On adult Sprague Dawley rats, we found the injection of A could result in a significant impair ment in spatial memory, a marked increase in the cellular level of iNOS and li pid peroxidation, and an apparent decrease in the expression of heme oxygen ase-1 (HO-1). By combining the treatment with A, resveratrol was able to con fer a significant improvement in spatial memory, and protect animals from A induced neurotoxicity. These neurological protection effects of resveratrol were associated with a reduction in the cellular levels of iNOS and lipid peroxidation and an increase in the production of HO-1. Moreover, the similar neurological and cellular response were also observed when A treatment was combined with the administration of a NOS inhibitor, N(G)-nitro-L-arginine methyl ester hy drochloride (L-NAME). These findings strongly implicate that iNOS is involved in the A-induced lipid peroxidation and HO-1 downregulation, and resveratrol protects animals from A-induced neurotoxicity by suppressing iNOS produc tion.

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