• The Korean Journal of Physiology and Pharmacology
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• v.15 no.6
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• pp.389-395
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• 2011

Repeated administration of psychostimulants such as cocaine leads to the development of behavioral sensitization. Extracellular signal-Regulated Kinase (ERK), an enzyme important for long-term neuronal plasticity, has been implicated in such effects of these drugs. Although the nucleus accumbens (NAcc) is the site mediating the expression of behavioral sensitization by drugs of abuse, the precise role of ERK activation in this site has not been determined. In this study we demonstrate that blockade of ERK phosphorylation in the NAcc by a single bilateral microinjections of PD98059 (0.5 or $2.0{\mu}g/side$), or U0126 (0.1 or $1.0{\mu}g/side$), into this site dose-dependently inhibited the expression of cocaine-induced behavioral sensitization when measured at day 7 following 6 consecutive daily cocaine injections (15 mg/kg, i.p.). Acute microinjection of either vehicle or PD98059 alone produced no different locomotor activity compared to saline control. Further, microinjection of PD98059 ($2.0{\mu}g/side$) in the NAcc specifically lowered cocaine-induced increase of ERK phosphorylation levels in this site, while unaffecting p-38 protein levels. These results indicate that ERK activation in the NAcc is necessary for the expression of cocaine-induced behavioral sensitization, and further suggest that repeated cocaine evokes neuronal plasticity involving ERK pathway in this site leading to long-lasting behavioral changes.

• Korean Journal of Poultry Science
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• v.28 no.2
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• pp.125-133
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• 2001

The goal of this paper was to examine the qualify zygote-acquiring method for in-vitro culture and the in-vitro culture method of the acquired zygote from a technological perspective. We have reported the results on the introduction of foreign DNAs using the described culturing method. After performing in-vitro and surrogate eggshell culture on a zygote acquired from the abdomen of a hen, 25.8% hatchability was acquired. After microinjecting foreign DNAs into the acquired zygote and performing in-vitro and surrogate eggshell culture using the same method, 13.1∼11.7% hatchability was acquired. Having compared the developments of the control subjects and the experimental subjects, the viability of the experimental subjects on the 4∼5th day of culturing was much lower compared to that of the control subjects. This is a result that shows that the microinjection process of foreign DNAs might have a negative effect on the existence of the embryo; therefore, various technical attempts should be made to minimize such negative effects. Having microinjected foreign DNAs into the zygote of a hen to produce transgenic chickens, 3 transgenic founders were Produced and 70 G1 progeny were produced as a result of the progeny test that had been performed to the present.

• Journal of Physiology & Pathology in Korean Medicine
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• v.29 no.5
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• pp.416-421
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• 2015

This study was performed to comparative investigate the distribution of primary sensory and motor neurons associated with Daereung(PC7) acupoints by using neural tracing technique. A total 16 SD rats were used in the present study. After anesthesia, the rats received microinjection of 6 ㎕ of cholera toxin B subunit(CTB) into the corresponding sites of the acupoints Daereung(PC7), in the human body for observing the distribution of the related primary sensory neurons in dorsal root ganglia(DRGs) and motor neurons in the spinal cord(C3∼T4) and sympathetic ganglia. Three days after the microinjection, the rats were anesthetized and transcardially perfused saline and 4% paraformaldehyde, followed by routine section of the DRGs, sympathetic chain ganglia(SCGs) and spinal cord. Labeled neurons and nerve fibers were detected by immunohistochemical method and observed by light microscope equipped with a digital camera. The labeled neurons were recorded and counted. From this research, the distribution of primary sensory and motor neurons associated with Daereung(PC7) acupoints were concluded as follows. Muscle meridian related Daereung(PC7) controlled by spinal segments of C5∼T1, C6∼T4, respectively.

• Applied Microscopy
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• v.22 no.2
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• pp.66-74
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• 1992

In an attempt to produce transgenic amphibia, bacteriophage ${\lambda}-DNA$ was microinjected into fertilized eggs of Xenopus laevis, and the effect on early embryogenesis and the ultrastructural behavior of exogenous DNA were investigated. The effect of microinjected gene on embryonic development showed differences according to the concentration of injected DNA and the incubation temperature. Various concentrations of ${\lambda}-DNA$ were tested. Among those, microinjection of 1-2 ng DNA dissolved in 20 nl TE buffer was not shown to disturb normal embryonic development and was recorded the highest survivability to the late tadpole stage (Stage 43); however, injection of increased concentrations of DNA than above provoked irregular cleavages or abnormal appearances, which resulted in reduced survivability. When the injected embryos were incubated at low temperatures (e.g., $12^{\circ}C$), 54.5% of the embryos developed to Stage 43, whereas 42.4% survived when incubated at room temperature. The survivability showed also differences according to the injection site. 58.0% of the embryos developed to Stage 43 when microinjected into the vegetal pole, whereas 44.9% survived when microinjected into the animal pole. To understand the structural fate or behavior of injected DNA a combined light and electron microscopical study was applied. The nucleus-like structure was observed in the ${\lambda}$ DNA-injected embryos, which was quite a similar to the interphase nuclei of normal Xenopus laevis. The nucleus-like structure showed the typical double-layered nuclear membrane and nuclear complexes; however, it consisted of unusual structures such as furrows of nuclear envelope into the nucleoplasm.

• Molecules and Cells
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• v.26 no.1
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• pp.81-86
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• 2008

RNA interference (RNAi) was performed on several essential genes in the pinewood nematode Bursaphelenchus xylophilus, which causes pine wilt disease. Double-stranded RNA (dsRNA) was delivered to larvae or adult worms by soaking, electroporation, or microinjection. Soaking and electroporation of L2-L3 stage worms in solutions containing dsRNA for essential genes induced over 25% lethality after 5 days, and gene-specific phenotypes were observed. This lethality agreed with significant reductions of the targeted transcripts, as assayed by reverse-transcription coupled with real time PCR. Microinjection was the most efficient route as measured by the hatching rate of F1 embryos, which was reduced by 46%. When adult worms were soaked in dsRNA, lethality was induced in the F1 larvae, revealing the persistence of knockdown phenotypes. The penetrance of the RNAi phenotypes for essential genes was relatively low but consistent, indicating that RNAi should be useful for studying the in vivo functions of B. xylophilus gene products.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.3
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• pp.275-281
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• 2015

Orthostatic hypotension is most common in elderly people, and its prevalence increases with age. Attenuation of the vestibulo-sympathetic reflex (VSR) is commonly associated with orthostatic hypotension. In this study, we investigated the role of glutamate on the vestibulo-solitary projection of the VSR pathway to clarify the pathophysiology of orthostatic hypotension. Blood pressure and expression of both pERK and c-Fos protein were evaluated in the nucleus tractus solitarius (NTS) after microinjection of glutamate into the medial vestibular nucleus (MVN) in conscious rats with sodium nitroprusside (SNP)-induced hypotension that received baroreceptor unloading via sinoaortic denervation (SAD). SNP-induced hypotension increased the expression of both pERK and c-Fos protein in the NTS, which was abolished by pretreatment with glutamate receptor antagonists (MK801 or CNQX) in the MVN. Microinjection of glutamate receptor agonists (NMDA or AMPA) into the MVN increased the expression of both pERK and c-Fos protein in the NTS without causing changes in blood pressure. These results indicate that both NMDA and AMPA receptors play a significant role in the vestibulo-solitary projection of the VSR pathway for maintaining blood pressure, and that glutamatergic transmission in this projection might play a key role in the pathophysiology of orthostatic hypotension.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.99-99
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• 2002

The hormone resistin is associated with typeII diabetes mellitus in rodent model. Resistin impairs glucose tolerance and insulin action. A new class of anti-diabetic drugs were called thiazolidinediones (TZDs) downregulates a resistin which is induced during adipocyte differentiation. But the connection between increased adiposity and resistin remains unknown. The objectives of this study was to clone a mouse resistin cDNA and to generate transgenic mice overexpressing mouse resistin gene. The 555 bp of mouse resistin was amplified from mob cDNAS by polymerase chain reaction (PCR) and cloned into pCR$\^$(R)/ 2.1 TOPO T-vector. Mouse resistin mRNA on the basis of Genbank sequence (acession no. AF323080). Then, the PCR product was cloned into pTargeT$\^$TM/ mammalian expression vector that has pCMV promoter and chimeric intron. Restriction enzyme analysis with BamH I and Not I was carried out to determine an orientation of the insert in the vector. The pCMV-mus/resistin gene was prepared from previous recombinant pTargeT$\^$TM/-mus/resistin by digestion of Bgl II, and has used for microinjection into pronuclei of one cell embryos. The microinjected embryos were transfered to pseudopregnant foster-mother. Mouse resistin expression was detected in transgenic F1 mice by Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR). Resistin gene expression mouse has heavier body weight which was measured higher level of plasma glucose than that of normal mouse. And in diet-induced experiments, the abdominal fat pads were isolated from each 24h starvation and re-feeding after fasting group mice that were assessed by RT-PCR analysis. In fasting group mice, resistin expression was higher than that of re-feeding group mice. This result suggests that the resistin gene overexpressing mice may be became to obesity and be useful as an animal disease model to be diabetes mellitus caused by insulin resistance of resistin.

• Proceedings of the Korean Society for Technology of Plasticity Conference
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• 2003.05a
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• pp.465-469
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• 2003

Microlens arrays were fabricated using a novel fabrication technology based on the exposure of a PMMA (Polymethylmethacrylate) sheet to deep X-rays and subsequent thermal treatment. X-ray irradiation causes the decrease of molecular weight of PMMA, which in turn decreases the glass transition temperature and consequently causes a net volume increase during the thermal cycle resulting in a swollen microlens. A new physical modeling and analyses for microlens formation were presented according to experimental procedure. A simple analysis based on the new model is found to be capable of predicting the shapes of microlens which depend on the thermal treatment. For the replication of microlens arrays having various diameters with different foci on the same surface, the hot embossing and the microinjection molding processes has been successfully utilized with a mold insert that is fabricated by Ni-electroplating based on a PMMA microstructure of microlenses. Fabricated microlenses showed good surface roughness with the order of 1 nm.

• Journal of Photoscience
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• v.9 no.2
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• pp.243-245
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• 2002

Pigment-dispersing factor (PDF) is an octadecapeptide distributed in the optic lobe and the brain in a variety of insect species. There are lines of evidence suggesting possible involvement of PDF in the insect circadian system. However, its physiological roles in the circadian time keeping mechanism have not been clearly defined. In this study, we have examined the phase shifting effects of Gryllus-PDF on the circadian locomotor rhythm in the cricket Gryllus bimaculatus of which circadian clock is located in the optic lobe. Phase shifts in the circadian activity rhythm were measured following microinjection of 22nl of vehicle (Ringer's solution) or O.lmM PDF into the optic lobe through the compound eye at various circadian times. The results showed that PDF induced phase shifts of the circadian clock in a phase-dependent manner, suggesting that it may play a role as an input signal for the circadian clock.

• Reproductive and Developmental Biology
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• v.32 no.4
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• pp.229-235
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• 2008

Silencing of Dicer1 by siRNA did not inhibit development up to the blastocyst stage, but decreased expression of selected transcription factors, including Oct-4, Sox2 and Nanog, suggesting that Dicer1 gene expression is associated with differentiation processes at the blastocyst stage (Cui et al., 2007). In order to get insights into genes which may be linked with microRNA system, we compared gene expression profiles in Gapdh and Dicer1 siRNA-microinjected blastocysts using the Applied Biosystem microarray technology. Our data showed that 397 and 737 out of 16354 genes were up- and down-regulated, respectively, following siRNA microinjection (p<0.05), including 24 up- and 28 down-regulated transcription factors. Identification of genes that are preferentially expressed at particular Dicer1 knock down embryos provides insights into the complex gene regulatory networks that drive differentiation processes in embryos at blastocyst stage.