• 제목/요약/키워드: in situ mRNA hybridization

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Ritanserin, a 5HT2/1C Receptor Antagonist, Does Not Block Cocaine-Induced Behavioral Alterations and zif268 mRNA Expression in the Striatum of the Rats

  • Choe, Eun-Sang;Kim, Jong-Yeon
    • The Korean Journal of Physiology and Pharmacology
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    • 제4권5호
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    • pp.355-359
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    • 2000
  • Cocaine induces immediate early gene expression and behavioral changes by blocking dopamine transporters in the terminals of nigrostriatal neurons in the striatum. The pharmacological role of serotonin 2/1C (5HT2/1C) receptors in cocaine-induced expression of zif268 (NGFI-A, egr1 and Krox-24) mRNA, a member of the zinc finger, was investigated using quantitative in situ hybridization histochemistry in vivo. Behavioral alterations induced by cocaine were also monitored in relation with blockade of the receptors. Systemic injection of ritanserin (1 mg/kg, s.c.), a 5HT2/1C receptor antagonist, did not reverse behavioral alterations and zif268 mRNA gene expression induced by 15 mg/kg cocaine, i.p., in the dorsal and ventral striatum. These data indicate that ritanserin-sensitive 5HT2/1C receptors are not necessary for cocaine-induced behavioral alterations and zif268 mRNA gene expression in the striatum.

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Streptococcus mutans의 gtfB 및 gtfC 유전자 발현에 대한 당의 영향 (THE EFFECTS OF SUGARS ON THE EXPRESSION OF GTFB AND GTFC MRNA)

  • 정혜진;김신;정진
    • 대한소아치과학회지
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    • 제34권2호
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    • pp.299-308
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    • 2007
  • Streptococcus mutans Ingbritt의 비수용성 글루칸을 합성하는 효소인 glucosyltransferase B 및 C의 mRNA 발현에 대한 각 당의 영향을 fluorescent in situ hybridization(FISH) 방법으로 관찰하여 다음의 결과를 얻었다. 1. 1% 설탕이 함유된 BHI 배지에서 S. mutans Ingbritt를 접종하여 배양할 때 배양 9시간 째 gtfB 및 gtfC유전자의 mRNA가 발현이 급격히 증가하였다. 2 BHI 액체배지에 설탕을 첨가한 경우 gtfB및 gtfC유전자의 mRNA가 발현되었으며, 10% 설탕을 첨가한 경우 1%와 5%보다 gtfB 및 gtfC 유전자의 발현이 감소되었다. 3. 1% 설탕이 첨가된 BHI 액체 배지에 당을 첨가한 경우 포도당은 10% 첨가하였을 때 gtfB 및 gtfC 유전자의 mRNA 발현이 감소하였고, 과당의 경우 1% 첨가한 경우 감소하여 5% 와 10%에서는 gtfB및 gtfC 유전자가 거의 발현되지 않았다. 4. 자일리톨의 경우 1%부터 gtfB및 gtfC유전자 mRNA 발현이 대조군보다 감소하였고, 5%와 10%에서는 두 유전자 모두 발현이 현저하게 감소하였다. 5. 유당의 경우 두 유전자의 발현에 거의 영향을 미치지 않는 것으로 관찰되었고, 솔비톨의 경우 각 농도에서 두 유전자의 발현이 대조군보다 감소하였다. 결론적으로 비수용성 글루칸 합성하는 gtfB및 gtfC 유전자의 mRNA 발현이 포도당, 과당 자일리톨 첨가에 의해 감소함을 알 수 있었다.

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Expression of the Multidrug Resistance Gene and its Product in Osteosarcomas of the Bone - Immunohistochemistry and In Situ Hybridization -

  • Park, Hye-Rim;Park, Yong-Koo
    • 대한골관절종양학회지
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    • 제3권1호
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    • pp.9-17
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    • 1997
  • Resistance to combination chemotherapy remains challenge in the treatment of osteosarcoma. One of the mechanisms of multiple drug resistance is an increased expression of the multidrug resistance gene(mdr1). Expression of the P-glycoprotein(mdr-1 gene product) was studied immunohistochemically and the mdr-1 gene by in situ hybridization in 33 osteosarcomas relating to various prognostic factors. Thirty cases out of 33 osteosarcomas(90.9%) showed positive cytoplasmic reactions with P-glycoprotein and nineteen instances(57.6%) were strong positive(2+). The older(>20 years) and female patients revealed more intense immunohistochemical reactions rather than those of the younger and male patients. Osteoblastic and chondroblastic osteosarcomas revealed more strong immunohistochemical reactions compared to fibroblastic types. There were no significant staining differences between the type of bony involvement, Broder's grade and the presence of necrosis. On follow-up, the mean survival rate was decreased in the strong positive group, however, this was not statistically significant. In situ hybridization for mdr-1 gene revealed positive signals in 22 cases out of 29 osteosarcomas(75.9%). Chemotherapy was done in 15 cases out of 28 patients(53.6%). The results of immunohistochemistry and in situ hybridization were not correlated with the protocols for chemotherapy. However, this result should be confirmed by a larger scale study about mdr1 mRNA expression.

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흰쥐 난소 및 부신에서 Steroidogenic Acute Regulatory Protein mRNA의 발현에 관한 연구 (Identification of Steroidogenic Acute Regulatory Protein mRNA in the Rat Ovary and Adrenal G land)

  • 김명옥
    • 한국발생생물학회지:발생과생식
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    • 제2권1호
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    • pp.39-43
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    • 1998
  • 스테로이드 호르몬의 합성은 콜레스테롤로부터 시작되고 Steroidogenic acute regulatory protein(StAR)은 스테로이드의 합성과정에서 콜레스테롤을 미토콘드리아의 안으로 신속하게 운반하는 역할을 한다. 스테로이드 호르몬은 난소, 부신, 고환에서 합성되며 본 연구에서는 흰쥐 난소와 부신에서 StAr mRNA의 발현 양상을 in situ hybridization 기법을 이용하여 조사하였다. 난소의 경우 StAR mRNa는 프로게스테론을 분비하는 황체에서 강한 발현을 보였고 엔드로겐을 분비하는 난포막세포에서도 약한 발현을 보였으며 에스트로겐을 분비하는 과립막세포에서는 발현되지 않았다. 황체에서도 황체의 발달 저도에 따라 차이를 보였고 성장한 황체에서는 강한 발현을 보인 반면, 퇴화하는 황체에서는 약한 발현을 보였다. 부신에서 StAR mRNA는 피질에서 강한 발현을 보였고 수질에서는 발현되지 않았다. 특히, 피질의 토리층에서보다 다발층과 그물층에서 강한 발현을 보였다. 난소와 부신에서 StAR mRNA는 스테로이드 호르몬의 종류에 따라서 발현 양상이 달랐고 스테로이드 호르몬 합성의 초기 단계에 관여함을 알 수 있었다.

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gtf 유전자 발현에 대한 xylitol의 영향 (THE EFFECT OF XYLITOL ON THE EXPRESSION OF GTF GENE)

  • 염정현;정진;정태성;김신
    • 대한소아치과학회지
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    • 제31권2호
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    • pp.304-313
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    • 2004
  • 천연의 당알콜계 감미료인 자일리톨을 사용하여 자당으로부터 글루캔을 합성하는 효소인 glucosyltransferase(GTF)의 mRNA 발현에 미치는 영향을 Fluorescent in situ hybridization(FISH)과 유식세포측정기를 이용하여 분석하여 다음의 결과를 얻었다. 1. FISH분석결과 1% 자당이 함유된 BHI 배지에 1%, 5%, 10% 자일리톨을 첨가하였을 때 gtfB, gtfC 및 gtfD mRNA 발현이 농도 의존적으로 억제되었고, gtfB 보다는 gtfC 및 gtfD의 발현이 더 크게 억제되었다. 2. 유식세포측정기로 분석한 결과 자당의 농도가 0.1%, 0.5%, 1%로 증가함에 따라 gtfB, gtfC 및 gtfD mRNA 발현이 증가하였다. 3. 유식세포측정기로 분석한 결과 자당이 1%함유되었을 때 1%, 5%, 10% 자일리톨을 첨가한 경우 gtfB, gtfC 및 gtfD mRNA발현이 유사하게 억제되었으며, 10% 자일리 톨을 첨가한 경우 gtfB, gtfC 및 gtfD mRNA발현이 가장 크게 억제되었다. 이상의 결과를 종합하면 자일리톨은 자당첨가에 의한 Streptococcus mutars의 gtf mRNA 발현을 억제하여 항우식효과에 기여하는 것으로 생각된다.

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미성숙과 성숙한 흰쥐 고환에서의 Steroidogenic acute regulatory protein mRNA의 발현 (Expression of steroidogenic acute regulatory protein mRNA in immature and adult rat testes)

  • 고필옥;곽수동
    • 대한수의학회지
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    • 제40권2호
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    • pp.229-236
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    • 2000
  • The synthesis of steroid hormone starts from cholesterol. Steroidogenic acute regulatory protein (StAR) acutely transfers cholesterol from the outer mitochondrial membrane to the inner in the early step of steroidogenesis. Many kinds of steroid hormone are mainly synthesized in adrenal grand, ovary, and testis. Among the steroid hormone, testosterone is synthesized in Leydig cells of the testis, the production of testosterone significantly increases in adult testis after puberty onset. Therefore, we think that the expression of StAR mRNA in testis will change according to the testicular development. The aim of this study is to determine the distribution of StAR mRNA in immature and adult rat testes and to confirm the functions of StAR in these testes. Thus, in situ hybridization was used in rat testes of the 2, 4, and 10 weeks of age. StAR mRNA was expressed in Leydig cells. Positive signals of StAR mRNA were weakly detected in Leydig cells of the 2 weeks of age. But, StAR mRNA was strongly expressed in Leydig cells of the 4 and 10 weeks of age, where steroidogenesis actively occur. In our results, the pattern of StAR mRNA expression was similar to the pattern of testosterone production in immature and adult rat testes. In conclusion, we can suggest that StAR acts as an important factor to regulate the synthesis of testosterone in Leydig cells of the rat testis.

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Developmental Patterns of mST3GaIV mRNA Expression in the Mouse: In Situ Hybridization using DIG-labeled RNA Probes

  • Ji, Min-Young;Lee, Young-Choon;Do, Su-Il;Nam, Sang-Yun;Jung, Kyu-Yong;Kim, Hyoung-Min;Park, Jong-Kun;Choo, Young-Kug
    • Archives of Pharmacal Research
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    • 제23권5호
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    • pp.525-530
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    • 2000
  • mST3GaIV synthesizes ganglioside GM3, the precursor for simple and complex a- and b- series gangliosides, and the expression and regulation of mST3GaIV (CMP-NeuAc: lactosylceramide $\alpha$2,3-sialyltransferase) activity is central to the production of almost all gangliosides, a class of glycosphingolipids implicated in variety of cellular processes such as transmembrane signaling, synaptic transmission, specialized membrane domain formation and cell-cell interactions. To understand the developmental expression of mST3GaIV in mice, we investigated the spatial and temporal expression of mST3GaIV mRNA during the mouse embryogenesis [embryonic (E) days; 19, E11, E13, E15] by in situ hybridization with digoxigenin-labeled RNA probes. All tissues from 19 and E11 were positive for mST3GaIV mRNA. On E13, mST3GaIV mRNA was expressed in various neural and non-neural tissues. In contrast to these, on E15, the telencephalon and liver produced a strong expression of mST3GaIV which was a quite similar to that of E13. In this stage, mST3GaIV mRNA was also expressed in some non-neural tissues. These data indicate that mST3GaIV is differently expressed at developmental stages of embryo, and this may be importantly related with regulation of organogenesis in mice.

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랫드 난소에서 난포 발달에 따른 DNA 결합 단백질 억제인자 (Inhibitor of DNA Binding Protein) Id1 and Id2 mRNA 발현 (Inhibitor of DNA Binding Protein (Id)1 and Id2 mRNA Expression on Folliculogenesis in Rat Ovary)

  • 황성수;김평희;고응규;양병철;성환후;민관식;윤종택
    • 한국수정란이식학회지
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    • 제23권3호
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    • pp.183-187
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    • 2008
  • This study was conducted to analyze the expression pattern of inhibitor of DNA binding proteins (Id)1 and Id2 mRNA on folliculogenesis in rat ovary. The ovaries were obtained from 27 days old Sprague-Dawley rat, fixed, dehydrated, and paraffin embedded. For in situ hybridization, anti-sense and sense Idl and Id2 cRNA probes were prepared and applied to the ovarian section. The ovarian sections were coated with NTB-2 emulsion. After that, the slides were developed and counterstained with hematoxylin and eosin staining. In oocytes, the hybridizational signals of Id1 mRNA were strong in primordial and primary follicles, however, there were no signals in that of atretic or preovulatory follicles. The Id2 mRNA signals were also strong in the oocytes of primordial, primary and secondary follicles. Interestingly, the Id2 mRNA was expressed specifically granulosa cells, but nor in oocyte or theca cells in dominant and preovulatory follicles. Based on these results, Id1 and Id2 mRNA was expressed specifically at follicle stages and follicular tissue and might be closely related with follicle development.

Mobile transposon-like element, clone MTi7:RNA interference를 이용한 역할 규명 (Mobile transposon-like element, clone MTi7: Finding its role(s) by RNA interference)

  • 박창은;신미라;전은현;조성원;이숙환;김경진;김남형;이경아
    • Clinical and Experimental Reproductive Medicine
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    • 제30권4호
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    • pp.299-307
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    • 2003
  • Objectives: The present study was conducted to evaluate the mobile transposon-like element, clone MTi7 (MTi7) expression in the mouse ovary and to determine its role(s) in the mouse oocytes by RNA interference (RNAi). Methods: MTi7 mRNA expression was localized by in situ hybridization in day5 and adult ovaries. Double stranded RNA (dsRNA) was prepared for c-mos, a gene with known function as control, and the MTi7. Each dsRNA was microinjected into the germinal vesicle (GV) stage oocytes then oocyte maturation and intracellular changes were evaluated. Results: In situ hybridization analysis revealed that MTi7 mRNA localized to the oocyte cytoplasm from primordial to preovulatory follicles. After dsRNA injection, we found 43-54% GV arrest of microinjected GV oocytes with 68%-90% decrease in targeted c-mos or MTi7 mRNA. Conclusions: This is the first report of the oocyte-specific expression of the MTi7 mRNA. From results of RNAi for MTi7, we concluded that the MTi7 is involved in the germinal vesicle breakdown in GV oocytes, and MTi7 may be implicated with c-mos for its function. We report here that RNAi provides an outstanding approach to study the function of a gene with unknown functions.