• Title/Summary/Keyword: In situ Hybridization Histochemistry

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Expression of Cdc25B mRNA in Duodenal Mucosa of Chicken

  • Qin, Junhui;Zhang, Hui;Bao, Huijun;Zhou, Qiang;Liu, Yi;Xu, Chunsheng;Chu, Xiaohong;Chen, Qiusheng
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.4
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    • pp.530-536
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    • 2010
  • Cdc25B is a mitotic regulator that might act as a starter phosphatase to initiate the positive feedback loop at the entry into mitotic (M) phase. In the present study, distribution of Cdc25B mRNA in duodenal mucosa of the chicken was demonstrated by means of in situ hybridization histochemistry (ISHH) using sense and antisense digoxigenin (DIG)-labeled RNA probes. The results showed that there were many labeled cells distributing in the duodenal mucosa of the adult chicken. Of these labeled cells, 81.60${\pm}$9.63% of Cdc25B mRNA positive cells was distributed in the basilar part and mid-portion of the intestinal gland and 36.21${\pm}$8.81% in the middle and basilar portion of villi of the small intestine of the chicken, respectively. Most of these labeled cells were positive in the regions of the stem cell and proliferation. The signals of ISHH decreased from basilar to upper part in the crypt of Lieberkuhn and weakened in the inferior villi of the duodenum. Moreover, the positive signals were both in the cytoplasm and cell nucleus. However, the labeled cells were negative in both the lamina muscularis mucosae and muscular layer. The results of ISHH suggested the existence of Cdc25B mRNA and vigorous proliferation activities in the duodenal mucosa of adult chicken, replenishing the cells which had sloughed off from the superior part of the villus. Our results provide some molecular evidence for a regular pattern of avian intestinal epitheliosis and functional partition and provide an approach to further study of the locations of Cdc25B in the chicken.

Amperozide Decreases Cocaine-Induced Increase in Behavior and Immediate Early Gene Expression in the Dorsal Striatum

  • Choe, Eun-Sang;Kim, Jong-Yeon
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.5
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    • pp.361-367
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    • 2000
  • Cocaine functions as indirect dopamine and serotonin (5-hydroxytryptamine, 5HT) agonists and induces genomic and behavioral alterations in the striatum. Previously we demonstrated that ritanserin, a 5HT2/1C receptor antagonist, is not responsible for cocaine-induced behavioral alterations and zif268 mRNA gene expression in the striatum (see the previous paper in this issue). In this study, it was hypothesized that dopamine and 5HT2/1C receptors are required for cocaine-induced behavioral alterations and c-fos and zif268 mRNA expression. This hypothesis was addressed by infusing amperozide which antagonizes both 5HT2/1C and dopamine receptors and was analyzed using the quantitative in situ hybridization histochemistry in vivo. Systemic injection of amperozide (5 mg/kg, s.c.) significantly blocked increase in behavior, c-fos and zif268 mRNA expression induced by 15 mg/kg cocaine, i.p., in the dorsal striatum. These data suggest that dopamine and 5HT2/1C receptors are necessary for cocaine-induced behavioral alterations and immediate early gene expression in the dorsal striatum.

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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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    • v.4 no.5
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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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Neural Tissue-Specific Epidermal Growth Factor (EGF)-like Domain Containing Protein, NELL2, Plays on Important Role in the Control Regulation of Puberty Onset in the Female Rat Hypothalamus

  • Ha, Chang-Man;Kang, Hae-Mook;Lee, Byung-Ju
    • Animal cells and systems
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    • v.4 no.4
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    • pp.367-373
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    • 2000
  • In the present study we determined if NELL2, a neural tissue-specific protein containing 6 epidermal growth factor (EGF)-like repeat domains, plays an important role in the regulation of puberty initiation in the rat hypothalamus. We origin811y found that NELL2 is a new estrogen-responsive gene in hypothalami derived from estrogen-sterilized and control rats using a PCR differential display. In the 40-day-old female rat hypothalamus, NELL2 was up-regulated by neonatal estrogen treatment. In situ hybridization histochemistry showed that NELL2 is very abundant in the ventromedial hypothalamic nucleus that is responsible for the control of sex behavior. NELL2 mRNA level in the medial basal hypothalamus showed a dramatic increase before female puberty onset, which suggests that NELL2 may be involved in the process regulating female puberty onset. We attemped to block NELL2 synthesis with intracerebroventricular injection of an antisense oligodeoxynucleotide (ODN) to the NELL2 mRNA, and examined its effect on the puberty onset of the female rat. The antisense ODN significantly delayed puberty initiation determined by vaginal opening. In summary, NELL2 may play an important role in the regulation of female puberty onset.

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Ethanol Suppressed the Expression of Steroidogenie Acute Regulatory Protein mRNA in the Prepubertal Rat Ovary (미성숙 흰쥐난소에서의 에탄올에 의한 Steroidogenic Acute Regulatory Protein 유전자 발현 억제)

  • Kang, Sang-Soo;Cho, Gyeong-Jae;Park, Wan-Sung
    • Development and Reproduction
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    • v.4 no.1
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    • pp.109-114
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    • 2000
  • The present study was undertaken to examine the effects of ethanol on the ovarian steroidogenic acute regulatory protein(StAR) gene expression during prepubertal and onset of puberty. From day 25, each rat began receiving either a control saline or ethanol. Animals were sacrificed on day 27 and 32, and their ovaries and blood were collected. In the present results, ethanol treatment significantly decreased serum luteinizing hormone contents at both time points. Uterine weights of ethanol-treated group were significantly lighter than control group at early time point while there was no noticeable discrepancy at late time point. Vaginal openings, a marker of onset of puberty, also clearly delayed in ethanol-treated group. Using an in situ hybridization histochemistry, we determined the expression of mRNAs encoding StAR. Ovaries from ethanol-treated rats showed a suppresed expression of StAR mRNA. These results demonstrate that ethanol can disturb the prepubertal ovarian function and onset of puberty, at least in part, through the inhibition of ovarian StAR gene expression.

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Modulation in NMDA and $GABA_A$ Receptor Expression after Cerebroventricular Infusion of Ginsenosides

  • Oh Seikwan;Kim Hack-Seang
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.96-112
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    • 2002
  • In the present study, we have investigated the effects of centrally administered ginsenoside Rc or Rgl on the modulation of NMDA receptor and $GABA_A$ receptor binding in rat brain. The NMDA receptor binding was analyzed by quantitative autoradiography using $[^3H]MK-801$ binding, and $GABA_A$ receptor bindings were analyzed by using $[^3H]muscimol\;and\;[^3H]flunitrazepam$ in rat brain slices. Rats were infused with ginsenoside Rc or Rg1 ($10\;{\mu}g/10{\mu}l/hr$, i.c.v.) for 7 days, through pre-implanted cannula by osmotic minipumps (Alzet, model 2ML), The levels of $[^3H]MK-801$ binding were highly decreased in part of cortex and cingulated by ginsenoside Rc and Rgl. The levels of $[^3H]muscimol$ binding were strongly elevated in almost all regions of frontal cortex by the treatment of ginseoside Rc but decreased by ginsenoside Rg 1. However, the $[^3H]flunitrazepam$ binding was not modulated by ginsenoside Rc or ginsenoside Rgl infusion. These results suggest that prolonged infusion of ginsenoside could differentially modulate $[^3H]MK-801\;and\;[^3H]muscimol$ binding in a region-specific manner. Also, we investigated the influence of centrally administered ginsenoside on the regulation of mRNA levels of the family of NMDA receptor subtypes (NR1, NR2A, NR2B, NR2C) by in situ hybridization histochemistry in the rat brain. The level of NR1 mRNA is significantly increased in temporal cortex, caudate putamen, hippocampus, and granule layer of cerebellum in Rgl-infused rats as compared to control group. The level of NR2A mRNA is elevated in the frontal cortex. In contrast, it was decreased in CAI area of hippocampus in Rgl-infused rats. However, there was no significant change of NR1 and NR2A mRNA levels in Rc-infused rats. The level of NR2B mRNA is elevated in cortex, caudate putamen, and thalamus in both Rc- and Rg-infused rats. In contrast, NR2B level is decreased in CA3 in Rgl-infused rats. The level of NR2C mRNA is increased in the granule layer of cerebellum in only Rg1 but not Rc infused rats. These results show that structure difference of ginsenoside may diversely affect the modulation of expression of NMDA receptor subunit mRNA after infusion into cerebroventricle in rats.

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