Non-radioactive detection of nerve growth factor receptor (NGFR) mRNA in rat brain using in situ hybridization histochemistry.

Springer, Joe E.; Robbins, Elaine; Gwag, Byoung Joo; Lewis, Michael; Baldino, Frank

doi:10.1177/39.2.1846159

Cited by 60 publications

(22 citation statements)

References 3 publications

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“…In preparation for hybridization, sections or smears were proteolyzed, refixed, and acetylated as described (Hafen and Levine, 1986;Springer et al, 1991). Hybridization was carried out for 12-16 h in the hybridization solution described by Springer et al (1991) at 55°C.…”

Section: In Situ Hybridizationmentioning

confidence: 99%

Evidence for channeled diffusion of pre-mRNAs during nuclear RNA transport in metazoans.

Zachar

Kramer

Mims

et al. 1993

The Journal of Cell Biology

141

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Abstract. We report studies using an enhanced experimental system to investigate organization of nuclear pre-mRNA metabolism. It is based on the powerful genetic system and polytene nuclei of Drosophila. We observe (at steady state) movement of a specific premRNA between its gene and the nuclear surface. This movement is isotropic, at rates consistent with diffusion and is restricted to a small nuclear subcompartment defined by exclusion from chromosome axes and the nucleolus. Bulk polyadenylated nuclear pre-mRNA precisely localizes in this same subcompartment indieating that most or all pre-mRNAs use the same route of intranuclear movement.In addition to association with nascent transcripts, snRNPs are coconcentrated with pre-mRNA in this subcompartment. In contrast to constitutive splices, at least one regulated splice occurs slowly and may undergo execution remotely from the site of pre-mRNA synthesis. Details of our results suggest that retention of incompletely spliced pre-mRNA is a function of the nuclear surface.We propose a simple model-based on channeled diffusion-for organization of intranuclear transport and metabolism of pre-mRNAs in polytene nuclei. We argue that this model can be generalized to all metazoan nuclei. METAZOAN nuclei contain specific substructures implicated in pre-mRNA metabolism (see, for example, Fakan and Puvion, 1980; Lerner et al., 1981; McConnell et al., 1987;Lawrence et al., 1989;Spector, 1990;Fu and Maniatis, 1990;Carter et al., 1991; CarmoFonseca et al., 1991; Huang and Spector, 1991; Shermoen and O'Farrell, 1991;Wu et al., 1991;Xing and Lawrence, 1991;Li and Bingham, 1991;Kopczynski and Muskavitch, 1992). The capacity to extend these provocative observations is currently constrained by lack of a suitable genetic system for dissection of structure-function relationships and by inadequate convenience (EM) or resolution (light microscopy) among the various techniques for analysis of three dimensional organization in highly complex, compact nuclei.We report development of a new experimental system that both augments physical analytical techniques and permits application of a powerful genetic system. Our approach exploits resources uniquely available in Drosophila. First ,,o500) of an individual interphase chromatid are tightly synapsed in register to produce a giant, polytene chromosome (for review see Ashburner, 1989). In contrast to most diploid interphase chromosomes, individual polytene chromosome arms are readily identifiable as discrete structures in intact nuclei. Moreover, the simple arrangement of Drosophila chromosome arms is retained in these giant nuclei. Thus, polytene nuclei provide an "exploded" view of a simple nucleus. Under these conditions the relatively limited resolution of rapid, convenient light microscopic techniques is sufficient to allow clear visualization of relationships between extrachromosomal features and sites of pre-mRNA synthesis, Third, molecular genetic tools are available permitting construction of genes abundantly transcribed in polytene...

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Section: In Situ Hybridizationmentioning

confidence: 99%

Evidence for channeled diffusion of pre-mRNAs during nuclear RNA transport in metazoans.

Zachar

Kramer

Mims

et al. 1993

The Journal of Cell Biology

141

View full text Add to dashboard Cite

show abstract

“…Digoxigenin-dUTPlabeled RNA probes were prepared with a DIG RNA Labeling Kit (Roche Applied Science, M a n n h e i m , G e r m a n y ) a c c o r d i n g t o t h e manufacturer's instructions. ISH was conducted using the standard procedure [13]. Signals were detected using an anti-digoxigenin-alkaline phosphatase-conjugated antibody (Roche Applied Science).…”

Section: In Situ Hybridizationmentioning

confidence: 99%

Expression Profiling of Mouse Placental Lactogen II and Its Correlative Genes Using a cDNA Microarray Analysis in the Developmental Mouse Placenta

Ishida

Ohashi

Kizaki

et al. 2007

Journal of Reproduction and Development

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Abstract. The placenta is a highly differentiated organ essential for embryonic growth and development. In order to search for key molecules that are associated with mouse placental lactogen II (mPL-II) gene expression, we applied mouse cDNA microarray analysis to RNAs extracted from placentae on days 10, 12, 14, 16 and 18 of pregnancy. Changes in gene expression were categorized between days 10 and 12, 12 and 14, 14 and 16 and 16 and 18 of pregnancy. After microarray analysis, which had a minimum detectable fold change for differential expression of 2, we selected 10 genes, Apoa2, Apoc2, Ceacam14, Creg1, Fmo1, Igf2, Slc2a1, Spink3, Spi1-1 and Tpbpa, exhibiting a expression pattern similar to the mPL-II gene. Furthermore, we performed real-time PCR analysis and in situ hybridization (ISH) to find correlative expression genes for the mPL-II gene. From these results, we identified a resemblance in gene expression between mPL-II and Igf2 and selected these genes for performance of double-fluorescence immunohistochemical staining. We colocalized these proteins in labyrinthine trophoblast cells. These results strongly suggest that the expression of mPL-II and Igf2 is highly related to placental development in mice. This large-scale identification of genes regulated during placentogenesis assists in further elucidation of the molecular basis of extraembryonic development and function.

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“…The protocol for in situ hybridization used in this study was based on that described previously [12]. The sections were pretreated with proteinase K (10 µ g / m l ) s o l u t i o n , 0 .…”

Section: In Situ Hybridizationmentioning

confidence: 99%

Cathepsin Gene Expression in Mouse Placenta during the Latter Half of Pregnancy

Ishida

Ono

Taguchi

et al. 2004

Journal of Reproduction and Development

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Abstract. Gene expressions and their interaction are complex and have not been definitely clarified in the placenta. To identify interactions of gene products previously not studied, we applied cDNA subtraction analyses to the placenta between days 12 and 16, days 12 and 14, days 14 and 16 of pregnancy. Among subtracted cDNAs cathepsin M, Q and R in PECs were specifically identified on days 14 and 16 pregnancy. All of these gene expressions exhibited a similar pattern to the mPL-II gene expression determined by northern blot and RT-PCR analyses. By means of in situ hybridization, these mRNAs were localized in the basal and labyrinth zones of the placenta on day 16 of pregnancy. Double staining studies of cathepsin Q or cathepsin R mRNA by in situ hybridization followed by immunohistochemical staining of mPL-II in the same section revealed that signals for cathepsin Q and cathepsin R mRNAs were colocalized in mPL-II immunopositive trophoblast cells in the basal and labyrinth zones of the placenta on day 16 of pregnancy. Possible association of cathepsins with mPL-II may play important roles in placental functions during the latter half of pregnancy in mice.

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Non-radioactive detection of nerve growth factor receptor (NGFR) mRNA in rat brain using in situ hybridization histochemistry.

Cited by 60 publications

References 3 publications

Evidence for channeled diffusion of pre-mRNAs during nuclear RNA transport in metazoans.

Evidence for channeled diffusion of pre-mRNAs during nuclear RNA transport in metazoans.

Expression Profiling of Mouse Placental Lactogen II and Its Correlative Genes Using a cDNA Microarray Analysis in the Developmental Mouse Placenta

Cathepsin Gene Expression in Mouse Placenta during the Latter Half of Pregnancy

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