Expression of tilapia prepro-melanin-concentrating hormone mRNA in hypothalamic and neurohypophysial cells

ABSTRACT Melanin-concentrating hormone (MCH) is a neuropeptide involved in background adaptation in teleost fish, and in multiple regulatory functions in mammals and fish. To study the expression of the MCH preprohormone (ppMCH) in teleosts, we first cloned a hypothalamic cDNA encoding the complete ppMCH of tilapia (Oreochromis mossambicus), and a cRNA probe derived from a 270 bp ppMCH cDNA fragment was used for the expression studies. The level of ppMCH mRNA expression in tilapia hypothalamus, measured by dot blot analysis, was significantly higher in fish adapted to a white background than in black-adapted animals, which is in accordance with the reported MCH plasma and tissue concentrations in fish. Northern blot analysis not only revealed a strong ppMCH mRNA signal in the hypothalamus, but also the presence of ppMCH mRNA in the neurointermediate lobe (NIL) of the pituitary. In situ hybridization and immunocytochemistry showed that ppMCH mRNA as well as MCH immunoreactivity are located in perikarya of two hypothalamic regions, namely in the nucleus lateralis tuberis (NLT) and the nucleus recessus lateralis (NRL). Quantitative analysis by dot blot hybridization revealed about eight times more ppMCH mRNA in the NLT than in the NRL and NIL of mature tilapias. ppMCH mRNA in the NIL could be localized to cell bodies of the neurohypophysis, which were also MCH immunoreactive.

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Changes in the levels of mRNAs for GH/prolactin/somatolactin family and Pit-1/GHF-1 in the pituitaries of pre-spawning chum salmon

Journal of Molecular Endocrinology ◽

10.1677/jme.0.0230189 ◽

1999 ◽

Vol 23 (2) ◽

pp. 189-198 ◽

Cited By ~ 30

Author(s):

S Taniyama ◽

T Kitahashi ◽

H Ando ◽

M Ban ◽

H Ueda ◽

...

Keyword(s):

Blot Analysis ◽

Chum Salmon ◽

Northern Blot Analysis ◽

Oncorhynchus Keta ◽

Upstream Migration ◽

Dot Blot ◽

Expression Of Genes ◽

Osmotic Change ◽

Key Points ◽

Dot Blot Analysis

Changes in the levels of pituitary mRNAs encoding GH, prolactin (PRL) and somatolactin (SL) were determined in pre-spawning chum salmon (Oncorhynchus keta) caught at a few key points along their homing pathway in 1994 and 1995. Furthermore, we analyzed relationships between expression of pituitary-specific POU homeodomain transcription factor (Pit-1/GHF-1) and GH/PRL/SL family genes. In 1994, seawater (SW) fish and matured fresh-water (FW) fish were sequentially captured at two points along their homing pathway, the coast and the hatchery. In addition to these two points, maturing FW fish were captured at the intermediate of the two points in 1995. The levels of hormonal mRNAs were determined by a quantitative dot blot analysis using single-stranded sense DNA as the standard. Relative levels of Pit-1/GHF-1 mRNAs were estimated by Northern blot analysis. In 1994, the levels of GH/PRL/SL family mRNAs except for PRL mRNA in the male FW fish were 1.8-4 times higher than those in the SW fish. In 1995, the level of PRL mRNA was somewhat sharply elevated in the maturing FW fish soon after entry into the FW environment, while that of SL mRNA was gradually increased during upstream migration from the coast to the hatchery. The levels of 3 kb Pit-1/GHF-1 mRNA in the FW fish were higher than those in the SW fish in both 1994 and 1995. The present results indicate that expression of genes for the GH/PRL/SL family and Pit-1/GHF-1 is coincidentally enhanced in homing chum salmon. Moreover, the present study suggests that expression of the SL gene is elevated with sexual maturation, whereas that of PRL gene is elevated with osmotic change during the final stages of spawning migration.

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Expression of the first calcitonin/CGRP gene in spontaneous and transplanted rat medullary thyroid carcinoma: a comparison of dot-blot analysis, in situ hybridization, and immunohistochemistry.

Journal of Histochemistry & Cytochemistry ◽

10.1177/40.11.1431062 ◽

1992 ◽

Vol 40 (11) ◽

pp. 1761-1767 ◽

Cited By ~ 4

Author(s):

M Denijn ◽

R A de Weger ◽

W den Otter ◽

J A van Unnik ◽

C J Lips

Keyword(s):

Gene Expression ◽

In Situ Hybridization ◽

Blot Analysis ◽

Transplanted Tumors ◽

Dot Blot ◽

Tissue Samples ◽

Medullary Thyroid ◽

Dot Blot Analysis ◽

I Gene

Calcitonin (CT) and calcitonin gene-related peptide (CGRP) are encoded by a single gene, the CALC-I gene. They are expressed in the thyroid and in the nervous system by alternative splicing of the pre-messenger RNA derived from the CALC-I gene. In medullary thyroid carcinoma (MTC), a malignancy derived from the calcitonin-producing C-cells in the thyroid, production of calcitonin and CGRP is a common feature. We investigated the CT and CGRP production of four spontaneous MTCs transplanted three to four times and 14 MTC lines transplanted for several years in WAG/Rij rats, a strain with hereditary MTC. The expression of CT and CGRP in the spontaneous and in the transplanted tumors was studied by means of RNA in situ hybridization (RISH), dot-blot analysis, and immunohistochemistry. A down-regulation of CT production in transplanted compared with spontaneous tumors was observed, but an inverse relation between CT and CGRP mRNA content in both spontaneous and transplanted tumors was not observed. In this study, RISH proved to be as sensitive as dot-blot analysis to detect gene expression in tissue samples. The different approaches of analyzing the gene expression in tissue samples (the cellular localization of gene expression by ISH vs the analysis of an extract of a total tissue sample with dot-blot analysis) showed that each technique is equal in value and that they are complementary to each other.

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Application of biotinylated oligonucleotide probes to the detection of pituitary hormone mRNA using Northern blot analysis, in situ hybridization at the light- and electron-microscope levels

The Histochemical Journal ◽

10.1007/bf00188075 ◽

1994 ◽

Vol 26 (10) ◽

pp. 771-777 ◽

Cited By ~ 1

Author(s):

Akira Matsuno ◽

Akira Teramoto ◽

Susumu Takekoshi ◽

Hirotoshi Utsunomiya ◽

Yoshitaka Ohsugi ◽

...

Keyword(s):

In Situ Hybridization ◽

Electron Microscope ◽

Northern Blot ◽

Blot Analysis ◽

Northern Blot Analysis ◽

Pituitary Hormone ◽

Oligonucleotide Probes

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Characterization of a fusion cDNA (RARA/myl) transcribed from the t(15;17) translocation breakpoint in acute promyelocytic leukemia

Molecular and Cellular Biology ◽

10.1128/mcb.12.2.800-810.1992 ◽

1992 ◽

Vol 12 (2) ◽

pp. 800-810

Author(s):

K S Chang ◽

S A Stass ◽

D T Chu ◽

L L Deaven ◽

J M Trujillo ◽

...

Keyword(s):

Cdna Library ◽

Acute Promyelocytic Leukemia ◽

Blot Analysis ◽

Blot Hybridization ◽

Promyelocytic Leukemia ◽

Dna Probe ◽

Translocation Breakpoint ◽

Chromosome 15 ◽

Dot Blot ◽

Mrna Species

A nonrandom chromosomal translocation breakpoint, t(15;17)(q22;q21), is found in almost all patients with acute promyelocytic leukemia (APL). Most of these breakpoints occur within the second intron of the retinoic acid receptor-alpha (RARA) gene. We screened a cDNA library of APL and have identified and sequenced a cDNA transcribed from the t(15;17) translocation breakpoint. The 5' end of cDNA p1715 consists of 503 bp of the RARA exon II sequence. A 1.76-kb cDNA without homology to any known gene available in GenBank was found truncated downstream. This cDNA sequence was assigned to chromosome 15 by dot blot hybridization of the flow cytometry-sorted chromosomes. We designate this fusion cDNA RARA/myl, which is different from myl/RARA reported by de The et al. (H. de The, C. Chomienne, M. Lanotte, L. Degos, and A. Dejean, Nature (London) 347:558-561, 1990). This result demonstrates that the two different types of hybrid mRNA can be transcribed from this breakpoint. We screened a non-APL cDNA library and identified a 2.8-kb myl cDNA. This cDNA is able to encode a polypeptide with a molecular weight of 78,450. Alternative splicing of the myl gene which resulted in myl proteins with different C terminals was found. Southern blot analysis of the genomic DNA isolated from 17 APL patients by using the myl DNA probe demonstrated that the myl gene in 12 samples was rearranged. Northern (RNA) blot analysis of RARA gene expression in two APL RNA samples showed abnormal mRNA species of 4.2 and 3.2 kb in one patient and of 4.8 and 3.8 kb in another patient; these were in addition to the normal mRNA species of 3.7 and 2.7-kb. The myl DNA probe detected a 2.6-kb abnormal mRNA in addition to the normal mRNA species of 3.2, 4.2, and 5.5 kb. Using the polymerase chain reaction, we demonstrated that both RARA/myl and myl/RARA were coexpressed in samples from three different APL patients. From this study, we conclude that the t(15;17) translocation breakpoint results in the transcription of two different fusion transcripts which are expected to be translated into fusion proteins.

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Dot Blot Analysis for Measuring Global N6-Methyladenosine Modification of RNA

Epitranscriptomics - Methods in Molecular Biology ◽

10.1007/978-1-4939-8808-2_20 ◽

2018 ◽

pp. 263-271 ◽

Cited By ~ 7

Author(s):

Arvindhan Nagarajan ◽

Radoslav Janostiak ◽

Narendra Wajapeyee

Keyword(s):

Blot Analysis ◽

Dot Blot ◽

Dot Blot Analysis

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bcr Rearrangement without juxtaposition of c-abl in chronic myelocytic leukemia.

Journal of Experimental Medicine ◽

10.1084/jem.162.6.2175 ◽

1985 ◽

Vol 162 (6) ◽

pp. 2175-2179 ◽

Cited By ~ 31

Author(s):

C R Bartram

Keyword(s):

Blot Analysis ◽

Southern Blot Analysis ◽

Gene Rearrangement ◽

Northern Blot Analysis ◽

Philadelphia Chromosome ◽

Leukemic Cells ◽

Chronic Myelocytic Leukemia ◽

Myelocytic Leukemia ◽

Genomic Recombination

Southern blot analysis detected a bcr gene rearrangement within leukemic cells of a Philadelphia chromosome-negative chronic myelocytic leukemia (CML) patient that led to transcription of a novel 7.3 kb bcr RNA species. Participation of the c-abl oncogene in this genomic recombination could be ruled out by in situ hybridization studies and Northern blot analysis.

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Cloning and widespread distribution of the rat rod-type cyclic nucleotide-gated cation channel

AJP Cell Physiology ◽

10.1152/ajpcell.1997.272.4.c1335 ◽

1997 ◽

Vol 272 (4) ◽

pp. C1335-C1344 ◽

Cited By ~ 57

Author(s):

C. Ding ◽

E. D. Potter ◽

W. Qiu ◽

S. L. Coon ◽

M. A. Levine ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

In Situ Hybridization ◽

Pineal Gland ◽

Northern Blot ◽

Blot Analysis ◽

Cyclic Nucleotide ◽

Northern Blot Analysis ◽

Chain Reaction ◽

Polymerase Chain

We used Northern blot analysis, ribonuclease protection assay (RPA), reverse transcriptase-polymerase chain reaction, and in situ hybridization to investigate the hypothesis that the CNG1 isoform of the cyclic nucleotide-gated nonselective cation channel may be widely distributed in tissues of the rat. A cDNA encoding the CNG1 isoform was isolated from rat eye and human retina, and partial sequences were isolated from rat pineal gland and human kidney. Northern blot analysis revealed a 3.1-kilobase (kb) CNG1 transcript in rat eye, pineal gland, pituitary, adrenal gland, and spleen, and a larger transcript of 3.5 kb was found in testis. RPA confirmed the identity of CNG1 mRNA in rat eye, lung, spleen, and brain. Polymerase chain reaction-based detection of the mRNA for CNG1 indicates that the channel is expressed in lower abundance in many other tissues, including thymus, skeletal muscle, heart, and parathyroid gland. The cellular distribution of CNG1 was further studied by in situ hybridization, which demonstrated expression of mRNA in lung, thymus, pineal gland, hippocampus, cerebellum, and cerebral cortex but not in heart or kidney.

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