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

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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Changes in expression of neurohypophysial hormone genes during spawning migration in chum salmon, Oncorhynchus keta

Journal of Molecular Endocrinology ◽

10.1677/jme.0.0180049 ◽

1997 ◽

Vol 18 (1) ◽

pp. 49-55 ◽

Cited By ~ 25

Author(s):

S Hiraoka ◽

H Ando ◽

M Ban ◽

H Ueda ◽

A Urano

Keyword(s):

Gene Expression ◽

Northern Blot ◽

Blot Analysis ◽

Chum Salmon ◽

Northern Blot Analysis ◽

Sea Water ◽

Spawning Migration ◽

Oncorhynchus Keta ◽

Males And Females ◽

Neuroendocrine Mechanism

ABSTRACT We analyzed changes in the hypothalamic levels of vasotocin (VT) and isotocin (IT) mRNA in chum salmon during spawning migration to the Ishikari river. The fish were caught at Atsuta, a fisherman's village facing the Ishikari bay, and at Chitose, an upstream branch of the Ishikari river. The former are referred to as sea water (SW) fish, and the latter as freshwater (FW) fish. The levels of VT and IT mRNA in the forebrains were determined by quantitative Northern blot analysis using single-stranded DNA with the same mRNA sequences as the standards. Levels of VT mRNA were higher in the FW males than the FW females, although no such difference was seen in the SW fish. Changes in the levels of VT mRNA were markedly different in males and females. In the males, no significant differences were seen in the levels of VT-I and VT-II mRNA between the SW and FW fish. However, in the females, the levels of VT mRNA in the FW fish were significantly lower than those in the SW fish. Changes in the levels of IT-I and IT-II mRNA were essentially similar in the males and females. These results suggest that the control of VT gene expression is different in males and females during spawning migration, although the neuroendocrine mechanism is not known.

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

Journal of Molecular Endocrinology ◽

10.1677/jme.0.0140199 ◽

1995 ◽

Vol 14 (2) ◽

pp. 199-207 ◽

Cited By ~ 28

Author(s):

D Gröneveld ◽

E R M Eckhardt ◽

A J M Coenen ◽

G J M Martens ◽

P H M Balm ◽

...

Keyword(s):

Blot Analysis ◽

Northern Blot Analysis ◽

Blot Hybridization ◽

White Background ◽

Dot Blot ◽

Expression Studies ◽

Melanin Concentrating Hormone ◽

Regulatory Functions ◽

Dot Blot Analysis

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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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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An Improved Method to Do Dot Blot Analysis

Analytical Biochemistry ◽

10.1006/abio.1994.1491 ◽

1994 ◽

Vol 222 (1) ◽

pp. 293-294 ◽

Cited By ~ 2

Author(s):

K.S. Saini ◽

B. Bhandari

Keyword(s):

Blot Analysis ◽

Improved Method ◽

Dot Blot ◽

Dot Blot Analysis

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Measurement of telomere DNA content by dot blot analysis

Nucleic Acids Research ◽

10.1093/nar/gkr235 ◽

2011 ◽

Vol 39 (12) ◽

pp. e84-e84 ◽

Cited By ~ 14

Author(s):

M. Kimura ◽

A. Aviv

Keyword(s):

Dna Content ◽

Blot Analysis ◽

Dot Blot ◽

Dot Blot Analysis

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Abstract P4-02-14: Digitalizing immunohistochemistry analysis (IHC) with quantitative dot blot analysis (QDB) by absolute quantification of biomarkers in FFPE specimens

10.1158/1538-7445.sabcs18-p4-02-14 ◽

2019 ◽

Author(s):

J Zhang ◽

W Zhang ◽

G Yu ◽

F Tang ◽

J Lv

Keyword(s):

Blot Analysis ◽

Absolute Quantification ◽

Dot Blot ◽

Immunohistochemistry Analysis ◽

Dot Blot Analysis

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Quantitative dot blot analysis (QDB), a versatile high throughput immunoblot method

Oncotarget ◽

10.18632/oncotarget.17236 ◽

2017 ◽

Vol 8 (35) ◽

pp. 58553-58562 ◽

Cited By ~ 14

Author(s):

Geng Tian ◽

Fangrong Tang ◽

Chunhua Yang ◽

Wenfeng Zhang ◽

Jonas Bergquist ◽

...

Keyword(s):

High Throughput ◽

Blot Analysis ◽

Dot Blot ◽

Dot Blot Analysis

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HSP-72 synthesis is promoted by increase in [Ca2+]i or activation of G proteins but not pHi or cAMP

AJP Cell Physiology ◽

10.1152/ajpcell.1994.267.1.c104 ◽

1994 ◽

Vol 267 (1) ◽

pp. C104-C114 ◽

Cited By ~ 47

Author(s):

J. G. Kiang ◽

F. E. Carr ◽

M. R. Burns ◽

D. E. McClain

Keyword(s):

Heat Shock ◽

G Proteins ◽

Blot Analysis ◽

Dot Blot ◽

Heat Shock Element ◽

The Family ◽

Shock Proteins ◽

Dot Blot Analysis ◽

The Relationship ◽

Acid Loading

The family of 70-kDa heat-shock proteins (HSP-70) is evolutionarily highly conserved and has been shown to enhance cell survival from thermal injury. This study characterized HSP-72 induction in human epidermoid A-431 cells exposed to 45 degrees C for 10 min and determined the relationship between HSP-72, intracellular pH (pHi), adenosine 3',5'-cyclic monophosphate (cAMP), G proteins, and intracellular cytosolic free Ca2+ concentration ([Ca2+]i). Heat shock induced HSP-72 production, which was dependent on both temperature and the duration of heating. This HSP-72 induction was confirmed by Western blot analysis. HSP-72 levels in cells that had been heated then returned to 37 degrees C were elevated at 2 h (1.5 +/- 0.1 x control), reached a maximum at 8 h (2.7 +/- 0.1 x control), and remained above baseline for up to 4 days. Levels of HSP-72 mRNA, determined by dot-blot analysis, reached a maximum at 2 h and returned to baseline within 8 h. Both actinomycin D and cycloheximide blocked HSP-72 induction. Because heating causes intracellular acidification and increases in cAMP and [Ca2+]i, we studied the effect of pHi, cellular cAMP, and [Ca2+]i on HSP-72 induction. The reduction of pHi to 6.9 by acid loading did not affect the basal level of HSP-72 in unheated cells. Treatment with pertussis toxin, cholera toxin, or forskolin, but not 8-bromo-cAMP, 3-isobutyl-1-methylxanthine, or N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide potentiated heat-induced HSP-72 production. Inhibition of the heat-induced increase in [Ca2+]i attenuated, but failed to completely block, heat-induced HSP-72 production, mRNA synthesis, and the heat-shock transcriptional factor-heat-shock element binding complex formation, which suggests there are Ca(2+)-dependent and -independent processes involved in HSP-72 synthesis. Our results show that an increase in [Ca2+]i or activation of G proteins, but not pHi and cAMP, enhances HSP-72 induction.

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New poly(A)+RNAs appear coordinately during the differentiation of Naegleria gruberi amebae into flagellates.

The Journal of Cell Biology ◽

10.1083/jcb.102.2.353 ◽

1986 ◽

Vol 102 (2) ◽

pp. 353-361 ◽

Cited By ~ 13

Author(s):

J Mar ◽

J H Lee ◽

D Shea ◽

C J Walsh

Keyword(s):

Blot Analysis ◽

Rna Synthesis ◽

In Vitro Translation ◽

Dot Blot ◽

Cross Hybridization ◽

Tubulin Mrna ◽

Dot Blot Analysis ◽

Rna Concentration ◽

Naegleria Gruberi

We have examined the nature of the requirement for RNA synthesis during the differentiation of Naegleria gruberi amebae into flagellates (Fulton, C., and C. Walsh, 1980, J. Cell Biol., 85:346-360) by looking for poly(A)+RNAs that are specific to differentiating cells. A cDNA library prepared from poly(A)+RNA extracted from cells 40 min after initiation of the differentiation (40-min RNA), the time when formation of flagella becomes insensitive to inhibitors of RNA synthesis, was cloned into pBR322. Recombinant clones were screened for sequences that were complementary to 40-min RNA but not to RNA from amebae (0-min RNA). Ten of these differentiation-specific (DS) plasmids were identified. The DS plasmids were found to represent at least four different poly(A)+RNAs based on cross-hybridization, restriction mapping, and Northern blot analysis. Dot blot analysis was used to quantify changes in DS RNA concentration. The four DS RNAs appeared coordinately during the differentiation. They were first detectable at 10-15 min after initiation, reached a peak at 70 min as flagella formed, and then declined to low levels by 120 min when flagella reached full length. The concentration of the DS RNAs was found to be at least 20-fold higher in cells at 70 min than in amebae. The changes in DS RNA concentration closely parallel changes in tubulin mRNA as measured by in vitro translation (Lai, E.Y., C. Walsh, D. Wardell, and C. Fulton, 1979, Cell, 17:867-878).

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