Sequence analysis of the turkey LH β subunit and its regulation by gonadotrophin-releasing hormone and prolactin in cultured pituitary cells

1995 ◽  
Vol 14 (1) ◽  
pp. 117-129 ◽  
Author(s):  
S You ◽  
L K Foster ◽  
J L Silsby ◽  
M E El Halawani ◽  
D N Foster
Keyword(s):  
Gene Expression ◽  
Amino Acid ◽  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Pituitary Cells ◽  
Β Subunit ◽  
Spontaneous Release ◽  
Gonadotrophin Releasing Hormone ◽  
Prl Gene

ABSTRACT cDNAs encoding the precursor molecule of the turkey LH β subunit (tLHβ) were cloned from a turkey pituitary cDNA library. The nucleotide sequence of the longest of two different tLHβ cDNA clones contained 592 bp, and included 23 bp of the 5′ untranslated region (UTR) and 92 bp of the 3′ UTR in addition to a 477 bp open reading frame that encoded a 39 amino acid leader polypeptide and a 120 amino acid mature apoprotein. Turkey and chicken LHβ sequences shared approximately 92 and 93% nucleotide and amino acid sequence similarities respectively. Northern blot analysis of total cellular anterior pituitary RNA showed that an approximate 800 base transcript hybridized to a 32P-labelled tLHβ cDNA probe. The gonadotrophin-releasing hormone (GnRH)-and prolactin (PRL)-regulated expression of LH and PRL in dispersed pituitary cells was determined by Northern blot analysis of tLHβ and PRL steady-state mRNA levels and by RIA analysis of secreted LH and PRL. GnRH-treated cells showed increased levels of both tLHβ mRNA and secreted LH, whereas mRNA and secreted levels of PRL did not change significantly. Cells treated with PRL showed lower levels of tLHβ and PRL mRNA as well as decreased release of LH and PRL. When cells were treated with both PRL and GnRH, increases in tLHβ mRNA and secreted levels of LH observed with GnRH alone were negated, whereas the decreases in mRNA and secreted levels of PRL observed with PRL alone were abrogated. These findings suggest that PRL can down-regulate tLHβ gene expression and spontaneous release of LH as well as autoregulate PRL gene expression and spontaneous release of PRL, while GnRH appears capable of modulating the effects of PRL-regulated LH and PRL gene expression and spontaneous release.

scholarly journals IL-8 and MCP Gene Expression and Production by LPS-Stimulated Human Corneal Stromal Cells

2012 ◽  
Vol 2012 ◽  
pp. 1-4 ◽  
Author(s):  
Roni M. Shtein ◽  
Susan G. Elner ◽  
Zong-Mei Bian ◽  
Victor M. Elner
Keyword(s):  
Gene Expression ◽  
Northern Blot ◽  
Stromal Cells ◽  
Blot Analysis ◽  
Time Course ◽  
Northern Blot Analysis ◽  
Statistical Significance ◽  
Dependent Manner ◽  
Corneal Inflammation ◽  
Chemotactic Protein

Purpose. To determine time course of effect of lipopolysaccharide (LPS) on production of interleukin-8 (IL-8) and monocyte chemotactic protein (MCP) by cultured human corneal stromal cells.Methods. Human corneal stromal cells were harvested from donor corneal specimens, and fourth to sixth passaged cells were used. Cell cultures were stimulated with LPS for 2, 4, 8, and 24 hours. Northern blot analysis of IL-8 and MCP gene expression and ELISA for IL-8 and MCP secretion were performed. ELISA results were analyzed for statistical significance using two-tailed Student'st-test.Results. Northern blot analysis demonstrated significantly increased IL-8 and MCP gene expression after 4 and 8 hours of exposure to LPS. ELISA for secreted IL-8 and MCP demonstrated statistically significant increases (P<0.05) after corneal stromal cell stimulation with LPS.Conclusions. This paper suggests that human corneal stromal cells may participate in corneal inflammation by secreting potent leukocyte chemotactic and activating proteins in a time-dependent manner when exposed to LPS.

Gene expression analysis of the pro-oestrous-stage rat uterus reveals neuroligin 2 as a novel steroid-regulated gene

2004 ◽  
Vol 16 (8) ◽  
pp. 763 ◽  
Author(s):  
Han-Seung Kang ◽  
Chae-Kwan Lee ◽  
Ju-Ran Kim ◽  
Seong-Jin Yu ◽  
Sung-Goo Kang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Oestrous Cycle ◽  
Mrna Levels ◽  
Rat Uterus ◽  
Ovx Rats

In the present study, differential gene expression in the uteri of ovariectomised (OVX) and pro-oestrous rats (OVX v. pro-oestrus pair) was investigated using cDNA expression array analysis. Differential uterine gene expression in OVX rats and progesterone (P4)-injected OVX rats (OVX v. OVX + P4 pair) was also examined. The uterine gene expression profiles of these two sets of animals were also compared for the effects of P4 treatment. RNA samples were extracted from uterine tissues and reverse transcribed in the presence of [α32P]-dATP. Membrane sets of rat arrays were hybridised with cDNA probe sets. Northern blot analysis was used to validate the relative gene expression patterns obtained from the cDNA array. Of the 1176 cDNAs examined, 23 genes showed significant (>two-fold) changes in expression in the OVX v. pro-oestrus pair. Twenty of these genes were upregulated during pro-oestrus compared with their expression in the OVX rat uterus. In the OVX v. OVX + P4 pair, 22 genes showed significant (>two-fold) changes in gene expression. Twenty of these genes were upregulated in the OVX + P4 animals. The genes for nuclear factor I–XI, afadin, neuroligin 2, semaphorin Z, calpain 4, cyclase-associated protein homologue, thymosin β-4X and p8 were significantly upregulated in the uteri of the pro-oestrus and OVX + P4 rats of both experimental pairs compared with the OVX rat uteri. These genes appear to be under the control of P4. One of the most interesting findings of the present study is the unexpected and marked expression of the neuroligin 2 gene in the rat uterus. This gene is expressed at high levels in the central nervous system and acts as a nerve cell adhesion factor. According to Northern blot analysis, neuroligin 2 gene expression was higher during the pro-oestrus and metoestrus stages than during the oestrus and dioestrus stages of the oestrous cycle. In addition, neuroligin 2 mRNA levels were increased by both 17β-oestradiol (E2) and P4, although P4 administration upregulated gene expression to a greater extent than injection of E2. These results indicate that neuroligin 2 gene expression in the rat uterus is under the control of both E2 and P4, which are secreted periodically during the oestrous cycle.

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

1997 ◽  
Vol 18 (1) ◽  
pp. 49-55 ◽  
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.

Northern blot analysis of nm23 gene expression in human lung cancer

1999 ◽  
Vol 11 (3) ◽  
pp. 188-191
Author(s):  
Liu Lun-xu ◽  
Zhou Qing-hua ◽  
Shi Ying-kang ◽  
Qin Yang ◽  
Sun Zhi-lin ◽  
...  
Keyword(s):  
Gene Expression ◽  
Lung Cancer ◽  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Human Lung ◽  
Human Lung Cancer ◽  
Nm23 Gene

scholarly journals Insulin-like growth factor 1 (IGF-1) gene expression in porcine ovarian tissue

1993 ◽  
Vol 73 (2) ◽  
pp. 253-257 ◽  
Author(s):  
S. T. Charlton ◽  
B. J. Cameron ◽  
D. R. Glimm ◽  
G. R. Foxcroft ◽  
J. J. Kennelly
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Northern Blot ◽  
Blot Analysis ◽  
Ribonucleic Acid ◽  
Northern Blot Analysis ◽  
Ovarian Tissue ◽  
Messenger Ribonucleic Acid ◽  
Polyadenylated Rna ◽  
Mrna Transcripts

A study was conducted to demonstrate IGF-1 gene expression in porcine ovarian tissue. Using northern blot analysis, IGF-1 messenger ribonucleic acid (mRNA) transcripts could not be consistently detected in total RNA. Authentic mRNA transcripts were detected in polyadenylated RNA-enriched RNA with sizes of 8.0, 3.6 and 2.3 bases, and possibly 0.8–1.1 kbases. Key words: Ovary, IGF-1, pig, mRNA

scholarly journals Evidence for the existence of a pristanoyl-CoA oxidase gene in man

1997 ◽  
Vol 325 (3) ◽  
pp. 593-599 ◽  
Author(s):  
Johannes C. T. VANHOOREN ◽  
Peter MARYNEN ◽  
Guy P. MANNAERTS ◽  
Paul P. VAN VELDHOVEN
Keyword(s):  
Amino Acid ◽  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Developmental Stages ◽  
Genomic Library ◽  
Immunoblot Analysis ◽  
Human Tissues ◽  
Oxidase Gene ◽  
Activity Measurements

In the rat, 2-methyl branched fatty acids and the bile acid intermediates di- and tri-hydroxycoprostanic acids are desaturated by pristanoyl-CoA oxidase and trihydroxycoprostanoyl-CoA oxidase respectively. In the human, these compounds are oxidized by a single enzyme, branched-chain acyl-CoA oxidase, which according to its amino acid sequence is the human homologue of rat trihydroxycoprostanoyl-CoA oxidase. Pristanoyl-CoA oxidase is apparently absent from human tissues as indicated by immunoblot analysis [Van Veldhoven, Van Rompuy, Fransen, de Béthune and Mannaerts (1994) Eur. J. Biochem. 222, 795–801] and Northern-blot analysis [Vanhooren, Fransen, de Béthune, Baumgart, Baes, Torrekens, Van Leuven, Mannaerts and Van Veldhoven (1996) Eur. J. Biochem. 239, 302–309] of human tissues. In this paper we present evidence, however, that at least the gene for pristanoyl-CoA oxidase is present in the human. A human liver cDNA encoding a protein of 700 amino acids, showing 75% amino acid identity with rat pristanoyl-CoA oxidase and harbouring a peroxisomal C-terminal-targeting signal (SKL), was isolated. Bacterial expression of the cDNA resulted in a fusion protein that was cross-reactive with antibodies directed against rat pristanoyl-CoA oxidase and the C-terminal SKL sequence. Screening of a genomic library with the isolated cDNA as a probe resulted in a genomic clone in which four introns were localized. By means of fluorescence in situhybridization the gene for human pristanoyl-CoA oxidase was mapped at chromosome position 4p15.3. We conclude that a gene for pristanoyl-CoA oxidase is present in the human genome. The gene appears to be expressed to such a low extent in liver that its mRNA cannot be detected by routine Northern-blot analysis and that its product remains undetected by standard immunoblotting or by enzyme activity measurements. We speculate that the gene may be expressed under special (e.g. certain developmental stages) conditions or in certain specialized tissues not examined thus far.

scholarly journals Synthesis and immunocytochemical localization of alpha 2u globulin in the duct cells of the rat submaxillary gland.

1982 ◽  
Vol 30 (12) ◽  
pp. 1293-1296 ◽  
Author(s):  
T Antakly ◽  
Y Laperche ◽  
P Feigelson
Keyword(s):  
Gene Expression ◽  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Submaxillary Gland ◽  
Immunocytochemical Localization ◽  
Duct Cells ◽  
Gland Duct ◽  
Convoluted Tubules ◽  
Globulin Gene

Alpha 2u globulin, a protein of unknown function so far believed to be synthesized exclusively in the male liver under multihormonal control, is now shown to be localized by immunocytochemistry in the granular convoluted tubules of the adult male submaxillary gland. In addition, using Northern blot analysis, we have shown specific alpha 2u globulin mRNA sequences in the RNA extracted from the submaxillary gland. Thus, it is evident that the protein is being synthesized therein. Alpha 2u globulin was also detected in the submaxillary gland duct cells of adult female and immature animals of both sexes, all of which are known not to synthesize alpha 2u globulin in their livers. The present data have established that alpha 2u globulin is synthesized in the rat submaxillary gland and indicate that the control of alpha 2u globulin gene expression in the rat liver and in the submaxillary gland is different.

Sign in / Sign up

Export Citation Format

Share Document