Growth hormone MRNA expression in rat embryo: An in situ RT-PCR visualization

1999 ◽  
Vol 91 (3) ◽  
pp. 279-279
Author(s):  
Sophie Recher ◽  
Anne Lambert ◽  
Mireille Raccurt ◽  
Hichem C. Mertani ◽  
Gérard Morel
Keyword(s):  
Growth Hormone ◽  
Mrna Expression ◽  
Rt Pcr ◽  
Rat Embryo ◽  
Growth Hormone Mrna

Endothelin receptor mRNA expression in renal medulla identified by in situ RT-PCR

1995 ◽  
Vol 269 (3) ◽  
pp. F449-F457 ◽  
Author(s):  
L. H. Chow ◽  
S. Subramanian ◽  
G. J. Nuovo ◽  
F. Miller ◽  
E. P. Nord
Keyword(s):  
Mrna Expression ◽  
Collecting Duct ◽  
Renal Medulla ◽  
Receptor Subtypes ◽  
Rt Pcr ◽  
Receptor Mrna ◽  
Pcr Products ◽  
Medullary Collecting Duct ◽  
Labeled Probe

Three subtypes of endothelin (ET) receptors have been identified by cDNA cloning, namely ET-RA, ET-RB, and ET-RC. In the current study the precise cellular distribution of the ET receptor subtypes in the renal medulla was explored by detecting the corresponding polymerase chain reaction (PCR)-amplified cDNAs by in situ reverse transcription (RT)-PCR. The PCR-amplified cDNAs were detected either by direct incorporation using digoxigenin-dUTP (dig-dUTP) as a nucleotide substrate in the PCR reaction or by in situ hybridization with the dig-dUTP-labeled probe. ET-RB mRNA was detected exclusively in the epithelial cells of the inner and outer medullary collecting duct. In contrast, ET-RA message was observed primarily in interstitial cells and pericytes of the vasae rectae in the outer and inner medulla. Southern blot analysis of PCR-amplified cDNAs reverse transcribed from extracted RNA of rat renal medulla confirmed the specificity of the RT-PCR products. ET-RC mRNA was not detected. We conclude that ET-RB is the major ET receptor found in rat renal medulla and is expressed exclusively on inner medullary collecting duct cells. The pattern of ET receptor mRNA expression described suggests different physiological actions for ET on the diverse cellular structures of the renal medulla.

scholarly journals Quantification of TGF-β1 mRNA along rat nephron in obstructive nephropathy

2001 ◽  
Vol 281 (3) ◽  
pp. F513-F521 ◽  
Author(s):  
Kyoichi Fukuda ◽  
Koji Yoshitomi ◽  
Taihei Yanagida ◽  
Masanori Tokumoto ◽  
Hideki Hirakata
Keyword(s):  
In Situ Hybridization ◽  
Mrna Expression ◽  
Transforming Growth Factor ◽  
Interstitial Fibrosis ◽  
Proximal Tubules ◽  
Obstructive Nephropathy ◽  
Renal Tubules ◽  
Rt Pcr ◽  
Tgf Β1

Unilateral ureteral obstruction (UUO) leads to interstitial fibrosis of the obstructed kidney, and transforming growth factor-β1 (TGF-β1) is thought to play an important role in this process. Although increased TGF-β1 mRNA expression in the obstructed kidney has been demonstrated, the source of the increased TGF-β1 remains to be elucidated. To determine the precise localization of TGF-β1 in the obstructed kidney, we examined TGF-β1 mRNA expression using in situ hybridization and competitive RT-PCR in rats with UUO. In situ hybridization demonstrated that TGF-β1 mRNA expression was preferentially increased in tubular epithelial cells and to a lesser degree in infiltrating macrophages in obstructed kidneys. Quantitative analysis using competitive RT-PCR in microdissected nephron segments revealed that levels of TGF-β1 mRNA in obstructed kidneys relative to control kidneys increased significantly in proximal tubules, thick ascending limbs of Henle, and distal convoluted tubules, whereas those in glomeruli and collecting ducts did not change significantly. Of the tubular segments, the proximal tubules appeared to predominantly contribute to increased TGF-β1 mRNA. Our findings suggest that renal tubules, particularly proximal tubules, are the main contributors to increased TGF-β1 mRNA expression in obstructed kidneys and to the subsequent interstitial fibrosis.

Malnutrition during lactation changes growth hormone mRNA expression in offspring at weaning and in adulthood

2007 ◽  
Vol 18 (2) ◽  
pp. 134-139 ◽  
Author(s):  
Egberto Gaspar de Moura ◽  
Patricia Cristina Lisboa ◽  
Claudia Mota Custódio ◽  
Maria Tereza Nunes ◽  
Kely de Picoli Souza ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Mrna Expression ◽  
Growth Hormone Mrna

Regulation of growth hormone expression by thyrotropin-releasing hormone through the pituitary-specific transcription factor Pit-1 in chicken pituitary

2004 ◽  
Vol 52 (4) ◽  
pp. 389-402 ◽  
Author(s):  
P. Van As ◽  
C. Careghi ◽  
V. Bruggeman ◽  
O. M. Onagbesan ◽  
S. Van der Geyten ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Mrna Expression ◽  
Developmental Stages ◽  
Pituitary Hormones ◽  
Thyroid Stimulating Hormone ◽  
Thyrotropin Releasing Hormone ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Releasing Hormone ◽  
Pou Domain

Pit-1 is a pituitary-specific POU-domain DNA binding factor, which binds to and trans-activates promoters of growth hormone- (GH), prolactin- (PRL) and thyroid stimulating hormone beta- (TSHβ) encoding genes. Pit-1 has been identified in several mammalian and avian species. Thyrotropin-releasing hormone (TRH) is located in the hypothalamus and it stimulates TSH, GH and PRL release from the pituitary gland. In the present study, we successfully developed a competitive RT-PCR for the detection of Pit-1 expression in the chicken pituitary, that was sensitive enough to detect picogram levels of Pit-1 mRNA. Applying this method, the effect of TRH injections on Pit-1 mRNA expression was determined in the pituitary of chick embryos and growing chicks. In both 18-day-old embryos and 10-day-old male chicks the Pit-1 mRNA expression was significantly increased following TRH injection, thereby indicating that the stimulatory effects of TRH on several pituitary hormones is mediated via its effect on Pit-1 expression. Therefore, a semi-quantitative RT-PCR method was used to detect possible changes in GH levels. TRH affected the GH mRNA levels at both developmental stages. These results, combined with the data on Pit-1 mRNA expression, indicate that Pit-1 has a role in mediating the stimulatory effects of TRH on pituitary hormones like GH.

scholarly journals Expression of Tandem P Domain K+ Channel, TREK-1, in the Rat Carotid Body

2006 ◽  
Vol 54 (4) ◽  
pp. 467-472 ◽  
Author(s):  
Y. Yamamoto ◽  
K. Taniguchi
Keyword(s):  
In Situ Hybridization ◽  
Mrna Expression ◽  
Carotid Body ◽  
Nerve Fibers ◽  
K Channel ◽  
Membrane Excitability ◽  
Rt Pcr ◽  
Glomus Cells ◽  
Afferent Nerve Endings

TREK-1 is one of the important potassium channels for regulating membrane excitability. To examine the distribution of TREK-1 in the rat carotid body, we performed RT-PCR for mRNA expression and in situ hybridization and immunohistochemistry for tissue distribution of TREK-1. RT-PCR detected mRNA expression of TREK-1 in the carotid body. Furthermore, in situ hybridization revealed the localization of TREK-1 mRNA in the glomus cells. TREK-1 immunoreactivity was mainly distributed in the glomus cells and nerve fibers in the carotid body. TREK-1 may modulate potassium current of glomus cells and/or afferent nerve endings in the rat carotid body.

scholarly journals Porcine endometrial and conceptus tissue kallikrein 1, 4, 11, and 14 gene expression

Reproduction ◽  
2006 ◽  
Vol 132 (6) ◽  
pp. 939-947 ◽  
Author(s):  
S C Fernando ◽  
J S Buck ◽  
M D Ashworth ◽  
J W Ross ◽  
R D Geisert ◽  
...  
Keyword(s):  
Gene Expression ◽  
Growth Factors ◽  
Mrna Expression ◽  
Estrous Cycle ◽  
Early Pregnancy ◽  
Tissue Remodeling ◽  
Uterine Epithelium ◽  
Tissue Kallikrein ◽  
Rt Pcr

Previous studies have suggested that the porcine endometrium may express several tissue kallikreins during the estrous cycle and early pregnancy. The present study investigated porcine endometrial and conceptus tissue kallikrein 1, 4, 11, and 14 mRNA expression during the estrous cycle and early pregnancy. Tissue kallikrein (KLK) gene expression was evaluated using quantitative RT-PCR and in situ hybridization. KLK1 expression was similar across the estrous cycle and early pregnancy, and localized to the endometrial luminal (L) and glandular (G) epithelium. KLK4 endometrial mRNA expression was greatest on days 0, 5, and 10 when compared with days 12, 15, and 17 of the estrous cycle and greater in cyclic compared with pregnant gilts. Expression of KLK4 was more intense in the stroma and uterine epithelium from days 0 to 10 of the estrous cycle. Endometrial KLK11 mRNA was not different between cyclic and pregnant gilts but the expression was greatest on days 10 and 12 compared with all other days evaluated. There was an increased intensity of KLK11 gene expression in the stratum compactum on day 10 of the estrous cycle and early pregnancy. Endometrial KLK14 mRNA expression was not detectable on days 5 and 10 but was expressed on days 0, 12, 15, and 17 of the estrous cycle and pregnancy. KLK14 expression was localized in the uterine L and G epithelium, and stroma throughout the endometrium after day 10. Conceptus KLK1 mRNA did not change from days 10 to 17 of gestation. However, conceptus KLK4, and 14 mRNA expression was greatest on day 10 with expression declining after day 14 of gestation. Expression of the various tissue kallikreins in the endometrium and conceptus during the estrous cycle and early pregnancy in the pig can serve in the activation of growth factors and tissue remodeling during the establishment of pregnancy.

Visualization of the expression of the mRNA of growth hormone by in situ RT-PCR

1998 ◽  
Vol 90 (3) ◽  
pp. 287-287
Author(s):  
Sophie Recher ◽  
Sylvie Ricard-Blum ◽  
Gérard Morel
Keyword(s):  
Growth Hormone ◽  
Rt Pcr

scholarly journals Expression of Hepatocyte Growth Factor mRNA in Rat Liver Cirrhosis Induced by N-nitrosodimethylamine as Evidenced by In Situ RT-PCR

2002 ◽  
Vol 50 (11) ◽  
pp. 1461-1468 ◽  
Author(s):  
Junko Hata ◽  
Etsuko Ikeda ◽  
Hiroshi Uno ◽  
Satoshi Asano
Keyword(s):  
Growth Factor ◽  
Hepatocyte Growth Factor ◽  
Mrna Expression ◽  
Kupffer Cells ◽  
Rt Pcr ◽  
Potent Inducer ◽  
Ito Cells ◽  
Rat Livers ◽  
Hepatocyte Growth

Hepatocyte growth factor (HGF) is a potent inducer of hepatocyte proliferation and is expressed during liver failure. In this study we used the in situ reverse transcriptase-polymerase chain reaction (RT-PCR) method to detect HGF mRNA expression in normal rat livers and cirrhotic rat livers induced by treatment with N-nitrosodimethylamine (DMN). In normal control livers, in situ RT-PCR detected HGF mRNA expression in Ito cells and Kupffer cells, both of which showed rounded morphologies. However, in the cirrhotic livers induced by DMN, HGF mRNA-positive cells were spindle-shaped and surrounded the hepatocytes located around the sinusoids. These cells appeared to be sinusoidal endothelial cells as well as Ito and Kupffer cells. Because it has been suggested that HGF expression is related to transforming growth factor-β (TGF-β) levels that may play an essential role in disease progression in cirrhotic livers, TGF-β mRNA expression in normal and cirrhotic livers was also compared using in situ RT-PCR. Our results confirmed that expression of TGF-β mRNA co-localized with HGF mRNA expression in the cirrhotic liver.

Estrogen Receptor-β mRNA Expression in Rat Ovary: Down-Regulation by Gonadotropins

1997 ◽  
Vol 11 (2) ◽  
pp. 172-182 ◽  
Author(s):  
Michael Byers ◽  
George G. J. M. Kuiper ◽  
Jan-Åke Gustafsson ◽  
Ok-Kyong Park-Sarge
Keyword(s):  
Estrogen Receptor ◽  
In Situ Hybridization ◽  
Mrna Expression ◽  
Granulosa Cells ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Adult Rats ◽  
Mature Adult ◽  
Preovulatory Follicles

Abstract We have examined the expression and regulation of the two estrogen receptor (ERα and ERβ) genes in the rat ovary, using Northern blotting, RT-PCR, and in situ hybridization histochemistry. Northern blotting results show that the ovary expresses both ERα and ERβ genes as single (∼6.5-kb) and multiple (ranging from ∼1.0-kb to ∼10.0-kb) transcripts, respectively. ERα mRNA is expressed at a level lower than ERβ mRNA in immature rat ovaries. This relationship appears unchanged between sexually mature adult rats and immature rats. In sexually mature adult rats undergoing endogenous hormonal changes, whole ovarian content of ERβ mRNA, as determined by RT-PCR, remained more or less constant with the exception of the evening of proestrus when ERβ mRNA levels were decreased. Examination of ERβ mRNA expression at the cellular level, by in situ hybridization, showed that ERβ mRNA is expressed preferentially in granulosa cells of small, growing, and preovulatory follicles, although weak expression of ERβ mRNA was observed in a subset of corpora lutea, and that the decrease in ERβ mRNA during proestrous evening is attributable, at least in part, to down-regulation of ERβ mRNA in the preovulatory follicles. This type of expression and regulation was not typical for ERα mRNA in the ovary. Although whole ovarian content of ERα mRNA was clearly detected by RT-PCR, no apparent modulation of ERα mRNA levels was observed during the estrous cycle. Examination of ERα mRNA expression at the cellular level, by in situ hybridization, showed that ERα mRNA is expressed at a low level throughout the ovary with no particular cellular localization. To further examine the potential role of the preovulatory pituitary gonadotropins in regulating ERβ mRNA expression in the ovary, we used immature rats treated with gonadotropins. In rats undergoing exogenous hormonal challenges, whole ovarian content of ERβ mRNA, as determined by RT-PCR, remained more or less unchanged after an injection of PMSG. In contrast, a subsequent injection of human CG (hCG) resulted in a substantial decrease in whole ovarian content of ERβ mRNA. In situ hybridization for ERβ mRNA shows that small, growing, and preovulatory follicles express ERβ mRNA in the granulosa cells. The preovulatory follicles contain ERβ mRNA at a level lower than that observed for small and growing follicles. In addition, there is an abrupt decrease in ERβ mRNA expression in the preovulatory follicles after hCG injection. The inhibitory effect of hCG on ERβ mRNA expression was also observed in cultured granulosa cells. Moreover, agents stimulating LH/CG receptor-associated intracellular signaling pathways (forskolin and a phorbol ester) readily mimicked the effect of hCG in down-regulating ERβ mRNA in cultured granulosa cells. Taken together, our results demonstrate that 1) the ovary expresses both ERα and ERβ genes, although ERβ is the predominant form of estrogen receptor in the ovary, 2) ERβ mRNA is localized predominantly to the granulosa cells of small, growing, and preovulatory follicles, and 3) the preovulatory LH surge down-regulates ERβ mRNA. These results clearly implicate the physiological importance of ERβ in female reproductive functions.

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