The quantification of ADAMTS expression in an animal model of cerebral ischemia using real‐time PCR

Tian, Yi; Zhang, P. B.; Xiao, Xue; Zhang, J. S.; Zhao, Junjie; Kang, Qianyan; Chen, X. L.; Qiu, Feifei; Liu, Y.

doi:10.1111/j.1399-6576.2006.01161.x

Cited by 26 publications

(21 citation statements)

References 25 publications

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“…Several reports have suggested the use of Rpl13a as a good candidate HKG in other tissues. Rpl13a expression was found to be unaltered in mouse prostate tissue explants, human prostate cancer (LNCaP) cells and human breast cancer (T-47D and ZR-75-1) cells following treatment with the histone deacetylase inhibitor Trichostatin-A and in a rat model of cerebral artery occlusion [23,24].…”

Section: Identification Of Appropriate Hkgs For Rt-pcr Studiesmentioning

confidence: 98%

Estrogen modulates expression of putative housekeeping genes in the mouse uterus

Schroder

Pelch

Nagel

2009

Endocr

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Estrogens regulate gene expression and cell proliferation in target tissues. In studies of estrogen-regulated gene expression, identification of appropriate housekeeping genes (HKGs), reference genes whose expression is not altered by treatment, is difficult. The goal of this study was to define HKGs unaltered by estrogen in the mouse uterus. Ovariectomized C57BL6 mice were dosed with 20 micrograms/kg ethinylestradiol and the uterus was collected at 6, 24, and 72 h later to bracket the biphasic time course of estrogen action in the rodent uterus. RNA was isolated, cDNA synthesized and equal amounts of cDNA were added to real-time PCR reactions. The expression of seven out of nine putative HKGs was altered by estrogen in the mouse uterus. Estrogen induced four gene expression profiles, expression of: (1) Actb and Hsp90ab1 were up-regulated early, (2) B2m and Gusb were up-regulated late, (3) Gapdh, Hprt1, and Ppia were up regulated at all time points, and (4) Rpl13a and 18srRNA were unaltered. This highlights the need to empirically determine the appropriate HKG for each experimental condition. Based on these results, we suggest using Rpl13a or 18srRNA as HKGs for xenoestrogen studies in the mouse uterus and as good candidates to test under different experimental conditions.

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Section: Identification Of Appropriate Hkgs For Rt-pcr Studiesmentioning

confidence: 98%

Estrogen modulates expression of putative housekeeping genes in the mouse uterus

Schroder

Pelch

Nagel

2009

Endocr

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“…Brevican cleavage is of particular relevance to nervous system physiology due to its involvement in synaptic plasticity (Yuan et al, 2002; Mayer et al, 2005) and glioma invasion (Matthews et al, 2000) (discussed in greater detail later in this review). Although substrates have not been identified, evidence suggests that ADAMTSs contribute to neurodegenerative disorders (i.e., ADAMTS-1 expression is increased in Alzheimer’s disease and Down syndrome) (Miguel et al, 2005) and cerebral ischemia (i.e., increased expression of ADAMTS-1, -4, -8, and -9) (Cross et al, 2006a; Tian et al, 2007b; Reid et al, 2009). Both ADAMTS-1 and ADAMTS-8 exert anti-angiogenic effects, but display differential temporal up-regulation in ischemia, with peak ADAMTS-1 expression at 24 hours post occlusion and peak ADMTS-8 expression after 3 days (Tian et al, 2007b); thus, each likely subserves different post-ischemia effects.…”

Section: Adam-tssmentioning

confidence: 99%

Metzincin Proteases and Their Inhibitors: Foes or Friends in Nervous System Physiology?

Rivera¹,

Khrestchatisky²,

Kaczmarek³

et al. 2010

J. Neurosci.

211

213

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Members of the metzincin family of metalloproteinases have long been considered merely degradative enzymes for extracellular matrix molecules. Recently, however, there has been growing appreciation for these proteinases and their endogenous inhibitors, tissue inhibitors of metalloproteinases (TIMPs), as fine modulators of nervous system physiology and pathology. Present all along the phylogenetic tree, in all neural cell types, from the nucleus to the synapse and in the extracellular space, metalloproteinases exhibit a complex spatiotemporal profile of expression in the nervous parenchyma and at the neurovascular interface. The irreversibility of their proteolytic activity on numerous biofactors (e.g., growth factors, cytokines, receptors, DNA repair enzymes, matrix proteins) is ideally suited to sustain structural changes that are involved in physiological or postlesion remodeling of neural networks, learning consolidation or impairment, neurodegenerative and neuroinflammatory processes, or progression of malignant gliomas. The present review provides a state of the art overview of the involvement of the metzincin/TIMP system in these processes and the prospects of new therapeutic strategies based on the control of metalloproteinase activity.The importance of proteolysis in tissue structure/function is reflected not only in the evolutionary conservation of protease genes in all kingdoms (e.g., from archaea and eubacteria to plants and animals) but also the genomic complexity of this protein class. The "degradome," the repertoire of proteases produced by cells, consists of at least 569 human, 629 rat, and 644 mouse proteases or protease-like proteins and homologs, whereas 156 human protease inhibitor genes have been identified (Puente et al., 2003). The proteases are classified into five major catalytic classes, including metalloproteinases and serine, cysteine, threonine, and aspartic proteinases, with the metalloproteinases representing the largest class (Fig. 1 A). The metzincin family of metalloproteinases is so named for the conserved Met residue at the active site and the use of a zinc ion in the enzymatic reaction. This family comprises matrix metalloproteinases (MMPs), a disintegrin and metalloproteinases (ADAMs), and ADAM proteases with thrombospondin motifs (ADAMTSs). Interest in MMPs began with the identification of an enzyme that contributes to tail resorption during tadpole metamorphosis (collagenase-1, MMP-1) (Gross and Lapiere, 1962) and increased on the discovery that these enzymes not only play a role in normal tissue remodeling but were upregulated in diverse human diseases, including chronic inflammatory disorders and cancer. MMPsMMPs, encoded by 24 human and 23 mouse genes, include secreted and membrane-associated members divided into four main subgroups according to their domain structure, including collagenases, stromelysins, gelatinases, and membrane-type MMPs (MT-MMPs) (Fig.

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“…The efficiency of the primers was confirmed by sequencing the conventional PCR products before applying for real-time PCR. The primers synthesized according to the sequences in previous reports were as follows: rat TRPV1 sense/antisense, 5Ј-GACATGCCACCCAGCAGG-3Ј/5Ј-TCAA-TTCCCACACACCTCCC-3Ј (Mezey et al, 2000); rat ␤-actin sense/antisense, 5Ј-TGACAGGATGCAGAA-GGAGA-3Ј/5Ј-TAGAGCCACCAATCCACACA-3Ј (Tian et al, 2007).…”

Section: Methodsmentioning

confidence: 99%

17- -Estradiol Enhanced Allodynia of Inflammatory Temporomandibular Joint through Upregulation of Hippocampal TRPV1 in Ovariectomized Rats

Wu¹,

Bi²,

Kou³

et al. 2010

Journal of Neuroscience

105

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Temporomandibular disorders (TMDs) predominantly affect reproductive female patients, with pain the most frequent complaint. Although estrogens are believed to play important roles in TMD pain, the mechanism underlying modulation of TMD pain by estrogens remains largely unknown. Accumulating evidence implies that the hippocampus is involved in sexual dimorphism of pain sensitivity. In this study, we investigated the hippocampal TRPV1 (transient receptor potential vanilloid 1) expression in ovariectomized rats that received 17-␤-estradiol substitution and found that 17-␤-estradiol enhanced the mechanical allodynia of inflamed temporomandibular joint (TMJ) induced by complete Freund's adjuvant. Real-time PCR and immunoblotting demonstrated that TMJ inflammation significantly induced hippocampal TRPV1 expression compared with the control group but failed to induce it in the ovariectomized rats that received no estradiol replacement. In addition, estradiol potentiated TMJ inflammation-induced hippocampal TRPV1 expression in a dose-dependent manner in the ovariectomized rats. In contrast, TRPV1 transcription in amygdala, prefrontal cortex, and thalamus was not affected by TMJ inflammation and estradiol. Immunostaining showed TRPV1 localized in the processes and cytoplasm of pyramidal neurons in CA1-CA3 regions of the hippocampus. Moreover, intrahippocampal injection of TRPV1 antagonists capsazepine and 5Ј-iodoresiniferatoxin into the CA1 region of the hippocampus significantly attenuated allodynia of inflamed TMJ in both nonovariectomized and ovariectomized rats that received estradiol replacement. Our results suggested that hippocampal TRPV1 can modulate central pain processing and estradiol may contribute to the sexual dimorphism of TMD pain sensitivity through upregulation of TRPV1 expression in the hippocampus.

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The quantification of ADAMTS expression in an animal model of cerebral ischemia using real‐time PCR

Abstract: The results suggest that RPL13A and cyclophilin are two appropriate housekeeping genes for the rat pMCAO model up to 21 days. ADAMTS1 mRNA levels increased, but ADAMTS8 decreased after pMCAO. Our data provide new insight into the mechanism of brain ischemia and self-repair after injury.

Cited by 26 publications

References 25 publications

Estrogen modulates expression of putative housekeeping genes in the mouse uterus

Estrogen modulates expression of putative housekeeping genes in the mouse uterus

Metzincin Proteases and Their Inhibitors: Foes or Friends in Nervous System Physiology?

17- -Estradiol Enhanced Allodynia of Inflammatory Temporomandibular Joint through Upregulation of Hippocampal TRPV1 in Ovariectomized Rats

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