| 包装 | 价格(元) |
| 1mg | 电议 |
| 5mg | 电议 |
Cell lines | |
Preparation Method | Full-length cDNAs encoding individual orphan GPCRs were subcloned into the pCDN mammalian expression vector. These plasmids were individually transfected into the human embryonic kidney (HEK) 293 cells, and stable transfectant cell lines established and maintained. For in vitro pharmacological assessments , CHO-K1 cells were stably transfected with human OX1R and OX2R expression vectors, and transfected cell lines were maintained . |
Reaction Conditions | 3 nmol /30 nmol |
Applications | The concentration of Orexin-A required to induce half-maximum response (EC50) was 30 nM. Obtained similar results in radioligand binding and [Ca2+]i transient assays performed with stably transfected HEK293 cells . Orexin A human, rat, mouse has high affinity for OX1R, with 38 nM IC50 and 34 nM EC50 values in the the [Ca2+]i transient assay. |
Animal models | C57BL/6J mice and OX1R+/+, OX1R-/-, OX2R+/+, OX2R-/- and OX1R-/-/OX2R-/- mice |
Preparation Method | Orexin A was injected by intracerebroventricular (ICV): the mice were gently restrained , injectors with a diameter of 0.15 mm (connected to Hamilton syringes by tubes) were introduced into the guide cannulae, and the animals were released in a cage. A total volume of 0.3 µl solution with 3 µg orexin A was then injected at a flow rate of 0.1 µl/min, controlled by a microinfusion pump. |
Dosage form | 0.3 µl solution with 3 µg orexin A /0.1 µl/min |
Applications | Orexin A is inhibited by almorexant specifically. The two known orexin receptors mediate sleep induction by almorexant and orexin A-induced locomotion. However, OX2R activation mediates locomotion induction by orexin A and antagonism of OX2R is sufficient to promote sleep in mice. |
| 产品描述 | Orexin A (human, rat, mouse), orchestrates diverse central and peripheral processes[1]. Orexin A (human, rat, mouse) is a specific, high-affinity agonist for G-protein-coupled receptor OX1R. Orexin A (human, rat, mouse) has a role in the regulation of feeding behavior. Orexin A (human, rat, mouse) is an effective anti-nociceptive and anti-hyperalgesic agent in mice and rats[2].Orexins first bind OXRs, which in turn activate at least three subtypes of G-proteins (Gq/11, Gi/o, and Gs) or other proteins (e.g., β-arrestin). These effectors subsequently regulate phospholipases, ion channels, and protein kinases, ultimately triggering the activation of various downstream signaling pathways[3,5]. Orexin-A induced a transient increase in [Ca2+]i in CHO/OX1R cells in a dose-dependent manner, but failed to induce detectable [Ca2+]i transients in mock transfected CHO cells. The calcium mobilization is likely caused by the activation of the Gq class of heterotrimeric G proteins[6].The calculated concentration of orexin-A required to induce half-maximum response (EC50) was 30 nM. The concentration of unlabeled Orexin-A required to displace 50% of specific radioligand binding (IC50) was 20 nM. Repeated competitive radioligand binding assays and [Ca2+]i transient dose-response studies using stably transfected CHO cells expressing the human OX2R cDNA. The results demonstrated that OX2R is indeed a high-affinity receptor for human orexin-B, with an IC50of 36 nM in the binding assay and an EC50of 60 nM in the [Ca2+]i transient assay . Orexin-A had high affinity for this receptor, with 38 nM IC50and 34 nM EC50values.These findings confirm that orexin-A is indeed a specific, high-affinity agonist for OX1R. The Orexin A concentration in cerebrospinal fluid (CSF) was abnormally low in seven of nine people with narcolepsy, implying that orexin transmission was deficient in these patients[7].In a later study, the same group reported a dramatic decrease in the CSF Orexin A levels in 32 of 38 successive narcolepsy-cataplexy cases[8].On the basis of these findings, they concluded that Orexin is deficient in most cases of human narcolepsy, suggesting possible diagnostic applications. Furthermore, the number of orexin neurons is reduced by 85%-95% in the LH of patients with narcolepsy[9]. Orexin mRNA and neuropeptide are completely absent in hypothalamus, pons and cortex of narcolepsy patients, and the secretory signal sequence of the orexin gene is deficient in the most serious cases of early onset narcolepsy[10].These observations further prove that narcolepsy is associated with deficiency in the orexin system.
The binding of orexins to orexin receptor type 1 (OX1R) or OX2R stimulates Gq or Gi subtypes, which subsequently induce the activation of phospholipase C (PLC), phospholipase A (PLA), phospholipase D (PLD) or Adenylyl cyclases (AC), ultimately resulting in an increase in cytosolic Ca2+and a downstream cascade response. In addition, OA binds OX1R and elevates Ca2+by activating nonselective cation channels (NSCCs)[11]. References: |
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