Mass spectrometric determination of prostanoids in rat hypothalamic paraventricular nucleus microdialysates.
The hypothalamic paraventricular nucleus (PVN) is one of the most important autonomic control centers in the brain. Several kinds of prostanoids, such as prostaglandin (PG) E2, are considered to act in the PVN as mediators of autonomic responses. In the present study, we used liquid chromatography ion trap tandem mass spectrometry (LC-ITMS(n)) to simultaneously quantify four prostanoids, thromboxane (Tx) B2, PGE2, PGD2 and 15-deoxy-∆(12,14) (15d)-PGJ2 in PVN microdialysates from urethane-anesthetized rats. The quantification limits were estimated to be 0.05ng/mL for TxB2, 0.025ng/mL for PGE2, 0.1ng/mL for PGD2, and 0.5ng/mL for 15d-PGJ2. The RSD% obtained from all prostanoids was <15%, indicating an acceptable level of reproducibility. LC-ITMS(n) analysis of rat PVN microdialysates revealed that TxA2 may play an important role in adrenomedullary outflow evoked by centrally administered N-methyl-d-aspartate, corticotrophin-releasing factor and glucagon-like peptide-1. This is the first study to use LC-ITMS(n) to analyze prostanoid levels in rat PVN microdialysates. This LC-ITMS(n) method will be useful for investigating the potential involvement of prostanoids in brain function.
Department of Pharmacology, Aichi Medical University Graduate School of Medicine, 1-1 Yazakokarimata, Nagakute, Aichi 480-1195, Japan; Department of Pharmacy, Aichi Medical University, 1-1 Yazakokarimata, Nagakute, Aichi 480-1195, Japan.
Masahiko
M
Tachi
Department of Pharmacology, Aichi Medical University Graduate School of Medicine, 1-1 Yazakokarimata, Nagakute, Aichi 480-1195, Japan. Electronic address: fumiokon@aichi-med-u.ac.jp.
Fumio
F
Kondo
Central Research Laboratory, Aichi Medical University School of Medicine, 1-1 Yazakokarimata, Nagakute, Aichi 480-1195, Japan.
Minoru
M
Fukayama
Central Research Laboratory, Aichi Medical University School of Medicine, 1-1 Yazakokarimata, Nagakute, Aichi 480-1195, Japan.
Kazuhiro
K
Yoshikawa
Department of Pharmacy, Aichi Medical University, 1-1 Yazakokarimata, Nagakute, Aichi 480-1195, Japan.
Katsuhiko
K
Matsuura
Department of Pharmacology, Aichi Medical University Graduate School of Medicine, 1-1 Yazakokarimata, Nagakute, Aichi 480-1195, Japan.
Shoshiro
S
Okada
D000758:Anesthesia; D000818:Animals; D002138:Calibration; D002395:Catecholamines / Q000097:blood; D002853:Chromatography, Liquid / Q000379:methods; D003346:Corticotropin-Releasing Hormone / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism*; D052216:Glucagon-Like Peptide 1 / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D017551:Microdialysis / Q000379:methods; D016202:N-Methylaspartate / Q000378:metabolism; D010286:Paraventricular Hypothalamic Nucleus / Q000378:metabolism*; D015230:Prostaglandin D2 / Q000031:analogs & derivatives* / Q000378:metabolism*; D011453:Prostaglandins / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D013929:Thromboxane B2 / Q000378:metabolism*; D014520:Urethane
C097240:15-deoxy-delta(12,14)-prostaglandin J2; D002395:Catecholamines; D011453:Prostaglandins; D014520:Urethane; D013929:Thromboxane B2; D016202:N-Methylaspartate; D052216:Glucagon-Like Peptide 1; D003346:Corticotropin-Releasing Hormone; D015232:Dinoprostone; D015230:Prostaglandin D2
Hypothalamic PVN; LC–ITMS; Microdialysis; Prostanoids