Neutral aminoaciduria in cystathionine β-synthase-deficient mice, an animal model of homocystinuria

Abstract

I. Neutral aminoaciduria in cystathionine ␤-synthase-deficient mice, an animal model of homocystinuria.

Quantification of Biological Thiols in the Plasma of a Homocystinuria Model with Cystathionine β-Synthase Deficiency Utilizing Hydrophilic Interaction Liquid Chromatography and Fluorescence Detection

“…Slight concentration differences in -Hetero and -WT mice between the present study and those in previous reports may be due to differences in age (15-18 days vs 21 days, respectively) and rearing conditions. Results of tCys and tGSH were also consistent with those of previous reports [5,11].…”

“…As the accumulation of Hcy caused by CBS deficiency should alter the amount of downstream thiol metabolites, a comprehensive analysis of biological thiols should offer useful information to understand the pathology of disease. However, only Hcy, Cys, and GSH have been measured in humans and animal models of homocystinuria [5][6][7][8][9][10][11]. Considering that γGluCys and CysGly are precursors and degradation products of GSH, respectively, simultaneous analysis of all the thiols mentioned above should offer helpful information to reveal how Hcy accumulation leads to diverse symptoms and to investigate suitable indicators of disease progression.…”

Hyperhomocysteinemia abrogates fasting-induced cardioprotection against ischemia/reperfusion by limiting bioavailability of hydrogen sulfide anions

“…CBS heterozygous knockout (Cbs +/− ) mice (B6.129P2-Cbs tm1Unc /J) [11] and NOS3 (nitric oxide synthase 3 (=endothelial NOS, eNOS)) homozygous knockout (Nos3 −/− ) mice (B6.129P2-Nos3 tm1Unc /J) [27] were purchased from the Jackson Laboratory (Bar Harbor, ME, USA). Because most Cbs −/− mice on a C57BL/6J background die before 4 weeks of age probably due to severe hepatic dysfunction [11,12,28,29], we used Cbs −/− mice on a C3H/HeJ background, which have been established after successive backcrossing and display improved survivability/ development growth [12,30]. Mice were allowed free access to water and a CE-2 standard dry rodent diet (CLEA Japan) that contained 0.44 % Met (ad libitum, AL) or a high-Met diet (CE-2+2.20 % Met) to induce dietary hyperhomocysteinemia [17].…”

“…Serum levels of total homocysteine (homocysteine and all its derivatives that give rise to the thiol homocysteine after reductive cleavage of disulfide bonds [33]) were measured using an Azwell auto homocysteine kit (Alfresa Pharma, Osaka, Japan) as described previously [14,29]. Both reduced glutathione and oxidized glutathione (GSH and GSSG, respectively) were measured using coulometric electrochemical detection as described previously [14], and GSH/total GSH (GSH + 2 × GSSG) molecular ratios were calculated.…”

DHHC7-mediated palmitoylation of the accessory protein barttin critically regulates the functions of ClC-K chloride channels

“…Next, we compared the distribution of ClC-K/barttin channels within a nephron. For detection of the ClC-K channels, we applied rabbit polyclonal antibody from Alomone Labs, which has long been accepted as a highly specific and fairly unambiguous tool for visualization of the ClC-K channel both in vitro and in vivo (28), (29), (30). For visualization of barttin, we used the goat polyclonal antibody sc-49611 from Santa Cruz.…”