Localization of a Mitochondrial Type of NADP-dependent Isocitrate Dehydrogenase in Kidney and Heart of Rat: An Immunocytochemical and Biochemical Study

Haraguchi, Celina M.; Mabuchi, Tadashi; Yokota, Sadaki

doi:10.1177/002215540305100210

Cited by 25 publications

(21 citation statements)

References 44 publications

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“…Rabbit anti-rat IgG was prepared by immunizing rabbits with purified rat IgG and affinity purifying the antibodies specific for rat IgG. Rabbit anti-mitochondrial isocitrate dehydrogenase (ICD1) has been described previously (Haraguchi et al 2003). Rabbit anti-histone H2B, rabbit anti-protein disulfide isomerase (PDI), and rabbit anti-a-tubulin were obtained from Cell Signaling Technology (Boston, MA), Sigma-Aldrich Japan (Tokyo), and Thermo Fisher Scientific (Fremont, CA), respectively.…”

Section: Antibodies and Related Probesmentioning

confidence: 99%

Expression of DDX25 in nuage components of mammalian spermatogenic cells: immunofluorescence and immunoelectron microscopic study

Onohara

Yokota

2011

Histochem Cell Biol

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The localization of DDX25/GRTH and gonadotropin-stimulated RNA helicase was studied in the spermatogenic cells of rat, mouse, and guinea pig by immunofluorescence and immunoelectron microscopy (IEM). Immunofluorescence studies identified four kinds of granular staining: (1) fine particles observed in meiotic cells; (2) small granules associated with a mitochondrial marker, appearing in pachytene spermatocytes after stage V; (3) short strands lacking the mitochondrial marker in late spermatocytes; and, (4) large irregularly shaped granules in round spermatids. IEM identified DDX25 signals in nine compartments: (1) fine dense particles in the meiotic cells; (2) intermitochondrial cement; (3) loose aggregates of 70-90 nm particles; (4) chromatoid bodies; (5) late chromatoid bodies; (6) satellite bodies; (7) granulated bodies; (8) mitochondria-associated granules; and, (9) reticulated bodies. Compartments (1) to (6) were previously classified into nuage while (7) to (9) were classified as nuage components by the present study. The results suggest that DDX25 functions in these nine compartments.

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Section: Antibodies and Related Probesmentioning

confidence: 99%

Expression of DDX25 in nuage components of mammalian spermatogenic cells: immunofluorescence and immunoelectron microscopic study

Onohara

Yokota

2011

Histochem Cell Biol

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“…It is unlike the bacterial enzyme, which is regulated by phosphorylation/dephosphorylation (25). The mNADP ϩ -ICDH isoform predominates (Ͼ95%) in the heart, where it is confined to cardiomyocytes (23,26,27). mNADP ϩ -ICDH exists as a homodimer of 413 amino acids per subunit with a molecular mass of 47 kDa and a pI of 9.0 (cf.…”

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confidence: 99%

Cardiac Mitochondrial NADP+-isocitrate Dehydrogenase Is Inactivated through 4-Hydroxynonenal Adduct Formation

Benderdour

Charron

deBlois

et al. 2003

Journal of Biological Chemistry

116

102

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Mitochondrial NADP؉ -isocitrate dehydrogenase activity is crucial for cardiomyocyte energy and redox status, but much remains to be learned about its role and regulation. We obtained data in spontaneously hypertensive rat hearts that indicated a partial inactivation of this enzyme before hypertrophy development. We tested the hypothesis that cardiac mitochondrial NADP ؉ -isocitrate dehydrogenase is a target for modification by the lipid peroxidation product 4-hydroxynonenal, an aldehyde that reacts readily with protein sulfhydryl and amino groups. This hypothesis is supported by the following in vitro and in vivo evidence. In isolated rat heart mitochondria, enzyme inactivation occurred within a few minutes upon incubation with 4-hydroxynonenal and was paralleled by 4-hydroxynonenal/ NADP ؉ -isocitrate dehydrogenase adduct formation. Enzyme inactivation was prevented by the addition of its substrate isocitrate or a thiol, cysteine or glutathione, suggesting that 4-hydroxynonenal binds to a cysteine residue near the substrate's binding site. Using an immunoprecipitation approach, we demonstrated the formation of 4-hydroxynonenal/NADP ؉ -isocitrate dehydrogenase adducts in the heart and their increased level (210%) in 7-week-old spontaneously hypertensive rats compared with control Wistar Kyoto rats. To the best of our knowledge, this is the first study to demonstrate that mitochondrial NADP ؉ -isocitrate dehydrogenase is a target for inactivation by 4-hydroxynonenal binding. Furthermore, the pathophysiological significance of our finding is supported by in vivo evidence. Taken altogether, our results have implications that extend beyond mitochondrial NADP ؉ -isocitrate dehydrogenase. Indeed, they emphasize the implication of post-translational modifications of mitochondrial metabolic enzymes by 4-hydroxynonenal in the early oxidative stress-related pathophysiological events linked to cardiac hypertrophy development.Mitochondrial dysfunction is considered to play a key role in the pathogenesis of cardiac hypertrophy and failure. Chronic alterations of fuel metabolism and oxidative stress status are factors that could impair the capacity of the mitochondria to fulfil their crucial role in energy production (1) and thereby contribute to the activation of signaling pathways governing cell death by apoptosis and/or necrosis (2, 3). Although the role of mitochondrial energy fuel deficits or of oxidative stress are often investigated separately, accumulating evidence indicates that these two factors are linked. For example, several metabolic enzymes can be inactivated through post-translational modifications by oxidative stress-related molecular components (4 -11). The latter molecules include oxygen-and nitrogen-derived reactive species or aldehydes produced from lipid peroxidation. Of specific interest to this study is 4-hydroxynonenal (HNE), 1 the major ␣-␤-unsaturated aldehyde formed from peroxidation of both -3 and -6 polyunsaturated fatty acids, whose formation is enhanced in hypertrophied and ischemic/ reperfused...

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“…Horseradish peroxidase (HRP)-labeled swine anti-guinea pig IgG was obtained from DAKO Japan (Tokyo). Rabbit anti-mitochondrial isocitrate dehydrogenase (ICD1) was described previously [24]. Rabbit anti-histone H2B, rabbit anti-protein disulfide isomerase (PDI), and rabbit anti-tubulin were obtained from Cell Signaling Technology (Boston, MA), Sigma-Aldrich Japan (Tokyo), and Thermo Fisher Scientific (Fremont, CA), respectively.…”

Section: Antibodiesmentioning

confidence: 99%

Expression and Localization of NANOS1 in Spermatogenic Cells during Spermatogenesis in Rat

Yokota¹,

Onohara²

2013

CellBio

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Expression and localization of NANOS1 in the spermatogenic cells of rat testis were studied by immunofluorescence and immunoelectron microscopy. Using immunofluorescence techniques, NANOS1 was localized in the cytoplasm and discrete granules of spermatocytes and spermatids. The staining intensity of NANOS1 signal varied depending upon the stage of the cycle of seminiferous epithelium. Double immunofluorescence staining with antibodies against NANOS1 and DDX4 showed that several DDX4-positive compartments of nuage were also stained for NONOS1. Immunoelectron microscopy revealed that the major subcellular localization sites for NANOS1 were the chromatoid body (CB) and satellite body (SB), and the minor sites were the rest of the nuage compartments, including the irregularly-shaped perinuclear granules (ISPG), and intermitochondrial cement (IMC). Non-nuage structures such as mitochondria-associated granules (MAG), granulated body (GB), and reticulated body (RB) were also labeled by theNANOS1 antibody. In addition, NANOS1 was found in the outer dense fibers of flagella of spermatids at steps 12-19, and in the head cap of late spermatids after step 15. These results suggest that NANOS1 is one of the nuage proteins and functions mainly in the CBs as well as in the cytoplasm. NANOS1 may also have additional functions in non-nuage structures such as MAGs, GBs and RBs.

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Localization of a Mitochondrial Type of NADP-dependent Isocitrate Dehydrogenase in Kidney and Heart of Rat: An Immunocytochemical and Biochemical Study

Cited by 25 publications

References 44 publications

Expression of DDX25 in nuage components of mammalian spermatogenic cells: immunofluorescence and immunoelectron microscopic study

Expression of DDX25 in nuage components of mammalian spermatogenic cells: immunofluorescence and immunoelectron microscopic study

Cardiac Mitochondrial NADP+-isocitrate Dehydrogenase Is Inactivated through 4-Hydroxynonenal Adduct Formation

Expression and Localization of NANOS1 in Spermatogenic Cells during Spermatogenesis in Rat

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