A General Method for Rapid Purification of Soluble Versions of Glycosyhosphatidylinositol-Anchored Proteins Expressed in Insect Cells: An Application for Human Tissue-Nonspecific Alkaline Phosphatase

Oda, Kimimitsu; Amaya, Yoshihiro; Fukushi-Irie, Mariko; Kinameri, Yasuko; Ohsuye, Kazuhiro; Kubota, Ichiro; Fujimura, Shinichi; Kobayashi, Jun

doi:10.1093/oxfordjournals.jbchem.a022505

Cited by 113 publications

(65 citation statements)

References 25 publications

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“…After inhibition of endogeneous peroxidase, sections were incubated with anti-TNALP rabbit polyclonal antibody (Oda et al 1999), and then with anti-rabbit IgG secondary antibody, for posterior visualization by the 3, 3'-diaminobenzidine method. Sections were counter-stained with methyl green, mounted with glycerin and photographed for successive TNALP-keratin double staining.…”

Section: Ihc For Tnalp Staining and Double Ihc For Tnalp-keratinmentioning

confidence: 99%

Hertwig’s epithelial root sheath cell behavior during initial acellular cementogenesis in rat molars

Yamamoto

Yamada

et al. 2014

Histochem Cell Biol

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This study was designed to examine developing acellular cementum in rat molars by immunohistochemistry, to elucidate (1) how Hertwig's epithelial root sheath disintegrates and (2) whether epithelial sheath cells transform into cementoblasts through epithelial-mesenchymal transition (EMT).Initial acellular cementogenesis was divided into three developmental stages, which can be seen in three different portions of the root: portion 1, where the epithelial sheath is intact; portion 2, where the epithelial sheath becomes fragmented; and portion 3, where acellular cementogenesis begins. Antibodies against three kinds of matrix proteinases, which degrade epithelial sheath-maintaining factors, including basement membrane and desmosomes, were used to investigate proteolytic activity of the epithelial sheath. Tissue non-specific alkaline phosphatase (TNALP) and keratin were used to investigate EMT. Epithelial sheath cells showed immunoreactivity for all three enzymes at fragmentation, which suggests that epithelial sheath disintegration is enzymatically mediated. Dental follicle cells and cementoblasts showed intense immunoreactivity for TNALP, and from portion 1 through to 3, the reaction extended from the alveolar bone-related zone to the root-related zone. Cells possessing keratin/TNALP double immunoreactivity were virtually absent. Keratin-positive epithelial sheath cells showed negligible immunoreactivity for TNALP, and epithelial cells did not appear to migrate to the dental follicle. Together, these findings suggest that a transition phenotype between epithelial cells and cementoblasts does not exist in the developing dental follicle, and hence that epithelial sheath cells do not undergo EMT during initial acellular cementogenesis.In brief, this study supports the notion that cementoblasts derive from the dental follicle. words

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Section: Ihc For Tnalp Staining and Double Ihc For Tnalp-keratinmentioning

confidence: 99%

Hertwig’s epithelial root sheath cell behavior during initial acellular cementogenesis in rat molars

Yamamoto

Yamada

et al. 2014

Histochem Cell Biol

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“…Seven-week old male wild-type and kl/kl mice (n=8 each, Japan CLEA, Tokyo, Japan) were used in this study, which followed the principles for care and research use of animals set by Hokkaido Sections were then incubated for 2-3 hr at room temperature (RT) with rabbit polyclonal antisera against human tissue nonspecific alkaline phosphatase (TNALPase) [36] diluted at 1:300 with 1%…”

Section: Tissue Preparationmentioning

confidence: 99%

Altered distribution of bone matrix proteins and defective bone mineralization in klotho-deficient mice

Sasaki

Hasegawa

Yamada

et al. 2013

Bone

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In an attempt to identify the histological properties of the klotho-deficient (kl/kl) bone matrix, bone mineralization and the localization of Ca 2+ -binding bone matrix proteins -osteocalcin, dentin matrix protein-1 (DMP-1) and matrix Gla protein (MGP) -were examined in kl/kl tibiae.While a widespread osteocalcin staining could be verified in the wild-type bone matrix, localization of the same protein in kl/kl tibiae seemed rather restricted to osteocytes with only a faint staining of the whole bone matrix. In wild-type mice, MGP immunoreactivity was present at the junction between the epiphyseal bone and cartilage, and at the insertion of the cruciate ligaments. In kl/kl mice, however, MGP was seen around the cartilaginous cores of the metaphyseal trabeculae and in the periphery of some cells of the bone surface. DMP-1 was identified in the osteocytic canalicular system of wild-type tibiae, but in kl/kl tibiae this protein was mostly found in the osteocytic lacunae and in the periphery of some cells of the bone surface.Mineralization of the kl/kl bone seemed somewhat defective, with broad unmineralized areas within its matrix. In these areas, mineralized osteocytes along with their lacunae and osteocytic cytoplasmic processes were found to have intense osteocalcin and DMP-1 staining. Taken together, it might be that the excessive production of Ca 2+ -binding molecules such as osteocalcin and DMP-1 by osteocytes concentrates mineralization around such cells, disturbing the completeness of mineralization in the kl/kl bone matrix. words

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“…For reducing nonspecific binding, 1% BSA (Serologicals Proteins Inc., Kankakee, IL) in PBS (1% BSA-PBS) was applied on the sections for 20 min. Sections were then incubated with antibodies against ALP, 23 DMP-1 (Takara Bio), and FGF-23 (R&D Systems, McKinley Place, NE) with 1% BSA-PBS at room temperature for 2 h and, after several washings in PBS, further incubated with HRP-or ALP-conjugated secondary antibodies (Chemicon International) for l h. For visualizing the immunoreaction, diaminobenzidine tetrahydrochloride or naphthol AS-BI was used as a substrate. For tartrate-resistant acid phosphatase detection, sections were incubated in a mixture of 8 mg of naphthol AS-BI phosphate (Sigma, St. Louis, MO), 70 mg of red violet LB salt (Sigma), and 50 mM L(ϩ) tartaric acid (0.76 g; Nacalai Tesque, Kyoto, Japan) diluted in 60 ml of a 0.1 M sodium acetate buffer (pH 5.0) for 20 min at 37°C.…”

Section: Bone Analysismentioning

confidence: 99%

Type IIc Sodium–Dependent Phosphate Transporter Regulates Calcium Metabolism

Segawa¹,

Onitsuka²,

Kuwahata³

et al. 2009

Journal of the American Society of Nephrology

115

111

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Primary renal inorganic phosphate (Pi) wasting leads to hypophosphatemia, which is associated with skeletal mineralization defects. In humans, mutations in the gene encoding the type IIc sodiumdependent phosphate transporter lead to hereditary hypophophatemic rickets with hypercalciuria, but whether Pi wasting directly causes the bone disorder is unknown. Here, we generated Npt2c-null mice to define the contribution of Npt2c to Pi homeostasis and to bone abnormalities. Homozygous mutants (Npt2c Ϫ/Ϫ ) exhibited hypercalcemia, hypercalciuria, and elevated plasma 1,25-dihydroxyvitamin D 3 levels, but they did not develop hypophosphatemia, hyperphosphaturia, renal calcification, rickets, or osteomalacia. The increased levels of 1,25-dihydroxyvitamin D 3 in Npt2c Ϫ/Ϫ mice compared with age-matched Npt2c ϩ/ϩ mice may be the result of reduced catabolism, because we observed significantly reduced expression of renal 25-hydroxyvitamin D-24-hydroxylase mRNA but no change in 1␣-hydroxylase mRNA levels. Enhanced intestinal absorption of calcium (Ca) contributed to the hypercalcemia and increased urinary Ca excretion. Furthermore, plasma levels of the phosphaturic protein fibroblast growth factor 23 were significantly decreased in Npt2c Ϫ/Ϫ mice. Sodium-dependent Pi co-transport at the renal brush border membrane, however, was not different among Npt2c ϩ/ϩ , Npt2c ϩ/Ϫ , and Npt2c Ϫ/Ϫ mice.In summary, these data suggest that Npt2c maintains normal Ca metabolism, in part by modulating the vitamin D/fibroblast growth factor 23 axis. 20: 104 -113, 200920: 104 -113, . doi: 10.1681 Inorganic phosphate (Pi) is an essential nutrient in terms of both cellular metabolism and skeletal mineralization. The kidney is a major regulator of Pi homeostasis and can increase or decrease its Pi reabsorptive capacity to accommodate Pi needs. Up to 70% of filtered Pi is reabsorbed in the proximal tubule in which sodium (Na)-dependent Pi transport systems in the brush border membrane (BBM) mediated the rate-limiting step in the overall Pi reabsorptive process. J Am Soc Nephrol

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A General Method for Rapid Purification of Soluble Versions of Glycosyhosphatidylinositol-Anchored Proteins Expressed in Insect Cells: An Application for Human Tissue-Nonspecific Alkaline Phosphatase

Cited by 113 publications

References 25 publications

Hertwig’s epithelial root sheath cell behavior during initial acellular cementogenesis in rat molars

Hertwig’s epithelial root sheath cell behavior during initial acellular cementogenesis in rat molars

Altered distribution of bone matrix proteins and defective bone mineralization in klotho-deficient mice

Type IIc Sodium–Dependent Phosphate Transporter Regulates Calcium Metabolism

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