Partial Characterization and Cellular Localization of Two Deoxyribonucleases in the Small Intestine of the Rat

1971 ◽  
Vol 49 (1) ◽  
pp. 38-43 ◽  
Author(s):  
M. W. Liebbrman ◽  
R. J. Sullivan ◽  
K. H. Shull ◽  
H. Liang ◽  
E. Farber
Keyword(s):  
Small Intestine ◽  
Pancreatic Duct ◽  
Cellular Localization ◽  
Deep Layer ◽  
Superficial Layer ◽  
Rat Small Intestine ◽  
Ph Optimum ◽  
Intestinal Crypt ◽  
Deoxyribonuclease I ◽  
Crypt Cells

We have partially characterized and localized two previously reported deoxyribonucleases from the rat small intestine. After separation of the crypt cells and muscle (the deep layer) from the villus cells (the superficial layer), the latter was found to contain a deoxyribonuclease I with a pH optimum around 6, and a molecular weight of 32 000 – 35 000. It was activated by Co2+, Mg2+, and Mn2+. Ligation of the pancreatic duct reduced the activity in intestinal extracts to about one-third of control levels. A deoxyribonuclease II with a pH optimum of 5.3–5.4 was found associated with the continuously dividing intestinal crypt cells. It was inhibited by Mg2+ and activated by EDTA. Ligation of the pancreatic duct was without effect on this enzyme. The deoxyribonuclease I is probably largely extracellular and serves a digestive function while the deoxyribonuclease II probably is related to intracellular DNA metabolism.

scholarly journals Vitamin A Up-Regulates Expression of Bone-Type Alkaline Phosphatase in Rat Small Intestinal Crypt Cell Line and Fetal Rat Small Intestine

1998 ◽  
Vol 128 (11) ◽  
pp. 1869-1877 ◽  
Author(s):  
Takeshi Nikawa ◽  
Kazuhito Rokutan ◽  
Kayo Nanba ◽  
Kaori Tokuoka ◽  
Shigetada Teshima ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Small Intestine ◽  
Vitamin A ◽  
Cell Line ◽  
Crypt Cell ◽  
Rat Small Intestine ◽  
Intestinal Crypt ◽  
Fetal Rat ◽  
Small Intestinal ◽  
Intestinal Crypt Cell

scholarly journals A novel marker glycoprotein for the microvillus membrane of surface colonocytes of rat large intestine and its presence in small-intestinal crypt cells.

1988 ◽  
Vol 106 (6) ◽  
pp. 1937-1946 ◽  
Author(s):  
S U Gorr ◽  
B Stieger ◽  
J A Fransen ◽  
M Kedinger ◽  
A Marxer ◽  
...  
Keyword(s):  
Small Intestine ◽  
Light And Electron Microscopy ◽  
Extraction Procedure ◽  
Distal Colon ◽  
Adult Rats ◽  
Intestinal Crypt ◽  
Phase Partitioning ◽  
Microvillus Membrane ◽  
Small Intestinal ◽  
Crypt Cells

Murine mAbs were produced against purified microvillus membranes of rat colonocytes in order to establish a marker protein for this membrane. The majority of antibodies binding to the colonic microvillus membrane recognized a single protein with a mean apparent Mr of 120 kD in both proximal and distal colon samples. The antigen is membrane bound as probed by phase-partitioning studies using Triton X-114 and by the sodium carbonate extraction procedure and is extensively glycosylated as assessed by endoglycosidase F digestion. Localization studies in adult rats by light and electron microscopy revealed the microvillus membrane of surface colonocytes as the principal site of the immunoreaction. The antigen was not detectable in kidney or liver by immunoprecipitation but was present in the small intestine, where it was predominantly confined to the apical membrane of crypt cells and much less to the microvillus membrane of differentiated enterocytes. During fetal development, the antigen appears first in the colon at day 15 and 1-2 d later in the small intestine. In both segments, it initially covers the whole luminal surface but an adult-like localization pattern develops soon after birth. The antibodies were also used to develop a radiometric assay for the quantification of the antigen in subcellular fractions of colonocytes in order to assess the validity of a previously developed method for the purification of colonic brush-border membranes (Stieger, B., A. Marxer, and H.P. Hauri. 1986. J. Membr. Biol. 91:19-31.). The results suggest that we have identified a valuable marker glycoprotein for the colonic microvillus membrane, which in adult rats may also serve as a marker for early differentiation of enterocyte progenitor cells in small-intestinal crypt cells.

scholarly journals Fibronectin synthesis by epithelial crypt cells of rat small intestine.

1978 ◽  
Vol 75 (11) ◽  
pp. 5548-5552 ◽  
Author(s):  
A. Quaroni ◽  
K. J. Isselbacher ◽  
E. Ruoslahti
Keyword(s):  
Small Intestine ◽  
Rat Small Intestine ◽  
Crypt Cells

scholarly journals Tryptophan 5-hydroxylase in rat intestine

1973 ◽  
Vol 131 (2) ◽  
pp. 375-380 ◽  
Author(s):  
T. Noguchi ◽  
M. Nishino ◽  
R. Kido
Keyword(s):  
Enzyme Activity ◽  
Small Intestine ◽  
Physiological Significance ◽  
Rat Small Intestine ◽  
Rat Intestine ◽  
Ph Optimum ◽  
Full Activity

Tryptophan 5-hydroxylase was partially purified from rat small intestine and characterized. The enzyme activity was mainly localized in the distal one-fourth of the small intestine. The enzyme required Fe2+, 2-amino-4-hydroxy-6,7-dimethyl-5,6,7,8-tetrahydropteridine and oxygen for full activity. The pH optimum of the reaction was 8.0. The hydroxylation rate of d-tryptophan by the enzyme was one-third that of l-tryptophan. l-Phenylalanine and l-tyrosine could not serve as substrates. The physiological significance of the enzyme is discussed.

scholarly journals The assay and partial characterization of macromolecular heparin depolymerase activity in rat small intestine

1979 ◽  
Vol 180 (3) ◽  
pp. 587-596 ◽  
Author(s):  
E Young ◽  
A A Horner
Keyword(s):  
Small Intestine ◽  
Total Radioactivity ◽  
Gel Chromatography ◽  
Rat Small Intestine ◽  
Ph Optimum ◽  
Tris Maleate ◽  
Two Peaks ◽  
And Storage ◽  
Second Peak

Homogenates of rat small intestine can depolymerize macromolecular rat skin heparin (RS heparin) to products similar in size to commercial heparin [Horner (1972) Proc. Natl. Acad. Sci. U.S.A. 69, 3469–3473]. This activity is attributed to an enzyme provisionally named ‘macromolecular heparin depolymerase’. An assay for macromolecular heparin depolymerase activity in rat small intestine has been developed, based on the action of the enzyme on 35S-labelled macromolecular RS heparin. The depolymerized products are separated into two peaks by gel chromatography through columns of Bio-Gel A-15m. The amount of label in the second peak, expressed as a percentage of the total radioactivity, is the index of enzyme activity. The pH optimum was found to be 6.0 and the temperature optimum 45 degrees C. The enzyme was shown to be most stable in 50mM-Tris/maleate buffer containing 1 mM-EDTA. Macromolecular heparin depolymerase activity measured as a function of time and substrate concentration produced curves typical of an enzymic reaction. Evidence was obtained demonstrating that the activity did not originate from bacteria in the intestine. Macromolecular heparin depolymerase activity was increased by dilution and storage at 7 degrees C for 24 h. This suggests that homogenates of rat small intestine contain an unstable inhibitor of the enzyme.

scholarly journals Crypt cell isolation in the small intestine of the mouse.

1977 ◽  
Vol 25 (7) ◽  
pp. 554-559 ◽  
Author(s):  
V C Kremski ◽  
L Varani ◽  
C DeSaive ◽  
P Miller ◽  
C Nicolini
Keyword(s):  
Small Intestine ◽  
Small Bowel ◽  
Epithelial Cells ◽  
Single Cell ◽  
Cell Isolation ◽  
Cell Suspensions ◽  
Crypt Cell ◽  
Intestinal Crypt ◽  
Successful Method ◽  
Crypt Cells

A successful method has been developed for isolating viable single cell suspensions of intestinal crypt cells from the small bowel of the mouse. The lumen of the intestine was perfused with a 0.2% trypsin solution that dissociated the lining epithelial cells. Crypt cell isolation, which proves to be extremely critical, occurred under optimal mechanical and chemical configurations about 75 min after the beginning of the procedure.

Intestinal Crypt Survival after X Irradiation of the Rat Small Intestine under Conditions of Radioprotection

1978 ◽  
Vol 74 (1) ◽  
pp. 129 ◽  
Author(s):  
Victoria L. Y. Tseng ◽  
Jerald W. Bybee ◽  
James W. Osborne
Keyword(s):  
Small Intestine ◽  
Rat Small Intestine ◽  
Intestinal Crypt ◽  
X Irradiation
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