Immunocytochemical localization of a calcium-binding protein in the rat duodenum

Marché, P.; Guern, Christian Le; Gassier, P.

doi:10.1007/bf00233554

Cited by 38 publications

(9 citation statements)

References 18 publications

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“…The baboon duodenum showed a positive reaction to antibodies to both the rat and mouse calbindin-D9Ks. The distribution of the stain was similar to that occurring in rat duodenum (Marche et al, 1979;Taylor et al, 1984;van Corven et al, 1985), although the intensity of staining was much weaker in the baboon duodenum. This result could be due to poor cross-reactivity between the antibodies to rat and mouse calbindin-D9K and the baboon antigen, or there may be lower levels of calbindin-D9K in the baboon enterocytes.…”

Section: Discussionsupporting

confidence: 70%

Immunohistochemical localization of calbindins (28K and 9K) in the tissues of the baboon Papio ursinus

1990

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An indirect immunoperoxidase procedure was used to detect the presence of calbindin-D28K and calbindin-D9K in the cerebellum, kidney, and duodenum of the baboon Papio ursinus. Antibodies to chick calbinding-D28K and to both rat and mouse calbindin-D9K were used. The cerebellum and kidney were shown to contain calbindin-D28K; the doudenum contained calbindin-D9K. In the cerebellum, positive staining was found in the Purkinje cells only; in the kidney, positive staining was found in the distal convoluted tubules, connecting tubules, and collecting tubules, extending deep into the medullary regions of the kidney. Staining in the duodenum was confined to the enterocytes of the villi, with no stain present in the crypt regions or goblet cells. Thus the baboon, a primate, contains the larger of the calbindins in both the cerebellum and kidney as does the human and monkey, but its distribution in the kidney is more generalized than that found in humans. The molecular weight of calbindin-D9K was found to be similar to that found in other animals. However, the calbindin-D28K from the baboon tissues appears to be slightly smaller than the protein found in other animals and may therefore be of similar size to the human calbindin-D28K (Mr 26,000).

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Section: Discussionsupporting

confidence: 70%

Immunohistochemical localization of calbindins (28K and 9K) in the tissues of the baboon Papio ursinus

1990

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“…This is established by the synthesis of the same ratio of proteins of similar molecular size in a cell-free translation mixture in the absence or presence of microsomal membranes [5]. This absence of processing is consistent with the intracellular localisation of 9-kDa cholecalcin within the duodenal absorptive cells [23] and Blobel's theory of precursors for secretory and membrane-inserted proteins [24]. If the larger protein is the result of a translational misreading of the first UGA codon in the rabbit reticulocyte lysate, its production should increase considerably when UGA-suppressor tRNA is added to the mixture.…”

Section: Discussionsupporting

confidence: 71%

“…9-kDa cholecalcin represents about 2% of the soluble protein of the duodenal mucosa [4] and is located in the absorptive cells [23], whereas 28-kDa cholecalcin is concentrated in the distal tubule of the kidney [31] and in the Purkinje cells of the cerebellum [32]. Despite the possible presence of a site-III-like structure in the 9-kDa cholecalcin mRNA [7], resembling the site I1 sequence of 28-kDa cholecalcin, Northern blots reveal no cross-hybridisation between probe cDNA pC109 and large RNA species from kidney and cerebellum.…”

Section: Discussionmentioning

confidence: 99%

Characterisation of rat 9‐kDa cholecalcin (CaBP) messenger RNA using a complementary DNA

Desplan

Brehier

Thomasset

1985

European Journal of Biochemistry

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The rat posesses two cholecalciferol-induced calcium-binding proteins, the cholecalcins (CaBP). The 9-kDa CaBP is mainly concentrated in the duodenum while 28-kDa CaBP is located in the kidney and cerebellum. The mRNA encoding 9-kDa CaBP has been characterised using the cloned cDNA, pC109, synthesised from rat duodenal 9-kDa CaBP mRNA J. Biol. Chem. 258, 13502-135051. Nucleotide sequence analysis of this cDNA shows the presence of two stop codons, TGA and TAG, at positions 207 and 271, respectively, of the 3' untranslated region. The cDNA-hybridised mRNA, isolated from rat duodenum, directs the cell-free synthesis of two proteins precipitable by antisera to 9-kDa intestinal CaBP. A major protein comigrates with 9-kDa CaBP whereas a minor product corresponds to a protein which is larger by 2000 Da. The minor protein appears to result from read-through of the 'leaky' UGA stop signal. No protein band which was immunoprecipitable with 28-kDa CaBP antiserum was detected when cDNA-hybridised mRNA from rat kidney and cerebellum was translated in a cell-free system. Northern blots show that the cDNA pC109 sequence hybridises to a homogeneous mRNA species 500-600 nucleotides long from rat duodenum. Larger mRNA species encoding 28-kDa CaBP are undetectable in rat kidney and cerebellum even under low stringency conditions. All these findings demonstrate that there is no cross-hybridisation between 9-kDa and 28-kDa CaBP mRNAs. Southern blot analysis of rat genomic DNA, that shows only one homologous 9-kDa gene, is consistent with these findings. Thus, all our data indicate that there are distinct genes coding for each rat cholecalcin.Cholecalcin, which is also known as calcium-binding protein (CaBP), is one of a group of intracellular proteins which bind calcium with high affinity ( K , = lo6 M-I) [l, 21 and its concentration is dependent upon 1,25-dihydroxycholeca1-ciferol (calcitriol) [3,4], the hormonal form ofvitamin D. Two forms have been identified in the rat [4]. A 28 000-M, molecule (28-kDa cholecalcin), found in kidney and cerebellum, has four calcium-binding sites, whereas a 9000-Mr protein (9-kDa cholecalcin) is mainly located in the duodenum and possesses two calcium-binding domains [l, 2, 41. The presence of two distinct mRNAs encoding either 9-kDa or 28-kDa cholecalcin was deduced from cell-free translation assays, since rat duodenal poly(A)-rich RNA directs the synthesis of 9-kDa cholecalcin whereas mRNAs isolated from kidney and cerebellum code for the 28-kDa cholecalcin [5]. We recently reported the molecular cloning of a cDNA fragment synthesised from rat duodenal mRNA encoding 9-kDa cholecalcin [6]. Nucleotide sequence analysis of a resulting 375-base-pair cDNA clone, pC109, allowed the assignment of 207 nucleotides in the coding region before the TGA stop codon [7] (Fig. 1). The deduced amino acid sequence for 9-kDa cholecdlcin contains the two 'EF hand' domains described by Moews and Kretsinger [8] corresponding to the calcium-binding sites I and I1 [7]. The homology observed between site I structure ...

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“…Stimulation of the animals with 1,25-(OH)z-& raised cellular uptake 12 hr later. All vitamin D-dependent proteins appear to be located inside the cell, either in the cytosol (CaBP) (Marche et al, 1979), in the Golgi apparatus (Weiser et al, 19811, the inner aspect of the brush border (Miller et al, 1979;Schachter and Kowarski, 1982), or of the basolateral membrane (Ghijsen and van Os, 1982). Induction by ~,~E P ( O H )~-D~ of CaBP and of the brush border calciumbinding protein in vitamin D-deficient animals has been demonstrated (Buckley and Bronner, 1980;Miller et al, 1979).…”

Section: Discussionmentioning

confidence: 99%

Calcium uptake by isolated rat intestinal cells

Bronner

Pansu

Bosshard

et al. 1983

Journal Cellular Physiology

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Intestinal cells were isolated by a combination of mechanical and enzymatic means, and their calcium uptake was assayed by a rapid filtration procedure. Calcium uptake was a time- and concentration-dependent process that was markedly elevated at 25 and 37 degrees C, as compared to 0 degree C. Cells isolated from rat duodenum exhibited higher uptakes than cells from jejunum, which in turn took up more calcium than cells from the ileum. Duodenal cells from vitamin D-deficient animals took up less calcium than cells from vitamin D-replete cells. In vivo vitamin D repletion with 1,25-dihydroxyvitamin D3 raised calcium uptake by duodenal cells from treated animals toward that of cells from replete rats. Furthermore, calcium uptake by duodenal cells from vitamin D-deficient animals approximated that of ileal cells from replete rats. These findings with isolated cells parallel prior findings of tissue calcium transport and suggest that cellular calcium uptake may be related to the saturable component of intestinal calcium absorption. Isolated intestinal cells may therefore constitute one experimental model for the study of transcellular calcium transport.

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Immunocytochemical localization of a calcium-binding protein in the rat duodenum

Cited by 38 publications

References 18 publications

Immunohistochemical localization of calbindins (28K and 9K) in the tissues of the baboon Papio ursinus

Immunohistochemical localization of calbindins (28K and 9K) in the tissues of the baboon Papio ursinus

Characterisation of rat 9‐kDa cholecalcin (CaBP) messenger RNA using a complementary DNA

Calcium uptake by isolated rat intestinal cells

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