Production and Properties of Vitamin‐D‐Induced mRNA for Chick Calcium‐Binding Protein

Spencer, Rosemary; Charman, M.; Emtage, J.S.; Lawson, David

doi:10.1111/j.1432-1033.1976.tb11127.x

Cited by 140 publications

(51 citation statements)

References 28 publications

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“…Rabbit polyclonal antibody against CaBP (diluted 1:10,000, gift of Dr. E. M. Lawson; Spencer et al, 1976) and mouse monoclonal antibody against CaBP (diluted 1:10,000, gift of Dr. W. Hunziker; Pinol et al, 1990) were used to study the survival and morphological changes of Purkinje cells, particularly their axons.…”

Section: Antibodiesmentioning

confidence: 99%

Purkinje Cell Survival and Axonal Regeneration Are Age Dependent: AnIn VitroStudy

1997

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Purkinje cells are among the most resistant neurons to axotomy and the most refractory to axonal regeneration. By using organotypic cultures, we have studied age-and environmentrelated factors implicated in Purkinje cell survival and axonal regeneration. Most Purkinje cells taken from 1-to 5-d-old rats, the period in which these neurons are engaged in intense synaptogenesis and dendritic remodeling, die 1 week after plating, whereas if cultured before or after this period, Purkinje cells survive, even in the absence of deep nuclear neurons, their postsynaptic targets. Cerebellar slices taken from 10-dold rats and kept in vitro for 1 week acquire a cellular composition resembling mature cerebellum. Their Purkinje cells are resistant to axotomy, but even when confronted with permissive environments (sciatic nerves or fetal cerebellar slices), their axons do not regenerate. In contrast, fetal rat and mouse Purkinje cells are able to regenerate their axons on mature cerebellar slices. This regeneration is massive, and the regrowing axons invade all cerebellar regions of the apposed mature slices, including white matter. These results show that Purkinje cell survival and axonal regeneration are age-related and independent from environmental constraints. Moreover, our observations suggest strongly that the onset of synaptogenesis of Purkinje cell axons could provide a signal to turn off their growth program and that, thereafter, permissive microenvironment alone is unable to reestablish such a program. Key words: axonal regeneration; neuronal survival; cerebellum; Purkinje cell maturation; cerebellar organotypic cultures; axonal growthMost adult neurons survive axotomy if the lesion occurs far from their cell bodies, whereas perinatal neurons often die even after far-away axotomy (for review, see Schwab and Bartholdi, 1996). Frequently, this perinatal cell death can be counteracted by neurotrophic factors or by embryonic tissue grafting (for review, see Schwab and Bartholdi, 1996). In addition, immature CNS neurons have a higher potential to regenerate their axons than adult CNS neurons (for review, see Schwab and Bartholdi, 1996). Nevertheless, if allowed to regrow into transplanted peripheral nerves, most adult CNS cut axons regenerate their lesioned processes (David and Aguayo, 1981;Aguayo, 1985). Furthermore, the application of blocking antibodies against CNS myelin inhibitory proteins favors regeneration of axotomized corticospinal axons after spinal cord injury (Schnell and Schwab, 1990;Schwab et al., 1993). These observations strongly support the hypothesis that environmental factors rather than the intrinsic inability of adult CNS axons to regenerate are the limiting factor in the failure of axonal regeneration.Adult Purkinje cells are among the most resistant neurons to axotomy and the most refractory to axonal regeneration: they do not regenerate their axons even in the presence of their embryonic targets (Rossi et al., 1995). These cells in addition are one of the rare categories of neurons whose axons hav...

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Section: Antibodiesmentioning

confidence: 99%

Purkinje Cell Survival and Axonal Regeneration Are Age Dependent: AnIn VitroStudy

1997

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“…3 could have arisen from cytoplasmic contamination. Preparation ofchick intestinalpolyribosomes andpolyribosomal-free cytoplasmic RNA Chick ileal mucosa was homogenized in rat liver cell sap and polyribosomes were prepared as described before (Spencer et al, 1976a).…”

Section: Nmentioning

confidence: 99%

“…One of the earliest responses of vitamin D-deficient chicks to the action of 1,25-dihydroxycholecalciferol is the synthesis in intestinal cells of calcium-binding protein as a consequence of the appearance of active mRNA for the protein on polyribosomes (Spencer et al, 1976a). In such chicks calcium-binding protein is first detectable as a nascent polypeptide chain on intestinal polyribosomes 2h after dosing with 1,25-dihydroxycholecalciferol.…”

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confidence: 99%

Stimulation of intestinal calcium-binding-protein mRNA synthesis in the nucleus of vitamin D-deficient chicks by 1,25-dihydroxycholecalciferol

Spencer

Charman

Lawson

1978

Biochemical Journal

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Stimulation of intestinal calcium transport by the hormone 1,25-dihydroxycholecalciferol appears to involve RNA transcriptions and the synthesis of new proteins. Although one of these proteins has been identified as calcium-binding protein, no RNA molecules specifically induced by the hormone in the nucleus have been identified. Nuclear RNA from intestine of vitamin D-deficient chicks before and at various time intervals after treatment with the hormone or cholecalciferol was tested for its ability to code for calcium-binding protein in a cell-free system. Calcium-binding-protein mRNA could only just be detected in the intestinal nuclei 2h after dosing with these steroids which is the same time that it was first observed in the polyribosomes. Thus 1,25-dihydroxycholecalciferol induces the production of new calcium-binding protein by stimulating the formation and rapid release from the nucleus ofnew mRNA molecules for this protein. Polyribosomal translation of the mRNA continued only as long as it was being synthesized, and the maximum rate of synthesis following a pulse dose of 125 ng of the hormone was the same as that observed after prolonged stimulation with cholecalciferol. The possibility that other 1,25-dihydroxycholecalciferol-dependent events may be occurring in the nucleus in the lag period between accumulation of the hormone in the intestine and the appearance of active calcium-binding-protein mRNA, and that these may ultimately control the synthesis of that mRNA, is discussed.

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“…The major RNA species, 2000 nucleotides long, corresponds approximately to the molecular mass of 700000 Da previously reported by Spencer et al [34] for 28-kDa cholecalcin mRNA. This different pattern as compared to the one of cDNA pC109-hybridised mRNA (Fig.…”

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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Production and Properties of Vitamin‐D‐Induced mRNA for Chick Calcium‐Binding Protein

Cited by 140 publications

References 28 publications

Purkinje Cell Survival and Axonal Regeneration Are Age Dependent: AnIn VitroStudy

Purkinje Cell Survival and Axonal Regeneration Are Age Dependent: AnIn VitroStudy

Stimulation of intestinal calcium-binding-protein mRNA synthesis in the nucleus of vitamin D-deficient chicks by 1,25-dihydroxycholecalciferol

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

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