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Query: EC:3.1.26.9 (
ribonuclease
)
6,589
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Double-labeling immunofluorescent histochemistry demonstrates that
calretinin
, a calcium-binding protein, coexists with calcitonin gene-related peptide, vasoactive intestinal peptide, and substance P in the fibers innervating the lamina propria of the rat intestinal villi. An acetylcholinesterase histochemical stain revealed that the majority of
calretinin
-containing cells in the myenteric ganglia were cholinergic and that about one half of the submucosal
calretinin
-containing cells colocalized with acetylcholinesterase. In situ hybridization studies confirmed the presence of
calretinin
mRNA in the dorsal root ganglia, and a
ribonuclease
protection assay verified the presence of
calretinin
message in the intestine. The coexistence of
calretinin
in calcitonin-gene-related-peptide-containing cells that also contained substance P and vasoactive intestinal polypeptide in the dorsal root ganglia suggest that these ganglia are the source of the quadruple colocalization within the sensory fibers of the villi. Although the function of
calretinin
in these nerves is unknown, it is hypothesized that the coexistence of three potent vasodilatory peptides influences the uptake of metabolized food products within the vasculature of the villi.
...
PMID:Quadruple colocalization of calretinin, calcitonin gene-related peptide, vasoactive intestinal peptide, and substance P in fibers within the villi of the rat intestine. 754 20
cDNA clones for
calretinin
, a member of the troponin-C family of calcium-binding proteins, were isolated from a cDNA library of the human colon carcinoma cell line WiDr. Sequence analysis revealed two forms of alternatively spliced
calretinin
mRNAs encoding C-terminally truncated proteins. Exon 7 was either spliced to exon 9 (delta 8) or to exon 10 (delta 8,9); both resulted in a frame shift and a translational stop at the second codon of exon 9 (delta 8), or at codon 15 of exon 10 (delta 8,9), respectively. The presence of delta 8 and delta 8,9
calretinin
mRNA in WiDr cells was confirmed using reverse-transcriptase PCR and sequence analysis of the amplicon, as well as by a
ribonuclease
protection assay. Co115/3 and three other human colon carcinoma cell lines were found, by reverse-transcriptase PCR to also contain delta 8,9
calretinin
mRNA. The truncated proteins were able to bind calcium, as evidenced by a calcium blot of the delta 8 form (
calretinin
-20k) and delta 8,9 form (
calretinin
-22k) expressed in Escherichia coli. Immunohistochemical staining using an antiserum specific for the novel C-terminus of
calretinin
-22k confirmed its presence in WiDr, Co115/3 and three additional colon carcinoma cell lines. The fact that alternative splicing of
calretinin
was found in five different cell lines suggests that alternatively spliced calretinins fulfill a physiological function.
...
PMID:Alternative splicing of calretinin mRNA leads to different forms of calretinin. 760 11
Calretinin
is an EF-hand calcium binding protein found predominantly in discrete sets of neurons in the central system, and in the sex hormone producing cells of the gonads.
Calretinin
mRNA levels were measured in discrete brain areas from vehicle and corticosterone treated rats (subcutaneous injections of 0, 0.1, 1, or 10 mg, 7 days) using a micropunch
ribonuclease
protection assay. Treatment with high dose corticosterone (10 mg) caused a 93% decrease in
calretinin
mRNA levels in the hypothalamic paraventricular nucleus compared to controls. Two other brain regions, the medial amygdaloid nucleus and the nucleus reuniens, demonstrated an approximately 40% decrease in
calretinin
mRNA following high dose corticosterone. In separate experiments, adrenalectomy and diurnal corticosterone variations had no effect on
calretinin
mRNA in the brain areas examined. In the testes, corticosterone treatment decreased
calretinin
protein in a dose dependent fashion (to 81%, 68%, and 39% of controls at doses of 10, 1, and 0.1 mg/day, respectively). Low dose corticosterone treatments decreased testicular but not neuronal
calretinin
mRNA, whereas high dose corticosterone reduced
calretinin
mRNA in testes and several discrete brain areas. This suggests that corticosterone's effects on brain
calretinin
may be due to its pathological effects, e.g. energy depletion of brain cells or interference with the normal support functions of glia.
...
PMID:Corticosterone effects on rat calretinin mRNA in discrete brain nuclei and the testes. 770 81
Calretinin
, a highly evolutionarily conserved E-F hand calcium binding protein, is expressed predominantly in neurons, with a few exceptions. The function of
calretinin
is not known. We demonstrate the expression of
calretinin
mRNA and protein in rat testes. Immunocytochemistry and in situ hybridization reveal that
calretinin
expression in testis is localized to the interstitial Leydig cells. Western blot and
ribonuclease
protection analyses show that
calretinin
protein and mRNA in testis is the same as that expressed in brain. It is suggested that
calretinin
may play a role in the production of testosterone.
...
PMID:Calretinin is expressed in the Leydig cells of rat testis. 791 40
A microdissection technique for quantitation of neurochemicals in discrete brain nuclei has been applied to quantitative measurement of mRNA. The method permits quantitation of low abundance mRNA from submilligram amounts of tissue (10-500 micrograms protein). Discrete nuclei and other regions of the brain are solubilized in concentrated guanidine thiocyanate solution, mRNA is directly hybridized with riboprobes, and detected with a
ribonuclease
protection assay. This method eliminates the necessity for RNA isolation from solid tissue. No assumptions regarding RNA recovery are necessary since tissue specimens are solubilized, hybridized and treated with
ribonuclease
in a single tube. We have determined the mRNA levels of
calretinin
, a predominantly neuron-specific calcium binding protein in microdissected nuclei and other regions of rat brain. For interassay comparison, measurement of sample protein and beta-actin mRNA permits normalization and quantitation in terms of these internal controls. The quantity of
calretinin
mRNA ranged from 281 +/- 35 fg/micrograms protein in the thalamic paraventricular nucleus to 2.3 +/- 0.5 fg/micrograms protein for the cerebral cortex. The
calretinin
/beta-actin ratios ranged from 79.9 +/- 9.3% to 1.3 +/- 0.1%, respectively. The combination of microdissection techniques with a lysate RNase protection assay: (1) establishes this technique as quantitative for detection of high and low abundance mRNAs from microdissected brain specimens; (2) bypasses the inefficiencies and uncertainties associated with isolating RNA; and (3) enables large numbers of determinations from discrete brain nuclei to be analyzed in 2 to 3 days.
...
PMID:Quantitative measurement of calretinin and beta-actin mRNA [correction of mRNAIN] in rat brain micropunches without prior isolation of RNA. 830 61