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Cloning and comparison of factor X from rhesus monkey (Macaca mulatta).

ABSTRACT: The reliability of the rhesus monkey as an important experimental animal depends on its genetic concordance with human. During our assessment of the rhesus monkey as a preclinical model for coagulation-related research, we cloned the full-length cDNA of rhesus monkey factor X (FX) and compared its genetic characteristics and coagulation activity with those of human FX. The full-length cDNA of rhesus monkey FX was 1683 bp in length, corresponding to 487 coding amino acids and sharing 94.71% nucleotide identity and 93.65% amino acid identity with human FX. When FX sequences from different animals were compared with that of human FX, rhesus monkey and baboon FX showed similar degrees of homology to human FX, which were less than that between human and chimpanzee FX sequences but remarkably higher than those of another 2 monkey species, bovine, pig, and rodents. Comparison of functional sites between human and rhesus monkey FX revealed high similarities between their amino acids sequences and 3-dimensional structures. The average coagulation activity of FX from 24 rhesus monkeys was in the normal range of that of healthy humans. The rhesus monkey therefore may be a suitable animal model for research addressing coagulation factor X.

SUBMITTER: Chen Y

PROVIDER: S-EPMC2771601 | biostudies-literature | 2009 Oct

REPOSITORIES: biostudies-literature

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Cloning and comparison of factor X from rhesus monkey (Macaca mulatta).

Chen Younan Y Qin Shengfang S Tan Weidong W Lu Yanrong Y Zhang Jie J Li Hongxia H Bu Hong H Cheng Jingqiu J

Comparative medicine 20091001 5

The reliability of the rhesus monkey as an important experimental animal depends on its genetic concordance with human. During our assessment of the rhesus monkey as a preclinical model for coagulation-related research, we cloned the full-length cDNA of rhesus monkey factor X (FX) and compared its genetic characteristics and coagulation activity with those of human FX. The full-length cDNA of rhesus monkey FX was 1683 bp in length, corresponding to 487 coding amino acids and sharing 94.71% nucle ...[more]

PMID: 19887032

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Project description:We demonstrated recently that frontal cortical expression of the Neuregulin (NRG) receptor ErbB4 is restricted to interneurons in rodents, macaques, and humans. However, little is known about protein expression patterns in other areas of the brain. In situ hybridization studies have shown high ErbB4 mRNA levels in various subcortical areas, suggesting that ErbB4 is also expressed in cell types other than cortical interneurons. Here, using highly-specific monoclonal antibodies, we provide the first extensive report of ErbB4 protein expression throughout the cerebrum of primates. We show that ErbB4 immunoreactivity is high in association cortices, intermediate in sensory cortices, and relatively low in motor cortices. The overall immunoreactivity in the hippocampal formation is intermediate, but is high in a subset of interneurons. We detected the highest overall immunoreactivity in distinct locations of the ventral hypothalamus, medial habenula, intercalated nuclei of the amygdala and structures of the ventral forebrain, such as the islands of Calleja, olfactory tubercle and ventral pallidum, and medium expression in the reticular thalamic nucleus. While this pattern is generally consistent with ErbB4 mRNA expression data, further investigations are needed to identify the exact cellular and subcellular sources of mRNA and protein expression in these areas. In contrast to in situ hybridization in rodents, we detected only low levels of ErbB4-immunoreactivity in mesencephalic dopaminergic nuclei but a diffuse pattern of immunofluorescence that was medium in the dorsal striatum and high in the ventral forebrain, suggesting that most ErbB4 protein in dopaminergic neurons could be transported to axons. We conclude that the NRG-ErbB4 signaling pathway can potentially influence many functional systems throughout the brain of primates, and suggest that major sites of action are areas of the "corticolimbic" network. This interpretation is functionally consistent with the genetic association of NRG1 and ERBB4 with schizophrenia.

Project description:GABAA receptors are composed of five subunits arranged around a central chloride channel. Their subunits originate from different genes or gene families. The majority of GABAA receptors in the mammalian brain consist of two ?-, two ?- and one ?- or ?-subunit. This subunit organization crucially determines the physiological and pharmacological properties of the GABAA receptors. Using immunohistochemistry, we investigated the distribution of 10 GABAA receptor subunits (?1, ?2, ?3, ?4, ?5, ?1, ?2, ?3, ?2, and ?) in the fore brain of three female rhesus monkeys (Macaca mulatta). Within the cerebral cortex, subunits ?1, ?5, ?2, ?3, and ?2 were found in all layers, ?2, ?3, and ?1 were more concentrated in the inner and outer layers. The caudate/putamen was rich in ?1, ?2, ?5, all three ?-subunits, ?2, and ?. Subunits ?3 and ?5 were more concentrated in the caudate than in the putamen. In contrast, ?1, ?2, ?1, ?2, ?2, and ? were highest in the pallidum. Most dorsal thalamic nuclei contained subunits ?1, ?2, ?4, ?2, ?3, and ?2, whereas ?1, ?3, ?1, and ?2 were most abundant in the reticular nucleus. Within the amygdala, subunits ?1, ?2, ?5, ?1, ?3, ?2, and ? were concentrated in the cortical nucleus, whereas in the lateral and basolateral amygdala ?1, ?2, ?5, ?1, ?3, and ?, and in the central amygdala ?1, ?2, ?3, and ?2 were most abundant. Interestingly, subunit ?3-IR outlined the intercalated nuclei of the amygdala. In the hippocampus, subunits ?1, ?2, ?5, ?2, ?3, ?2, and ? were highly expressed in the dentate molecular layer, whereas ?1, ?2, ?3, ?5, ?1, ?2, ?3, and ?2 were concentrated in sector CA1 and the subiculum. The distribution of GABAA receptor subunits in the rhesus monkey was highly heterogeneous indicating a high number of differently assembled receptors. In most areas investigated, notably in the striatum/pallidum, amygdaloid nuclei and in the hippocampus it was more diverse than in the rat and mouse indicating a more heterogeneous and less defined receptor assembly in the monkey than in rodent brain.

Dataset Information

Cloning and comparison of factor X from rhesus monkey (Macaca mulatta).

Publications

Cloning and comparison of factor X from rhesus monkey (Macaca mulatta).

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