Biomedical subjects

J L Cao

Publications and source records attributed to J L Cao.

5 recordsLinked to original sources

PubMed2001

[Intrathecal injection of NOS antisense oligonucleotides inhibits the increase of NMDA1AR mRNA expression in the spinal cord and brainstem of morphine-withdrawal rats].

Methods of reverse transcription polymerase chain reaction (RT-PCR), intrathecal injection and antisense drugs were used to study the effects of nitric oxide (NO) on the scores of morphine-withdrawal syndrome and the expression of NMDA1AR mRNA in rat spinal cord and brainstem. Intrathecal injection of NOS antisense oligonucleotides (AS-ONs) significantly decreased the scores of morphine-withdrawal symptoms. The effect of nNOS AS-ONs was greater than that of eNOS AS-ONs. The expression of NMDA1AR mRNA in the spinal cord and brainstem increased in morphine-dependent rats and increased to a greater extent in morphine-withdrawal rats. Intrathecal injection of nNOS AS-ONs significantly inhibited the increased expression of NMDA1AR mRNA in the spinal cord and brainstem of morphine-withdrawal rats. Intrathecal injection of eNOS antisense oligonucleotides inhibited the expression of NMDA1AR mRNA in the spinal cord of morphine-withdrawal rats, but did not in the brainstem. It is suggested that NO mediates morphine-withdrawal reaction and participates in modulating the expression of NMDA1AR mRNA in morphine-withdrawal rats.

Animals↗

PubMed2001

[NO involvement in mediation of spinal neuron sensitization in morphine-withdrawal rats].

The effects of nitric oxide (NO) on the activation of spinal cord neurons were studied using immunocytochemistry, intrathecal injection and antisense oligonucleotides (AS-ONs) techniques in morphine-withdrawal rats. Acute administration of naloxone and chronic administration of morphine changed neither the expression of Fos-LI and NADPH-d positive neurons nor the expression of Fos/NADPH-d double-labeled neurons in the spinal cord of rats. Fos-LI, NADPH-d positive and Fos/NADPH-d double-labeled neurons were increased significantly in number in morphine-withdrawal rats and they were observed in all the laminae of the spinal cord. Intrathecal injection of L-NA, nNOS antisense oligonucleotides significantly inhibited the expression of Fos-LI in the spinal cord and decreased the scores for morphine-withdrawal symptoms in morphine-withdrawal rats, but not in nNOS-S group. The results suggest that NO mediates the spinal neurons sensitization in morphine-withdrawal rats.

Animals↗

PubMed2000

[Increased expression of formalin-induced Fos and NADPH-d positive neurons in the spinal cord of morphine-tolerant rats].

Fos immunocytochemistry, NADPH-d histochemistry and Fos/NADPH-d double-labeling method were used to study the changes in formalin-induced Fos, NADPH-d positive and Fos/NADPH-d double-labeled neurons in the spinal cord of morphine-tolerant rats. Formalin-induced Fos-like immunoreactivity (Fos-LI) was located in the superficial laminae and neck of ipsilateral spinal cord. Acute administration of morphine decreased the expression of Fos-LI in non-tolerant rats, while the expression of Fos-LI was significantly increased in morphine-tolerant rats. Fos-LI was distributed not only in the whole laminae of the ipsilateral spinal cord but also in the contralateral spinal cord. Acute administration of morphine was ineffective in decreasing the expression of Fos-LI in morphine-tolerant rats. Morphine tolerance increased the expression of formalin-induced NADPH-d positive neurons in the superficial laminae of spinal dorsal horn. A few formalin-induced Fos/NADPH-d double-labeled neurons were detected in the superficial laminae of spinal dorsal horn of non-tolerant rats. In morphine-tolerant rats, on the other hand, formalin-induced Fos/NADPH-d double-labeled neurons were increased and distributed in the whole laminae of the ipsilateral spinal cord and the contralateral superficial spinal cord. It is suggested that NO is involved in the increase of formalin-induced Fos-LI in the spinal cord of morphine-tolerant rats and may play an important role in the development of morphine tolerance.

Animals↗

PubMed1991

Conversion of a mitochondrial gene for mammalian cytochrome c oxidase subunit II into its universal codon equivalent and expression in vivo and in vitro.

To begin to assess the independent structural and functional characteristics of the mitochondrially encoded subunits of mammalian cytochrome c oxidase, we have converted the cloned mitochondrial gene for rat subunit II (coxII) into its universal codon equivalent (ucoxII) by oligonucleotide-directed, site-specific mutagenesis. This involved synthesizing 12 oligodeoxynucleotides to achieve the 13 ATA to ATG and the 5 TGA to TGG changes needed. To express ucoxII in Escherichia coli, we used a number of different expression vectors in which the promoters and ribosome-binding sequences of the messenger RNA were varied. While ucoxII alone was expressed at a low level, a striking increase in the level of expression resulted when the ucoxII gene was fused to other E. coli genes. The COXII peptide was identified by proteolytic digestion, partial sequencing, and reaction with specific antisera. A cro-beta-galactosidase-COXII fusion protein has been purified, characterized, and used to produce polyclonal antibodies to the COXII peptide. The ucoxII gene was also expressed in a cell-free translation system and in Xenopus oocytes, yielding a nondenatured, membrane-associated peptide with the same apparent molecular weight as authentic subunit II. In oocytes and in a reticulocyte lysate in vitro system supplemented with microsomal membranes, the protein is glycosylated and coisolates with the washed membrane fraction. In both cases, the COXII peptide is soluble under mild conditions in a nonionic detergent and is precipitable by antibodies to subunit II. The production of subunit II in the in vitro translation system is stimulated as strongly by addition of soybean phospholipid vesicles as by microsomal membranes, providing further evidence of membrane insertion and stabilization.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

PubMed1990

Specific sequences downstream from -6 are not essential for proper and efficient in vitro utilization of the Escherichia coli lactose promoter.

A series of deletion mutants of the wild-type Escherichia coli lactose promoter, with endpoints at +25, +19, +14, +1 and -6 (relative to the start of transcription at +1), was constructed and the deleted DNA replaced with non-lac DNA. These mutants were used to show that no specific DNA sequences downstream from -6 are required for efficient promoter utilization in vitro. In all cases transcription is dependent on the presence of the catabolite activator protein (CAP) and cAMP, and begins at +1 at a level indistinguishable from that at the wild-type promoter. A set of lac DNA fragments deleted to -6 was constructed, having an A, C, G or T residue at +1 and heterologous DNA downstream. These synthetic promoters allow systematic testing of the effect of the initiating nucleotide on the transcription process. Again, transcription occurs mainly from +1, at a level similar to the normal wild-type level. No substantial differences between these promoters are observed in the rates of formation of stable complexes, in the degree of complex formation, in the rate at which polymerase "escapes" from the complex or in abortive transcription products. Equivalent results are seen with a related set of constructs based on the CAP-insensitive lac UV5 promoter. Thus, lac promoter sequences including consensus hexamers at -10 and -35, plus the spacer region between them, provide specificity and efficiency both in initiation of transcription by RNA polymerase and in CAP-polymerase interactions. A question as to whether there is a third RNA polymerase binding site at lac, in addition to the known overlapping P1 and P2 regions, was not unambiguously answered. However, if a "P3" site does exist, it must lie between P1 and P2. Alternatively, the variety of polymerase interactions at wild-type lac may reflect different structural states of the enzyme. The results presented here indicate that DNA downstream from -6 plays little part in determining the conformation of the enzyme at the lactose promoter.

Base Sequence↗