Primary structure and expression of a sodium channel characteristic of denervated and immature rat skeletal muscle

The alpha subunit of a voltage-sensitive sodium channel characteristic of denervated rat skeletal muscle was cloned and characterized. The cDNA encodes a 2018 amino acid protein (SkM2) that is homologous to other recently cloned sodium channels, including a tetrodotoxin (TTX)-sensitive sodium channel from rat skeletal muscle (SkM1). The SkM2 protein is no more homologous to SkM1 than to the rat brain sodium channels and differs notably from SkM1 in having a longer cytoplasmic loop joining domains 1 and 2. Steady-state mRNA levels for SkM1 and SkM2 are regulated differently during development and following denervation: the SkM2 mRNA level is highest in early development, when TTX-insensitive channels predominate, but declines rapidly with age as SkM1 mRNA increases; SkM2 mRNA is not detectable in normally innervated adult skeletal muscle but increases greater than 100-fold after denervation; rat cardiac muscle has abundant SkM2 mRNA but no detectable SkM1 message. These findings suggest that SkM2 is a TTX-insensitive sodium channel expressed in both skeletal and cardiac muscle.     

甘肃东乡几种早中新世哺乳动物化石

本文记述了在甘肃东乡椒子沟发现的几种早中新世哺乳动物化石:Gomphotherium sp、Dzungariotheriura orgosense、Rhinocerotidae gen. indet.和Paraentelodon macrognathus sp. nov.,着重讨论了全北区猪齿兽类的系统关系。根据化石的性质判断,东乡椒子沟化石点的地质时代应与欧洲Burdigalian早期,亦即MN_3相当或稍早。笔者将原定为上新世临夏组的第一、二岩性段划出,命名为椒子沟组,其时代为早中新世。     

人体小卫星DNA探针的制备

根据人体小卫星DNA核心顺序,化学合成长23碱基寡核苷酸探针,筛选人体基因组文库,旨在获得能用作遗传分析探针的小卫星顺序。结果得到15个含小卫星的阳性重组子。随机取其一(C_(35.9))作探针,试做群体分析。所有个体均可检出多条杂交带。其中某些带具有多态性。在一定检测条件下,检出的DNA图谱在有限的个体内具有个体特异性。结果表明筛选文库得到的小卫星顺序可用于小卫星多态性的检测。其它小卫星探针的筛选和应用性研究正在进行。     

c-kit protein, a transmembrane kinase: identification in tissues and characterization.

The proto-oncogene c-kit encodes a transmembrane kinase which is related to the receptors for colony-stimulating factor type 1 and platelet-derived growth factor, as well as to the immunoglobulin superfamily. Antibodies specific for the kinase domain of the P80 gag-kit protein of the Hardy-Zuckerman 4 feline sarcoma virus were prepared. These kit-specific antibodies were used to identify and characterize the c-kit protein in cat brain tissue. The c-kit protein product displays an autophosphorylating activity in immune complex kinase assays, and, in turn, this activity was used to identify the c-kit protein in different tissues. In cat brain, a single 145-kilodalton (kDa) glycoprotein was detected. Its N-linked carbohydrates were found to be sensitive to digestion with the endoglycosidases (neuraminidase, endoglycosidase F, and endoglycosidase H), indicating hybrid and/or complex and high-mannose structures. A partial purification of the c-kit protein was achieved by wheat germ agglutinin affinity chromatography, and the autophosphorylating activity of the partially purified c-kit protein was characterized and found to be specific for tyrosine. The kit antibodies cross-react with the murine c-kit protein product, and variant c-kit proteins in different mouse tissues were identified, with sizes of about 145 kDa (brain), 160 kDa (spleen), and 150 kDa (testis).     

Oligo-riboprobes. Tools for in situ hybridisation

In situ hybridisation detection of mRNAs using riboprobes has become a widely used technique. However, the identification of cells producing closely-related yet distinct mRNAs is difficult with the usual size probes. Moreover, it is not always easy to obtain the required cDNA essential for cRNA probe synthesis. To avoid these problems, we have used synthetic oligodeoxynucleotides to generate short, single stranded RNA probes ("oligo-riboprobes"). These probes can be labelled to very high (10(9) cpm/micrograms) specific activity and can be prepared for any published nucleotide sequence. We have used these probes to localise beta (preprotachykinin) PPT mRNA producing neurons in rat hypothalamus and bowel. The results were compared to that obtained with cRNA probes generated from beta preprotachykinin cDNA.     

Expression of glutathione peroxidase I gene in selenium-deficient rats.

We have characterized a cDNA pGPX1211 encoding rat glutathione peroxidase I. The selenocysteine in the protein corresponded to a TGA codon in the coding region of the cDNA, similar to earlier findings in mouse and human genes, and a gene encoding the formate dehydrogenase from E. coli, another selenoenzyme. The rat GSH peroxidase I has a calculated subunit molecular weight of 22,155 daltons and shares 95% and 86% sequence homology with the mouse and human subunits, respectively. The 3'-noncoding sequence (greater than 930 bp) in pGPX1211 is much longer than that of the human sequences. We found that glutathione peroxidase I mRNA, but not the polypeptide, was expressed under nutritional stress of selenium deficiency where no glutathione peroxidase I activity can be detected. The failure of detecting any apoprotein for the glutathione peroxidase I under selenium deficiency and results published from other laboratories supports the proposal that selenium may be incorporated into the glutathione peroxidase I co-translationally.     

Prostaglandin I2 and the constricted maternal ovine placenta: effects of local infusion and placental age

We have tested the hypotheses that systemic responses to the infusion of prostaglandin I2 may have masked the ability of this substance to dilate the maternal placenta and that the inability of prostaglandin I2 to dilate the maternal near-term placenta may be a function of placental age. Regional blood flows were measured with radioactive microspheres. In 8 near-term sheep the control flows were measured and angiotensin II (AII) infusion was begun at 5 micrograms/min and continued for the duration of the experiments. At t = 15 min, regional blood flows were again measured. Prostaglandin I2 was then infused via a retrograde uterine arterial catheter at 10 micrograms/min. At t = 30 min, the flows were again measured. At this time the infusion of prostaglandin I2 was stopped and at t = 45 min the blood flows were measured for the last time. AII increased the resistance of all tissues examined. The blood pressure increased with AII and did not change thereafter. The non-placental uterine tissue served by the retrograde catheter dilated with prostaglandin I2. The placental tissue had an initial resistance of 59 +/- 6 mmHg.ml-1.min.g which increased to 98 +/- 22 mmHg.ml-1.min.g with the infusion of AII (P less than 0.05). This resistance remained constant at 82 +/- 19 mmHg.ml-1.min.g with the administration of prostaglandin I2 and did not change after prostaglandin I2 was removed. The local application of prostaglandin I2 in the presence of AII induced vasoconstriction caused vasodilatation in the nonplacental vessels but could not change the AII induced constriction in the placental vasculature.(ABSTRACT TRUNCATED AT 250 WORDS)