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1.
Journal of Korean Society of Endocrinology ; : 518-527, 1997.
Article in Korean | WPRIM | ID: wpr-55249

ABSTRACT

The estrogen receptor (ER) is present in a wide variety of mammalian tissues and is required for the physiological responses of estrogen, including estrogen-induced tissue-specific changes in gene expression. But most of our knowledge on the regulation of ER mRNA levels comes from in vivo steroid replacement experiments or cancer cell lines that express the ER. Thus the present study was attempted to determine 1) the anterior pituitary ER mRNA levels during rat estrous cycle 2) if estradiol itself directly modulates the ER mRNA levels in cultured rat anterior pituitary using RT-PCR method. In rats with 4 day estrous cycle, the ER mRNA levels in anterior pituitary gland reached to maximum at proestrus 11:00h just before serum estradiol concentration showed the highest. From then, the ER mRNA levels gradually declined during the rest of the proestrus. On the other hands, in cultured rat anterior pituitary cells, the ER mRNA levels were significantly decreased by the treatment of estradiol. These results indicate that the surge of estradiol was proceeded by the increase in pituitary ER mRNA levels during the proestrus and in cultured anterior pituitary cells, estrogen might be involved in the down-regulation of the ER mRNA levels.


Subject(s)
Animals , Rats , Cell Line , Down-Regulation , Estradiol , Estrogens , Estrous Cycle , Gene Expression , Hand , Pituitary Gland, Anterior , Proestrus , RNA, Messenger
2.
Korean Journal of Anatomy ; : 109-120, 1997.
Article in Korean | WPRIM | ID: wpr-651462

ABSTRACT

Recently, surmountable amounts of genes are being cloned without information about them and it has become neccessary to develop new techniques for discovering genes with more informaiion like as chromosomal location and possible functions. We have developed one such a method and applied it to search for genes that may be related with the neural development. The mRNAs were extracted from cerebral cortex of 18 week old human fetus, cDNAs were made by reverse transcription from these mRNAs and Uni-amp cDNAs having Uni-amp adapters at both ends were made for subsequent PCR. To observe the distribution of the Uni-amp cDNAs on the chromosome, fluorescent in situ hybridization was performed with biotin labeled Uni-amp cDNAs. Among the chromosome bands showing strong hybridization with the cDNAs, 7q22 was microdissected from the chromosome hybridized with unlabeled Uni-amp cDNAs and amplified by PCR with Uni-amp primers. These amplified cDNA fragment were subcloned to vectors and the nucleic acid sequences were analysed. As a result, 46 different clones were obtained. They were categorized as 12 clones of well characterized genes, 14 clones showing low homology with known genes, 13 clones of simply registered uncharacterized human cDNAs, 7 clones of unknown genes. In situ hybridization histochemistry of 34 novel genes, except 12 known genes, were performed on developing and adult rat tissue sections to see the tissue specificity and developmental expression of these genes. The expression of several novel genes were restricted to the nervous system. From these results, it may be suggested that our technique is very useful to clone the genes expressed in the developing human braine with confirmed chromosomal location. In addition, this cloning technique can be used to discover the new genes related with neural development in combination with functional screening methods.


Subject(s)
Adult , Animals , Humans , Rats , Biotin , Brain , Cerebral Cortex , Clone Cells , Cloning, Organism , DNA, Complementary , Fetus , In Situ Hybridization , In Situ Hybridization, Fluorescence , Mass Screening , Microdissection , Nervous System , Organ Specificity , Polymerase Chain Reaction , Reverse Transcription , RNA, Messenger
3.
Korean Journal of Anatomy ; : 397-410, 1997.
Article in Korean | WPRIM | ID: wpr-654905

ABSTRACT

Kainic acid[KA], a glutamic acid analogue, has been widely used as a excitotoxin in the study of neurotoxicity due to its ability to induce strong excitation and to increase intracellular calcium concentration of the mature central nervous system[CNS] neurons. However, it is not well known whether KA is also strongly cytotoxic to the neurons of the postnatal brain. We have injected KA into rats at different developmental stages and have investigated the changes in the expression of c-fos[transcriptional factor and a marker of neuronal activity], heat shock proetin 72[hsp 72, a neuronal injury marker], and glial fibrillary acidic protein[GFAP, a neuronal injury marker] mRNAs, which are known to be increased in KA-induced neurotoxicity, and glyceraldehyde 3-phosphate dehydrogenase [GAPDH, a house keeping gene] mRNAs with in situ hybridization histochemistry using specific riboprobes. The expression of c-fos mRNA was first identified in the CA3 area of hippocampus from 6hr after KA treatment in P7 rats. The c-fos mRNA-expressing area and the level of expression was gradually increased from P7 to adult. Hsp 72 mRNA was first expressed in the dentate gyrus and hippocampus from 6hr after KA treatment in P2l rats. In the adult rats, hsp 72 mRNA was broadly expressed in the brain at 2hr after KA treatment. The increase of GFAP mRNA expression was first identified in Pl4 rat brain from 6hrs after KA treatment, and by the development of brain it tends to appear earlier. The expression of GAPDH mRNA, however, did not show changes after KA treatment except for the adult rats showing a slight decrease at 12hr after KA treatment. These results suggest that KA may offer different level of cytotoxicity to the developing neurons by their developmental status and the difference may be correlated with the completion of synaptogenesis and increase of KA receptor.


Subject(s)
Adult , Animals , Humans , Rats , Brain , Calcium , Dentate Gyrus , Glial Fibrillary Acidic Protein , Glutamic Acid , Glyceraldehyde 3-Phosphate , Hippocampus , Hot Temperature , HSP70 Heat-Shock Proteins , In Situ Hybridization , Kainic Acid , Neurons , Neurotoxins , Oxidoreductases , RNA, Messenger , Shock
4.
Korean Journal of Obstetrics and Gynecology ; : 1567-1575, 1997.
Article in Korean | WPRIM | ID: wpr-208199

ABSTRACT

Follicle stmulating hormone ( FSH ) consist of alpha and beta subunits, which are encoded by se-parate genes. Pituitary release of FSH appears to be regulated by the hypothalamic GnRH and the gonadal steroid hormones. In addition, inhibin and follistatin produced by the gonad have been known to inhibit FSH secretion selectively. However, little is known about their regulation of the biosynthesis of FSH subunits at transcriptional and posttranscriptional levels. In the pre-sent study, we studied the time course of changes in alpha and FSH beta subunit mRNA concentrati-ons after castration and the effects of ovarian steroids of changes in alpha and FSH beta subunit mRNA concentrations after castration and the effects of ovarian steroids on alpha and FSH beta subunit mRNA in ovariectomized rats in order to determine Whether FSH subunit synthesis is modulated at the pretranslational levels, and whether synthesis and secretion are differently regulated. Results are as follows : 1. The time course of the rise in the steady state alpha subunit and FSH beta subunit mRNA levels were observed after ovariectomy, which paralleled the increases in serum and pituitary FSH concentrations. The time course experiments revealed differences in the patterns of alpha and FSH beta subunit mRNA responses, the rise in FSH beta subunit mRNA levels being more pro- minent than the rise in alpha subunit mRNA. 2. FSH beta mRNA levels were negatively regulated by the single injection of progesterone but not by estradiol, suggesting that FSH beta subunit mRNA seemed to be more sensitive to ne-gative feedback by progesterone than estradiol. Similar results were obtained by the continuous treatment of ovarian steroids for 1~4 days, but inhibition was more prominent with continuous treatment. It is, therefore, concluded that estradiol and progesterone inhibit the synthesis of FSH at the pretranslational level by modulating the steady state levels of alpha and FSH beta subunit mRNA, progesterone effect being more promiment than that of estradiol and alpha and FSH beta subunit are regulated in a different manner.


Subject(s)
Animals , Female , Rats , Castration , Estradiol , Follicle Stimulating Hormone , Follicle Stimulating Hormone, beta Subunit , Follistatin , Gonadal Steroid Hormones , Gonadotropin-Releasing Hormone , Gonads , Inhibins , Ovariectomy , Progesterone , RNA, Messenger , Steroids
5.
Journal of the Korean Pediatric Society ; : 595-601, 1992.
Article in Korean | WPRIM | ID: wpr-93820

ABSTRACT

No abstract available.


Subject(s)
Humans , Diagnosis , Growth Hormone
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