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      • 2D-LC Characterization of Comb-Shaped Polymers Using Isotope Effect

        Ahn, Seonyoung,Im, Kyuhyun,Chang, Taihyun,Chambon, Pierre,Fernyhough, Christine M. American Chemical Society 2011 ANALYTICAL CHEMISTRY - Vol.83 No.11

        <P>A rigorous molecular characterization of comb-shaped polystyrene (PS) was carried out taking advantage of its molecular structure, a normal hydrogenous backbone, and deuterated side chains. Normal phase LC (NPLC) can separate the comb PS species well according to their molecular weight. Nonetheless, it cannot distinguish the backbone from the side chains and the differently structured polymers having a similar molecular weight, e.g, a single backbone comb and a coupled backbone comb with fewer side chains. In contrast to NPLC, the hydrogenous polymer is retained longer than the deuterated counterpart in reversed phase LC (RPLC). When the isotope sensitivity of RPLC is taken advantage of, the comb PS is cross fractionated by NPLC and RPLC, and a two-dimensional mapping with respect to the backbone chain length and the number of branches is fully established.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/ancham/2011/ancham.2011.83.issue-11/ac2005907/production/images/medium/ac-2011-005907_0004.gif'></P>

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        Estrogen receptor alpha-induced cholecystokinin type A receptor expression in the female mouse pituitary.

        Kim, Hyun Joon,Gieske, Mary C,Hudgins, Susan,Kim, Beob Gyun,Krust, Andree,Chambon, Pierre,Ko, Chemyong Journal of Endocrinology, Ltd. [etc.] 2007 The Journal of endocrinology Vol.195 No.3

        <P>Estrogen plays a critical role in inducing LH surge. In the pituitary, estrogen receptor alpha (ERalpha) mediates the action of estrogen, while the downstream pathway of ERalpha activation is yet to be elucidated. Here, we report the finding that cholecystokinin type A receptor (CCK-AR) is an ERalpha downstream gene in the mouse anterior pituitary. In the cycling mouse pituitary, the expression of CCK-AR mRNA is markedly higher in the afternoon of proestrus compared with metestrus. Both ovariectomy (OVX) and null mutation of the ERalpha gene completely abolish CCK-AR mRNA expression. Injection of 17beta-estradiol to OVX wild-type mice induces recovery of CCK-AR mRNA expression to levels observed at proestrus, but no such recovery is induced in OVX ERalpha knockout mice. The same pattern of estrogen dependency in inducing CCK-AR mRNA expression was seen in cultured primary anterior pituitary cells, indicating that estrogen directly acts on pituitary cells to induce CCK-AR expression. Immunohistological analysis revealed that more than 80% of gonadotrophs express CCK-AR in the afternoon of proestrus. To test whether CCK-AR mediated the sensitizing effect of estrogen in GnRH-induced LH secretion, primary pituitary cells were primed with estrogen followed by treatment with GnRH in the presence or absence of lorglumide, a CCK-AR antagonist. While both groups secreted LH upon GnRH treatment, lorglumide treatment significantly decreased LH secretion. Taken together, this study finds CCK-AR to be an ERalpha downstream gene in the pituitary and suggests that CCK-AR may play a role in the estrogen sensitization of the pituitary response to GnRH.</P>

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        Theca-specific estrogen receptor-alpha knockout mice lose fertility prematurely.

        Lee, Sungeun,Kang, Dong-Wook,Hudgins-Spivey, Susan,Krust, Andree,Lee, Eun-Young,Koo, Youngbum,Cheon, Yongpil,Gye, Myung Chan,Chambon, Pierre,Ko, ChemYong Association for the Study of Internal Secretions 2009 Endocrinology Vol.150 No.8

        <P>Estrogen receptor-alpha (Esr1) mediates estrogen action in regulating at all levels of the hypothalamic-pituitary-ovarian axis. Whereas the importance of Esr1 in hypothalamus and pituitary has been demonstrated by loss of fertility in the neuron- and pituitary-specific Esr1 knockout mice, whether Esr1 plays a critical role in the ovary remains to be determined. In the ovary, Esr1 is mainly expressed in the theca/interstitial cells and germinal epithelium and thus is believed to mediate estrogen action in these cells. In this study, we assessed the importance of Esr1 in the ovarian theca cells in regulating female reproduction. The Cre-LoxP approach was used to selectively delete the Esr1 gene in the theca cells, and the reproductive consequence of the deletion was measured. Adolescent theca-specific Esr1 knockout (thEsr1KO) mice (<4 months of age) are fertile and cycling. However, they begin to display an erratic pattern of estrous cycles and become infertile before they reach the age of 6 months. The ovaries of thEsr1KOmice (>or=4 months) have fewer corpora lutea but more antral follicles than the age-matching wild-type mice. The numbers of 17-hydroxylase-expressing cells are largely increased in the interstitium of the thEsr1KO mouse ovary. Interestingly, whereas basal levels of serum testosterone and FSH were mildly elevated, LH level was either markedly lower or undetectable in the thEsr1KO mice. When superstimulated by exogenous gonadotropins, thEsr1KO mice released significantly fewer oocytes that wild-type littermates and developed multiple hemorrhagic cysts. Taken together, this study demonstrates that theca Esr1 plays a critical role in regulating female reproduction.</P>

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