+publisher:"Colorado State University" +contributor:("Clay, Colin")

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Colorado State University

1. De Lille, Alexandra. Physical and molecular characteristics of day 75 nuclear transfer cloned bovine conceptuses.

Degree: PhD, Biomedical Sciences, 2012, Colorado State University

URL: http://hdl.handle.net/10217/67429

► This study was designed to measure fetal and placental characteristics in bovine day 75 nuclear transfer and control pregnancies. Responses included mRNA concentration of the… (more)

▼ This study was designed to measure fetal and placental characteristics in bovine day 75 nuclear transfer and control pregnancies. Responses included mRNA concentration of the insulin-like growth factor (IGF) system [IGF-1, IGF-2, IGF1R, IGF2R, IGFBBP-1, -2, -3] and the vascular endothelial growth factor (VEGF) system [VEGF, PlGF, VEGF1R, and VEGF2R]. Fetal attrition of the cloned pregnancies up to day 75 was high (89%, 63 out of 71 frozen embryos transferred; 8 of 16 cloned conceptuses present on day 30 survived to day 75, as did 5 of 5 controls). No significant differences in mean weights of large and medium placentomes were observed between 8 clones and 5 controls. However, the variance of mean weight of large placentomes was greater in clones than in controls; one gestation had placentomes six standard deviations larger than controls. Interestingly, the mean umbilical cord weight/length ratio was significantly greater for clones (P < 0.05). Mean fetal length, fetal weight, fetal weight/length index and mean weights for heart, brain, liver, kidneys and the mean brain/liver index did not differ between cloned and control day 75 conceptuses, but numbers per group were limited. Northern blot analysis, revealed the presence of three transcripts of 3.7kb, 2.2kb and 1.7kb for VEGF and one 1.7 kb transcript for PlGF mRNA in the cotyledons and allantochorion of day 45 cloned and control gestations. All three VEGF bands were present in both cloned and control day 75 cotyledons and caruncles, but the PlGF transcript was barely detectable, except for the cotyledons of one clone. mRNA for all of genes studied could be detected with real time PCR in day 75 cotyledons and caruncles, and fetal livers contained mRNA for all IGF's and IGFBP's evaluated. In all placentomal tissues, PlGF mRNA concentration was 100-fold less than VEGF mRNA, which seems to be the driving force for placentomal vascularization at day 75. There was a trend for a reduction by half of the PlGF mRNA concentration in caruncle of clones vs. controls (P = 0.06). VEGF2R (KDR) mRNA was abundant, but VEGF1R (Flt-1), was only present in very low concentrations; our primer set did not distinguish between soluble versus membrane bound receptor mRNA for VEGF1R. Four of the cloned conceptuses contained substantially less cotyledonary IGF1R mRNA than the other clones and controls. IGFBP-3 mRNA concentrations were very high in placentomes; IGFBP-1 and -2 mRNA concentration on the other hand was very low for clones and controls. mRNA for IGFBP-1, -2, -3, however, was abundant in day 75 fetal livers, while IGF-1 mRNA was scarce in this tissue. Fetal livers from cloned pregnancies contained 4-fold more IGF-2 mRNA than controls (P<0.01). We observed that liver IGF-2 mRNA concentration and liver weight increased with weight of the largest placentome; in clones these increases were associated with a decrease in cotyledonary IGF-2 mRNA, while the opposite occurred with controls. Interestingly, there was a trend to lower IGF2R mRNA concentrations (P = 0.09), and… Advisors/Committee Members: Seidel, George (advisor), Anthony, Russell (committee member), Clay, Colin (committee member), Garry, Franklyn (committee member).

Subjects/Keywords: nuclear transfer cloning; insulin like growth factor; placenta; angiogenesis; bovine

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APA (6^th Edition):

De Lille, A. (2012). Physical and molecular characteristics of day 75 nuclear transfer cloned bovine conceptuses. (Doctoral Dissertation). Colorado State University. Retrieved from http://hdl.handle.net/10217/67429

Chicago Manual of Style (16^th Edition):

De Lille, Alexandra. “Physical and molecular characteristics of day 75 nuclear transfer cloned bovine conceptuses.” 2012. Doctoral Dissertation, Colorado State University. Accessed April 15, 2021. http://hdl.handle.net/10217/67429.

MLA Handbook (7^th Edition):

De Lille, Alexandra. “Physical and molecular characteristics of day 75 nuclear transfer cloned bovine conceptuses.” 2012. Web. 15 Apr 2021.

Vancouver:

De Lille A. Physical and molecular characteristics of day 75 nuclear transfer cloned bovine conceptuses. [Internet] [Doctoral dissertation]. Colorado State University; 2012. [cited 2021 Apr 15]. Available from: http://hdl.handle.net/10217/67429.

Council of Science Editors:

De Lille A. Physical and molecular characteristics of day 75 nuclear transfer cloned bovine conceptuses. [Doctoral Dissertation]. Colorado State University; 2012. Available from: http://hdl.handle.net/10217/67429

Colorado State University

2. Cleys, Ellane Rachael. Androgen signaling in the placenta.

Degree: PhD, Biomedical Sciences, 2014, Colorado State University

URL: http://hdl.handle.net/10217/83721

► Placental estrogen signaling is known to regulate placental trophoblast function and differentiation. However, the role of placental androgen signaling has never been investigated, despite the… (more)

▼ Placental estrogen signaling is known to regulate placental trophoblast function and differentiation. However, the role of placental androgen signaling has never been investigated, despite the rise of maternal serum androgens throughout gestation. Recent findings have shown increased maternal serum androgen in patients with the placental induced disorder preeclampsia. Preeclampsia, a maternal hypertension and proteinuria condition instigated by insufficient trophoblast differentiation and invasion into maternal spiral arteries, is also associated with increased placental expression of androgen receptor and an increased risk of incidence in patients with polymorphisms in androgen receptor that decrease androgen signaling. These findings suggest a crucial role for placental androgen signaling. Moreover, research investigating androgen's role in cancer progression has shown that many androgen responsive genes regulate cell proliferation, differentiation to invasive phenotypes, and tissue vascularization, all processes necessary for normal placental development. Androgen signaling in tumor tissues is further regulated by androgen receptor complexes with histone lysine demethylases. These complexes are recruited to androgen response elements in DNA and dynamically regulate histone tail modifications for transcription initiation. This led us to the overall hypothesis that (1) androgen signaling in trophoblast cells is important for placental development, and (2) androgen receptor complexes with histone lysine demethylases in the placenta to regulate vascularization, growth and invasion factors in trophoblast cells. To test this hypothesis, we utilized a prenatal androgenization ewe model as well as human first trimester placental samples and immortalized human trophoblast cell lines. Using the prenatal androgenized ewe model, we report for the first time expression of histone lysine demethylases in the placenta. Furthermore, we showed androgen receptor complexes with histone lysine demethylases and is recruited to an androgen response elements in the 5'untranslated flanking sequence of vascular endothelial growth factor in the sheep placenta. We also report that histone lysine demethylase are present in human first trimester syncytiotrophoblast and complex with androgen receptor in immortalized trophoblasts. Additionally, we demonstrated that androgen receptor complexes with histone lysine demethylases are also present in choriocarcinoma ACH-3P and BeWo cells. Dihydrotestosterone treatment in these cells led to down-regulation of androgen responsive genes, specifically KDM3A and MMP2. Inhibition of androgen receptor through flutamide treatment altered mRNA levels for genes regulating vascularization, including HIF1α, PPARα, and PPARy. Hypoxia also decreased CYP19 levels, however, further investigation is needed to confirm dihydrotestosterone and flutamide effect on protein expression in trophoblast cells. These data suggest that histone lysine demethylases complex with androgen receptor to regulate androgen responsive… Advisors/Committee Members: Bouma, Gerrit (advisor), Clay, Colin (advisor), Tobet, Stuart (committee member), Di Pietro, Santiago (committee member).

Subjects/Keywords: histone lysine demethylases; trophoblast; preeclampsia; androgen; placenta

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APA (6^th Edition):

Cleys, E. R. (2014). Androgen signaling in the placenta. (Doctoral Dissertation). Colorado State University. Retrieved from http://hdl.handle.net/10217/83721

Chicago Manual of Style (16^th Edition):

Cleys, Ellane Rachael. “Androgen signaling in the placenta.” 2014. Doctoral Dissertation, Colorado State University. Accessed April 15, 2021. http://hdl.handle.net/10217/83721.

MLA Handbook (7^th Edition):

Cleys, Ellane Rachael. “Androgen signaling in the placenta.” 2014. Web. 15 Apr 2021.

Vancouver:

Cleys ER. Androgen signaling in the placenta. [Internet] [Doctoral dissertation]. Colorado State University; 2014. [cited 2021 Apr 15]. Available from: http://hdl.handle.net/10217/83721.

Council of Science Editors:

Cleys ER. Androgen signaling in the placenta. [Doctoral Dissertation]. Colorado State University; 2014. Available from: http://hdl.handle.net/10217/83721

Colorado State University

3. Urias Castro, Christian. Hypothalamic concentration of kisspeptin and GnRH during breeding season (BS) and non breeding season (NBS) in sheep.

Degree: PhD, Biomedical Sciences, 2016, Colorado State University

URL: http://hdl.handle.net/10217/173370

► The kisspeptin system has emerged as an important regulator of mammalian reproduction. In ewes, kisspeptin neurons are located in specific hypothalamic regions such as the… (more)

▼ The kisspeptin system has emerged as an important regulator of mammalian reproduction. In ewes, kisspeptin neurons are located in specific hypothalamic regions such as the preoptic area (POA) and medio-basal hypothalamus (MBH) which include the arcuate nucleus (ARC). A specific radioimmunoassay (RIA) for the quantification of hypothalamic kisspeptin was developed to test the hypothesis that estradiol decreases the production of kisspeptin during the NBS in the MBH in addition to other forebrain areas that harbor kisspeptin neurons and/or axons such as the POA, the anterior hypothalamic area (AHA), and the median eminence (ME). The kisspeptin RIA results indicated that the concentrations of kisspeptin per milligram of tissue were decreased during NBS in the MBH and the POA with a tendency for lower kisspeptin concentrations observed in the AHA. Likewise, the total content of kisspeptin was decreased in the MBH and POA during the NBS, with a similar tendency for lower content of kisspeptin observed in the AHA during the NBS. Supporting the notion that kisspeptin modulates secretion of GnRH at the level of the ME, a positive correlation between these neuropeptides was observed during the BS in this region. It may be, that kisspeptin neurons are relevant for the seasonal regulation of GnRH and LH secretion exerted by estradiol, since the GnRH neurons do not express estrogen receptor alpha (ERα) which is the relevant ER subtype for the regulation of the hypothalamic pituitary gonadal axis (GnRH/LH pulsatility). We investigated if the negative feedback exerted by estradiol during the NBS, promoting a decrease in the concentrations of kisspeptin in the ARC, can be blocked by the intracerebral ventricular (ICV) administration of an estradiol antagonist (ICI) to promote an increase in LH pulsatility. As expected, in ewes that received the ICI treatment an increase in the average number of LH pulses was observed. The increased frequency in LH pulsatility was probably a consequence of eliminating estradiol inhibitory actions over ARC kisspeptin neurons which send axonal projections to the ME and promote the release of GnRH, and thus LH. Interestingly, the ME is a circumventricular organ (CVO) located outside of the blood brain barrier (BBB). Ovariectomized ewes were immunized against kisspeptin and antiserum to kisspeptin generated. The antibodies to kisspeptin were intended to eliminate kisspeptin release from the ME and consequently block kisspeptin input to GnRH axon terminals. The blockade of kisspeptin input to GnRH axon terminals was intended to inhibit the release of GnRH and hence LH. When compared to controls, ewes immunized against kisspeptin tended to have lower average and basal secretion of LH. The lack of significant decrease observed in immunized ewes suggests that higher titers to kisspeptin could be needed to fully suppress GnRH and hence LH pulsatility. Still, the tendency for lower levels of LH observed in immunized ewes suggest that kisspeptin release from the ME is relevant for the modulation of the… Advisors/Committee Members: Nett, Terry (advisor), Clay, Colin (advisor), Han, Hyng Chul (committee member), Bouma, Gerrit (committee member).

Subjects/Keywords: kisspeptin; radio innuno assay; kisspeptin antibodies; estrogen receptor

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Urias Castro, C. (2016). Hypothalamic concentration of kisspeptin and GnRH during breeding season (BS) and non breeding season (NBS) in sheep. (Doctoral Dissertation). Colorado State University. Retrieved from http://hdl.handle.net/10217/173370

Chicago Manual of Style (16^th Edition):

Urias Castro, Christian. “Hypothalamic concentration of kisspeptin and GnRH during breeding season (BS) and non breeding season (NBS) in sheep.” 2016. Doctoral Dissertation, Colorado State University. Accessed April 15, 2021. http://hdl.handle.net/10217/173370.

MLA Handbook (7^th Edition):

Urias Castro, Christian. “Hypothalamic concentration of kisspeptin and GnRH during breeding season (BS) and non breeding season (NBS) in sheep.” 2016. Web. 15 Apr 2021.

Vancouver:

Urias Castro C. Hypothalamic concentration of kisspeptin and GnRH during breeding season (BS) and non breeding season (NBS) in sheep. [Internet] [Doctoral dissertation]. Colorado State University; 2016. [cited 2021 Apr 15]. Available from: http://hdl.handle.net/10217/173370.

Council of Science Editors:

Urias Castro C. Hypothalamic concentration of kisspeptin and GnRH during breeding season (BS) and non breeding season (NBS) in sheep. [Doctoral Dissertation]. Colorado State University; 2016. Available from: http://hdl.handle.net/10217/173370

Colorado State University

4. Drennan, Meggan L. L-type calcium channel-dependent signaling impacts GnRH receptor function and intercellular communication in cultured gonadotropes.

Degree: MS(M.S.), Biomedical Sciences, 2020, Colorado State University

URL: http://hdl.handle.net/10217/212068

► The hypothalamic-pituitary-gonadal (HPG) axis is a negative feedback biological system critical in fertility, reproduction and development. Gonadotropin-releasing hormone (GnRH) is first released by the hypothalamus… (more)

▼ The hypothalamic-pituitary-gonadal (HPG) axis is a negative feedback biological system critical in fertility, reproduction and development. Gonadotropin-releasing hormone (GnRH) is first released by the hypothalamus and binds to GnRH receptors (GnRH-R) on gonadotrope cells of the anterior pituitary gland where the receptors must mediate a variety of pulsatile signals. The gonadotropin hormones, luteinizing hormone (LH) and follicle stimulating hormone (FSH), are subsequently released by the pituitary and act upon the ovaries and testes, further producing gonadal steroids to be circulated throughout the body. GnRH pulse frequency and amplitude determine successful gonadotropin release, which is ultimately regulated by the GnRH-R. The GnRH-R is a heterotrimeric G-protein coupled 7-transmembrane domain receptor with Gα, β, and γ subunits. Ligand binding initiates an intracellular cascade that leads to a global increase of cytosolic calcium concentration by way of calcium influx through voltage-gated calcium (Cav) channels, and intracellular calcium release from endoplasmic reticulum (ER) stores. Gonadotropes depend on intracellular calcium concentration to carry out their specific physiological function, such as transcription of gonadotropin subunits, hormone biosynthesis and release. Calcium flux is a normal and important aspect of cellular function, including cell-cell communication. Calcium oscillations have been well documented in multiple cell types, with different patterns being induced with distinct treatments. Observations in this line of research include the following: different oscillatory patterns lead to different physiological outcomes, the rate at which internal calcium is secreted from the ER can greatly impact these patterns, and IP3 receptor clustering on the ER results in localized changes in calcium concentration rather than a marked global difference, implicating a spatial stochasticity. These oscillations have shown evidence of paracellular coupling at gap junctions, as well as synchrony following extracellular diffusion. Chapter two of this thesis details experiments investigating calcium oscillations using a membrane-targeted calcium indicator. Immortalized αT3-1 cells were transfected with a membrane-targeted GCaMP and TIRF microscopy was used to capture fluorescent calcium activity. Cells were treated with GnRH as well as various pharmaceutical treatments that would exploit L-type Cav channel function and manipulate normal intracellular calcium release. An array of observations was recorded. Qualitatively, there was an overall increase in calcium activity in the majority of cells after GnRH treatment. Drug-induced inhibition of calcium influx and intracellular calcium release diminished calcium activity entirely. Further, synchronized activity was captured among several cell groups, showing both pre-established synchrony and GnRH-induced synchronized peaking. Further research should be conducted to better understand the full mechanism underlying these behavioral responses, but these experiments… Advisors/Committee Members: Amberg, Gregory (advisor), Clay, Colin (committee member), Garrity, Deborah (committee member), Kelp, Nicole (committee member).

Subjects/Keywords: GnRH; gonadotropin-releasing hormone; receptor; gonadotrope; calcium oscillation; pituitary

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APA (6^th Edition):

Drennan, M. L. (2020). L-type calcium channel-dependent signaling impacts GnRH receptor function and intercellular communication in cultured gonadotropes. (Masters Thesis). Colorado State University. Retrieved from http://hdl.handle.net/10217/212068

Chicago Manual of Style (16^th Edition):

Drennan, Meggan L. “L-type calcium channel-dependent signaling impacts GnRH receptor function and intercellular communication in cultured gonadotropes.” 2020. Masters Thesis, Colorado State University. Accessed April 15, 2021. http://hdl.handle.net/10217/212068.

MLA Handbook (7^th Edition):

Drennan, Meggan L. “L-type calcium channel-dependent signaling impacts GnRH receptor function and intercellular communication in cultured gonadotropes.” 2020. Web. 15 Apr 2021.

Vancouver:

Drennan ML. L-type calcium channel-dependent signaling impacts GnRH receptor function and intercellular communication in cultured gonadotropes. [Internet] [Masters thesis]. Colorado State University; 2020. [cited 2021 Apr 15]. Available from: http://hdl.handle.net/10217/212068.

Council of Science Editors:

Drennan ML. L-type calcium channel-dependent signaling impacts GnRH receptor function and intercellular communication in cultured gonadotropes. [Masters Thesis]. Colorado State University; 2020. Available from: http://hdl.handle.net/10217/212068

Colorado State University

5. Gates, Katherine C. Role of proline rich 15 in trophoblast cell development, The.

Degree: PhD, Biomedical Sciences, 2012, Colorado State University

URL: http://hdl.handle.net/10217/67920

► Maintenance of pregnancy in eutherian mammals requires a sophisticated and tightly regulated program of gene expression in order to develop a fully functional placenta. This… (more)

▼ Maintenance of pregnancy in eutherian mammals requires a sophisticated and tightly regulated program of gene expression in order to develop a fully functional placenta. This transient organ mediates nutrient and gas exchange between the mother and fetus while protecting the fetus from the maternal immune system. Deviations from the normal regulation of gene expression during early pregnancy can lead to early embryonic loss as well as dysfunctional placentation, which can cause significant maternal and fetal morbidity and mortality. Proline rich 15 (PRR15) is a low molecular weight nuclear protein expressed by the trophoblast during early gestation in several mammalian species, including humans, mice, cattle, sheep, and horses. Immunohistochemistry revealed localization of PRR15 to the trophectoderm and extraembryonic endoderm of day 15 sheep conceptuses. In humans, PRR15 is localized in the nuclei of both first and second trimester trophoblast cells. Additional research has shown increased PRR15 transcription in colorectal cancers with mutations in the adenomatous polyposis coli (Apc) protein, suggesting a link to the Wnt signaling pathway. PRR15 mRNA concentrations increase when trophoblast cells, both sheep (oTR) and human (ACH-3P), are cultured on Matrigel, a basement membrane matrix. The expression profile in the sheep conceptus during pregnancy revealed a rise in PRR15 mRNA concentrations during the period of conceptus elongation with a peak in expression at day 16 of gestation, followed by a decline to day 30 of gestation. This peak coincides with a halt in elongation of the conceptus, and the initial period of apposition to the uterine luminal epithelium. Lentiviral-mediated knockdown of PRR15 in ovine trophectoderm at the blastocyst stage led to demise of the embryo by day 15 of gestation. This provides compelling evidence that PRR15 is a critical factor during this precarious window of development when initial attachment and implantation begin. The first aim of this research was to determine the effect of PRR15 deficiency on trophoblast gene expression, as well as trophoblast proliferation and survival. The human first trimester trophoblast cell line, ACH-3P, was infected with control lentivirus (LL3.7) and lentivirus expressing a short hairpin (sh)RNA to target PRR15 mRNA for degradation, resulting in a 68% decrease in PRR15 mRNA (p<0.01). Microarray analysis of these cell lines revealed differential expression of genes related to cancer, focal adhesion, and p53 signaling. We selected 21 genes for validation of mRNA levels by quantitative real-time RT-PCR, 18 (86%) of which gave results consistent with the microarray analysis, with similar direction and magnitude fold changes. This included significant up-regulation of GDF15, a cytokine increased in pregnancies with preeclampsia. GDF15 mRNA concentrations were examined more extensively during early ovine gestation, which revealed that GDF15 was low during peak PRR15 expression, then increased significantly at day 30 when PRR15 was nearly undetectable.… Advisors/Committee Members: Anthony, Russell V. (advisor), Clay, Colin (committee member), Duval, Dawn L. (committee member), Hansen, Thomas R. (committee member).

Subjects/Keywords: implantation; microarray; placenta; proline rich 15; transcription; trophoblast

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Gates, K. C. (2012). Role of proline rich 15 in trophoblast cell development, The. (Doctoral Dissertation). Colorado State University. Retrieved from http://hdl.handle.net/10217/67920

Chicago Manual of Style (16^th Edition):

Gates, Katherine C. “Role of proline rich 15 in trophoblast cell development, The.” 2012. Doctoral Dissertation, Colorado State University. Accessed April 15, 2021. http://hdl.handle.net/10217/67920.

MLA Handbook (7^th Edition):

Gates, Katherine C. “Role of proline rich 15 in trophoblast cell development, The.” 2012. Web. 15 Apr 2021.

Vancouver:

Gates KC. Role of proline rich 15 in trophoblast cell development, The. [Internet] [Doctoral dissertation]. Colorado State University; 2012. [cited 2021 Apr 15]. Available from: http://hdl.handle.net/10217/67920.

Council of Science Editors:

Gates KC. Role of proline rich 15 in trophoblast cell development, The. [Doctoral Dissertation]. Colorado State University; 2012. Available from: http://hdl.handle.net/10217/67920

Colorado State University

6. Dang, An Khanh. Regulation of local L-type calcium channel signaling in anterior pituitary gonadotropes.

Degree: PhD, Biomedical Sciences, 2017, Colorado State University

URL: http://hdl.handle.net/10217/185697

► The binding of gonadotropin-releasing hormone (GnRH) to its receptor initiates signaling cascades in gonadotropes which result in enhanced luteinizing hormone (LH) and follicle stimulating hormone… (more)

▼ The binding of gonadotropin-releasing hormone (GnRH) to its receptor initiates signaling cascades in gonadotropes which result in enhanced luteinizing hormone (LH) and follicle stimulating hormone (FSH) biosynthesis and secretion. Most dramatic is the sharp rise in LH secretion ("LH surge") that precedes and is necessary for follicular maturation and ovulation. Ca2+ influx activates mitogen-activated protein kinases (MAPKs) which lead to increased transcription of LH and FSH genes. Interestingly, previous research suggests that two MAPK signaling pathways, ERK and JNK, are activated by either Ca2+ influx through L-type Ca2+ channels or by global Ca2+ signals originating from intracellular stores, respectively. These discrete Ca2+ sources for divergent signaling cascades provides a mechanism in which gonadotropes can decode different pathways for appropriate gonadotropin release during various stages of the ovulatory cycle. However, direct evidence supporting an underlying subplasmalemmal local Ca2+ signaling through L-type Ca2+ channels distinct from intracellular Ca2+ was lacking. Here we used a combination of electrophysiology and total internal reflection fluorescence (TIRF) microscopy to visualize discrete sites of Ca2+ influx (Ca2+ sparklets) in gonadotrope-derived αT3-1 cells in real time. These localized GnRH-induced Ca2+ influxes are mediated by L-type Ca2+ channels and important for downstream ERK activation. In addition, precise structural and molecular elements to create a microenvironment suitable for localized subplasmalemmal L-type Ca2+ channel signaling was necessary for gonadotrope function, in which GnRH-dependent stimulation of L-type Ca2+ channel influx was found to require PKC and a dynamic actin cytoskeleton. More recently, we have further elucidated molecular mechanisms modulating localized L-type Ca2+ channel influx. Reactive oxygen species (ROS) are cognate signaling molecules that mediate cell function, but their role in regulating Ca2+ in gonadotropes is unknown. We have explored GnRH regulation of both NADPH oxidase complexes and mitochondrial sources of ROS and assessed ROS modulation of L-type Ca2+ channel activity in gonadotropes. We identified GnRH-induced spatially localized ROS "puncta" in αT3-1 cells, and ROS increased local Ca2+ channel activity in both αT3-1 cells and primary mouse gonadotropes. In addition, GnRH increased mitochondrial oxidation activity at the subplasmalemmal surface and mitochondrial ROS increased localized L-type Ca2+ channel influx. Also, active L-type Ca2+ channels were associated with subplasmalemmal mitochondria. Taken together, this dissertation explored the first direct evidence for localized L-type Ca2+ channel signaling in αT3-1 cells and elucidated signaling mechanisms in gonadotropes. Specifically, cellular organization via an intact cytoskeletal platform and ROS regulated L-type Ca2+ channel sparklet activity that are important for the downstream ERK activation and gonadotropin gene expression that regulates reproduction. Advisors/Committee Members: Amberg, Greg (advisor), Clay, Colin (advisor), Tamkun, Michael (committee member), Navratil, Amy (committee member), Duval, Dawn (committee member).

Subjects/Keywords: gonadotrope; reactive oxygen species; GnRH; reproductive endocrinology; L-type calcium channel

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Dang, A. K. (2017). Regulation of local L-type calcium channel signaling in anterior pituitary gonadotropes. (Doctoral Dissertation). Colorado State University. Retrieved from http://hdl.handle.net/10217/185697

Chicago Manual of Style (16^th Edition):

Dang, An Khanh. “Regulation of local L-type calcium channel signaling in anterior pituitary gonadotropes.” 2017. Doctoral Dissertation, Colorado State University. Accessed April 15, 2021. http://hdl.handle.net/10217/185697.

MLA Handbook (7^th Edition):

Dang, An Khanh. “Regulation of local L-type calcium channel signaling in anterior pituitary gonadotropes.” 2017. Web. 15 Apr 2021.

Vancouver:

Dang AK. Regulation of local L-type calcium channel signaling in anterior pituitary gonadotropes. [Internet] [Doctoral dissertation]. Colorado State University; 2017. [cited 2021 Apr 15]. Available from: http://hdl.handle.net/10217/185697.

Council of Science Editors:

Dang AK. Regulation of local L-type calcium channel signaling in anterior pituitary gonadotropes. [Doctoral Dissertation]. Colorado State University; 2017. Available from: http://hdl.handle.net/10217/185697

Colorado State University

7. Ruggeri, Elena. Analysis of equine zygote development after intracytoplasmic sperm injection.

Degree: PhD, Biomedical Sciences, 2016, Colorado State University

URL: http://hdl.handle.net/10217/173335

► Intracytoplasmic sperm injection (ICSI) is an established and widely used method to achieve oocyte fertilization in equine reproductive assisted technologies. However, not all the oocytes… (more)

▼ Intracytoplasmic sperm injection (ICSI) is an established and widely used method to achieve oocyte fertilization in equine reproductive assisted technologies. However, not all the oocytes fertilized by ICSI undergo cleavage and develop into viable embryos. Limited knowledge on equine zygote development after ICSI is available, and reasons why developmental failure occurs after ICSI have been only partially studied and need further investigation. Fertility decline and early embryo loss is associated with maternal aging in the mare, and it is concomitant with reduced oocyte quality. Relatively little is known about the effect of maternal aging and zygote developmental failure or success in the mare. Effects of in vitro maturation of the oocyte or zygote development in the mare still need to be clarified and further studied. The overall objective of this dissertation was to study equine zygote development after ICSI using confocal microscopy. Objectives were to: (1) compare cytoskeletal and nuclear changes during progression of equine zygote development after ICSI for in vivo versus in vitro matured oocytes; (2) compare changes in cytoskeletal and chromosomal configurations after ICSI between oocytes from young and old mares to define maternal-aging related alterations; (3) determine cytoskeletal and nuclear alterations associated with fertilization failure in ICSI-produced presumptive zygotes in young and old mares; (4) determine cell-aging and cell donor-aging effects on cytoskeleton and chromatin configurations. Specifically, in our studies we evaluated the tubulin and actin cytoskeleton, chromatin, and kinetochores/centromeres. Immunostaining and confocal imaging of the equine zygotes was performed using a spinning disk confocal microscope. After ICSI, five distinct events of development were observed with no major differences over time whether oocytes matured in vivo or in vitro. Oocytes matured in vivo appeared to reach the pronucleus stage earlier after ICSI compared to in vitro matured oocytes. Abnormal phenotypes associated with fertilization failure were more significant in oocytes matured in vitro than in vivo. When ICSI was performed in oocytes from young and old mares, similar stages of zygote development were observed, and the number of zygotes reaching the pronucleus stage was similar between the two age groups. Nucleolus like bodies, sites of ribosomal RNA involved in embryonic genome activation, were observed only in zygotes at the pronucleus stage from young mares; no nucleolus-like bodies were observed in pronuclei of zygotes from old mares. Pronuclei morphology, based on CREST staining, and DNA localization, also differed between pronuclei of young and old mares. Actin vesicles were observed significantly more often within zygotes from old mares compared to young mares during all stages of zygote developmental progression. When potential zygotes were analyzed after failure of cleavage after ICSI, actin vesicles were greater in area, perimeter and number in oocytes from old mares than those… Advisors/Committee Members: Carnevale, Elaine (advisor), Clay, Colin (advisor), Albertini, David (committee member), DeLuca, Jennifer (committee member), Seidel, George (committee member).

Subjects/Keywords: cytoskeleton; intracytoplasmic sperm injection; zygote; equine; confocal microscopy; maternal aging

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Ruggeri, E. (2016). Analysis of equine zygote development after intracytoplasmic sperm injection. (Doctoral Dissertation). Colorado State University. Retrieved from http://hdl.handle.net/10217/173335

Chicago Manual of Style (16^th Edition):

Ruggeri, Elena. “Analysis of equine zygote development after intracytoplasmic sperm injection.” 2016. Doctoral Dissertation, Colorado State University. Accessed April 15, 2021. http://hdl.handle.net/10217/173335.

MLA Handbook (7^th Edition):

Ruggeri, Elena. “Analysis of equine zygote development after intracytoplasmic sperm injection.” 2016. Web. 15 Apr 2021.

Vancouver:

Ruggeri E. Analysis of equine zygote development after intracytoplasmic sperm injection. [Internet] [Doctoral dissertation]. Colorado State University; 2016. [cited 2021 Apr 15]. Available from: http://hdl.handle.net/10217/173335.

Council of Science Editors:

Ruggeri E. Analysis of equine zygote development after intracytoplasmic sperm injection. [Doctoral Dissertation]. Colorado State University; 2016. Available from: http://hdl.handle.net/10217/173335

Colorado State University

8. Hergenreder, Joanna R. Serum exosome profile as related to early pregnancy status in the mare.

Degree: MS(M.S.), Animal Sciences, 2011, Colorado State University

URL: http://hdl.handle.net/10217/46736

► During early pregnancy in the mare the conceptus and mare must communicate in order to establish and maintain pregnancy. This coordinate communication is most pronounced… (more)

▼ During early pregnancy in the mare the conceptus and mare must communicate in order to establish and maintain pregnancy. This coordinate communication is most pronounced between days 12 and 16 post-ovulation, at which time the conceptus is highly mobile throughout the uterus preventing endometrial prostaglandin F2α release and subsequent luteolysis. The mechanism behind successful establishment and maintenance of pregnancy in the mare is currently unknown. Recently, cell-secreted vesicles, called exosomes, were detected in high amounts in serum of pregnant women. Exosomes are 50-100 nm vesicles containing bioactive materials such as mRNA, miRNA, and protein. Exosomes can mediate immune-responses through membrane protein interaction and delivery of bioactive products into cells. Interestingly, exosomes have been described in various body fluids, including urine, breast milk, and serum. We hypothesized that exosomes are present in serum in the mare and that their relative amount differs with pregnancy status. To test this hypothesis, we determined the presence and relative amount of exosomes in serum of pregnant and non-pregnant mares. Serum samples were obtained from mares in a cross-over design, with each mare serving as both a pregnant treatment and non-mated control (n=3/day). Blood samples were obtained by jugular venipuncture on days 12, 14, 16, and 18 post-ovulation. Serum was removed, snap frozen, and stored at -80°C. Exosome isolation, for flow-cytometry and transmission electron microscopy (TEM), was performed using ExoQuick (System Biosciences, Inc.), a precipitation solution designed to isolate exosomes from fluids. After exosome isolation, samples were analyzed using flow cytometry with 100 nm sized beads as an internal control and a counting bead standard for relative amount determination. Flow cytometry analysis revealed the presence of exosomes in serum of both pregnant and non-pregnant mares in variable amounts. Furthermore, analysis revealed the presence of two distinct size populations, one of smaller exosomes (< 100 nm) previously undescribed, which were more abundant in mare serum from day 12 of pregnancy, and the second of the expected 100 nm size at each day examined. TEM analysis validated the results from the flow cytometry as each population, determined by size and granularity, was visually characterized. Along with the 100 nm and slightly smaller sized vesicles, TEM also revealed the presence of vesicles slightly larger than 100 nm, with small amounts of vesicles ∼200 nm in size, indicating the presence of exosomes as well as microvesicles. Therefore, we conclude that exosomes are present in mare serum and further characterization of such populations can provide clues about the intercellular mode of communication in early pregnancy. Advisors/Committee Members: Bruemmer, Jason E. (advisor), Bouma, Gerrit J. (committee member), Veeramachaneni, D. N. Rao (committee member), Clay, Colin McKeown (committee member), Han, Hyungchul (committee member).

Subjects/Keywords: microvesicles; intercellular communication; mare; early pregnancy; exosomes

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Hergenreder, J. R. (2011). Serum exosome profile as related to early pregnancy status in the mare. (Masters Thesis). Colorado State University. Retrieved from http://hdl.handle.net/10217/46736

Chicago Manual of Style (16^th Edition):

Hergenreder, Joanna R. “Serum exosome profile as related to early pregnancy status in the mare.” 2011. Masters Thesis, Colorado State University. Accessed April 15, 2021. http://hdl.handle.net/10217/46736.

MLA Handbook (7^th Edition):

Hergenreder, Joanna R. “Serum exosome profile as related to early pregnancy status in the mare.” 2011. Web. 15 Apr 2021.

Vancouver:

Hergenreder JR. Serum exosome profile as related to early pregnancy status in the mare. [Internet] [Masters thesis]. Colorado State University; 2011. [cited 2021 Apr 15]. Available from: http://hdl.handle.net/10217/46736.

Council of Science Editors:

Hergenreder JR. Serum exosome profile as related to early pregnancy status in the mare. [Masters Thesis]. Colorado State University; 2011. Available from: http://hdl.handle.net/10217/46736

Colorado State University

9. Seabrook, Jill L. Role of LIN28 in the molecular regulation of placenta development and function, The.

Degree: PhD, Biomedical Sciences, 2013, Colorado State University

URL: http://hdl.handle.net/10217/79438

To view the abstract, please see the full text of the document. Advisors/Committee Members: Winger, Quinton A. (advisor), Clay, Colin M. (committee member), Bouma, Gerrit J. (committee member), DeLuca, Jennifer G. (committee member).

Subjects/Keywords: LIN28; trophoblast; syncytiotrophoblast; placenta

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Seabrook, J. L. (2013). Role of LIN28 in the molecular regulation of placenta development and function, The. (Doctoral Dissertation). Colorado State University. Retrieved from http://hdl.handle.net/10217/79438

Chicago Manual of Style (16^th Edition):

Seabrook, Jill L. “Role of LIN28 in the molecular regulation of placenta development and function, The.” 2013. Doctoral Dissertation, Colorado State University. Accessed April 15, 2021. http://hdl.handle.net/10217/79438.

MLA Handbook (7^th Edition):

Seabrook, Jill L. “Role of LIN28 in the molecular regulation of placenta development and function, The.” 2013. Web. 15 Apr 2021.

Vancouver:

Seabrook JL. Role of LIN28 in the molecular regulation of placenta development and function, The. [Internet] [Doctoral dissertation]. Colorado State University; 2013. [cited 2021 Apr 15]. Available from: http://hdl.handle.net/10217/79438.

Council of Science Editors:

Seabrook JL. Role of LIN28 in the molecular regulation of placenta development and function, The. [Doctoral Dissertation]. Colorado State University; 2013. Available from: http://hdl.handle.net/10217/79438

Colorado State University

10. Donner, Nina Caroline. Behavioral effects of estrogen receptor beta acting locally to regulate the expression of tryptophan hydroxylase 2 (THP2) in serotonergic neurons of the dorsal raphe nuclei.

Degree: MS(M.S.), Biomedical Sciences, 2008, Colorado State University

URL: http://hdl.handle.net/10217/6393

► Affective disorders often involve serotonin (5-HT)-related dysfunctions and are twice as common in women than men. Interactions between estrogen and the brain 5-HT system have… (more)

▼ Affective disorders often involve serotonin (5-HT)-related dysfunctions and are twice as common in women than men. Interactions between estrogen and the brain 5-HT system have long been proposed to contribute to sex differences in mood and anxiety disorders, but the mechanisms underlying this phenomenon have yet to be revealed. Estrogen signaling is mediated by two different receptors termed estrogen receptor alpha and estrogen receptor beta. While estrogen receptor alpha (ERalpha) has mainly reproductive responsibilities, in brain, estrogen receptor beta (ERbeta) has been shown to attenuate anxiety- and despair-like behaviors in rodent models. However, little is known about ERbeta regulation of function in the brainstem raphe nuclei. The raphe nuclei are the main 5-HT system of the brain, and projections from the dorsal raphe nuclei (DRN) innervate many important forebrain and limbic areas. The work presented in this thesis addressed the possibility that ERbeta may be involved in the regulation of 5-HT gene expression specifically in DRN neurons. My studies examined the effects of systemic versus local, intracerebral application of the selective ERbeta agonist diarylpropionitrile (DPN) and the nonselective ERligandestradiol (E) on tryptophan hydroxylase 2 (TPH2) mRNA expression within the DRN of female rats. TPH2 is the brain-specific, rate-limiting enzyme catalyzing 5-HT synthesis, and is expressed in every 5-HT neuron. Thus, it provides an excellent tool to assess the capacity for 5-HT production with the DRN. In these studies, TPH2 mRNA expression was assessed via in situ hybridization. In addition, relevant behavioral parameters were tested in all animals to evaluate each compound’s effect on two closely related, but yet different mental states, anxiety-like and despair-like behavior. Both, chronic systemic and chronic local DPN administration to ovariectomized (OVX) female rats significantly enhanced TPH2 mRNA expression in mid- and caudal subregions of the DRN after 8 days of treatment. Respective controls received systemic vehicle (27% hydroxypropyl-beta-cyclodextrin) or blank control pellets. Local application of DPN caused a stronger effect than systemic drug delivery. Chronic local delivery of E (0.5 μM) increased TPH2 mRNA expression in the same subregions of the DRN as did DPN, but its overall effect was weaker compared to the selective ERbeta agonist. Interestingly, while systemic DPN-administration confirmed the anxiolytic nature of ERbeta in two separate anxiety tests (elevated plus maze and open field test), the effect was lost when DPN was delivered locally. However, local DPN- as well as E-treatment both resulted in attenuated despair-like behavior, as measured in the forced-swim test. Chapter 3 describes the experimental design, results and interpretation of these studies in depth. Taken together, my data indicate that local actions of ERbeta agonist onto DRN neurons are sufficient to decrease despair-like behavior, whereas ERbeta stimulation of other brain regions is necessary to alter… Advisors/Committee Members: Handa, Robert J. (advisor), Tjalkens, Ronald (committee member), Clay, Colin McKeown (committee member), Tobet, Stuart (committee member).

Subjects/Keywords: TPH2; tryptophan hydroxylase 2; DRN; dorsal raphe nuclei; ERbeta; estrogen receptor beta; brain 5-HT system; Estrogen – Agonists; Serotonin; Sensory receptors; Neurons

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Donner, N. C. (2008). Behavioral effects of estrogen receptor beta acting locally to regulate the expression of tryptophan hydroxylase 2 (THP2) in serotonergic neurons of the dorsal raphe nuclei. (Masters Thesis). Colorado State University. Retrieved from http://hdl.handle.net/10217/6393

Chicago Manual of Style (16^th Edition):

Donner, Nina Caroline. “Behavioral effects of estrogen receptor beta acting locally to regulate the expression of tryptophan hydroxylase 2 (THP2) in serotonergic neurons of the dorsal raphe nuclei.” 2008. Masters Thesis, Colorado State University. Accessed April 15, 2021. http://hdl.handle.net/10217/6393.

MLA Handbook (7^th Edition):

Vancouver:

Donner NC. Behavioral effects of estrogen receptor beta acting locally to regulate the expression of tryptophan hydroxylase 2 (THP2) in serotonergic neurons of the dorsal raphe nuclei. [Internet] [Masters thesis]. Colorado State University; 2008. [cited 2021 Apr 15]. Available from: http://hdl.handle.net/10217/6393.

Council of Science Editors:

Donner NC. Behavioral effects of estrogen receptor beta acting locally to regulate the expression of tryptophan hydroxylase 2 (THP2) in serotonergic neurons of the dorsal raphe nuclei. [Masters Thesis]. Colorado State University; 2008. Available from: http://hdl.handle.net/10217/6393

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