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55. Control of the Anterior Pituitary Cell Lineage Regulator POU1F1 by the Stem Cell Determinant Musashi.

Author

Allensworth-James, Melody, Banik, Jewel, Odle, Angela, Hardy, Linda, Lagasse, Alex, Moreira, Ana Rita Silva, Bird, Jordan, Thomas, Christian L, Avaritt, Nathan, Kharas, Michael G, Lengner, Christopher J, Byrum, Stephanie D, MacNicol, Melanie C, Childs, Gwen V, and MacNicol, Angus M

Subjects
STEM cells, LEPTIN receptors, SOMATOTROPIN, NUCLEOTIDE sequence, TRANSCRIPTION factors
Abstract

The adipokine leptin regulates energy homeostasis through ubiquitously expressed leptin receptors. Leptin has a number of major signaling targets in the brain, including cells of the anterior pituitary (AP). We have previously reported that mice lacking leptin receptors in AP somatotropes display growth hormone (GH) deficiency, metabolic dysfunction, and adult-onset obesity. Among other targets, leptin signaling promotes increased levels of the pituitary transcription factor POU1F1, which in turn regulates the specification of somatotrope, lactotrope, and thyrotrope cell lineages within the AP. Leptin's mechanism of action on somatotropes is sex dependent, with females demonstrating posttranscriptional control of Pou1f1 messenger RNA (mRNA) translation. Here, we report that the stem cell marker and mRNA translational control protein, Musashi1, exerts repression of the Pou1f1 mRNA. In female somatotropes, Msi1 mRNA and protein levels are increased in the mouse model that lacks leptin signaling (Gh -CRE Lepr -null), coincident with lack of POU1f1 protein, despite normal levels of Pou1f1 mRNA. Single-cell RNA sequencing of pituitary cells from control female animals indicates that both Msi1 and Pou1f1 mRNAs are expressed in Gh -expressing somatotropes, and immunocytochemistry confirms that Musashi1 protein is present in the somatotrope cell population. We demonstrate that Musashi interacts directly with the Pou1f1 mRNA 3′ untranslated region and exerts translational repression of a Pou1f1 mRNA translation reporter in a leptin-sensitive manner. Musashi immunoprecipitation from whole pituitary reveals coassociated Pou1f1 mRNA. These findings suggest a mechanism in which leptin stimulation is required to reverse Musashi-mediated Pou1f1 mRNA translational control to coordinate AP somatotrope function with metabolic status. [ABSTRACT FROM AUTHOR]

Published
2021
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56. The Importance of Leptin to Reproduction.

Author

Childs, Gwen V, Odle, Angela K, MacNicol, Melanie C, and MacNicol, Angus M

Subjects
LEPTIN, GONADOTROPIN, MESSENGER RNA, GONADOTROPIN releasing hormone, KISSPEPTIN neurons, LUTEINIZING hormone releasing hormone receptors, GROWTH factors
Abstract

A healthy nutritional state is required for all aspects of reproduction and is signaled by the adipokine leptin. Leptin acts in a relatively narrow concentration range: too much or too little will compromise fertility. The leptin signal timing is important to prepubertal development in both sexes. In the brain, leptin acts on ventral premammillary neurons which signal kisspeptin (Kiss1) neurons to stimulate gonadotropin releasing hormone (GnRH) neurons. Suppression of Kiss1 neurons occurs when agouti-related peptide neurons are activated by reduced leptin, because leptin normally suppresses these orexigenic neurons. In the pituitary, leptin stimulates production of GnRH receptors (GnRHRs) and follicle-stimulating hormone at midcycle, by activating pathways that derepress actions of the messenger ribonucleic acid translational regulatory protein Musashi. In females, rising estrogen stimulates a rise in serum leptin, which peaks at midcycle, synchronizing with nocturnal luteinizing hormone pulses. The normal range of serum leptin levels (10-20 ng/mL) along with gonadotropins and growth factors promote ovarian granulosa and theca cell functions and oocyte maturation. In males, the prepubertal rise in leptin promotes testicular development. However, a decline in leptin levels in prepubertal boys reflects inhibition of leptin secretion by rising androgens. In adult males, leptin levels are 10% to 50% of those in females, and high leptin inhibits testicular function. The obesity epidemic has elucidated leptin resistance pathways, with too much leptin in either sex leading to infertility. Under conditions of balanced nutrition, however, the secretion of leptin is timed and regulated within a narrow level range that optimizes its trophic effects. [ABSTRACT FROM AUTHOR]

Published
2021
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61. Association of Gnrhr mRNA With the Stem Cell Determinant Musashi: A Mechanism for Leptin-Mediated Modulation of GnRHR Expression

Author

Odle, Angela K, primary, Beneš, Helen, additional, Melgar Castillo, Andrea, additional, Akhter, Noor, additional, Syed, Mohsin, additional, Haney, Anessa, additional, Allensworth-James, Melody, additional, Hardy, Linda, additional, Winter, Benjamin, additional, Manoharan, Ragul, additional, Syed, Raiyan, additional, MacNicol, Melanie C, additional, MacNicol, Angus M, additional, and Childs, Gwen V, additional

Published
2017
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64. Evasion of regulatory phosphorylation by an alternatively spliced isoform of Musashi2

Author

MacNicol, Melanie C., primary, Cragle, Chad E., additional, McDaniel, F. Kennedy, additional, Hardy, Linda L., additional, Wang, Yan, additional, Arumugam, Karthik, additional, Rahmatallah, Yasir, additional, Glazko, Galina V., additional, Wilczynska, Ania, additional, Childs, Gwen V., additional, Zhou, Daohong, additional, and MacNicol, Angus M., additional

Published
2017
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65. Gametes in Paradise–How the Oviduct Epithelial Microenvironment Supports and Protects Against Maternal Stress.

Author

Childs, Gwen V

Abstract

The author comments on how the oviduct epithelial microenvironment supports and protects against maternal stress in gametes. Topics discussed include an overview of how ovarian steroids stimulate epithelial remodeling during the reproductive cycle, significance of the ablation of progesterone (P4) or estradiol (E2) receptors, and effect of emotional and physical stresses on adrenal glucocorticoids.

Published
2023
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74. Anterior Pituitary Leptin Expression Changes in Different Reproductive States: Stimulation, in vitro, by Gonadotropin Releasing Hormone (GnRH)

Author

Akhter, Noor, Johnson, Brandy W., Crane, Christopher, Iruthayanathan, Mary, Zhou, Yi-Hong, Kudo, Akihiko, and Childs, Gwen V.

Subjects
Leptin, Male, endocrine system, digestive, oral, and skin physiology, Estrous Cycle, Article, Rats, Gonadotropin-Releasing Hormone, Rats, Sprague-Dawley, Pituitary Gland, Anterior, Pregnancy, Animals, Lactation, Pregnancy, Animal, Female, RNA, Messenger, reproductive and urinary physiology, hormones, hormone substitutes, and hormone antagonists
Abstract

This study was designed to learn more about the changes in expression of rat anterior pituitary (AP) leptin during the estrous cycle. QRT-PCR assays of cycling rat AP leptin mRNA showed 2-fold increases from metestrus to diestrus followed by an 86% decrease on the morning of proestrus. Percentages of leptin cells increased in proestrus and pregnancy to 55-60% of AP cells. Dual labeling for leptin proteins and growth hormone (GH) or gonadotropins showed that the rise in leptin protein-bearing cells from diestrus to proestrus was mainly in GH cells. Only 10-20% of leptin cells in male or cycling female rats coexpress gonadotropins. In contrast, 50-73% of leptin cells from pregnant or lactating females coexpress gonadotropins and only 19% coexpress GH, indicating plasticity in the distribution of leptin. Leptin cells expressed GnRH receptors, and estrogen and GnRH together increased the coexpression of leptin mRNA and gonadotropins. GnRH increased cellular leptin proteins three to four times and mRNA 9.8 times in proestrous rats and stimulated leptin secretion in cultures from diestrous, proestrous, and pregnant rats. These regulatory influences, and the high expression of AP leptin during proestrus and pregnancy, suggest a supportive role for leptin during key events involved with reproduction.

Published
2006

80. Simultaneous Identification of a Specific Gene Protein Product and Transcript Using Combined Immunocytochemistry and In Situ Hybridization with Non-Radioactive Probes

Author

Childs, Gwen V.

Subjects
pituitary hormones, electron microscopy, avidin-biotin cytochemistry, immunoperoxidase cytochemistry, cRNA, oligoprobe, In situ hybridization, Biology, messenger RNA (mRNA)
Abstract

Simultaneous identification of messenger RNA (mRNA) and proteins in the same cells or tissues is a valuable tool to help the cell biologist evaluate the cell secretory cycle. Some cells may produce the mRNA and delay the production of the proteins. Alternatively, the proteins may be rapidly secreted. Other cells may produce both in sequence within the same time frame. Because of this difference, some cells can only be identified by their mRNA product. Others may have both products. This presentation describes a non-radioactive approach to the detection of both products with dual-peroxidase labeling protocols in use in this laboratory since 1983. The first detection system uses biotinylated cRNA probes or oligoprobes in in situ hybridization along with antisera to biotin to detect the hybrid. The detection system is amplified by 2-3 layers of anti-biotin, second antibody (made against the anti-biotin) and streptavidin conjugated to horseradish peroxidase. After the mRNA is detected with a blue-black substrate (nickel intensified diaminobenzidine), the antigens are detected with immunoperoxidase techniques and orange-amber substrate. The in situ hybridization protocol can also be used at the electron microscopic level. Trouble shooting and control protocols are also described. This approach has been shown to be valuable for detection of pituitary hormones, growth factors mRNAs and antigens.

Published
1996

82. The Somatotrope as a Metabolic Sensor: Deletion of Leptin Receptors Causes Obesity

Author

Childs, Gwen V., primary, Akhter, Noor, additional, Haney, Anessa, additional, Syed, Mohsin, additional, Odle, Angela, additional, Cozart, Michael, additional, Brodrick, Zachary, additional, Gaddy, Dana, additional, Suva, Larry J., additional, Akel, Nisreen, additional, Crane, Christopher, additional, Benes, Helen, additional, Charlesworth, Amanda, additional, Luque, Raul, additional, Chua, Streamson, additional, and Kineman, Rhonda D., additional

Published
2010
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93. In Situ Hybridization with Nonradioactive Probes.

Author

Walker, John M., Darby, Ian A., and Childs, Gwen V.

Abstract

In situ hybridization takes advantage of the reaction between two complementary single-stranded nucleic acid molecules. These molecules bind by means of hydrogen bonding of complementary base pairs. Hybridization performed in situ includes techniques that detect these hybrids in a cell or tissue section by cytochemistry. [ABSTRACT FROM AUTHOR]

Published
2000
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96. Secretory Mechanisms in Opiocortin Cells during Cold Stress.

Author

TEXAS UNIV MEDICAL BRANCH AT GALVESTON, Childs, Gwen V., TEXAS UNIV MEDICAL BRANCH AT GALVESTON, and Childs, Gwen V.

Abstract

The goals of the study were to characterize the sequence of events involved in the hypothalamic-pituitary responses to cold stress. In the hypothalamus, studies focused on Arginine vasopressin neurons and changes in expression of mRNA following cold and novel environment exposure. In the pituitary, studies focused on AVP and CRH target cells, including corticotropes, thyrotropes, and intermediate lobe cells. Changes in size, granulation, expression of mRNA, and binding sites for AVP and CRH were detected. In vitro studies of second messenger effects or. CRH and AVP binding provided more clues about mechanisms of action. Finally, a new cell type that changed to a multihormonal corticothyrotrope under the influence of AVP was described. It could provide an efficient means of stimulating both the thyroid and the adrenal. Studies implicating locally produced epidermal growth factor (EGF) as another modulatory agent in the stress response were also done because EGF stimulates corticotropes. Cells producing EGF increased expression of EGF mRNA after exposure to cold. In summary, these studies identified four critical hormones that interact with each other in the control of pituitary responses to cold. These included: AVP CRH, throtropin in releasing hormone (TRH) and epidermal growth factor. corticotropes, thyrotropes, cold stress, anterior pituitary, paraventricular nucleus , arginine vasopressin, adrenocorticotropin, Corticotropin releasing hormone, in situ hybridization, immunocytochemistry, image analysis, RIA, rats.

Published
1992

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