scholarly journals 219.Analysis of ovarian macrophage populations using macrophage-specific green fluorescent protein (GFP) transgenic mice

2004 ◽  
Vol 16 (9) ◽  
pp. 219
Author(s):  
C. E. Haynes ◽  
R. J. Norman ◽  
R. L. Robker
Keyword(s):  
Transgenic Mice ◽  
Hormone Receptors ◽  
Ovarian Function ◽  
Fluorescent Protein ◽  
Immune Cell ◽  
Ovarian Tissue ◽  
Gene Promoter ◽  
Peritoneal Macrophages ◽  
Reproductive Tissues ◽  
Green Fluorescent

Macrophages represent a major immune cell type in reproductive tissues and are thought to regulate multiple aspects of reproduction, including ovarian function. We have previously shown distinctive phenotypes and functions of ovarian macrophages such that many immunological mediators, such as cytokines and hormone receptors, are uniquely regulated within these cells across the oestrus cycle. In order to isolate macrophages from ovarian tissue by fluorescence activated cell sorting (FACS), we acquired transgenic mice (from DA Hume, Institute for Molecular Bioscience, University of Queensland) which express GFP exclusively in macrophages (1). In these mice GFP is expressed under direction of the c-fms gene promoter, which encodes the receptor for colony-stimulating factor-1 (CSF-1R), a major macrophage growth factor. Using flow cytometry we confirmed that 95% of peritoneal macrophages express GFP and 88% co-express GFP and the classical macrophage marker F4/80. The distribution of GFP+ macrophages in tissues was co-localized with macrophage markers F4/80 and major histocompatibility complex class II (MHCII) by immunohistochemistry using phycoerythrin (PE)-labelled antibodies. The liver, uterus and oviduct exhibited many GFP+ cells in characteristic macrophage distributions. Furthermore, GFP fluorescence was tightly co-localized with PE fluorescence of either F4/80 or MHCII, indicating that CSF-1R is expressed in the macrophages of these tissues. In contrast, macrophages in the ovary were positive for F4/80 and MHCII, but rarely expressed GFP. Thus unlike macrophages of other reproductive tissues, ovarian macrophages do not consistently express CSF-1R. In ovaries from gonadotrophin-primed immature females, GFP was not expressed in macrophages (F4/80+/MHCII+) surrounding follicles but was detected in macrophages within the regressing corpus luteum. Thus CSF-1R is a hormonally regulated gene, expressed only in specific subsets of ovarian macrophages suggesting that CSF-1 controls functional activities of ovarian macrophages at specific stages of the ovarian cycle. (1) Sasmono, R. T., et al. (2003) Blood 101, 1155–1163.

Transgenic mice with green fluorescent protein-labeled pancreatic β-cells

2003 ◽  
Vol 284 (1) ◽  
pp. E177-E183 ◽  
Author(s):  
Manami Hara ◽  
Xiaoyu Wang ◽  
Toshihiko Kawamura ◽  
Vytas P. Bindokas ◽  
Restituto F. Dizon ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Transgenic Mice ◽  
Cell Biology ◽  
Cell Function ◽  
Fluorescent Protein ◽  
Gene Promoter ◽  
Β Cells ◽  
Β Cell ◽  
Β Cell Function ◽  
Green Fluorescent

We have generated transgenic mice that express green fluorescent protein (GFP) under the control of the mouse insulin I gene promoter (MIP). The MIP-GFP mice develop normally and are indistinguishable from control animals with respect to glucose tolerance and pancreatic insulin content. Histological studies showed that the MIP-GFP mice had normal islet architecture with coexpression of insulin and GFP in the β-cells of all islets. We observed GFP expression in islets from embryonic day E13.5 through adulthood. Studies of β-cell function revealed no difference in glucose-induced intracellular calcium mobilization between islets from transgenic and control animals. We prepared single-cell suspensions from both isolated islets and whole pancreas from MIP-GFP-transgenic mice and sorted the β-cells by fluorescence-activated cell sorting based on their green fluorescence. These studies showed that 2.4 ± 0.2% ( n = 6) of the cells in the pancreas of newborn (P1) and 0.9 ± 0.1% ( n = 5) of 8-wk-old mice were β-cells. The MIP-GFP-transgenic mouse may be a useful tool for studying β-cell biology in normal and diabetic animals.

283. Regulated expression of the c-fms receptor characterises distinct ovarian macrophage populations

2005 ◽  
Vol 17 (9) ◽  
pp. 118
Author(s):  
K. H. Van der Hoek ◽  
C. E. Minge ◽  
R. J. Norman ◽  
R. L. Robker
Keyword(s):  
Cell Biology ◽  
Ovarian Function ◽  
Fluorescent Protein ◽  
Immune Cell ◽  
Inflammatory Responses ◽  
Gene Promoter ◽  
Specific Gene ◽  
Corpora Lutea ◽  
Regulated Expression ◽  
Classically Activated

Macrophages represent a major immune cell type in reproductive tissues and are thought to regulate multiple aspects of reproduction, including ovarian function. We have previously shown a distinctive phenotype of ovarian thecal macrophages present around the preovulatory follicle, such that secreted cytokines are uniquely regulated within these cells across the oestrus cycle. C-fms is a macrophage-specific gene that encodes the receptor for colony-stimulating factor-1 (CSF-1), and that regulates macrophage proliferation, differentiation and migration, as well as pro-inflammatory responses.1,2 We acquired transgenic mice (from DA Hume, Institute for Molecular Bioscience, University of Queensland) that express green fluorescent protein (GFP) exclusively in macrophages under direction of the c-fms gene promoter.3 In ovaries from these animals we have previously reported that macrophages constitutively positive for macrophage markers, F4/80 and MHCII, exhibited spatially regulated expression of GFP (c-fms); being GFP+ within the stroma surrounding small follicles, particularly atretic follicles, but GFP– in theca surrounding preovulatory follicles and healthy corpora lutea (CL), further reinforcing the concept that these macrophages are not classically activated but have a unique resident phenotype. Further examination of the GFP+ ovarian macrophage population has revealed that the highest levels of GFP expression were in macrophages associated with TUNEL+ regressing CL and, even though CSF-1 typically induces proliferation, the GFP+ macrophages within the regressing CL did not incorporate BrdU label nor express cyclin D1. This indicates that in the murine ovary c-fms expression may not regulate ovarian macrophage proliferation or migration but more likely represents a subset of classically activated ovarian macrophages that are actively differentiating or phagocytically active. (1)Fixe P and Praloran V (1998) Cytokine 10, 32–37.(2)Pixley FJ and Stanley ER (2004) Trends in Cell Biology 14, 628–38.(3)Sasmono RT et al. (2003) Blood 101, 1155–63.

High level Purkinje cell specific expression of green fluorescent protein in transgenic mice

2001 ◽  
Vol 115 (6) ◽  
pp. 455-464 ◽  
Author(s):  
Xulun Zhang ◽  
Stephan L. Baader ◽  
Feng Bian ◽  
Wolfgang Müller ◽  
John Oberdick
Keyword(s):  
Green Fluorescent Protein ◽  
Transgenic Mice ◽  
Purkinje Cell ◽  
Fluorescent Protein ◽  
Specific Expression ◽  
Green Fluorescent ◽  
High Level

scholarly journals Uterine Cells Improved Ovarian Function in a Murine Model of Ovarian Insufficiency

2019 ◽  
Vol 26 (12) ◽  
pp. 1633-1639 ◽  
Author(s):  
Andres Reig ◽  
Ramanaiah Mamillapalli ◽  
Alexis Coolidge ◽  
Joshua Johnson ◽  
Hugh S. Taylor
Keyword(s):  
Stem Cells ◽  
Young Women ◽  
Murine Model ◽  
Ovarian Function ◽  
Fluorescent Protein ◽  
Primary Ovarian Insufficiency ◽  
Age Group ◽  
Ovarian Dysfunction ◽  
Ovarian Insufficiency ◽  
Green Fluorescent

Primary ovarian insufficiency (POI) is defined as ovarian dysfunction in women younger than 40 years. It affects 1% of the women in this age-group and can occur iatrogenically after chemotherapy. Stem cells have been used in attempt to restore ovarian function in POI. In particular, endometrial mesenchymal stem cells (eMSCs) are easily obtainable in humans and have shown great potential for regenerative medicine. Here, we studied the potential for uterine cell (UC) suspensions containing eMSCs to improve ovarian function in a murine model of chemotherapy-induced POI. Green fluorescent protein (GFP)-labeled UC or phosphate-buffered solution (PBS) was delivered intravenously after chemotherapy. There was a significant increase in oocytes production and serum anti-Müllerian hormone concentrations after 6 weeks, as well as a 19% higher body mass in UC-treated mice. Similarly, we observed an increased number of pups in mice treated with UC than in mice treated with PBS. None of the oocytes or pups incorporated GFP, suggesting that there was no contribution of these stem cells to the oocyte pool. We conclude that treatment with UC indirectly improved ovarian function in mice with chemotherapy-induced POI. Furthermore, our study suggests that endometrial stem cell therapy may be beneficial to young women who undergo ovotoxic chemotherapy.

BAC transgenic mice express enhanced green fluorescent protein in central and peripheral cholinergic neurons

2006 ◽  
Vol 27 (3) ◽  
pp. 391-397 ◽  
Author(s):  
Yvonne N. Tallini ◽  
Bo Shui ◽  
Kai Su Greene ◽  
Ke-Yu Deng ◽  
Robert Doran ◽  
...  
Keyword(s):  
Nervous System ◽  
Green Fluorescent Protein ◽  
Transgenic Mice ◽  
Peripheral Nervous System ◽  
Enhanced Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Cholinergic Neurons ◽  
Autonomic Nerves ◽  
Organ Systems ◽  
Green Fluorescent

The peripheral nervous system has complex and intricate ramifications throughout many target organ systems. To date this system has not been effectively labeled by genetic markers, due largely to inadequate transcriptional specification by minimum promoter constructs. Here we describe transgenic mice in which enhanced green fluorescent protein (eGFP) is expressed under the control of endogenous choline acetyltransferase (ChAT) transcriptional regulatory elements, by knock-in of eGFP within a bacterial artificial chromosome (BAC) spanning the ChAT locus and expression of this construct as a transgene. eGFP is expressed in ChATBAC-eGFP mice in central and peripheral cholinergic neurons, including cell bodies and processes of the somatic motor, somatic sensory, and parasympathetic nervous system in gastrointestinal, respiratory, urogenital, cardiovascular, and other peripheral organ systems. Individual epithelial cells and a subset of lymphocytes within the gastrointestinal and airway mucosa are also labeled, indicating genetic evidence of acetylcholine biosynthesis. Central and peripheral neurons were observed as early as 10.5 days postcoitus in the developing mouse embryo. ChATBAC-eGFP mice allow excellent visualization of all cholinergic elements of the peripheral nervous system, including the submucosal enteric plexus, preganglionic autonomic nerves, and skeletal, cardiac, and smooth muscle neuromuscular junctions. These mice should be useful for in vivo studies of cholinergic neurotransmission and neuromuscular coupling. Moreover, this genetic strategy allows the selective expression and conditional inactivation of genes of interest in cholinergic nerves of the central nervous system and peripheral nervous system.

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