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https://www.readbyqxmd.com/read/28100033/relaxin-3-inputs-target-hippocampal-interneurons-and-deletion-of-hilar-relaxin-3-receptors-in-floxed-rxfp3-mice-impairs-spatial-memory
#1
M Haidar, G Guèvremont, C Zhang, Rad Bathgate, E Timofeeva, C M Smith, A L Gundlach
Hippocampus is innervated by γ-aminobutyric acid (GABA) 'projection' neurons of the nucleus incertus (NI), including a population expressing the neuropeptide, relaxin-3 (RLN3). In studies aimed at gaining an understanding of the role of RLN3 signaling in hippocampus via its Gi/o -protein-coupled receptor, RXFP3, we examined the distribution of RLN3-immunoreactive nerve fibres and RXFP3 mRNA-positive neurons in relation to hippocampal GABA neuron populations. RLN3-positive elements were detected in close-apposition with a substantial population of somatostatin (SST)- and GABA-immunoreactive neurons, and a smaller population of parvalbumin- and calretinin-immunoreactive neurons in different hippocampal areas, consistent with the relative distribution patterns of RXFP3 mRNA and these marker transcripts...
January 18, 2017: Hippocampus
https://www.readbyqxmd.com/read/28098199/optogenetic-modulation-of-urinary-bladder-contraction-for-lower-urinary-tract-dysfunction
#2
Jae Hong Park, Jin Ki Hong, Ja Yun Jang, Jieun An, Kyu-Sung Lee, Tong Mook Kang, Hyun Joon Shin, Jun-Kyo Francis Suh
As current clinical approaches for lower urinary tract (LUT) dysfunction such as pharmacological and electrical stimulation treatments lack target specificity, thus resulting in suboptimal outcomes with various side effects, a better treatment modality with spatial and temporal target-specificity is necessary. In this study, we delivered optogenetic membrane proteins, such as channelrhodopsin-2 (ChR2) and halorhodopsin (NpHR), to bladder smooth muscle cells (SMCs) of mice using either the Cre-loxp transgenic system or a viral transfection method...
January 18, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28090567/the-rna-polymerase-iii-subunit-polr3b-is-required-for-the-maintenance-of-small-intestinal-crypts-in-mice
#3
Julia Kieckhaefer, Sabina Lukovac, Diana Z Ye, Dolim Lee, Danielle J Beetler, Michael Pack, Klaus H Kaestner
BACKGROUND & AIMS: The continuously self-renewing mammalian intestinal epithelium, with high cellular turnover, depends on adequate protein synthesis for its proliferative capacity. RNA polymerase III activity is closely related to cellular growth and proliferation. Here, we studied the role of Polr3b, a large RNA polymerase III subunit, in the mammalian intestinal epithelium. METHODS: We derived mice with an intestinal epithelium-specific hypomorphic mutation of the Polr3b gene, using VillinCre-mediated gene ablation...
November 2016: Cellular and Molecular Gastroenterology and Hepatology
https://www.readbyqxmd.com/read/28078648/generation-of-a-floxed-allele-of-the-mouse-microrna-200-clusters
#4
Hongyan Ren, Cong Tao, Kui Li, Yanzhen Bi, Xinmin Zheng
MicroRNA-8 and its target gene, u-shaped (ush), regulate body size in Drosophila. As the mouse homolog of the Drosophila miR-8, whether the miR-200 family has similar functions and how they perform their regulatory roles in body size control is unknown. In order to discover the biological function of the miR-200s in vivo, we generated mice lacking the miR-200b/200a/429 gene cluster using the Cre/loxp system. miR-200b/200a/429 null mutant mice were viable, fertile, and showed normal development.
January 11, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/28076846/effect-of-trp53-gene-deficiency-on-brain-injury-after-neonatal-hypoxia-ischemia
#5
Ana A Baburamani, Kristina S Sobotka, Regina Vontell, Carina Mallard, Veena G Supramaniam, Claire Thornton, Henrik Hagberg
Hypoxia-ischemia (HI) can result in permanent life-long injuries such as motor and cognitive deficits. In response to cellular stressors such as hypoxia, tumor suppressor protein p53 is activated, potently initiating apoptosis and promoting Bax-dependent mitochondrial outer membrane permeabilization. The aim of this study was to investigate the effect of Trp53 genetic inhibition on injury development in the immature brain following HI. HI (50 min or 60 min) was induced at postnatal day 9 (PND9) in Trp53 heterozygote (het) and wild type (WT) mice...
January 5, 2017: Oncotarget
https://www.readbyqxmd.com/read/28063344/reducing-persistent-polyomavirus-infection-increases-functionality-of-virus-specific-memory-cd8-t-cells
#6
Qingsong Qin, Matthew Lauver, Saumya Maru, Eugene Lin, Aron E Lukacher
Mouse polyomavirus (MuPyV) causes a smoldering persistent infection in immunocompetent mice. To lower MuPyV infection in acutely and persistently infected mice, and study the impact of a temporal reduction in viral loads on the memory CD8 T cell response, we created a recombinant MuPyV in which a loxP sequence was inserted into the A2 strain genome upstream of the early promoter and another loxP sequence was inserted in cis into the intron shared by all three T antigens. Using mice transgenic for tamoxifen-inducible Cre recombinase, we demonstrated that reduction in MuPyV load during persistent infection was associated with differentiation of virus-specific CD8 T cells having a superior recall response...
January 4, 2017: Virology
https://www.readbyqxmd.com/read/28060852/contribution-of-vascular-cells-to-neointimal-formation
#7
Falei Yuan, Dong Wang, Kang Xu, Jixian Wang, Zhijun Zhang, Li Yang, Guo-Yuan Yang, Song Li
The de-differentiation and proliferation of smooth muscle cells (SMCs) are widely accepted as the major contributor to vascular remodeling. However, recent studies indicate that vascular stem cells (VSCs) also play an important role, but their relative contribution remains to be elucidated. In this study, we used genetic lineage tracing approach to further investigate the contribution of SMCs and VSCs to neointimal thickening in response to endothelium denudation injury or artery ligation. In vitro and in vivo analysis of MYH11-cre/Rosa-loxP-RFP mouse artery showed that SMCs proliferated at a much slower rate than non-SMCs...
2017: PloS One
https://www.readbyqxmd.com/read/28057770/neuroendocrine-integration-of-nutritional-signals-on-reproduction
#8
Maggie Corr Evans, Greg Anderson
Reproductive function in mammals is energetically costly and therefore tightly regulated by nutritional status. To enable this integration of metabolic and reproductive function, information regarding peripheral nutritional status must be relayed centrally to the gonadotropin-releasing hormone (GnRH) neurons that drive reproductive function. The metabolically relevant hormones leptin, insulin and ghrelin have been identified as key mediators of this 'metabolic control of fertility'. However, the neural circuitry through which they act to exert their control over GnRH drive remains incompletely understood...
January 5, 2017: Journal of Molecular Endocrinology
https://www.readbyqxmd.com/read/28053125/efficient-mapping-of-transgene-integration-sites-and-local-structural-changes-in-cre-transgenic-mice-using-targeted-locus-amplification
#9
Carol Cain-Hom, Erik Splinter, Max van Min, Marieke Simonis, Monique van de Heijning, Maria Martinez, Vida Asghari, J Colin Cox, Søren Warming
Cre/LoxP technology is widely used in the field of mouse genetics for spatial and/or temporal regulation of gene function. For Cre lines generated via pronuclear microinjection of a Cre transgene construct, the integration site is random and in most cases not known. Integration of a transgene can disrupt an endogenous gene, potentially interfering with interpretation of the phenotype. In addition, knowledge of where the transgene is integrated is important for planning of crosses between animals carrying a conditional allele and a given Cre allele in case the alleles are on the same chromosome...
January 3, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28052921/the-circadian-oscillator-of-the-cerebral-cortex-molecular-biochemical-and-behavioral-effects-of-deleting-the-arntl-clock-gene-in-cortical-neurons
#10
Tenna Bering, Mikkel Bloss Carstensen, Gitta Wörtwein, Pia Weikop, Martin Fredensborg Rath
A molecular circadian oscillator resides in neurons of the cerebral cortex, but its role is unknown. Using the Cre-LoxP method, we have here abolished the core clock gene Arntl in those neurons. This mouse represents the first model carrying a deletion of a circadian clock component specifically in an extrahypothalamic cell type of the brain. Molecular analyses of clock gene expression in the cerebral cortex of the Arntl conditional knockout mouse revealed disrupted circadian expression profiles, whereas clock gene expression in the suprachiasmatic nucleus was still rhythmic, thus showing that Arntl is required for normal function of the cortical circadian oscillator...
January 3, 2017: Cerebral Cortex
https://www.readbyqxmd.com/read/28049473/enhanced-heterologous-protein-productivity-by-genome-reduction-in-lactococcus-lactis-nz9000
#11
Duolong Zhu, Yuxin Fu, Fulu Liu, Haijin Xu, Per Erik Joakim Saris, Mingqiang Qiao
BACKGROUND: The implementation of novel chassis organisms to be used as microbial cell factories in industrial applications is an intensive research field. Lactococcus lactis, which is one of the most extensively studied model organisms, exhibits superior ability to be used as engineered host for fermentation of desirable products. However, few studies have reported about genome reduction of L. lactis as a clean background for functional genomic studies and a model chassis for desirable product fermentation...
January 3, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28029195/expression-of-tamoxifen-inducible-cre-recombinase-in-lcn5-creer-t2-transgenic-mouse-caput-epididymis
#12
Juan Xu, Guangxin Yao, Yanfei Ru, Shengsong Xie
The epididymis, which connects the testis to vas deferens, plays a crucial role regulating sperm maturation and fertilization. Here, a tamoxifen-inducible CreER(T2) recombinase transgenic mouse was generated to study the function of genes in the caput epididymis using the Cre/LoxP system, which is driven by the 1.8-kb Lcn5 promoter (Lcn5-CreER(T2) ). Both CRE recombinase and ER(T2) mRNA were specifically expressed in the caput epididymis, beginning at postnatal Day 30 and increasing thereafter. Crossing these Lcn5-CreER(T2) transgenic mice with Rosa26;mT/mG reporter mice, which express membrane-bound GFP (mGFP) only after CRE is active at its genetic locus, resulted in the presence of GFP only in the middle/distal caput epididymis after tamoxifen induction...
December 28, 2016: Molecular Reproduction and Development
https://www.readbyqxmd.com/read/28024880/embryonic-ablation-of-neuronal-vgf-increases-energy-expenditure-and-reduces-body-weight
#13
Cheng Jiang, Wei-Jye Lin, Masato Sadahiro, Andrew C Shin, Christoph Buettner, Stephen R Salton
Germline ablation of VGF, a secreted neuronal, neuroendocrine, and endocrine peptide precursor, results in lean, hypermetabolic, and infertile adult mice that are resistant to diet-, lesion-, and genetically-induced obesity and diabetes (Hahm et al., 1999, 2002). To assess whether this phenotype is predominantly driven by reduced VGF expression in developing and/or adult neurons, or in peripheral endocrine and neuroendocrine tissues, we generated and analyzed conditional VGF knockout mice, obtained by mating loxP-flanked (floxed) Vgf mice with either pan-neuronal Synapsin-Cre- or forebrain alpha-CaMKII-Cre-recombinase-expressing transgenic mice...
December 20, 2016: Neuropeptides
https://www.readbyqxmd.com/read/28011173/neuronal-expression-of-a-thyroid-hormone-receptor-%C3%AE-mutation-alters-mouse-behaviour
#14
S Richard, N Aguilera, M Thévenet, O Dkhissi-Benyahya, F Flamant
In humans, alterations in thyroid hormone signalling are associated with mood and anxiety disorders, but the neural mechanisms underlying such association are poorly understood. The present study investigates the involvement of neuronal thyroid hormone receptor α (TRα) in anxiety, using mouse genetics and Cre/loxP technology to specifically alter TRα signalling in neurons. We evaluated the behaviour of mice expressing a dominant negative, neuron-specific mutation of TRα (TRα(AMI)/Cre3 mice), using the elevated-plus maze, light-dark box and open-field tests...
December 21, 2016: Behavioural Brain Research
https://www.readbyqxmd.com/read/28011085/self-excising-cre-mutant-lox-marker-recycling-system-for-multiple-gene-integrations-and-consecutive-gene-deletions-in-aspergillus-oryzae
#15
Silai Zhang, Akihiko Ban, Naoki Ebara, Osamu Mizutani, Mizuki Tanaka, Takahiro Shintani, Katsuya Gomi
In this study, we developed a self-excising Cre/loxP-mediated marker recycling system with mutated lox sequences to introduce a number of biosynthetic genes into Aspergillus oryzae. To construct the self-excising marker cassette, both the selectable marker, the Aspergillus nidulans adeA gene, and the Cre recombinase gene (cre), conditionally expressed by the xylanase-encoding gene promoter, were designed to be located between the mutant lox sequences, lox66 and lox71. However, construction of the plasmid failed, possibly owing to a slight expression of cre downstream of the fungal gene promoter in Escherichia coli...
December 20, 2016: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/27999416/dermal-delivery-of-constructs-encoding-cre-recombinase-to-induce-skin-tumors-in-pten-loxp-loxp-braf-ca-mice
#16
Marcel A Deken, Ji-Ying Song, Jules Gadiot, Adriaan D Bins, Paula Kroon, Inge Verbrugge, Christian U Blank
Current genetically-engineered mouse melanoma models are often based on Tyr::CreER(T2)-controlled MAPK pathway activation by the BRAF(V600E) mutation and PI3K pathway activation by loss of PTEN. The major drawback of these models is the occurrence of spontaneous tumors caused by leakiness of the Tyr::CreER(T2) system, hampering long-term experiments. To address this problem, we investigated several approaches to optimally provide local delivery of Cre recombinase, including injection of lentiviral particles, DNA tattoo administration and particle-mediated gene transfer, to induce melanomas in Pten(LoxP/LoxP);Braf(CA/+) mice lacking the Tyr::CreER(T2) allele...
December 20, 2016: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/27979828/conditional-genetic-deletion-of-ano1-in-interstitial-cells-of-cajal-impairs-ca2-transients-and-slow-waves-in-adult-mouse-small-intestine
#17
John Malysz, Simon J Gibbons, Siva A Saravanaperumal, Peng Du, Seth T Eisenman, Chike Cao, Uhtaek Oh, Dieter Saur, Sabine Klein, Tamas Ordog, Gianrico Farrugia
Myenteric plexus interstitial cells of Cajal (ICC-MY) in the small intestine are Kit(+) electrical pacemakers that express the Ano1/TMEM16A Ca(2+)-activated Cl(-) channel, whose functions in the GI tract remain incompletely understood. In this study, an inducible Cre-LoxP-based approach was used to advance the understanding of Ano1 in ICC-MY of adult mouse small intestine. Kit(CreERT2/+);Ano1(Fl/Fl) mice were treated with tamoxifen or vehicle, and small intestines (mucosa-free) examined. Quantitative RT-PCR demonstrated ~50% reduction in Ano1 mRNA in intestines of conditional knock-outs (cKOs) compared to vehicle-treated controls...
December 15, 2016: American Journal of Physiology. Gastrointestinal and Liver Physiology
https://www.readbyqxmd.com/read/27976675/cyst-wall-protein-1-is-fundamental-for-golgi-like-organelle-neogenesis-and-cyst-wall-biosynthesis-in-giardia-lamblia
#18
Jacqueline A Ebneter, Sally D Heusser, Elisabeth M Schraner, Adrian B Hehl, Carmen Faso
The genome of the protozoan parasite Giardia lamblia is organized in two diploid nuclei, which has so far precluded complete analysis of gene function. Here we use a previously developed Cre/loxP-based knock-out and selection marker salvage strategy in the human-derived isolate WB-C6 to eliminate all four copies of the Cyst-Wall-Protein-1 locus (CWP1). Because these loci are silenced in proliferating trophozoites and highly expressed only in encysting cells, CWP1 ablation allows functional characterization of a conditional phenotype in parasites induced to encyst...
December 15, 2016: Nature Communications
https://www.readbyqxmd.com/read/27943405/n-ice-plasmids-for-generating-n-terminal-3%C3%A2-%C3%A3-%C3%A2-flag-tagged-genes-that-allow-inducible-constitutive-or-endogenous-expression-in-saccharomyces-cerevisiae
#19
Yueping Zhang, Nina D Serratore, Scott D Briggs
PCR-mediated homologous recombination is a powerful approach to introduce epitope tags into the chromosomal loci at the N-terminus or the C-terminus of targeted genes. Although strategies of C-terminal epitope tagging of target genes at their loci are simple and widely used in yeast, C-terminal epitope tagging is not practical for all proteins. For example, a C-terminal tag may affect protein function or a protein may get cleaved or processed resulting in the loss of the epitope tag. Therefore, N-terminal epitope tagging may be necessary to resolve these problems...
December 12, 2016: Yeast
https://www.readbyqxmd.com/read/27943201/monosynaptic-tracing-in-developing-circuits-using-modified-rabies-virus
#20
Laura Cocas, Gloria Fernandez
An attenuated rabies virus that expresses fluorescent protein has made it possible to analyze retrograde (presynaptic) monosynaptic connections in vivo. By combining attenuated rabies virus with a Cre-loxP based system to target cells in a subtype-specific fashion, it is possible to examine neuronal input in vivo onto any class of neuron, in development and in the mature brain. We describe here the methods to amplify deletion mutant, pseudotyped rabies virus, selectively target cells of interest using genetic and viral approaches, as well as the stereotaxic procedures required to target neuronal subtypes of interest in vivo...
2017: Methods in Molecular Biology
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