3.Commonly used cre transgenic mice and their applications in hematopoietic system.
Lu-Yun PENG ; Tao CHENG ; Wei-Ping YUAN
Journal of Experimental Hematology 2014;22(5):1442-1447
Cre-lox recombination system consists of two elements: Cre recombinase enzyme and lox sites. Cre recombinase can recombine the lox site sequences by specifically detecting and cutting them. The direction and position of lox sites determine the functional effects of Cre enzyme such as deletion, inversion or chromosomal translocation. The hematopoietic system of mouse consists of multi-lineages and various developmental stage hematopoietic cells that are differentiated from hematopoietic stem cells (hematopoietic stem cells, HSC). The hematopoietic stem cells are maintained in the bone marrow microenvironment (niche). Currently, a variety of floxed conditional-knockout mice, recognized by Cre-lox recombination system, are used for the study of the hematopoietic system. This review summarizes the commonly used Cre transgenic mice and their applications in the study of hematopoietic system.
Animals
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Hematopoietic Stem Cells
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cytology
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metabolism
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Integrases
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Mice
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Mice, Transgenic
4.Correlation Analysis of Molecularly-Defined Cortical Interneuron Populations with Morpho-Electric Properties in Layer V of Mouse Neocortex.
Jun-Wei CAO ; Xiao-Yi MAO ; Liang ZHU ; Zhi-Shuo ZHOU ; Shao-Na JIANG ; Lin-Yun LIU ; Shu-Qing ZHANG ; Yinghui FU ; Wen-Dong XU ; Yong-Chun YU
Neuroscience Bulletin 2023;39(7):1069-1086
Cortical interneurons can be categorized into distinct populations based on multiple modalities, including molecular signatures and morpho-electrical (M/E) properties. Recently, many transcriptomic signatures based on single-cell RNA-seq have been identified in cortical interneurons. However, whether different interneuron populations defined by transcriptomic signature expressions correspond to distinct M/E subtypes is still unknown. Here, we applied the Patch-PCR approach to simultaneously obtain the M/E properties and messenger RNA (mRNA) expression of >600 interneurons in layer V of the mouse somatosensory cortex (S1). Subsequently, we identified 11 M/E subtypes, 9 neurochemical cell populations (NCs), and 20 transcriptomic cell populations (TCs) in this cortical lamina. Further analysis revealed that cells in many NCs and TCs comprised several M/E types and were difficult to clearly distinguish morpho-electrically. A similar analysis of layer V interneurons of mouse primary visual cortex (V1) and motor cortex (M1) gave results largely comparable to S1. Comparison between S1, V1, and M1 suggested that, compared to V1, S1 interneurons were morpho-electrically more similar to M1. Our study reveals the presence of substantial M/E variations in cortical interneuron populations defined by molecular expression.
Mice
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Animals
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Neocortex/physiology*
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Mice, Transgenic
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Interneurons/physiology*
5.Transgenic mice produced by intracytoplasmic sperm injection.
Ming LI ; A-Yong YAN ; Hui YAO ; Xiao-Rong AN ; Ke-Mian GOU
Chinese Journal of Biotechnology 2006;22(5):784-788
In our previous study, normal and fertile mice were successful produced from oocytes following intracytoplasmic sperm injection (ICSI). In the present study, the possibility of producing transgenic embryos and offspring with this procedure was evaluated. After freezing-thawed once using HEPES-CZB medium without cryoprotectants, the cauda sperm from KM fertile male were exposed to the circular or linear pEGFP-N1 DNA for 1 min and then co-injected into metaphase II oocytes of B6D2F1 strain. When the zygotes with two pronuclei were cultured in CZB medium to day 3.5, 39.1% (9/23) of them, derived from oocytes co-injected with sperm head and pEGFP-N1 plasmid DNA, were expressed GFP protein. After transfer of the ICSI embryos with two pronuclei from co-injection of sperm head and foreign DNA, seven recipients delivered 30 pups (23.8%, 30/126). Southern blot results revealed that three of sixteen offspring integrated with GFP and neomycin genes together (18.8 %). Interestingly, all of them were produced from oocytes co-injected sperm head and linear DNA (33.3%, 3/9), while none of seven ICSI offspring integrated either GFP or neomycin gene in the group of co-injection of sperm head and circular plasmid DNA. These results indicated that the high efficiency of transgenic mouse could be produced by ICSI. It may be shown that linear DNA is more easily to integrate into host genome than circular DNA when ICSI was used to produce transgenic animals.
Animals
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Male
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Mice
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Mice, Inbred C57BL
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Mice, Inbred DBA
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Mice, Transgenic
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genetics
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Sperm Injections, Intracytoplasmic
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methods
6.Reproductive performance of genetically engineered mice housed in different housing systems.
Shikha YADAV ; Inderjeet YADAV ; Kunal PRATAP ; Pradeep Kumar TIWARI ; Vijay Pal SINGH
Laboratory Animal Research 2017;33(2):68-75
The genetically engineered mice require special husbandry care and are mainly housed in Individually Ventilated Cage (IVC) systems and Static Micro Isolator Cages (SMIC) to minimize the risk for spreading undesirable microorganisms. However, the static micro isolation cage housing like SMIC are being replaced with IVC systems in many facilities due to a number of benefits like a higher density housing in limited space, better protection from biohazards and allergens and decreased work load due to decreased frequency of cage changing required in this system. The purpose of this study was to examine the reproductive performance of genetically engineered mice housed in individually ventilated cages (IVC) and Static Micro Isolator Cages (SMIC). When the B6C3-Tg (APPswe, PSEN1dE9) 85Dbo/Mmjax transgenic mice were housed in these two housing systems, the number of litters per dam, number of pups born per dam and number of pups weaned per dam were found to be slightly higher in the IVC as compared to the SMIC but the difference was not significant (P<0.05). In case of Growth Associated Protein 43 (GAP-43) knockout mice, the number of litters born per dam and the number of pups born per dam were marginally higher in the IVC as compared to those housed in SMIC but the difference was not significant (P<0.05). Only the number of pups weaned per dam were found to be significantly higher as compared to those housed in the SMIC system at P<0.05.
Allergens
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Animals
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GAP-43 Protein
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Hazardous Substances
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Housing*
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Mice*
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Mice, Knockout
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Mice, Transgenic
7.Breeding of transgenic mice expressing human tau isoform with P301L mutation and identification of homozygous transgenic mice.
Yan-yan WANG ; Ru-zhui CHEN ; Xiao-nani ZHU ; Jing LIU ; Zhi-hui LI ; Xiu-juan LIU ; Zhi-hui LI ; Xin NA ; Shan-shan LIANG ; Guo-guang QIU ; Wei ZHANG ; Hai WANG ; Xue-lan WANG
Chinese Journal of Applied Physiology 2012;28(3):221-224
OBJECTIVETo establish homozygous transgenic mouse strain expressing human tau isoform with P301L mutation.
METHODSFive transgenic mice expressing human tau isoform with P301L mutation were obtained by microinjection into male nuclei. Homozygote and hemizygote were identified by PCR and real-time fluorescent quantitative PCR.
RESULTSNinety five homozygous transgenic mice were selected, and the results indicated that homozygous transgenic mice were superior to hemizygote in simulating the changes of biological characteristics.
CONCLUSIONExogenous gene tau is able to stably transmit to next generation and the combination of SYBR Green real-time fluorescent quantitative PCR with the traditional mating is a fast, reliable and economical way to screen homozygous and hemizygous transgenic mice.
Animals ; Female ; Homozygote ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Microinjections ; Mutation ; tau Proteins ; genetics
8.Chronic Exposure to Hypoxia Inhibits Myelinogenesis and Causes Motor Coordination Deficits in Adult Mice.
Lin CHEN ; Shu-Yu REN ; Rui-Xue LI ; Kun LIU ; Jing-Fei CHEN ; Yu-Jian YANG ; Yong-Bin DENG ; Han-Zhi WANG ; Lan XIAO ; Feng MEI ; Fei WANG
Neuroscience Bulletin 2021;37(10):1397-1411
Exposure to chronic hypoxia is considered to be a risk factor for deficits in brain function in adults, but the underlying mechanisms remain largely unknown. Since active myelinogenesis persists in the adult central nervous system, here we aimed to investigate the impact of chronic hypoxia on myelination and the related functional consequences in adult mice. Using a transgenic approach to label newly-generated myelin sheaths (NG2-CreER
Animals
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Clemastine
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Hypoxia/complications*
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Mice
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Mice, Inbred C57BL
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Mice, Transgenic
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Myelin Sheath
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Oligodendroglia
9.Effect of Glomerular Cyclooxygenase-2 Overexpression on Podocyte Injury Induced by Puromycin in Mice.
Korean Journal of Nephrology 2007;26(2):127-136
PURPOSE: The aim of this study is to investigate the effect of glomerular cyclooxygenase-2 (COX-2) overexpression on podocyte injury by puromycin aminonucleoside (PAN) in nephrin-driven COX-2 transgenic mice. METHODS: We administrated PAN intravenously on day-1 (15 mg/100 g body weight) and day-3 (30 mg/100 g body weight) in wild type (male B6/D2 mice) and COX-2 transgenic mice. An additional group received a selective COX-2 inhibitor (SC58236) with PAN. The animals from each group were sacrificed at the end of day-3 and 10. We investigated albuminuria, foot process effacement and glomerular COX-2 and nephrin expression. RESULTS: PAN induced albuminuria only in COX-2 transgenic mice, with the peak on day-3. Selective COX-2 inhibition significantly reduced albuminuria. EM examination demonstrated foot process effacement, which was improved partially by selective COX-2 inhibition, in COX-2 transgenic mice treated with PAN on day-3. Glomerular COX-2 expression increased significantly on day-3 in COX-2 transgenic mice treated with PAN, whereas expression of nephrin showed a tendency to decrease on day-3. These changes of expression of COX-2 and nephrin were partly attenuated by selective COX- 2 inhibition. Unlike COX-2 transgenic mice, wild-type mice did not show any changes even after PAN treatment. CONCLUSION: In COX-2 transgenic mice, PAN induced albuminuria, podocyte injury and up-regulation of glomerular COX-2, which were ameliorated by selective COX-2 inhibitor. These results suggest that COX-2 may play an important role in increasing susceptibility of podocytes to injury and selective COX-2 inhibition may ameliorate podocyte injury.
Albuminuria
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Animals
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Cyclooxygenase 2*
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Foot
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Mice*
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Mice, Transgenic
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Podocytes*
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Puromycin Aminonucleoside
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Puromycin*
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Up-Regulation
10.Forebrain NMDA receptor 2B subunit over-expression has no influence on anxiety behaviors of mice..
Shi-Jia LI ; Min-Hua XU ; Chun-Xia LI ; Tian-Shu ZHOU ; Bo MENG ; Bing MEI
Acta Physiologica Sinica 2009;61(3):272-278
It has been known that the glutamate transmission system and N-methyl-D-aspartate receptor (NMDA-R) were possibly related to anxiety processes. Although anxiety symptom can be relieved by NMDA-R antagonists and partial agonists treatment, the functions of NMDA-R and its subunits in anxiety behaviors remain unclear. We used forebrain specific NR2B over-expression mice to examine whether the increase of NR2B subunit level would induce anxiety behaviors. The results indicated that the juvenile (3-5 months old), middle-aged (8-10 months old) and old (19-22 months old ) NR2B transgenic mice showed no significant difference in open field test and elevated plus maze test as compared with the control mice. Capillary electrophoresis of monoamine neurotransmitter in subregions of forebrain revealed no significant difference between transgenic and control mice of 16-18 months age. These results suggest that the increase of NR2B expression and followed NR1 and NR2A expression augmentations in the forebrain have no significant effect on anxiety-related behaviors in mice.
Animals
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Anxiety
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metabolism
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Mice
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Mice, Transgenic
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Prosencephalon
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metabolism
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Receptors, N-Methyl-D-Aspartate
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metabolism