Lentivirus‐Mediated Expression of MxA in Chicken Spermatogonial Stem Cells (English)

in Reproduction in Domestic Animals ; 45 , 5 ; e131-e137
Reproduction in Domestic Animals

The aim of this study was to construct a lentivirus vector that carries the human myxovirus‐resistant A (MxA) anti‐virus gene for efficiently infecting rooster spermatogonial stem cells (SSCs). A lentiviral vector carrying the enhanced green fluorescent protein (EGFP) and MxA fusion gene (EGFP‐MxA) was constructed using TOPO technology in this study. Enhanced green fluorescent protein‐MxA was inserted in the right orientation as determined by restriction enzyme digest. No gene recombination in the vector occurred. After infecting 293FT cells, EGFP‐MxA fusion protein was expressed as granular green fluorescence characteristic of EGFP‐MxA expression, suggesting that the TOPO expression vector was properly constructed and the fusion protein expressed correctly. The EGFP‐MxA recombinant lentivirus was packaged by cotransfecting 293FT cells with EGFP‐MxA and the packaging plasmids. We also purified SSCs from testicle tissues from 25‐day‐old roosters for infection with the EGFP‐MxA recombinant virus. After infecting rooster SSCs with the recombinant virus for 72 h, EGFP‐MxA expression was detected by EGFP expression. Enhanced green fluorescent protein‐MxA expression in SSCs was further confirmed at the transcription level by RT‐PCR and at the protein level by immunocytochemistry. Staining with Hoechst 33342 showed that infected SSCs differed from the sertoli cells. Similar to stem cells, SSCs were positive for alkaline phosphoric acid enzyme and for stage‐specific embryonic antigen‐1 stem cell factor. The results demonstrated that the recombinant virus made in our study can infect rooster SSCs efficiently to express the anti‐virus protein MxA, establishing the basis of transferring MxA into SSCs to obtain virus‐resistant, genetically‐modified roosters.

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Table of contents – Volume 45, Issue 5

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755
Effects of N‐acetyl‐cysteine and N‐acetyl‐cysteine‐amide Supplementation on In Vitro Matured Porcine Oocytes
Whitaker, BD / Knight, JW | 2010
760
Morpho‐physical Recording of Bovine Conceptus (Bos indicus) and Placenta from Days 20 to 70 of Pregnancy
Assis Neto, AC / Pereira, FTV / Santos, TC / Ambrosio, CE / Leiser, R / Miglino, MA | 2010
773
Influence of Ovulation Status, Seasonality and Embryo Transfer Method on Development of Cloned Porcine Embryos
Koo, OJ / Kang, JT / Kwon, DK / Park, HJ / Lee, BC | 2010
779
Oocyte Morphology from Primordial to Early Tertiary Follicles of Yak
Yu, SJ / Yong, YH / Cui, Y | 2010
786
Efficiency of SCNT Buffalo (Bubalus bubalis) Embryos in Different Culture Medium and Analysis of mRNA Expression of Insulin‐like Growth Factors During Embryogenesis
Pandey, A / Gupta, SC / Singh, N / Rana, JS / Gupta, N | 2010
796
Participation of Opioid Peptides in Sucking‐induced Oxytocin and Prolactin Secretions in Lactating Goats
Bruna, FA / De Di Nasso, EG / Soaje, M / Deis, RP / Carón, RW | 2010
803
The Effect of Parity of the Dam on Sexual Maturation, Serum Concentrations of Metabolic Hormones and the Response to Luteinizing Hormone Releasing Hormone in Bull Calves
Bagu, ET / Davies, KL / Epp, T / Arteaga, A / Barrett, DM / Duggavathi, R / Barth, A / Rawlings, NC | 2010
811
Association between Myeloperoxidase Concentration in Equine Frozen Semen and Post‐Thawing Parameters
Ponthier, J / Franck, T / Detilleux, J / Mottart, E / Serteyn, D / Deleuze, S | 2010
817
Relationships between Early Foal Health, Future Performance and Their Dams Reproductive Health
Hemberg, E / Kindahl, H / Lundeheim, N / Einarsson, S | 2010
821
Expression of Prostate Glycoconjugates in the Stallion and Castrated Horse
Parillo, F / Mancuso, R / Vullo, C / Catone, G | 2010
832
The Effect of Long‐term In Vivo Culture in Bovine Oviduct and Uterus on the Development and Cryo‐tolerance of In Vitro Produced Bovine Embryos
Havlicek, V / Kuzmany, A / Cseh, S / Brem, G / Besenfelder, U | 2010
838
Effect of Lycopene on Cytoplasmic Maturation of Porcine Oocytes In Vitro
Watanabe, H / Okawara, S / Bhuiyan, MMU / Fukui, Y | 2010
846
Effects of Ethanol on Synthesis of Prostaglandin F2α in Bovine Females
De Barros, FRO / Bertan, CM / Dias, LOP / Fantini, DA / Ayres, GR / Marques, VB / Miguez, PHP / Binelli, M | 2010
851
Synergistic Effect of Porcine Follicular Fluid and Dibutyryl Cyclic Adenosine Monophosphate on Development of Parthenogenetically Activated Oocytes from Pre‐Pubertal Gilts
Nascimento, AB / Albornoz, MS / Che, L / Visintin, JA / Bordignon, V | 2010
860
Influence of Superovulatory Protocols on In Vitro Production of Nellore (Bos indicus) Embryos
Monteiro, FM / Ferreira, MMG / Potiens, JR / Eberhardt, BG / Trinca, LA / Barros, CM | 2010
865
Alterations of Spindle and Microfilament Assembly in Aged Cat Oocytes
Jin, Y‐X / Cui, X‐S / Yu, X‐F / Han, Y‐J / Kong, I‐K / Kim, N‐H | 2010
872
Endometrial Population of Oestrogen Receptors Alpha and Beta and Progesterone Receptors A and B During the Different Phases of the Follicular Wave of Llamas (Lama glama)
Bianchi, CP / Sahlin, L / Meikle, A / Masironi, B / Cavilla, MV / Aba, MA | 2010
881
13,14‐Dihydro‐15‐Keto Prostaglandin F2α Release in Response to Oxytocin Challenge Early Post‐Partum in Anoestrous Nelore Cows Submitted to Temporary Calf Removal and Progesterone Priming
Satrapa, RA / Pinheiro, VG / Ereno, RL / Membrive, CMB / Piagentini, M / Binelli, M / Barros, CM | 2010
888
Follicle Growth and Oocyte Developmental Competence in Cows With Liver Damage
Iwata, H / Tanaka, H / Kanke, T / Sakaguchi, Y / Shibano, K / Kuwayama, T / Monji, Y | 2010
896
Fertility Time Trends in Dairy Herds in Northern Portugal
Rocha, A / Martins, A / Carvalheira, J | 2010
900
Allotransplantation of Transgenic Mouse Ovaries Expressing Enhanced Green Fluorescent Protein under the Control of the Murine Phosphoglycerate Kinase 1 Promoter
Wu, HT / Chou, CK / Hung, YC / Yu, CK | 2010
907
Transplacental Transfer of Iron in the Water Buffalo (Bubalus bubalis): Uteroferrin and Erythrophagocytosis
Pereira, FTV / Braga, FC / Burioli, KC / Kfoury Jr, JR / Oliveira, LJ / Papa, PC / Carvalho, AF / Ambrósio, CE / Bazer, FW / Miglino, MA | 2010
915
Expression of Apoptosis Regulatory Genes and Incidence of Apoptosis in Different Morphological Quality Groups of In Vitro‐produced Bovine Pre‐implantation Embryos
Melka, MG / Rings, F / Hölker, M / Tholen, E / Havlicek, V / Besenfelder, U / Schellander, K / Tesfaye, D | 2010
922
Effects of FSH and LH on Steroid Production by Buffalo (Bubalus bubalis) Granulosa Cells Cultured In Vitro Under Serum‐Free Conditions
Shanmugam, M / Pandita, S / Palta, P | 2010
927
Ghrelin and Leptin Did Not Improve Meiotic Maturation of Porcine Oocytes Cultured In Vitro
Suzuki, H / Sasaki, Y / Shimizu, M / Matsuzaki, M / Hashizume, T / Kuwayama, H | 2010
931
Effect of Sodium Nitroprusside, a Nitric Oxide Donor, and Aminoguanidine, a Nitric Oxide Synthase Inhibitor, on In Vitro Development of Buffalo (Bubalus bubalis) Embryos
Saugandhika, S / Kumar, D / Singh, MK. / Shah, R / Anand, T / Chauhan, MS / Manik, RS / Singla, S K / Palta, P | 2010
934
Late Abscess Formation Caused by Silk Suture Following Hysterectomy in a Female Dog
Boza, S / Lucas, X / Zarelli, M / Soler, M / Belda, E / Agut, A | 2010
937
Factors Affecting the Readiness and Preparation of Replacement Heifers in Tropical Breeding Environments
Maquivar, M / Galina, CS | 2010
e1
Effect of Cooling Rate to 5°C, Straw Size and Farm on Fertilizing Ability of Cryopreserved Rabbit Sperm
Mocé, E / Lavara, R / Vicente, JS | 2010
e8
Cloprostenol in Equine Reproductive Practice: Something More Than a Luteolytic Drug
Cuervo‐Arango, J / Newcombe, JR | 2010
e12
Effects of Different Oocyte Activation Procedures on Development and Gene Expression of Porcine Pre‐Implantation Embryos
Cervera, RP / Silvestre, MA / Martí, N / García‐Mengual, E / Moreno, R / Stojkovic, M | 2010
e21
An Inter‐Subspecies Cloned Buffalo (Bubalus bubalis) Obtained by Transferring of Cryopreserved Embryos via Somatic Cell Nuclear Transfer
Yang, BZ / Yang, CY / Li, RC / Qin, GS / Zhang, XF / Pang, CY / Chen, MT / Huang, FX / Li, Z / Zheng, HY et al. | 2010
e26
Expression of Progesterone Receptor in the Utero‐tubal Junction After Intra‐uterine and Deep Intra‐uterine Insemination in Sows
Tummaruk, P / Tienthai, P / Manee‐In, S / Srisuwatanasagul, S | 2010
e32
Ion, Protein, Phospholipid and Energy Substrate Content of Oviduct Fluid During the Oestrous Cycle of Buffalo (Bubalus bubalis)
Vecchio, D / Neglia, G / Di Palo, R / Campanile, G / Balestrieri, ML / Giovane, A / Killian, G / Zicarelli, L / Gasparrini, B | 2010
e40
In Vitro Effect of the Reproductive Hormones on the Oxidative Burst Activity of Polymorphonuclear Leucocytes from Cows: A Flow Cytometric Study
Chaveiro, A / Moreira da Silva, F | 2010
e46
Synchronizing Cell Cycle of Goat Fibroblasts by Serum Starvation Causes Apoptosis
Dalman, A / Eftekhari‐Yazdi, P / Valojerdi, MR / Shahverdi, A / Gourabi, H / Janzamin, E / Fakheri, R / Sadeghian, F / Hasani, F | 2010
e54
The Possible Involvement of Salsolinol and Hypothalamic Prolactin in the Central Regulatory Processes in Ewes During Lactation
Górski, K / Romanowicz, K / Herman, A / Molik, E / Gajewska, A / Tomaszewska‐Zaremba, D / Misztal, T | 2010
e61
The Pattern of Embryonic Fixation and its Relationship to Pregnancy Loss in Thoroughbred Mares
Sharma, S / Davies Morel, MCG / Dhaliwal, GS / Dadarwal, D | 2010
e68
Effects of Retinoids on the In Vitro Development of Capra Hircus Embryos to Blastocysts in Two Different Culture Systems
Chiamenti, A / Aguiar Filho, CR / Freitas Neto, LM / Chaves, RM / Paula‐Lopes, FF / Lima, PF / Gonçalves, PBD / Oliveira, MAL | 2010
e73
Effect of Dimethyl Sulfoxide on Cell Cycle Synchronization of Goldfish Caudal Fin Derived Fibroblasts Cells
Choresca, CH Jr / Koo, OJ / Hong, SG / Oh, HJ / Gomez, DK / Kim, JH / Lee, BC / Park, SC | 2010
e78
Neurogenic Differentiation of EGFP Gene Transfected Amniotic Fluid‐Derived Stem Cells from Pigs at Intermediate and Late Gestational Ages
Zheng, Y‐M / Zhao, X‐E / An, Z‐X | 2010
e83
Expression Pattern of Apoptotic Genes in Vitrified‐Thawed Bovine Oocytes
Anchamparuthy, VM / Pearson, RE / Gwazdauskas, FC | 2010
e91
Follicular, Oocyte and Embryo Features Related to Metabolic Status in Primiparous Lactating does Fed with High‐Fibre Rearing Diets
Arias‐Álvarez, M / García‐García, RM / Rebollar, PG / Nicodemus, N / Millán, P / Revuelta, L / Lorenzo, PL | 2010
e101
Pronuclear Embryo Yield in Canindé and Saanen Goats for DNA Microinjection
Moura, RR / Lopes‐Junior, ES / Teixeira, DIA / Serova, IA / Andreeva, LE / Melo, LM / Freitas, VJF | 2010
e107
Evaluation of a Neck Mounted 2‐Hourly Activity Meter System for Detecting Cows About to Ovulate in Two Paddock‐Based Australian Dairy Herds
Hockey, CD / Morton, JM / Norman, ST / McGowan, MR | 2010
e118
Poor Prediction Value of Sperm Head Morphometry for Fertility and Litter Size in Rabbit
Marco‐Jiménez, F / Vicente, J‐S / Lavara, R / Balasch, S / Viudes‐de‐Castro, M‐P | 2010
e124
Evaluation of Prolactin Receptor (PRLR) as Candidate Gene for Male Fertility in Hanoverian Warmblood Horses
Giesecke, K / Hamann, H / Sieme, H / Distl, O | 2010
e131
Lentivirus‐Mediated Expression of MxA in Chicken Spermatogonial Stem Cells
Liu, L / He, P / Cai, K / Zhang, Y / Li, J / Cao, F / Ding, Z / Zhang, N | 2010
e138
Effect of Embryo Density on In vitro Developmental Characteristics of Bovine Preimplantative Embryos with Respect to Micro and Macroenvironments
Hoelker, M / Rings, F / Lund, Q / Phatsara, C / Schellander, K / Tesfaye, D | 2010
e146
Histological Study of Canine Mammary Gland During the Oestrous Cycle
Santos, M / Marcos, R / Faustino, AMR | 2010
e155
Comparison of Post‐Thaw DNA Integrity of Boar Spermatozoa Assessed with the Neutral Comet Assay and Sperm‐Sus Halomax Test Kit
Fraser, L / Parda, A / Filipowicz, K / Strzeżek, J | 2010
e161
Induction of Abortion with Aglepristone in Cats on Day 45 and 46 After Mating
Georgiev, P / Bostedt, H / Goericke‐Pesch, S / Dimitrov, M / Petkov, P / Stojanthev, K / Tsoneva, V / Wehrend, A | 2010
e168
Effects of a Progesterone‐Based Oestrous Synchronization Protocol in 51‐ to 57‐Day Postpartum High‐Producing Dairy Cows
Garcia‐Ispierto, I / López‐Gatius, F / Bech‐Sàbat, G / Yániz, JL / Angulo, E / Maris, C / Floc’h, S / Martino, A | 2010
e174
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Smolinska, N / Kaminski, T / Siawrys, G / Przala, J | 2010
e184
Intermediate Filament Keratin Dynamics During Oocyte Maturation Requires Maturation/M‐Phase Promoting Factor and Mitogen‐Activated Protein Kinase Kinase Activities in the Hamster
Kabashima, K / Matsuzaki, M / Suzuki, H | 2010
e189
In Vitro Maturation of Dromedary (Camelus dromedarius) Oocytes: Effect of Different Protein Supplementations and Epidermal Growth Factor*
Wani, NA / Wernery, U | 2010
e194
Do Cytoplasmic Lipid Droplets Accumulate in Immature Oocytes from Over‐Conditioned Repeat Breeder Dairy Heifers?
Awasthi, H / Saravia, F / Rodríguez‐Martínez, H / Båge, R | 2010
e199
Two Unusual Cases of Canine Prostatitis: Prostatitis in a Castrated Dog and Preputial Oedema in an Intact Male
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e201
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e207
Effect of Early Postpartum Ovulation on Fertility in Dairy Cows
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e212
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e217
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e221
Epidemiology of Bovine Venereal Campylobacteriosis: Geographic Distribution and Recent Advances in Molecular Diagnostic Techniques
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