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Voltages up to 600V did not affect cryopreserved bovine spermatozoa on capillary-type electroporation.

التفاصيل البيبلوغرافية
العنوان: Voltages up to 600V did not affect cryopreserved bovine spermatozoa on capillary-type electroporation.
المؤلفون: Blödorn EB; Laboratório de Genômica Estrutural, Programa de Pós-Graduação em Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil., Domingues WB; Laboratório de Genômica Estrutural, Programa de Pós-Graduação em Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil., Komninou ER; Laboratório de Biotecnologia do Cancer, Programa de Pós-Graduação em Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil., Daneluz L; Laboratório de Genômica Estrutural, Programa de Pós-Graduação em Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil., Dellagostin EN; Laboratório de Genômica Estrutural, Programa de Pós-Graduação em Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil., Weege A; Laboratório de Genômica Estrutural, Programa de Pós-Graduação em Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil., Varela AS Junior; Laboratótio de Reprodução Animal Comparada, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande, Rio Grande, RS, Brazil., Corcini CD; Laboratótio de Reprodução Animal Comparada, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande, Rio Grande, RS, Brazil., Collares TV; Laboratório de Biotecnologia do Cancer, Programa de Pós-Graduação em Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil., Campos VF; Laboratório de Genômica Estrutural, Programa de Pós-Graduação em Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil. Electronic address: campos.vinicius@ufpel.edu.br.
المصدر: Reproductive biology [Reprod Biol] 2018 Dec; Vol. 18 (4), pp. 416-421. Date of Electronic Publication: 2018 Sep 13.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Elsevier Urban & Partner Country of Publication: Poland NLM ID: 101160559 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2300-732X (Electronic) Linking ISSN: 1642431X NLM ISO Abbreviation: Reprod Biol Subsets: MEDLINE
أسماء مطبوعة: Publication: <2013- > : Olsztyn, Poland : Elsevier Urban & Partner
Original Publication: Olsztyn, Poland : Polish Society for Biology of Reproduction, c2001-
مواضيع طبية MeSH: Semen Preservation*, Electroporation/*methods , Spermatozoa/*physiology, Animals ; Cattle ; Cryopreservation ; Male ; Semen Analysis ; Sperm Motility/physiology
مستخلص: Physical methods such as electroporation have been used to improve the DNA uptake efficiency of sperm cells. This study aims to develop an efficient capillary-type electroporation method for incorporation of exogenous DNA into bovine cryopreserved sperm cells with minimal detrimental effects for later use in SMGT. Electroporation of the samples was performed in 2 different groups (with 1 μg of DNA and without DNA transfection) and under five different voltages: 500 V, 600 V, 700 V, 800 V and 900 V. Non-electroporated sperm cells (with and without DNA) were used as control. Kinetics parameters were determined using computer assisted semen analyses, whereas membrane integrity, fluidity, mitochondrial function and DNA uptake were evaluated by flow cytometry. Results revealed that all tested voltages reduced electroporated sperm motility (P < 0.05) when compared to the control (non-electroporated cells). Mitochondrial function results showed no statistical difference among groups. Similarly, groups electroporated with lower (500 V, 600 V and 700 V) voltages showed no difference in cell membrane integrity and fluidity. Groups electroporated at higher voltages (800 V and 900 V) demonstrated negative effects in cells membrane integrity when compared to other groups and control. Also, all electroporated groups demonstrated significant higher percentages of transfected sperm cells when compared to the control group (P < 0.05). Under the recommendation of using voltages up to 600 V, this method represents a safe and efficient alternative for electroporation of bovine spermatozoa.
(Copyright © 2018 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier B.V. All rights reserved.)
فهرسة مساهمة: Keywords: Capillary-type electroporation; SMGT; Sperm; Transfection; Transgenesis
تواريخ الأحداث: Date Created: 20180918 Date Completed: 20190110 Latest Revision: 20190110
رمز التحديث: 20231215
DOI: 10.1016/j.repbio.2018.09.003
PMID: 30220550
قاعدة البيانات: MEDLINE
الوصف
تدمد:2300-732X
DOI:10.1016/j.repbio.2018.09.003