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转基因山羊中的激素诱导泌乳

Hormonal induced lactation in transgenic goats.

作者信息

Cammuso C, Porter C, Nims S, Gaucher D, Melican D, Bombard S, Hawkins N, O'Coin A, Ricci C, Brayman C, Buzzell N, Ziomek C, Gavin W

机构信息

Genzyme Transgenics Corporation, Framingham, MA 01702, USA.

出版信息

Anim Biotechnol. 2000;11(1):1-17. doi: 10.1080/10495390009525943.

Abstract

The aim of this study was to hormonally induce lactation in prepubertal, nulliparous, and male goats both transgenic and non-transgenic. Analysis of milk quality, recombinant protein expression levels, total amount of recombinant protein produced, and the affect on long-term reproductive capability was assessed. Fifty-one goats (Saanen, Alpine, and Toggenburg), male and non-pregnant females, 2-31 months of age, either non-transgenic or transgenic were evaluated with a total of 10 transgenes (constructs) represented. Animals were given estradiol (0.25 mg/kg, i.m.) and progesterone (0.75 mg/kg, i.m.) on days 1, 3, 5, 7, 9, 11 and 13, while prednisilone (0.4 mg/kg, i.m.) was administered on days 14-16 with mammary massage occurring daily from day 5 onward. Forty of 51 animals, (36 of 38 females and 4 of 13 males) produced milk with total volumes in the 30-day experiment, ranging from 20 microl to 530 mls per day, or approximately 500 microl to 6.8 liters total. Milk composition was analyzed for various parameters (total protein, fat content, total solids and somatic cell count) with no significant differences found between induced and natural milk. Expression levels of recombinant proteins from transgenic animals that were analyzed during the induced lactation, and subsequently during normal lactations, were found to have no significant differences. Total amount of recombinant protein produced was evaluated at different expression levels with no statistical significance seen. While over 90% of the females placed in the regimen became pregnant, there was a correlation between increased age at time of induction and an increase in number of breedings, or reproductive cycles needed to establish a pregnancy after induction. For males, 100% placed in the regimen settled females after hormonal induction of lactation. Semen quality was evaluated prior to, during, and after hormonal treatments. Semen volume and sperm number did not differ; however, for a small percentage of males, there was a decrease in sperm and post thaw motility after hormonal treatments. These levels returned to normal within 4-5 weeks. Subsequent natural lactations showed total milk volumes within breed standards. These findings indicate that hormonal induction of lactation in the caprine species is a viable alternative to pregnancy for initiating lactation and milk production, does not adversely impact reproductive performance long-term, and can benefit the early assessment of recombinant proteins produced in a transgenic founder program.

摘要

本研究的目的是通过激素诱导使青春期前、未生育的转基因和非转基因雄性及雌性山羊泌乳。评估了奶品质、重组蛋白表达水平、重组蛋白总产量以及对长期繁殖能力的影响。对51只山羊(萨能山羊、阿尔卑斯山羊和吐根堡山羊)进行了评估,包括雄性和未怀孕的雌性,年龄在2至31个月之间,有非转基因和转基因的,共代表10种转基因(构建体)。在第1、3、5、7、9、11和13天给动物注射雌二醇(0.25毫克/千克,肌肉注射)和孕酮(0.75毫克/千克,肌肉注射),在第14至16天注射泼尼松龙(0.4毫克/千克,肌肉注射),从第5天起每天进行乳腺按摩。51只动物中有40只(38只雌性中的36只和13只雄性中的4只)在30天的实验中产出了奶,每天的总量在20微升至530毫升之间,或30天总量约为500微升至6.8升。分析了奶的各种参数(总蛋白、脂肪含量、总固体和体细胞计数),诱导泌乳的奶和自然泌乳的奶之间未发现显著差异。对转基因动物在诱导泌乳期间及随后正常泌乳期间的重组蛋白表达水平进行分析,未发现显著差异。评估了不同表达水平下重组蛋白的总产量,未发现统计学意义上的差异。虽然超过90%接受该方案的雌性怀孕了,但诱导时年龄的增加与配种次数的增加或诱导后怀孕所需的繁殖周期数之间存在相关性。对于雄性,100%接受该方案的雄性在激素诱导泌乳后使雌性受孕。在激素治疗前、治疗期间和治疗后评估了精液质量。精液体积和精子数量没有差异;然而,对于一小部分雄性,激素治疗后精子数量和解冻后活力有所下降。这些水平在4至5周内恢复正常。随后的自然泌乳显示奶总产量在品种标准范围内。这些发现表明,在山羊物种中通过激素诱导泌乳是启动泌乳和产奶的一种可行替代怀孕的方法,不会对长期繁殖性能产生不利影响,并且有利于在转基因奠基计划中对产生的重组蛋白进行早期评估。

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