Name: determination of reproductive competence by confirming pubertal onset and performing a fertility assay in mice and rats
Uploaded: 2018-10-13T15:00:00
Description: Watch this Scientific Journal Video about Determination of Reproductive Competence by Confirming Pubertal Onset and Performing a Fertility Assay in Mice and Rats at JoVE.com

我感谢提交人为本出版物的基础而作出的初步工作。多亏了 Aitor 老板, 珍娜. 瑞安和艾瑞卡 l. 舍勒帮助准备手稿。多亏了杰西卡. 苍利和奥斯汀下巴的技术援助手稿。H.M.H. 是由尤尼斯肯尼迪国家儿童健康 & 人类发展研究所的国家卫生研究院的奖励编号 R00HD084759 支持。

此处描述的所有方法均已通过密歇根州立大学的机构动物护理和使用委员会批准, 并按照实验室动物的护理和使用指南进行。
1. 确定青春期发作
<ol><li>遵循服装的制度准则, 至少, 有必要穿干净的实验室大衣和干净的手套。总是用干净的手套来处理老鼠。</li>
	<li>通过在桌子上放置一个垫子来准备工作区域。<ol>
<li>将干净的鼠标保持架顶部与网格朝向上朝上。</li>
		<li>在工作?...

<ol>
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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Leptin+accelerates+the+onset+of+puberty+in+normal+female+mice.">Leptin accelerates the onset of puberty in normal female mice.</a> Journal of Clinical Investigation. 99 (3), 391-395 (1997).</li><li>Bohlen, T. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+short-day+photoperiod+delays+the+timing+of+puberty+in+female+mice+via+changes+in+the+kisspeptin+system.">A short-day photoperiod delays the timing of puberty in female mice via changes in the kisspeptin system.</a> Frontiers in Endocrinology. 9 (FEB), 1-9 (2018).</li><li>Shahab, M., Mastronardi, C., Seminara, S. B., Crowley, W. F., Ojeda, S. R., Plant, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+hypothalamic+GPR54+signaling:+A+potential+mechanism+for+initiation+of+puberty+in+primates.">Increased hypothalamic GPR54 signaling: A potential mechanism for initiation of puberty in primates.</a> Proceedings of the National Academy of Sciences. , (2005).</li><li>Hoffmann, H. M., Mellon, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+small+population+of+hypothalamic+neurons+govern+fertility:+the+critical+role+of+VAX1+in+GnRH+neuron+development+and+fertility+maintenance.">A small population of hypothalamic neurons govern fertility: the critical role of VAX1 in GnRH neuron development and fertility maintenance.</a> Neuroscience communications. 2, 5-9 (2016).</li><li>Navarro, V. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+Neurokinin+B+in+the+Control+of+Female+Puberty+and+Its+Modulation+by+Metabolic+Status.">Role of Neurokinin B in the Control of Female Puberty and Its Modulation by Metabolic Status.</a> Journal of Neuroscience. 32 (7), 2388-2397 (2012).</li><li>Hoffmann, H. M., Tamrazian, A., Xie, H., P&#233;rez-Mill&#225;n, M. I., Kauffman, A. S., Mellon, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterozygous+deletion+of+ventral+anterior+homeobox+(Vax1)+causes+subfertility+in+mice.">Heterozygous deletion of ventral anterior homeobox (Vax1) causes subfertility in mice.</a> Endocrinology. 155 (10), 4043-4053 (2014).</li><li>Kauffman, A. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Kisspeptin+Receptor+GPR54+Is+Required+for+Sexual+Differentiation+of+the+Brain+and+Behavior.">The Kisspeptin Receptor GPR54 Is Required for Sexual Differentiation of the Brain and Behavior.</a> Journal of Neuroscience. 27 (33), 8826-8835 (2007).</li><li>Teles, M. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brief+report:+A+GPR54-activating+mutation+in+a+patient+with+central+precocious+puberty.">Brief report: A GPR54-activating mutation in a patient with central precocious puberty.</a> New England Journal of Medicine. , (2008).</li><li>Korenbrot, C. C., Huhtaniemi, I. T., Weiner, R. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preputial+separation+as+an+external+sign+of+pubertal+development+in+the+male+rat.">Preputial separation as an external sign of pubertal development in the male rat.</a> Biology of reproduction. , (1977).</li><li>Gaytan, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+validation+of+a+method+for+precise+dating+of+female+puberty+in+laboratory+rodents:+The+puberty+ovarian+maturation+score+(Pub-Score).">Development and validation of a method for precise dating of female puberty in laboratory rodents: The puberty ovarian maturation score (Pub-Score).</a> Scientific Reports. 7 (March), 1-11 (2017).</li><li>Caligioni, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+reproductive+status/stages+in+mice.">Assessing reproductive status/stages in mice.</a> Current Protocols in Neuroscience. , 1-11 (2010).</li><li>Mayer, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Timing+and+completion+of+puberty+in+female+mice+depend+on+estrogen+receptor+-signaling+in+kisspeptin+neurons.">Timing and completion of puberty in female mice depend on estrogen receptor -signaling in kisspeptin neurons.</a> Proceedings of the National Academy of Sciences. 107 (52), 22693-22698 (2010).</li><li>McLean, A. C., Valenzuela, N., Fai, S., Bennett, S. A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Performing+Vaginal+Lavage,+Crystal+Violet+Staining,+and+Vaginal+Cytological+Evaluation+for+Mouse+Estrous+Cycle+Staging+Identification.">Performing Vaginal Lavage, Crystal Violet Staining, and Vaginal Cytological Evaluation for Mouse Estrous Cycle Staging Identification.</a> Journal of Visualized Experiments. (67), 4-9 (2012).</li><li>Hedrich, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Laboratory Mouse. , (2012).</li><li>Hoffmann, H. M., Trang, C., Gong, P., Kimura, I., Pandolfi, E. C., Mellon, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deletion+of+Vax1+from+Gonadotropin-Releasing+Hormone+(GnRH)+Neurons+Abolishes+GnRH+Expression+and+Leads+to+Hypogonadism+and+Infertility.">Deletion of Vax1 from Gonadotropin-Releasing Hormone (GnRH) Neurons Abolishes GnRH Expression and Leads to Hypogonadism and Infertility.</a> Journal of Neuroscience. 36 (12), 3506-3518 (2016).</li><li>Sloboda, D. M., Howie, G. J., Pleasants, A., Gluckman, P. D., Vickers, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pre-+and+postnatal+nutritional+histories+influence+reproductive+maturation+and+ovarian+function+in+the+rat.">Pre- and postnatal nutritional histories influence reproductive maturation and ovarian function in the rat.</a> PLoS ONE. , (2009).</li><li>Manual, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Breeding+Strategies+for+Maintaining+Colonies+of+Laboratory+Mice.">Breeding Strategies for Maintaining Colonies of Laboratory Mice.</a> Management. , (2007).</li><li>Kennedy, G. C., Mitra, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Body+weight+and+food+intake+as+initiating+factors+for+puberty+in+the+rat.">Body weight and food intake as initiating factors for puberty in the rat.</a> The Journal of Physiology. , (1963).</li><li>Sisk, C. L., Foster, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neural+basis+of+puberty+and+adolescence.">The neural basis of puberty and adolescence.</a> Nature Neuroscience. 7 (10), 1040-1047 (2004).</li><li>Nelson, J. F., Karelus, K., Felicio, L. S., Johnson, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+influences+on+the+timing+of+puberty+in+mice.">Genetic influences on the timing of puberty in mice.</a> Biology of reproduction. , (1990).</li><li>Nelson, J. F., Felicio, L. S., Randall, P. K., Sims, C., Finch, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+longitudinal+study+of+estrous+cyclicity+in+aging+C57BL/6J+mice:+I.+Cycle+frequency,+length+and+vaginal+cytology.">A longitudinal study of estrous cyclicity in aging C57BL/6J mice: I. Cycle frequency, length and vaginal cytology.</a> Biology of reproduction. , (1982).</li><li>Falconer, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Weight+and+age+at+puberty+in+female+and+male+mice+of+strains+selected+for+large+and+small+body+size.">Weight and age at puberty in female and male mice of strains selected for large and small body size.</a> Genetical Research. , (1984).</li><li>Rodriguez, I., Araki, K., Khatib, K., Martinou, J. C., Vassalli, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+vaginal+opening+is+an+apoptosis-dependent+process+which+can+be+prevented+by+the+overexpression+of+Bcl2.">Mouse vaginal opening is an apoptosis-dependent process which can be prevented by the overexpression of Bcl2.</a> Developmental Biology. , (1997).</li><li>Lomniczi, A., Wright, H., Ojeda, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epigenetic+regulation+of+female+puberty.">Epigenetic regulation of female puberty.</a> Frontiers in Neuroendocrinology. 36, 90-107 (2015).</li><li>Selmanoff, M. K., Goldman, B. D., Ginsburg, B. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+changes+in+serum+luteinizing+hormone,+follicle+stimulating+hormone+and+androgen+levels+in+males+of+two+inbred+mouse+strains.">Developmental changes in serum luteinizing hormone, follicle stimulating hormone and androgen levels in males of two inbred mouse strains.</a> Endocrinology. 100 (1), 122-127 (1977).</li><li>Larder, R., Clark, D. D., Miller, N. L. G., Mellon, P. 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小鼠的整体健康对于成功的生育力测定21至关重要。在进行生育试验时, 不每天对老鼠进行身体检查是很重要的, 因为这会引起压力。进一步避免频繁的笼型变化, 因为这些也有压力。理想的笼型改变每周不超过1-2 次。暗相照射对夜间啮齿动物的繁殖有负面影响。在黑暗的时间里, 不要在繁殖室开灯。如果在天黑时需要进入房间, 请使用昏暗的红光照明。另一个压力源是在谢切诺夫...

通过青春期的过渡需要达到性成熟和生殖能力。成年期的青春期过渡和生育能力的维持由生殖轴调节, 也称为下丘脑-垂体-性腺轴 (图 1)。青春期开始和维持生育的时间是严格的内部和环境因素, 以增加后代和父母的生存几率1,2。该议定书提供了一种非侵入性的方法来确定小鼠和大鼠的青春期发作, 以确认在建立生育研究以评估生殖能力之前的性行为成熟度。
生育研究是在性成熟的动物进行的, 可以在动物经历青春期后开始。在青春期开始之前, 生殖轴是静止的, 并且性成熟的关键驱动因素, 促性腺激素释放激素 (促性腺素), 释放到垂体不足以启动青春期 (图 1)。青春期发作是一个复杂的过程, 导致增加促性腺激素释放在中位隆起。促性腺激素促进促黄体素 (LH) 和卵泡刺激激素 (FSH) 分泌从垂体, 两种激素对性腺成熟和生殖功能至关重要 (图 1)3,4,5.
对生殖轴的侮辱导致生育能力下降, 也可以促进或延缓青春期发作。已知影响青春期发作时间和生殖能力的条件包括接触内分泌干扰化学品6,7, 增加/减少体重1,8, 变化日长2,9和遗传突变10,11,12,13,14,15。
性成熟的开始是一个关键步骤, 需要在建立生育试验之前完成。通过包皮分离、阴道开放和第一次发情确定青春期发作的好处是这些程序的非侵入性特征, 因为它们不需要收集或牺牲动物16,17。
在青春期发作确定后, 正确地建立生育研究将提供有关生殖轴完整性的重要信息, 并且通常有生成实验动物的第二个优势进一步研究 (细化)18. 本议定书所述的生育率研究设置可以检测男性和女性生殖能力方面的次要和重大缺陷。评估的关键参数包括 1) 时间到第一个垃圾, 2) 在给定的时间段内生成的凋落物的数量和 3) 垃圾大小。最后, 还包括为确定生育障碍原因而进行的后续研究类型的建议。
所述的协议是指小鼠, 代表数据反映了转基因小鼠所做的工作。但是, 所有包含的协议在大鼠中同样有效。

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			<td height="40" style="height:40px;width:216px;">Sterile Cotton Balls</td>
			<td style="width:167px;">Fisher</td>
			<td style="width:280px;">22456885</td>
			<td style="width:429px;"></td>
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		<tr height="40">
			<td height="40" style="height:40px;">Surface protector</td>
			<td>Fisher</td>
			<td>1420637</td>
			<td style="width:429px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Light meter</td>
			<td>VWR</td>
			<td>21800-014</td>
			<td style="width:429px;"></td>
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		<tr height="40">
			<td height="40" style="height:40px;">Methylene blue&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>M9140</td>
			<td style="width:429px;"></td>
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		<tr height="40">
			<td height="40" style="height:40px;">Microscope Slides</td>
			<td>Genesee Scientific</td>
			<td>29-101</td>
			<td style="width:429px;"></td>
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			<td height="40" style="height:40px;width:216px;">Optimouse rack with cages</td>
			<td>AnimalCare systems</td>
			<td>C89100</td>
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			<td height="40" style="height:40px;">Water Bottle Basket&#160;</td>
			<td>AnimalCare systems</td>
			<td>C61011</td>
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		<tr height="40">
			<td height="40" style="height:40px;">Filtered Cage Tops</td>
			<td>AnimalCare systems</td>
			<td>C78210</td>
			<td></td>
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		<tr height="40">
			<td height="40" style="height:40px;">Optimice Standard Feeder</td>
			<td>AnimalCare systems</td>
			<td>C40100SG</td>
			<td style="width:429px;"></td>
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		<tr height="40">
			<td height="40" style="height:40px;">Cage Card Holder</td>
			<td>AnimalCare systems</td>
			<td>C43251</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Cage Cards</td>
			<td>AnimalCare systems</td>
			<td>M52010</td>
			<td></td>
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		<tr height="40">
			<td height="40" style="height:40px;">Bottle Assambley</td>
			<td>AnimalCare systems</td>
			<td>C79122P</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Bed R&#39;Nest Nesting</td>
			<td style="width:167px;">The Andersons</td>
			<td style="width:280px;">BRN4WSR</td>
			<td style="width:429px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">1/8&#34; Corn Cob bedding&#160;</td>
			<td style="width:167px;">The Andersons</td>
			<td style="width:280px;">8B</td>
			<td style="width:429px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Standard mouse chow</td>
			<td>Teklad</td>
			<td style="width:280px;">7904 (7004)</td>
			<td style="width:429px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Scale</td>
			<td>VWR</td>
			<td style="width:280px;">10205-004</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Polypropylene Beaker</td>
			<td>Fisher</td>
			<td>14-955-111F</td>
			<td></td>
		</tr>
	</tbody>
</table>

determination of reproductive competence by confirming pubertal onset and performing a fertility assay in mice and rats

对生殖能力的评估对于理解治疗或基因操作对生殖轴的影响至关重要, 也称为下丘脑-垂体-性腺轴。生殖轴是环境和内部输入的关键集成商, 适应生育条件, 有利于繁殖。在开始对小鼠和大鼠的生育研究之前, 性成熟度被评估为排除了观察到的生殖表型是由延迟或缺席青春期发作引起的可能性。本议定书描述了一种非侵入性的方法来评估男性青春期发作通过确定包皮分离, 并在女性通过阴道开放和第一次发情。在确认青春期完成和性成熟的成就后, 可以开始生育研究。该程序描述了小鼠和大鼠的最佳育种条件, 如何建立生育研究, 以及评估和确定治疗或基因删除是否影响生育能力的参数。

所提供的结果来自两个不同的转基因小鼠模型, 其中转录因子腹前同源盒 1 (Vax1) 已被删除在整个身体上的一个等位基因, 这里称为杂合子小鼠 (13), 或Vax1已有条件地删除在促性腺激素的神经元22, 这里称为条件 KO (cKO)。在建立生育研究之前, 确认所有小鼠青春期发作是很重要的。
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Watch this Scientific Journal Video about Determination of Reproductive Competence by Confirming Pubertal Onset and Performing a Fertility Assay in Mice and Rats at JoVE.com

Determination of Reproductive Competence by Confirming Pubertal Onset and Performing a Fertility Assay in Mice and Rats

在小鼠和大鼠中确认青春期发作和执行生育力测定生殖能力

许多治疗和基因突变影响性成熟和生育的时间。本议定书描述了一种非侵入性的方法来评估小鼠和大鼠青春期发作之前, 建立一个生育研究在有性成熟的动物。

Assessment of reproductive competence is critical for understanding the impact of a treatment or genetic manipulation on the reproductive axis, also ...

这种方法可以帮助回答生殖领域的关键问题，例如如何通过预注射分离、阴道开放以及在进行生育研究之前建立第一个雌激素来确定小鼠和小鼠的青春期发病。该技术的主要优点是，通过使用证明的非侵入性方法建立青春期发作，可以跟踪和进行性成熟动物的生育研究。这项技术的影响延伸到诊断生殖缺陷，如基因突变或治疗易受影响生殖能力，如内分泌破坏化学品造成的。虽然这种方法可以提供对小鼠生殖能力的洞察，但它也可以应用于其他模型系统，如大鼠和仓鼠。这种方法的视觉演示至关重要，因为它需要良好和细腻的动物处理技能。正确握住小鼠，以进行以下肿瘤前分离和阴道开口的视觉识别尤为重要。要准备工作区，请将垫子放在桌子上。然后放置一个干净的鼠标笼顶部与网格朝上，在垫子上。接下来，在工作区中设置比例。将干净的 500 至 1000 毫升烧杯放在秤上，然后进行皮。然后在工作区旁边放置一个工作表以记录数据。在此之后，将鼠标笼放在工作区中。打开笼子，将盖子、水瓶和食物架放在桌子上。将鼠标握在尾巴上时，请放在干净的鼠标笼顶部。在保持尾部的保持的同时，允许鼠标探索笼子顶部的网格。轻轻地向后拉尾巴，接近另一只手靠近颈部和耳朵的皮肤。用拇指和食指抓住肩膀和颈部之间的松弛皮肤。然后用剩下的手指抓住背部的皮肤，轻轻按压鼠标。用自由手，轻轻地推皮周围的阴道背。在预吸分离时，预发皮肤向后滑动，露出青光的性气。检查后，记录是否存在预处理分离。在此之后，称量鼠标并记录其重量。然后将鼠标重新引入外壳笼，将烧杯放在笼子地板上，让鼠标自己退出。在工作区放置棉球、大烧杯和一瓶无菌水。将无菌水倒入烧杯中。然后，用烧杯中的无菌水加湿棉球，并放在干净的笼子顶部旁边。将鼠标笼放在工作区中。然后将鼠标移到笼子顶部，握住它的尾巴底部。轻轻拉上鼠标的尾巴，鼓励鼠标向前移动。在此之后，抬起尾巴，同时用三根手指支撑臀部。用加湿的棉球轻轻清洁外阴，然后检查外阴以确定阴道是否完全打开。记录阴道开口是否发生。然后称鼠标，然后回到家里的笼子里。准备一个干净的老鼠笼与盖子，水，食物，床上用品和筑巢材料，每对繁殖的老鼠。然后填写笼卡，并填写必要的信息。将准备好的床上用品笼放在干净的桌子上，然后慢慢将戴手套的手放入鼠标笼中，并放在笼子里一小段时间。用戴手套的手慢慢接近老鼠。当老鼠跑过手，抓住一只老鼠的尾巴在拇指和食指之间，然后抬起鼠标的尾巴。鼠标可以由尾巴保持，而无需身体的支持长达两到三秒钟。将小鼠放在准备好的繁殖笼中，然后在确保雌性小鼠有食物、水和筑巢材料后关闭笼子。运用该方法，研究了两种不同的转基因小鼠模型。阴道开放和预注射分离发生在相同的年龄和重量在异质小鼠和对照小鼠。相反，阴道开放和预分娩分离在有条件的淘汰女性和男性中显著延迟，并随着体重增加。在4.5周大时，雌性对等小鼠和有条件的淘汰小鼠的体重相当，而有条件的淘汰雄性则略重于对组。这表明，有条件敲除小鼠阴道开放和预分离的延迟与体重增加的延迟没有关系。青春期发作结束后，生育研究开始于10至16周。雄性小鼠和雌性异卵小鼠均为亚育鼠。在三个月的研究中，异质雌性产生的垃圾较少，雄性小鼠和雌性异卵小鼠产生的垃圾较少。为了说明在建立生育分析之前确认青春期发病的重要性，在有条件的敲孔小鼠中评估了生殖能力。在70天的生育研究中，无论是有条件的淘汰男性还是女性都没有产生任何垃圾。在尝试此程序时，重要的是要记住小心处理小鼠，并在接近和处理动物时保持冷静。按照这个程序， 其他方法， 如测量循环性激素水平可以回答其他问题， 如不孕不育或其他亚不育是由异常睾酮， 雌激素， 卵泡刺激激素， 和叶黄素激素水平.不要忘记，由于有咬人和过敏的风险，与老鼠一起工作可能是危险的。为了减少接触，在执行此过程时，应始终采取预防措施，如戴上手套和实验室外套，以及小心处理小鼠。

Research

JoVE Journal

Biology

Manual Restraint and Common Compound Administration Routes in Mice and Rats

手动约束和常见的化合物在小鼠和大鼠的给药途径

Being able to safely and effectively restrain mice and rats is an important part of conducting research.  Working confidently and humanely with mice and ...

科研

生物学

Diagnostic Necropsy and Selected Tissue and Sample Collection in Rats and Mice

剖检诊断，并选择在大鼠和小鼠的组织和样品收集

There are multiple sample types that may be collected from a euthanized animal in order to help diagnose or discover infectious agents in an animal ...

Ovariectomy and 17&beta;-estradiol Replacement in Rats and Mice: A Visual Demonstration

Ovariectomy and 17ÃƒÅ½Ã‚Â²-estradiol Replacement in Rats and Mice: A Visual Demonstration

在大鼠和小鼠的卵巢切除和17β-雌二醇更换：一个可视化示范

Estrogens are a family of female sexual hormones with an exceptionally wide spectrum of effects. When rats and mice are used in estrogen research they are ...

A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells

快速，特异微孔板检测为内和细胞外抗坏血酸在培养细胞的测定

Vitamin C (ascorbate) plays numerous important roles in cellular metabolism, many of which have only come to light in recent years. For instance, within ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

在造血干细胞和祖细胞的成年小鼠颅骨骨髓体内四维跟踪

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

In Vivo Microinjection and Electroporation of Mouse Testis

In Vivo Microinjection and Electroporation of Mouse Testis

在小鼠睾丸活体显微注射和电穿孔

This video and article contribution gives a comprehensive description of microinjection and electroporation of mouse testis in vivo. This particular ...

En Face Detection of Nitric Oxide and Superoxide in Endothelial Layer of Intact Arteries

En Face Detection of Nitric Oxide and Superoxide in Endothelial Layer of Intact Arteries

一氧化氮和超氧化物歧化酶恩的人脸检测在完整的动脉内皮细胞层

Endothelium-derived nitric oxide (NO) produced from endothelial NO-synthase (eNOS) is one of the most important vasoprotective molecules in cardiovascular ...

The Mouse Isolated Perfused Kidney Technique

鼠标离体肾技术

The mouse isolated perfused kidney (MIPK) is a technique for keeping a mouse kidney under ex vivo conditions perfused and functional for 1 hr. This is a ...

Determination of Sialic Acids in Liver and Milk Samples of Wild-type and CMAH Knock-out Mice.

Determination of Sialic Acids in Liver and Milk Samples of Wild-type and CMAH Knock-out Mice.

测定野生型肝脏和牛奶样品中的唾液酸<em&gt; CMAH</em&gt;敲出小鼠

CMAH (cytidine monophosphate-N-acetylneuraminic acid hydroxylase) is responsible for the oxidation of cytidine monophosphate-N-acetylneuraminic acids in ...

Reproductive Techniques for Ovarian Monitoring and Control in Amphibians

两栖动物卵巢监测和控制的生殖技术

Ovarian control and monitoring in amphibians require a multi-faceted approach. There are several applications that can successfully induce reproductive ...