<meta charset="utf8"></head><body>探索母体运动对大鼠后代健康的益处

<ol>
	<li>Hanson, M. A., Gluckman, P. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+developmental+conditioning+of+later+health+and+disease:+Physiology+or+pathophysiology.">Early developmental conditioning of later health and disease: Physiology or pathophysiology.</a> Physiol Rev. 94 (4), 1027-1076 (2014).</li><li>Calkins, K., Devaskar, S. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fetal+origins+of+adult+disease.">Fetal origins of adult disease.</a> Curr Probl Pediatr Adolesc Health Care. 41 (6), 158-176 (2011).</li><li>Chavatte-Palmer, P., Tarrade, A., Rousseau-Ralliard, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diet+before+and+during+pregnancy+and+offspring+health:+The+importance+of+animal+models+and+what+can+be+learned+from+them.">Diet before and during pregnancy and offspring health: The importance of animal models and what can be learned from them.</a> Int J Environ Res Public Health. 13 (6), 586 (2016).</li><li>Lapehn, S., Paquette, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+placental+epigenome+as+a+molecular+link+between+prenatal+exposures+and+fetal+health+outcomes+through+the+dohad+hypothesis.">The placental epigenome as a molecular link between prenatal exposures and fetal health outcomes through the dohad hypothesis.</a> Curr Environ Health Rep. 9 (3), 490-501 (2022).</li><li>Shrestha, A., Prowak, M., Berlandi-Short, V. M., Garay, J., Ramalingam, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maternal+obesity:+A+focus+on+maternal+interventions+to+improve+health+of+offspring.">Maternal obesity: A focus on maternal interventions to improve health of offspring.</a> Front Cardiovasc Med. 8, 696812 (2021).</li><li>Haire-Joshu, D., Tabak, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preventing+obesity+across+generations:+Evidence+for+early+life+intervention.">Preventing obesity across generations: Evidence for early life intervention.</a> Annu Rev Public Health. 37, 253-271 (2016).</li><li>Kusuyama, J., Alves-Wagner, A. B., Makarewicz, N. S., Goodyear, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+maternal+and+paternal+exercise+on+offspring+metabolism.">Effects of maternal and paternal exercise on offspring metabolism.</a> Nat Metab. 2 (9), 858-872 (2020).</li><li>Blaize, A. N., Pearson, K. J., Newcomer, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+maternal+exercise+during+pregnancy+on+offspring+chronic+disease+susceptibility.">Impact of maternal exercise during pregnancy on offspring chronic disease susceptibility.</a> Exerc Sport Sci Rev. 43 (4), 198-203 (2015).</li><li>Hinman, S. K., Smith, K. B., Quillen, D. M., Smith, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exercise+in+pregnancy:+A+clinical+review.">Exercise in pregnancy: A clinical review.</a> Sports Health. 7 (6), 527-531 (2015).</li><li>Acog committee. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physical+activity+and+exercise+during+pregnancy+and+the+postpartum+period:+Acog+committee+opinion,+number+804.">Physical activity and exercise during pregnancy and the postpartum period: Acog committee opinion, number 804.</a> Obstet Gynecol. 135 (4), e178-e188 (2020).</li><li>Poole, D. 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=Guidelines+for+animal+exercise+and+training+protocols+for+cardiovascular+studies.">Guidelines for animal exercise and training protocols for cardiovascular studies.</a> Am J Physiol Heart Circ Physiol. 318 (5), H1100-H1138 (2020).</li><li>Beleza, J., 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=Self-paced+free-running+wheel+mimics+high-intensity+interval+training+impact+on+rats'+functional,+physiological,+biochemical,+and+morphological+features.">Self-paced free-running wheel mimics high-intensity interval training impact on rats' functional, physiological, biochemical, and morphological features.</a> Front Physiol. 10, 593 (2019).</li><li>August, P. M., Rodrigues, K. D. S., Klein, C. P., Dos Santos, B. G., Matt&#233;, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+gestational+exercise+practice+and+litter+size+reduction+on+maternal+care.">Influence of gestational exercise practice and litter size reduction on maternal care.</a> Neurosci Lett. 741, 135454 (2021).</li><li>Li, 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=Prenatal+exercise+reprograms+the+development+of+hypertension+progress+and+improves+vascular+health+in+shr+offspring.">Prenatal exercise reprograms the development of hypertension progress and improves vascular health in shr offspring.</a> Vascul Pharmacol. 139, 106885 (2021).</li><li>Li, 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=Exercise+during+pregnancy+enhances+vascular+function+via+epigenetic+repression+of+ca(v)1.2+channel+in+offspring+of+hypertensive+rats.">Exercise during pregnancy enhances vascular function via epigenetic repression of ca(v)1.2 channel in offspring of hypertensive rats.</a> Life Sci. 231, 116576 (2019).</li><li>Musial, B., 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=Exercise+alters+the+molecular+pathways+of+insulin+signaling+and+lipid+handling+in+maternal+tissues+of+obese+pregnant+mice.">Exercise alters the molecular pathways of insulin signaling and lipid handling in maternal tissues of obese pregnant mice.</a> Physiol Rep. 7 (16), e14202 (2019).</li><li>Dos Santos, 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=Resistance+exercise+was+safe+for+the+pregnancy+and+offspring's+development+and+partially+protected+rats+against+early+life+stress-induced+effects.">Resistance exercise was safe for the pregnancy and offspring's development and partially protected rats against early life stress-induced effects.</a> Behav Brain Res. 445, 114362 (2023).</li><li>Barnes, M. D., Heaton, T. L., Goates, M. C., Packer, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intersystem+implications+of+the+developmental+origins+of+health+and+disease:+Advancing+health+promotion+in+the+21st+century.">Intersystem implications of the developmental origins of health and disease: Advancing health promotion in the 21st century.</a> Healthcare. 4 (3), 45 (2016).</li><li>Bakrania, B. A., George, E. M., Granger, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+preeclampsia:+Investigating+pathophysiology+and+therapeutic+targets.">Animal models of preeclampsia: Investigating pathophysiology and therapeutic targets.</a> Am J Obstet Gynecol. 226, S973-S987 (2022).</li><li>Ferrari, N., 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=Exercise+during+pregnancy+and+its+impact+on+mothers+and+offspring+in+humans+and+mice.">Exercise during pregnancy and its impact on mothers and offspring in humans and mice.</a> J Dev Orig Health Dis. 9 (1), 63-76 (2018).</li><li>Lerman, L. O., 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=Animal+models+of+hypertension:+A+scientific+statement+from+the+american+heart+association.">Animal models of hypertension: A scientific statement from the american heart association.</a> Hypertension. 73 (6), e87-e120 (2019).</li><li>Mangwiro, Y. 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=Exercise+initiated+during+pregnancy+in+rats+born+growth+restricted+alters+placental+mtor+and+nutrient+transporter+expression.">Exercise initiated during pregnancy in rats born growth restricted alters placental mtor and nutrient transporter expression.</a> J Physiol. 597 (7), 1905-1918 (2019).</li><li>Kim, H., Lee, S. H., Kim, S. S., Yoo, J. H., Kim, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+maternal+treadmill+running+during+pregnancy+on+short-term+memory+and+hippocampal+cell+survival+in+rat+pups.">The influence of maternal treadmill running during pregnancy on short-term memory and hippocampal cell survival in rat pups.</a> Int J Dev Neurosci. 25 (4), 243-249 (2007).</li><li>Harris, J. E., 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=Exercise-induced+3'-sialyllactose+in+breast+milk+is+a+critical+mediator+to+improve+metabolic+health+and+cardiac+function+in+mouse+offspring.">Exercise-induced 3'-sialyllactose in breast milk is a critical mediator to improve metabolic health and cardiac function in mouse offspring.</a> Nat Metab. 2 (8), 678-687 (2020).</li><li>Zhang, Y., 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=Maternal+exercise+represses+nox4+via+sirt1+to+prevent+vascular+oxidative+stress+and+endothelial+dysfunction+in+shr+offspring.">Maternal exercise represses nox4 via sirt1 to prevent vascular oxidative stress and endothelial dysfunction in shr offspring.</a> Front Endocrinol. 14, 1219194 (2023).</li><li>Shan, 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=Maternal+exercise+upregulates+the+DNA+methylation+of+agtr1a+to+enhance+vascular+function+in+offspring+of+hypertensive+rats.">Maternal exercise upregulates the DNA methylation of agtr1a to enhance vascular function in offspring of hypertensive rats.</a> Hypertens Res. 46 (3), 654-666 (2023).</li></ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Caliper</td>
			<td>Mitutoyo Measuring Instruments (Shanghai) Co., Ltd</td>
			<td>530-101 N15</td>
			<td>Fetus dissect</td>
		</tr>
		<tr>
			<td>Circular water bucket</td>
			<td>Naliya</td>
			<td>N/A</td>
			<td>Rat swimming</td>
		</tr>
		<tr>
			<td>Experimental Surgical Instrument&#160;</td>
			<td>Shenzhen RWD Life Technology Co., LTD</td>
			<td>SP0001-G</td>
			<td>Fetus dissect</td>
		</tr>
		<tr>
			<td>Glass Microscope Slides</td>
			<td>Jiangsu Shitai experimental equipment Co., LTD</td>
			<td>80312-3161</td>
			<td>Vaginal smear</td>
		</tr>
		<tr>
			<td>Glass petri dish</td>
			<td>Merck Life Technologies</td>
			<td>BR455751-10EA</td>
			<td>Offspring rearing</td>
		</tr>
		<tr>
			<td>Hairdryer</td>
			<td>Panasonic</td>
			<td>EH-WNE5H</td>
			<td>Post-swimming care</td>
		</tr>
		<tr>
			<td>Information Card</td>
			<td>Zhongke Life Science</td>
			<td>SS3</td>
			<td>Rat mating</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Shenzhen RWD Life Technology Co., LTD</td>
			<td>R510-22-10</td>
			<td>Fetus dissect</td>
		</tr>
		<tr>
			<td>Light microscope</td>
			<td>Olympus</td>
			<td>IX71-F22PH</td>
			<td>Vaginal smear</td>
		</tr>
		<tr>
			<td>Mating cage</td>
			<td>Zhongke Life Science</td>
			<td>SS3</td>
			<td>Rat mating</td>
		</tr>
		<tr>
			<td>Medical Absorbent Cotton</td>
			<td>Hongxiang Sanitary Materials Co., LTD</td>
			<td>N/A</td>
			<td>The pregnant rats are anticipating giving birth</td>
		</tr>
		<tr>
			<td>Rodent Anesthesia Machine, Gas Anesthesia</td>
			<td>Shenzhen RWD Life Technology Co., LTD</td>
			<td>R500IE</td>
			<td>Fetus dissect</td>
		</tr>
		<tr>
			<td>Rodent breeding feed</td>
			<td>Beijing Huafukang Biotechnology Co., LTD</td>
			<td>1032</td>
			<td>Pregnant rat feeding</td>
		</tr>
		<tr>
			<td>Rodent maintenance feed</td>
			<td>Beijing Huafukang Biotechnology Co., LTD</td>
			<td>1022</td>
			<td>Offspring rearing</td>
		</tr>
		<tr>
			<td>Rodent sunflower seeds</td>
			<td>Jolly</td>
			<td>JP241</td>
			<td>Nutritional supplement</td>
		</tr>
		<tr>
			<td>Soundproof cotton</td>
			<td>Kufu Medical Instrument</td>
			<td>N/A</td>
			<td>The pregnant rats are anticipating giving birth</td>
		</tr>
		<tr>
			<td>Sterile Cotton Swab (2 mm diameter)</td>
			<td>Kufu Medical Instrument</td>
			<td>N/A</td>
			<td>Vaginal smear</td>
		</tr>
		<tr>
			<td>Sterile gauze</td>
			<td>Kufu Medical Instrument</td>
			<td>N/A</td>
			<td>Fetus dissect</td>
		</tr>
		<tr>
			<td>Stroke-physiological Saline Solution (0.9% NaCl)</td>
			<td>Shandong Hualu Pharmaceutical Co., LTD</td>
			<td>N/A</td>
			<td>Vaginal smear</td>
		</tr>
		<tr>
			<td>Thermometer</td>
			<td>Beekman organism</td>
			<td>N/A</td>
			<td>The monitoring of rat swimming</td>
		</tr>
		<tr>
			<td>Towels</td>
			<td>Grace</td>
			<td>N/A</td>
			<td>Post-swimming care</td>
		</tr>
		<tr>
			<td>Transparent plastic container</td>
			<td>Naliya</td>
			<td>N/A</td>
			<td>Swimming adaptive training</td>
		</tr>
		<tr>
			<td>Ultraviolet light</td>
			<td>Merck Life Technologies</td>
			<td>Z169633-1EA</td>
			<td>Post-swimming care</td>
		</tr>
		<tr>
			<td>Water heater</td>
			<td>Haier</td>
			<td>EC6001-Q6S</td>
			<td>Rat swimming</td>
		</tr>
		<tr>
			<td>Weight Scale</td>
			<td>Electronlc Acale</td>
			<td>JM.A10001</td>
			<td>Body weight measurement</td>
		</tr>
		<tr>
			<td>Wistar Rats</td>
			<td>Vital river Laboratory Animal Technology Co., LTD</td>
			<td>N/A</td>
			<td>Experiment</td>
		</tr>
	</tbody>
</table>

swimming exercise protocol and care methods for pregnant rats

Read this Scientific Article Protocol about Author Spotlight: Enhancing Offspring Health with Maternal Exercise During Pregnancy at JoVE.com

<meta charset="utf8"></head><body>作者聚焦：怀孕期间通过母体运动增强大鼠的后代健康

怀孕大鼠的游泳锻炼方案和护理方法

The developmental origins of health and disease concept highlights the impact of early environments on chronic non-communicable diseases like diabetes, ...

swimming-exercise-protocol-and-care-methods-for-pregnant-rats

Research

JoVE Journal

Developmental Biology

Swimming Exercise Protocol and Care Methods for Pregnant Rats

470 次观看.  Beijing Sport University. 我们的研究描述了大鼠的繁殖方法。产前游泳训练产生大鼠和繁殖后护理方案，这将对运动生理学和围产期生物学的研究人员有很大帮助。随着怀孕的进展和大鼠腹部的肿胀，治疗和定期锻炼可能会导致腹部反复接触坚硬的表面。相比之下，游泳运动方案中的水环境提供浮力，降低了怀孕动物因运动而受伤的风险。母亲肥胖会导致后代心血管和...

视频: 作者聚焦：怀孕期间通过母体运动增强大鼠的后代健康

Understanding Early Organogenesis Using a Simplified In Situ Hybridization Protocol in Xenopus

Organogenesis is the study of how organs are specified and then acquire their specific shape and functions during development. The Xenopuslaevis embryo is very useful for studying organogenesis because their large size makes them very suitable for identifying organs at the earliest steps in organogenesis. At this time, the primary method used for identifying a specific organ or primordium is whole mount in situ hybridization with labeled antisense RNA probes specific to a gene that is expressed in the organ of interest. In addition, it is relatively easy to manipulate genes or signaling pathways in Xenopus and in situ hybridization allows one to then assay for changes in the presence or morphology of a target organ. Whole mount in situ hybridization is a multi-day protocol with many steps involved. Here we provide a simplified protocol with reduced numbers of steps and reagents used that works well for routine assays. In situ hybridization robots have greatly facilitated the process and we detail how and when we utilize that technology in the process. Once an in situ hybridization is complete, capturing the best image of the result can be frustrating. We provide advice on how to optimize imaging of in situ hybridization results. Although the protocol describes assessing organogenesis in Xenopus laevis, the same basic protocol can almost certainly be adapted to Xenopus tropicalis and other model systems.

Understanding Early Organogenesis Using a Simplified In Situ Hybridization Protocol in Xenopus

The Xenopus laevis embryo continues to be exceptionally useful in the study of early development due to its large size and ease of manipulation. A simplified protocol for whole mount in situ hybridization protocol is provided that can be used in the identification of specific organs in this model system.

了解早期器官使用简体<em&gt;原位</em&gt;杂交协议<em&gt;爪</em

Organogenesis is the study of how organs are specified and then acquire their specific shape and functions during development.  The Xenopuslaevis embryo ...

科研

发育生物学

Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements

Skeletal muscle mitochondria play a specific role in many disease pathologies. As such, the measurement of oxygen consumption as an indicator of mitochondrial function in this tissue has become more prevalent. Although many technologies and assays exist that measure mitochondrial respiratory pathways in a variety of cells, tissue and species, there is currently a void in the literature in regards to the compilation of these assays using isolated mitochondria from mouse skeletal muscle for use in microplate based technologies. Importantly, the use of microplate based respirometric assays is growing among mitochondrial biologists as it allows for high throughput measurements using minimal quantities of isolated mitochondria. Therefore, a collection of microplate based respirometric assays were developed that are able to assess mechanistic changes/adaptations in oxygen consumption in a commonly used animal model. The methods presented herein provide step-by-step instructions to perform these assays with an optimal amount of mitochondrial protein and reagents, and high precision as evidenced by the minimal variance across the dynamic range of each assay.

The methods presented provide step-by-step instructions for the performance of a collection of microplate based respirometric assays using isolated mitochondria from minimal quantities of mouse skeletal muscle. These assays are able to measure mechanistic changes/adaptations in mitochondrial oxygen consumption in a commonly used animal model.

使用分离线粒体从小鼠骨骼肌进行高通量微孔板呼吸测量最小数量

Skeletal muscle mitochondria play a specific role in many disease pathologies. As such, the measurement of oxygen consumption as an indicator of ...

Methods for Analyzing the Impacts of Natural Uranium on In Vitro Osteoclastogenesis

Uranium has been shown to interfere with bone physiology and it is well established that this metal accumulates in bone. However, little is known about the effect of natural uranium on the behavior of bone cells. In particular, the impact of uranium on osteoclasts, the cells responsible for the resorption of the bone matrix, is not documented. To investigate this issue, we have established a new protocol using uranyl acetate as a source of natural uranium and the murine RAW 264.7 cell line as a model of osteoclast precursors. Herein, we detailed all the assays required to test uranium cytotoxicity on osteoclast precursors and to evaluate its impact on the osteoclastogenesis and on the resorbing function of mature osteoclasts. The conditions we have developed, in particular for the preparation of uranyl-containing culture media and for the seeding of RAW 264.7 cells allow to obtain reliable and highly reproductive results. Moreover, we have optimized the use of software tools to facilitate the analysis of various parameters such as the size of osteoclasts or the percentage of resorbed matrix.

Methods for Analyzing the Impacts of Natural Uranium on In Vitro Osteoclastogenesis

Uranium is known to affect bone metabolism. Here, we present a protocol aimed at investigating the effect of natural uranium exposure on the viability, the differentiation, and the function of osteoclasts, the cells in charge of bone resorption.

天然铀对体外破影响的分析方法

Uranium has been shown to interfere with bone physiology and it is well established that this metal accumulates in bone. However, little is known about ...

Streamlined 3D Cerebellar Differentiation Protocol with Optional 2D Modification

Reducing the complexity and cost of differentiation protocols is important for researchers. This interest fits with concerns about possible unintended effects that extrinsic patterning factors might introduce into human pluripotent stem cell (hPSC) models of brain development or pathophysiology, such as masking disease phenotype. Here, we present two cerebellar differentiation protocols for hPSCs, designed with simpler startup method, fewer patterning factors, and less material requirements than previous protocols. Recently, we developed culture procedures, which generate free-floating 3-dimensional (3D) products consistent with other brain &#34;organoid&#34; protocols, including morphologies relevant to modeling brain development such as sub/ventricular zone- and rhombic lip-like structures. The second uses an adherent, 2D monolayer procedure to complete differentiation, which is shown capable of generating functional cerebellar neurons, as products are positive for cerebellar-associated markers, and exhibit neuron-like calcium influxes. Together, these protocols offer scientists a choice of options suited to different research purposes, as well as a basic model for testing other types of streamlined neural differentiations.

We describe a simplified 3D differentiation protocol for hPSCs, using defined medium and reduced growth factors, capable of generating cell aggregates with early neuroepithelial structures and positive for cerebellar-associated markers, as well as an optional 2D modification for differentiating cells as a monolayer to generate functional neurons.

简化的3D 小脑鉴别协议, 可选2D 修改

Reducing the complexity and cost of differentiation protocols is important for researchers. This interest fits with concerns about possible unintended ...

Methods for the Study of Regeneration in Stentor

Cells need to be able to regenerate their parts to recover from external perturbations. The unicellular ciliate Stentor coeruleus is an excellent model organism to study wound healing and subsequent cell regeneration. The Stentor genome became available recently, along with modern molecular biology methods, such as RNAi. These tools make it possible to study single-cell regeneration at the molecular level. The first section of the protocol covers establishing Stentor cell cultures from single cells or cell fragments, along with general guidelines for maintaining Stentor cultures. Culturing Stentor in large quantities allows for the use of valuable tools like biochemistry, sequencing, and mass spectrometry. Subsequent sections of the protocol cover different approaches to inducing regeneration in Stentor. Manually cutting cells with a glass needle allows studying the regeneration of large cell parts, while treating cells with either sucrose or urea allows studying the regeneration of specific structures located at the anterior end of the cell. A method for imaging individual regenerating cells is provided, along with a rubric for staging and analyzing the dynamics of regeneration. The entire process of regeneration is divided in three stages. By visualizing the dynamics of the progression of a population of cells through the stages, the heterogeneity in regeneration timing is demonstrated.

Methods for the Study of Regeneration in Stentor

The giant ciliate, Stentor coeruleus, is an excellent system to study regeneration and wound healing. We present procedures for establishing Stentor cell cultures from single cells or cell fragments, inducing regeneration by cutting cells, chemically inducing the regeneration of membranellar band and oral apparatus, imaging, and analysis of cell regeneration.

Stentor再生的研究方法

Cells need to be able to regenerate their parts to recover from external perturbations. The unicellular ciliate Stentor coeruleus is an excellent model ...

Analysis of Congenital Heart Defects in Mouse Embryos Using Qualitative and Quantitative Histological Methods

Congenital heart defects (CHD) are the most common type of birth defect in humans, affecting up to 1% of all live births. However, the underlying causes for CHD are still poorly understood. The developing mouse constitutes a valuable model for the study of CHD, because cardiac developmental programs between mice and humans are highly conserved. The protocol describes in detail how to produce mouse embryos of the desired gestational stage, methods to isolate and preserve the heart for downstream processing, quantitative methods to identify common types of CHD by histology (e.g., ventricular septal defects, atrial septal defects, patent ductus arteriosus), and quantitative histomorphometry methods to measure common muscular compaction phenotypes. These methods articulate all the steps involved in sample preparation, collection, and analysis, allowing scientists to correctly and reproducibly measure CHD.

In this protocol, we describe procedures to qualitatively and quantitatively analyze developmental phenotypes in mice associated with congenital heart defects.

使用定性和定量组织方法对小鼠胚胎先天性心脏缺陷的分析

Congenital heart defects (CHD) are the most common type of birth defect in humans, affecting up to 1% of all live births. However, the underlying causes ...

Is My Mouse Pregnant? High-Frequency Ultrasound Assessment

The mouse is the mammalian animal model of choice for many human diseases and biological processes. Developmental biology often requires staged-pregnant mice to determine evolving processes at various timepoints. Moreover, optimal and efficient breeding of model mice requires an assessment of timed pregnancies. Most commonly, mice are mated overnight, and the presence of a vaginal plug is determined; however, the positive predictive value of this technique is suboptimal, and one needs to wait to know if the mouse is truly pregnant. High-resolution ultrasound biomicroscopy is an effective and efficient tool for imaging: 1) Whether a mouse is pregnant; 2) What gestational stage the mouse has reached; and 3) Whether there are intrauterine losses. In addition to the embryos and fetuses, the investigator must also recognize common artifacts in the abdominal cavity so as not to mistake these for a gravid uterus. This article provides a protocol for imaging along with illustrative examples.

High-resolution ultrasound can help streamline experiments requiring timed-pregnant mice by determining the state of pregnancy, gestational age, and pregnancy losses. Presented here is a protocol to illustrate methods to assess mouse pregnancies as well as potential pitfalls (image artifacts) that may mimic pregnancy.

我的老鼠怀孕了吗？高频超声波评估

The mouse is the mammalian animal model of choice for many human diseases and biological processes. Developmental biology often requires staged-pregnant ...

Protocol for Human Blastoids Modeling Blastocyst Development and Implantation

A model of the human blastocyst formed from stem cells (blastoid) would support scientific and medical advances. However, its predictive power will depend on its ability to efficiently, timely, and faithfully recapitulate the sequences of blastocyst development (morphogenesis, specification, patterning), and to form cells reflecting the blastocyst stage. Here we show that na&#239;ve human pluripotent stem cells cultured in PXGL conditions and then triply inhibited for the Hippo, transforming growth factor- &#946;, and extracellular signal-regulated kinase pathways efficiently undergo morphogenesis to form blastoids (&gt;70%). Matching with developmental timing (~4 days), blastoids unroll the blastocyst sequence of specification by producing analogs of the trophoblast and epiblast, followed by the formation of analogs of the primitive endoderm and the polar trophoblasts. This results in the formation of cells transcriptionally similar to the blastocyst (&gt;96%) and a minority of post-implantation analogs. Blastoids efficiently pattern by forming the embryonic-abembryonic axis marked by the maturation of the polar region (NR2F2+), which acquires the specific potential to directionally attach to hormonally stimulated endometrial cells, as in utero. Such a human blastoid is a scalable, versatile, and ethical model to study human development and implantation in vitro.

A protocol outlining the formation of human blastoids that efficiently, timely, and sequentially generate blastocyst-like cells.

人爆破模拟胚发育和植入的协议

A model of the human blastocyst formed from stem cells (blastoid) would support scientific and medical advances. However, its predictive power will depend ...

In Ovo and Ex Ovo Methods to Study Avian Inner Ear Development

The inner ear perceives sound and maintains balance using the cochlea and vestibule. It does this by using a dedicated mechanosensory cell type known as the hair cell. Basic research in the inner ear has led to a deep understanding of how the hair cell functions, and how dysregulation can lead to hearing loss and vertigo. For this research, the mouse has been the pre-eminent model system. However, mice, like all mammals, have lost the ability to replace hair cells. Thus, when trying to understand cellular therapies for restoring inner ear function, complementary studies in other vertebrate species could provide further insights. The auditory epithelium of birds, the basilar papilla (BP), is a sheet of epithelium composed of mechanosensory hair cells (HCs) intercalated by supporting cells (SCs). Although the anatomical architecture of the basilar papilla and the mammalian cochlea differ, the molecular mechanisms of inner ear development and hearing are similar. This makes the basilar papilla a useful system for not only comparative studies but also to understand regeneration. Here, we describe dissection and manipulation techniques for the chicken inner ear. The technique shows genetic and small molecule inhibition methods, which offer a potent tool for studying the molecular mechanisms of inner ear development. In this paper, we discuss in ovo electroporation techniques to genetically perturb the basilar papilla using CRIPSR-Cas9 deletions, followed by dissection of the basilar papilla. We also demonstrate the BP organ culture and optimal use of culture matrices, to observe the development of the epithelium and the hair cells.

In Ovo and Ex Ovo Methods to Study Avian Inner Ear Development

The chick is a cost-effective, accessible, and widely available model organism for a variety of studies. Here, a series of protocols is detailed to understand the molecular mechanisms underlying avian inner ear development and regeneration.

在蛋 和 卵外 研究中研究鸟类内耳发育的方法

The inner ear perceives sound and maintains balance using the cochlea and vestibule. It does this by using a dedicated mechanosensory cell type known as ...

Methods for Studying Uterine Contributions to Pregnancy Establishment in an Ovariectomized Mouse Model

For pregnancy to be established, a viable blastocyst must successfully interact with a receptive uterine lining (endometrium) to facilitate implantation and placenta formation and enable ongoing pregnancy. The limitations to pregnancy success caused by embryonic defects are well known and have been largely overcome in recent decades with the rise of in vitro fertilization (IVF) and assisted reproductive technologies. As yet, however, the field has not overcome the limitations caused by an inadequately receptive endometrium, thus resulting in stagnating IVF success rates. Ovarian and endometrial functions are closely intertwined, as hormones produced by the ovary are responsible for the endometrium's menstrual cyclicity. As such, when using rodent models of pregnancy, it can be difficult to ascertain whether an observed result is due to an ovarian or uterine deficit. To overcome this, an ovariectomized mouse model was developed with embryo transfer or artificial decidualization to allow the study of uterine-specific contributions to pregnancy. This article will provide instructions on how to perform ovariectomy and offer insights into various techniques for supplying exogenous hormones to support successful artificial decidualization or pregnancy following embryo transfer from healthy donors. These techniques include subcutaneous injection, slow-release pellets, and osmotic mini pumps. The key advantages and disadvantages of each method will be discussed, enabling researchers to choose the best study design for their specific research question.

Pregnancy establishment is a dynamic process involving complex embryo and uterine crosstalk. The precise contributions of the maternal uterine environment to these processes remain an active area of investigation. Here, detailed protocols are provided to aid in designing in vivo animal models to address these research questions.

研究卵巢切除小鼠模型中子宫对妊娠建立的贡献的方法

For pregnancy to be established, a viable blastocyst must successfully interact with a receptive uterine lining (endometrium) to facilitate implantation ...