This work was supported by the National Natural Science Foundation of China (32071174, 32200941, 32371183, and 31771312).

All methods described here have been approved by the animal ethical committee at Beijing Sport University and carried out in compliance with the National Institutes of Health (NIH) guidelines and the Chinese animal protection laws and institutional guidelines.
1. Mating and feeding rats
<ol>
	<li>Purchase specific pathogen-free (SPF) rated 10-week-old Wistar female rats and 11-week-old Wistar male rats.</li>
	<li>Implement adaptive feeding: Feed the Wistar rats ad libitu...

Exploring Maternal Exercise for the Well-Being of Rat Offspring

<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. 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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=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. 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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. 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This study aims to establish a feasible exercise program for pregnant rats, including detailed rat mating methods, swimming training protocols, and research methods to evaluate physiological indicators in fetuses. We have developed a prenatal swimming exercise training model through practice, with corresponding solutions to potential issues that may arise in the protocol (Table 1). This protocol will make it more practical and assist researchers in establishing prenatal exercise models.
<p class="jov...

The Developmental Origins of Health and Disease (DOHaD) concept emerged over the last two decades, which has shown that environmental influences during early development affect the risk of later pathophysiological processes associated with chronic non-communicable disease (NCD), especially diabetes, cardiovascular disease, and some types of cancer1. During the most plastic phase of fetal development, where offspring are exposed to the intrauterine environment, the gene and environment interaction and uteroplacental perfusion are crucial factors of fetal reprogramming2. Previous studies have predominantly utilized animal models to investigate the detrimental effects, such as maternal undernutrition, overnutrition, or obesity during pregnancy, and prenatal exposure to chemicals or hypoxia, on fetal development and long-term phenotypic effects in the offspring3. This produced low birth weight and later growth and produced adverse effects on offspring, including elevated blood pressure, dyslipidemia, and insulin resistance4. Since the rate of overweight and obesity is rapidly increasing among women of reproductive age, establishing effective interventions to prevent the intergenerational transmission of these deleterious maternal disorders is urgently needed, which has an important impact on population health5,6.
Exercise has long been recognized as an important preventive therapeutic for type 2 diabetes, hypertension, and several other diseases7. Exercise during pregnancy has beneficial effects for the mother and confers beneficial effects on offspring, thus reducing the maternal transmission of disease to offspring8. Several years of research have demonstrated exercise&#39;s safety and profound benefits, including substantial reductions in common pregnancy conditions such as gestational diabetes mellitus9,10. Regarding maternal and fetal safety, conducting exercise training and post-nursing protocols is challenging.
David11 et al. described animal exercise models applicable to cardiovascular health research. The three commonly used exercise models, voluntary wheel running, motorized treadmills, and swimming, have advantages and disadvantages. The voluntary wheel running closely mimics the locomotor behavior of rats, enabling them to move according to their circadian rhythm. However, this sort of exercise needs more precise regulation of intensity and duration. The motorized treadmill can control exercise intensity, volume, and duration, but sometimes it needs electric stimulation, which may trigger physical and psychological stress in the animals. Swimming is widely used in rodent studies due to the inherent swimming ability of rats, which can also avoid electric stimulation or mechanical damage to their feet and tails12. In studies on prenatal exercise models13,14, swimming is a commonly used exercise modality. As pregnancy progresses and the abdomen of the rat swells, treadmill and wheel running exercises may cause repeated contact of the abdomen with hard surfaces. In contrast, the water environment in swimming provides buoyancy, reducing the risk of exercise-related injuries for pregnant animals.
This protocol presents the rat breeding methods, how to perform the swimming training during pregnancy, and the post-breeding nursing protocols for pregnant rats after training. This pregnant swimming training model can be applied to a wide range of rat and mouse species, which may thus provide a useful rodent model for future investigation using animal-based models to study the mechanisms regulating the beneficial effects of maternal exercise on offspring health and the benefits of exercise across generations.

<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

The developmental origins of health and disease concept highlights the impact of early environments on chronic non-communicable diseases like diabetes, cardiovascular disease, and cancer. Studies using animal models have investigated how maternal factors such as undernutrition, overnutrition, obesity, and exposure to chemicals or hypoxia affect fetal development and offspring health, leading to issues like low birth weight, high blood pressure, dyslipidemia, and insulin resistance. Given the increasing prevalence of overweight and obesity among reproductive-age women, effective interventions are critical. Maternal exercise during pregnancy has emerged as a key intervention, benefiting both mother and offspring and reducing the risk of disease. This study compares the differences of three exercise models on pregnant rats: voluntary wheel running, motorized treadmills, and swimming. Swimming is the most beneficial option due to its safe and controlled intensity levels. This protocol details the rat breeding methods, swimming training during pregnancy, and post-breeding nursing protocols. This model, suitable for various rat and mouse species, is useful for studying the benefits of maternal exercise on offspring health and intergenerational wellness.

Author Spotlight: Enhancing the Offspring Health in Rats with Maternal Exercise During Pregnancy

Swimming Exercise Protocol and Care Methods for Pregnant Rats

Our research delineates rat-breeding methods. Prenatal swimming training produces, and the post-breeding nursing protocols for rats, which will be of good help to the researchers of exercise physiology and the perinatal biology. As pregnancy progresses and the abdomen of the rat swells, treatment and the regularly exercises may cause repeated contact of the abdomen with hard surfaces.In contrast, the water environment in swimming exercise protocol provides buoyancy, reducing the risk of exercise-related injuries for pregnant animals. Maternal obesity can lead to an increase in incidents of cardiovascular and metabolic diseases in offspring. In the future, when we apply this swimming exercise protocol to a high-fat diet, induce the obese, pregnant mice to explore the apt in a tic mag design.and find a new way to curb the intergenerational inheritance of obesity-related diseases.

In the rat breeding methods section (Figure 1), we determine the estrous cycle of female rats by making vaginal secretion slides to increase the success rate of breeding. The rats&#39; estrous cycle lasts 4-5 days, including proestrus, estrus, metestrus, and diestrus. Female rats in metestrus or diestrus exhibit behaviors such as distancing themselves from males. At the same time, those in estrus are willing to mate, and ovulation in female rats mostly occurs at the end of estrus. During pro...

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

Here, we present a protocol to delineate rat breeding methods, swimming training procedures, and post-breeding nursing protocols for pregnant rats after training. This protocol provides an animal model for studying the effects of maternal exercise during pregnancy on offspring and its underlying mechanisms.

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

Mating and Feeding Rats for Maternal Exercise Training to Study Offspring Health

To begin, label the microscope slides with the date and identification details of the rat. Between 6:00 and 7:00 p.m. grasp the base of the female rat's tail with the thumb and forefinger while applying pressure from the other fingers onto the lumbar vertebra of the rat.To collect vaginal secretions, moisten the cotton swab with saline solution. After lifting the rat tail, insert the tip of the swab gently into the vagina and roll it. Then, roll the swab tip across the slide.And locate the cells under the microscope using the 10X objective. After recording the estrus cycle stage, place the female rat along with a male rat in the mating cage between 7:00 to 8:00 p.m. On the second day between 7:00 and 8:00 a.m.if a copulatory plug is found at the base of the mating cage, examine the vaginal secretion smear of the female rat, as demonstrated earlier. If pregnancy is confirmed, isolate the female rat in a separate cage. On gestation day 20, replace the pregnant rat's cage with a clean one.And place medical absorbent cotton in the cage. Supply sufficient water and food for seven days. And position the cage in a quiet location.On the 21st day postpartum, distinguish the gender of the offspring rats. In females, the distance between the anus and the external genitalia is shorter than it is for males. Place the male and female offspring in two separate cages.

Prenatal Rat Swimming to Explore Offspring Health Benefits

To begin, obtain female rats for adaptation training. Then, grasp the tip of the rat's tail, lift it, and place the rat in a circular bucket filled with water. Throughout the 15 minute adaptation training, monitor the water temperature every five minutes, and add warm water to maintain the temperature.After the training, wash the rat fur in clean water, tempered at approximately 34 degrees Celsius, and transfer the rat to a cage with towels. Using a hand dryer, dry its fur thoroughly. Weigh the pregnant rat every day before training.For swimming training sessions, place the rat in water, taken in a circular bucket, and let it swim while maintaining the water temperature at about 34 degrees Celsius. Following the exercise, dry the rat, and provide it with five grams of rodent sunflower seeds as a nutritional supplement. Thoroughly clean and disinfect all equipment with ultraviolet light, including water buckets and towels.

Assessing the Physical Traits of Fetuses Obtained from Swim-Trained Pregnant Rats

To begin, obtain a swim-trained euthanized pregnant rat. Using a surgical knife, make an incision in the lower abdomen to expose the uterus completely. With forceps and scissors, carefully open the uterus to reveal the fetuses and placentas.Gently peel away the surrounding amnion, preventing any harm to the fetuses. Sequentially, separate fetuses and placentas by their anatomical positions. Using sterile gauze, clean the fetus to eliminate any blood and mucous.Then, weigh the fetus and the placenta. Next, observe the distance between the anus and genitalia. And document the gender, as well as the total number of male and female fetuses.Unfold the curled up fetus. And using a caliper, measure the length while the fetus is lying flat.

372 Views.  Beijing Sport University.  Here, we present a protocol to delineate rat breeding methods, swimming training procedures, and post-breeding nursing protocols for pregnant rats after training. This protocol provides an animal model for studying the effects of maternal exercise during pregnancy on offspring and its underlying mechanisms.

Exploring Maternal Exercise for the Well-Being of Rat Offspring

Summary