For more information or to watch the webinar, visit the following link: https://insidescientific.com/webinar/application-of-treadmill-exercise-using-mice-as-a-model-for-studying-the-influence-of-maternal-exercise-on-offspring-metabolic-reprogramming/ Epidemiological studies show the beneficial effects of exercise during pregnancy in alleviating birth-related complications and improving neonatal metabolic health, whereas the underlying molecular mechanisms remain unclear. A protocol for maternal exercise (ME) of mice facilitates mechanistic exploration based on the regimen of the exercise guidelines for pregnant women. Treadmill exercise allows precise control of exercise intensity and duration compared to voluntary wheel running exercise, which leads to reduced variations among individual mouse within treatments and facilitates translation into ME in humans. In this webinar, Dr. Son explains the exercise protocol for studying the effects of ME on fetal/offspring health. He discusses his findings on how ME enhances placental vascularization/development and fetal brown fat/muscle development. He also elaborates on how the exercise protocol can be useful for standardizing maternal treadmill exercise using mice as an experimental model. Key Topics Include: Treadmill exercise protocol for mice Application of forced exercise protocol for offspring metabolic health Offspring brown fat/muscle development in response to maternal exercise

1. Copyright 2022. All Rights Reserved. Contact Presenter for Permission
Application of Treadmill Exercise
Using Mice as a Model for
Studying the Influence of
Maternal Exercise on Offspring
Metabolic Reprogramming
Junseok Sun, PhD
OB/GYN and Reproductive Sciences
University of Maryland School of Medicine
Assistant Professor

2. Application of Treadmill Exercise using Mice
: As a Model for Studying Influence of Maternal Exercise
on Offspring Metabolic Reprogramming
Jun Seok Son, Ph.D.
InsideScientific Webinar
April 5th, 2022

3. Fetal/Offspring Metabolic Reprogramming
Exercise
Healthy adult
Adult with metabolic
dysfunction
Infant
Nutrition

4. Interscapular
Perirenal
Brown adipose tissue
(BAT)
Skeletal muscle
Fetal Metabolic Reprogramming: BAT and muscle

5. Brown adipose tissue
(BAT)
 Mitochondria ↑
 Energy consumption ↑
(thermogenesis)
White adipose tissue
(WAT)
VS.
 Mitochondria ↓
 Energy storage ↑
BAT versus WAT: mitochondrial density

6. Mitochondrion: as a power station for energy generation
Glycolysis
Krebs
Lipolysis
Glucose
TAG
ATP
β-oxidation Power station

7. Does brown/beige fat thermogenesis regulate metabolism?
(UCP1-dependent manner)
Intermembrane space
Mitochondrial matrix
H+
H+
H+ H+
H+
H+
H+ H+
H+
H+ H+
H+
H+
H+
H+
H+ H+
H+
H+
H+
H+
H+
H+
H+
Heat
ATP
I
II
III IV UCP1
NADH
NAD+ FADH2
FAD+
e-
O2
2H2O

8. UCP1-independent thermogenesis?
(Gamu & Tupling, Nat Med, 2017)

9. The roles of brown and beige fat
Thermogenesis
 UCP1-dependent
 UCP1-independent
Metabolism
 Glucose (glycolysis)
 Fatty acids (lipolysis)
 Mitochondria
Secretion of molecules
 Peptides
 Lipids
 miRNAs
 …

10. BAT
Exercise effects on Brown fat development
Thermogenesis
(non-shivering)
Intermembrane space
Mitochondrial matrix
H+
H+
H+ H+
H+
H+
H+ H+
H+
H+ H+
H+
H+
H+
H+
H+ H+
H+
H+
H+
H+
H+
H+
H+
Heat
ATP
I
II
III IV UCP1
NADH
NAD+ FADH2
FAD+
e-
O2
2H2O
UCP1-dependent thermogenesis
Exercise training

11. White adipocyte
Exercise effects on Browning WAT
Inguinal WAT
Browning
Whitening
Beige adipocyte
Inguinal WAT

12. How to mediate brown/beige fat thermogenesis?
(Short- and Long-term Effects of Exercise)
(Egan & Zierath, Cell Metab, 2013; Physical Activity Guidelines for Americans, 2nd Edition)

13. Maternal exercise effects: A systemic & meta-analysis
(Chen et al., Sports Med, 2021)
(Maternal normal weight)
(Maternal obesity)

14. Exercise protocol for pregnant animals
Wheel
Running Ex
Voluntary wheel running program
Carter et al., 2012
Am J Physiol Endocrinol Metab
Carter et al., 2013
Med Sci Sports Exerc
Laker et al., 2014
Diabetes
Stanford et al., 2015
Diabetes
Eclarinal et al., 2016
FASEB J
Stanford et al., 2017
Diabetes
Kusuyama et al., 2021
Cell Metab
…
Treadmill exercise program
Quiclet et al., 2016
Am J Physiol Endocrinol Metab
Son et al., 2019
J Physiol
Son et al., 2020
Sci Adv
Son et al., 2020
Cell Rep
Falcao-Tebas et al., 2020
J Physiol
Son et al., 2022
eBioMedicine
…

15. Pros and Cons of Treadmill Exercise: versus voluntary exercise
Treadmill Ex
Forced exercise
Pros
 Control exercise duration and intensity
 Reduce the variation of exercise effects
among mice
Cons
 Induce stress
Wheel
Running Ex
Voluntary exercise
VS.
Pros
 Reduce stress
Cons
 Cannot control exercise during and intensity
 Induce large variation of exercise duration
and intensity among mice

16. Exercise Guidelines for Pregnant Women
(Zavorsky & Longo, Sports Med, 2011)

17. Trimester-dependent Exercise Intensity
(Chae et al., Bio-Protocol, 2020)
https://bio-protocol.org/e3838

18. Treadmill exercise training for mice
(Chae et al., Bio-Protocol, 2020)
https://bio-protocol.org/e3838

19. M-Ctrl_HFD M-Ex_HFD
Female
Male
Liver (Oil Red O staining)
 Protects liver steatosis
 Reduces adipose tissue mass
(Son et al., Science Advances, 2020)
Maternal exercise prevents offspring hepatic lipid steatosis
Glucose tolerance
Body weight

20. Offspring Metabolic Reprogramming: Brown/beige fat
M-Ctrl_HFD M-Ex_HFD
Female
Male
M-Ctrl_HFD M-Ex_HFD
Female
Male
(Son et al., Science Advances, 2020)

21. Fetal Metabolic Reprogramming: Brown fat
M-Ctrl M-Ex
(Son et al., Unpublished data)
Exercise during pregnancy (daily)

22. How does ME mediate fetal reprogramming?

23. Fetal Metabolic Reprogramming: Role of placenta
Placentokines
(Placenta + cytokine)
Interscapular
Brown adipose tissue
(BAT)
(Chae et al., FEBS Journal, 2021)
Skeletal muscle
Reprogramming

24. Myokines
Muscle
Adipokines
Adipose tissue
Exercise-induced
secretion
Exerkines
(Chae et al., FEBS Journal, 2021)
Placentokines? Exerkines?
Placentokines
Placenta

25. Maternal Exercise
-induced placentokines
Liver
SOD3 SOD3
in liver
- Demethylation of glucose
metabolic genes ↑
Others
- Glucose tolerance ↑
- Insulin sensitivity ↑
BAT
Apelin
Apelin in muscle
- Mitochondrial biogenesis ↑
- Demethylation of Pgc1a ↑
in BAT
- Fat thermogenesis ↑
- Mitochondrial biogenesis ↑
- Demethylation of Prdm16 ↑
Others
- Glucose homeostasis ↑
- Insulin resistance ↓
- Metabolic capacity (OCRs) ↑
Adiponectin
Adiponectin
- Fetal weight ↓
- Fetal glucose level ↓
Irisin
Irisin
- Fetal growth
- Preterm birth
(Chae et al., FEBS Journal, 2021)
Placentokines: As mediators for fetal reprogramming

26. ME-mediated apelin in fetal reprogramming?

27. Maternal exercise elevates circulating apelin levels
(Castan-Laurell et al., Endocrine, 2011; Son et al., Science Advances, 2020)
Role of apelin in metabolism

28. Fetal side
of placenta
(Labyrinth
zone)
Fetal tissues
Hypothesis 2
Hypothesis 3
Hypothesis 1
From maternal
circulation
Where is apelin secreted and released from?

29. Apelin expression in
labyrinth zone
(Son et al., Science Advances, 2020)
[Hypothesis 2]
 Placental labyrinth expresses apelin by exercise
during pregnancy.
[Hypothesis 3]
 Fetal brown fat and muscle express apelin by
maternal exercise.
Placenta as an endocrine organ

30. Apelin injection during pregnancy (daily)
Fetal Metabolic Reprogramming: Brown fat
(Son et al., Science Advances, 2020)

31. Fetal Metabolic Reprogramming: Muscle
(Son et al., Cell Reports, 2020)

32. Muscle-based thermogenesis
(Son et al., eBioMedicine, 2022)

33. Maternal exercise enhances offspring muscle NST
Maternal Obesity
(Muscular non-shivering
thermogenesis)
Sarcolipin
Maternal
Exercise
(Son et al., eBioMedicine, 2022)

34. Muscle-based non-shivering thermogenesis
(Son et al., eBioMedicine, 2022)
Gene expression
Protein levels

35. Muscle-based non-shivering thermogenesis
(SLN-dependent/UCP3-dependent)
(Son et al., eBioMedicine, 2022)

36. Summary
1) Placentokines
(Placenta + cytokine)
3) Sarcolipin-dependent
thermogenesis Improved offspring
metabolic health
 Glucose tolerance
 Insulin sensitivity
Maternal
Exercise
2) UCP1-dependent
thermogenesis

37. Acknowledgements
Collaborators (PIs)
Min Du, PhD (WSU)
Zhihua Jiang, PhD (WSU)
Mei-Jun Zhu, PhD (WSU)
Hongyang Wang, PhD (Shanghai Academy)
Previous lab members (WSU)
Jeanene M. de Avila, MS
Song Ah Chae, MS
Yanting Chen, PhD
Xiangdong Liu, MS
Liang Zhao, PhD
Qiyu Tian, MS
Yao Gao, MS

38. Conclusions
 Maternal exercise reprograms fetal brown fat and muscle development via
hormones secreted from placenta.
 Apelin supplementation during pregnancy partially mirrors the beneficial
effects of maternal exercise on fetal brown fat and muscle mitochondrial
biogenesis and thermogenesis.
 Exercise-dependent placenta-derived hormones may be the therapeutic
targets to overcome pediatric metabolic dysfunction.

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