Activation of adenosine monophosphate–activated protein kinase (AMPK) enhances energy metabolism, motility, and fertilizing ability of cryopreserved spermatozoa in domestic cat model

  • Paweena ThuwanutEmail author
  • Pierre Comizzoli
  • Kamthorn Pruksananonda
  • Kaywalee Chatdarong
  • Nucharin Songsasen
Gamete Biology



Increasing intracellular energy storage by chemically activating adenosine monophosphate–activated protein kinase (AMPKα) prior to sperm cryopreservation may improve post-thawed sperm function. Using the domestic cat as a biomedical model, the objectives were to (1) confirm the expression of AMPKα and its regulatory kinases in epididymal spermatozoa and (2) assess the influence of AMPK activator, 5′-aminoimidasole-4-carboxamide-1-β-d-ribofuranoside (AICAR) on epididymal sperm function before and after cryopreservation.


In study I, sperm samples of different qualities were obtained from cauda epididymides of domestic cats and evaluated for AMPKα expression. In study II, epididymal spermatozoa were equilibrated for either 30 or 60 min in the presence of 0 (control), 0.5, 2.0, and 5.0 mM AICAR and sperm functions were assessed before and after cryopreservation. In study III, epididymal spermatozoa were treated as in study II and evaluated for AMPKα signaling protein expressions (phospho-AMPKα Thr172 and GLUT1) as well as ATP levels.


AMPKα protein expression was higher in high-motility vs poor-motility samples. Thirty-minute equilibration with 0.5 mM AICAR improved motion characteristics and fertilizing ability of cryopreserved sperm to the control. Increased expressions of phospho-AMPKα Thr172 and GLUT1 as well as intracellular ATP level were confirmed in sperm samples equilibrated with 0.5 or 2.0 mM AICAR for 30 min.


Presence and role of AMPKα protein in cat regulating sperm function were demonstrated before and after cryopreservation. Findings could be used to potentially enhance cryopreserved sperm function in sub-fertile men.


AMPKα protein Energy regulation Felids Non-rodent model Sperm functions 



The authors are thankful for Dr. Em-Orn Olanrattamanee for statistical analysis.

Funding information

The present study was financially supported by (I) The Thailand Research Fund (Contract No. MRG 5980231) and (II) Research Unit of Reproductive Medicine and Fertility Preservation, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Division of Reproductive Medicine, Department of Obstetrics-Gynecology, Faculty of MedicineChulalongkorn UniversityBangkokThailand
  2. 2.Research Unit of Reproductive Medicine and Fertility Preservation, Faculty of MedicineChulalongkorn UniversityBangkokThailand
  3. 3.Smithsonian Conservation Biology InstituteVirginia and Washington DCUSA
  4. 4.Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary ScienceChulalongkorn UniversityBangkokThailand

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