E-Poster Presentation ESA-SRB-ANZBMS 2021

Leydig Cell Glucocorticoid Receptor is Required to Maintain Steroidogenesis in Adulthood (#509)

Anne-Louise Gannon 1 , Annalucia Darbey 1 , Grace Chensee 1 , Natalie Reed 1 , Shanu Parameswaran 1 , Ben Lawrence 1 , Sarah Smith 1 , Lee B Smith 1 , Diane Rebourcet 1
  1. The University of Newcastle, Callaghan, NSW, Australia

Glucocorticoids are used in a wide range of clinical settings, including asthma, eczema, arthritis, Crohn's disease, mental health conditions, and, more recently, COVID-19. An estimated 1 in 2 Australians have received steroid treatment, and this prevalence is likely to increase[1]. 

Androgens are essential for life-long health and well-being[2]. Disruptions to the production or action of androgens are associated with many chronic pathologies and metabolic disorders. Glucocorticoids, exerting their action via the glucocorticoid receptor (GR), can regulate androgen biosynthesis by blocking key enzymes required for their production in the androgen-producing cells in the testis, Leydig cells[3]. Despite glucocorticoid action on androgen production being well documented, how they regulate testis function is unknown.

To establish the role of GR-signalling in the testis in adulthood, we utilised a novel technique to rapidly generate cell-specific knockouts using an adeno-associated virus to deliver Cre recombinase to Leydig cells specifically. Leydig cell GR knockout mice were generated via injection of Adeno-Associated Virus serous type-9 (AAV-9)[4] carrying either GFP (control) or Cre recombinase into the interstitial compartment of the testis of GR floxed mice. Mice were injected in adulthood and were collected following one round of spermatogenesis.

Our preliminary data show that interstitial delivery of Cre recombinase via AAV-9, is an effective and cell-specific method to ablate GR from Leydig cells. Analysis of blockade of GR signalling in Leydig cells in this model shows markedly suppressed luteinising hormone receptor (Lhcgr) and steroidogenic enzymes required for normal androgen production. This important new data suggests that GR-signalling plays a physiological role in normal testis function and potentially fertility. These novel findings provide a timely and previously unavailable opportunity to elucidate the role of GR-signalling in testis function and its ability to influence LC function and androgen production.

  1. Barnes PJ. Glucocorticosteroids: current and future directions. Br J Pharmacol. 2011;163(1):29-43. Epub 2011/01/05. doi: 10.1111/j.1476-5381.2010.01199.x. PubMed PMID: 21198556; PubMed Central PMCID: PMCPMC3085866.
  2. Aki Murashimaa SK, Axel Thomsonc, Gen Yamadaa,. Androgens and mammalian male reproductive tract development. Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms. 2014.
  3. Tilbrook A, Turner A, Clarke I. Effects of stress on reproduction in non-rodent mammals: the role of glucocorticoids and sex differences. Reviews of reproduction. 2000;5(2):105-13.
  4. Darbey A, Rebourcet D, Curley M, Kilcoyne K, Jeffery N, Reed N, et al. A comparison of in vivo viral targeting systems identifies adeno‐associated virus serotype 9 (AAV9) as an effective vector for genetic manipulation of Leydig cells in adult mice. Andrology. 2021;9(1):460-73.