Immunohistochemical study of the ubiquitin–nuclear factor-κB pathway in the endometrium of the baboon (Papio anubis) with and without endometriosis
Romina S. Ilad A D F , Steven D. Fleming A , Christopher R. Murphy B and Asgerally T. Fazleabas C EA Department of Obstetrics and Gynaecology at Westmead Hospital, The University of Sydney, Westmead, NSW 2145, Australia.
B Department of Anatomy and Histology, The University of Sydney, Sydney, NSW 2006, Australia.
C Center for Women’s Health and Reproduction, Department of Obstetrics and Gynecology, University of Illinois, Chicago, IL 60612-7313, USA.
D Present address: Graduate School of Medicine, Building 28, The University of Wollongong, Wollongong, NSW 2522, Australia.
E Present address: Department of Obstetrics and Gynaecology and Reproductive Biology, Michigan State University, College of Human Medicine, Grand Rapids, MI 49503, USA.
F Corresponding author. Email: ri536@uow.edu.au
Reproduction, Fertility and Development 22(7) 1118-1130 https://doi.org/10.1071/RD08086
Submitted: 28 April 2008 Accepted: 11 March 2010 Published: 20 August 2010
Abstract
The aim of the present study was to conduct a semiquantitative immunohistochemical investigation into the levels of intermediary proteins within the nuclear factor (NF)-κB pathway throughout the menstrual cycle in a non-human primate, namely the baboon (Papio anubis), with and without endometriosis. Formalin-fixed eutopic (n = 2–4) and ectopic (n = 6–7) endometrial tissues from baboons at the mid-luteal phase were embedded in paraffin and examined for NF-κB pathway components (i.e. IκB kinase (IKK) α, IKKβ, phosphorylated (phospho-) IκBα and phospho-NF-κB p65 subunit), ubiquitin, 19S proteasome and the NF-κB activator tumour necrosis factor (TNF)-α. Similarly, endometrial tissues from baboons at the late follicular, mid-luteal and menses phase (n = 2–4) were investigated to determine the levels of these proteins throughout the menstrual cycle. Cytoplasmic stromal IKKα and glandular 19S proteasome immunostaining was elevated in the ectopic endometrium, whereas levels of ubiquitin, phospho-p65, IKKβ, TNF-α and nuclear 19S proteasome were similar in the eutopic and ectopic endometrium. A significant decrease in phospho-IκBα nuclear immunostaining was observed within glandular cells of the ectopic endometrium. In the eutopic endometrium, IKKα, ubiquitin and 19S proteasome immunostaining was elevated in different phases of the menstrual cycle, whereas levels of phospho-p65, IKKβ, phospho-IκBα and TNF-α remained unchanged. We have demonstrated that, in the baboon endometriosis model, levels of IKKα immunostaining are elevated, whereas those of phospho-IκBα are reduced, consistent with the hypothesis that excessive NF-κB activity plays a role in reducing ectopic endometrial apoptosis, which contributes to the pathophysiology of endometriosis. Further studies are required to confirm a causal association between elevated IKKα levels and reduced endometrial apoptosis.
Additional keyword: IκB kinase.
Acknowledgements
This research was supported by the National Institute of Child Health and Human Development and the National Institutes of Health through cooperative agreement (U54 HD 40093 to A.T.F.) as part of the Specialised Cooperative Centres Program in Reproduction and Infertility Research. The authors thank students and staff from the Department of Obstetrics and Gynaecology at The University of Illinois at Chicago, particularly Patty Mavrogianis, Prajna Banarjee and Kevin Jackson, for their technical advice and sectioning of baboon tissues. The Charitable Trust at Westmead Hospital is acknowledged for covering the travel associated with the use of specialist facilities at Chicago.
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