Expression of endothelial and inducible nitric oxide synthases is modulated in the endometrium of cyclic and early pregnant mares
H. Welter A , H. Bollwein B , F. Weber C , S. Rohr C and R. Einspanier D EA Physiologie, Zentralinstitut für Ernährungs- und Lebensmittelforschung (ZIEL), Wissenschaftszentrum Weihenstephan (WZW), TU München, Weihenstephaner Berg 3, D-85350 Freising, Germany.
B Clinic for Cattle, Bischofsholer Damm 15, School for Veterinary Medicine, 30173 Hanover, Germany.
C Clinic of Veterinary Gynaecology and Obstetrics, Faculty of Veterinary Medicine, University of Munich, Munich, Germany.
D Institute of Veterinary Biochemistry, Free University of Berlin, Oertzenweg 19b, D-14163 Berlin, Germany.
E To whom correspondence should be addressed. email: einspani@zedat.fu-berlin.de
Reproduction, Fertility and Development 16(7) 689-698 https://doi.org/10.1071/RD03103
Submitted: 11 November 2003 Accepted: 20 September 2004 Published: 9 December 2004
Abstract
The expression of the endothelial and inducible nitric oxide synthases (eNOS and iNOS, respectively) was examined in the endometrium of cyclic and pregnant mares by real-time polymerase chain reaction and immunohistology. The concentration of eNOS mRNA varied throughout the oestrous cycle, with significantly higher transcripts on Day 5 of the oestrous cycle (P < 0.05), whereas iNOS transcription did not change significantly over time (P > 0.05). In early pregnant mares both eNOS and iNOS mRNA increased between Days 12 and 15 (P < 0.05). In cyclic mares, eNOS protein was detected immunocytochemically in endometrial epithelia, the basement membrane, the endothelial layer and smooth muscle cells of the vasculature. Using immunocytochemical methods, iNOS protein was undetectable in the endometrium of cyclic mares but could be demonstrated in pregnant mares. Endometrial epithelia of pregnant mares were immunopositive for both proteins with a more intense labelling for iNOS. Thus, the present study describes for the first time the modulation and spatial distribution of eNOS and iNOS expression during the oestrous cycle and early pregnancy, suggesting that ovarian steroids are differently involved in the regulation of each NOS. Localisation of eNOS protein in endometrial epithelia and various vascular components indicates that this isoform may be involved in the regulation of endometrial cyclicity. The presence and increase of both forms of NOS during early gestation suggest a role for them in the control of endometrial vascular bed and glandular activity to provide a suitable microenvironment for successful pregnancy.
Extra keywords: endothelial nitric oxide synthase, immunohistology, inducible nitric oxide synthase, real-time polymerase chain reaction.
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