Received: 18 July 2022 | Accepted: 26 May 2023 DOI: 10.1111/eve.13847_1
OR IG INA L A R T I C L E
Image analysis quantification of sperm acrosome- associated 3 protein expression in domesticated and free- roaming equine ovaries
Mikaela K. Lee | Victoria M. Strandberg | Brynley C. Cozzi | Ann M. Ramsey | Kendall A. Evanchak | Michelle A. Kutzler
Department of Animal and Rangeland Sciences , Oregon State University , Corvallis , Oregon , USA
Correspondence : Victoria M. Strandberg Email:
strandbv@oregonstate.edu
SUMMARY
Background : In the United States, the carrying capacity of the land has been overwhelmed with the wild horse and burro populations. Identification of a long- term contra- ception method is needed to control growing populations as current methods cannot quell these growing numbers. Sperm acrosome- associated 3 protein (SPACA3) is a protein expressed in both male and female reproductive systems. SPACA3 may prove to be a viable contraception target in horses in immunosterilant research. Objectives : The objective of the current study was to eval- uate and compare ovarian immunoexpression of SPACA3 between domesticated and free- roaming mares. This in- formation may lead to the development of a SPACA3 im- munosterilant
to reduce free- roaming horse and burro
populations. Study design : Non- randomised comparative study design. Methods : Routine immunohistochemistry was performed on ovarian sections from domesticated ( n = 8) and free- roaming mares ( n = 8). At least three representative images of each fol- licle stage (primordial, primary, secondary and tertiary) from each ovary were digitally captured and analysed using Image J software. Expression of SPACA3 was quantified in cellular locations (granulosa or theca) specific to each follicle stage: primordial, primary, secondary and tertiary. Results : Domesticated horses had higher SPACA3 expression compared to free- roaming mares in granulosa cells of primor- dial
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( p = 0.0022), primary ( p < 0.001), secondary ( p = 0.025) © 2023 EVJ Ltd.
and tertiary follicles ( p = 0.0078). In theca cells, domesticated horses had higher SPACA3 expression in tertiary follicles ( p = 0.022), but not secondary follicles ( p = 0.089). The highest SPACA3 expression was observed in the granulosa cells of ter- tiary follicles ( p = 0.022). Main limitations : Varying background colours of representa- tive images taken of ovaries, leading to slight technical issues in Image J software. Conclusions : This is the first report quantifying SPACA3 immu- noexpression during folliculogenesis and describing the spatial and temporal expression of SPACA3 in the equine ovary. This information may lead to the development of a SPACA3 immu- nosterilant to reduce free- roaming horse and burro populations.
K E YWORDS horse , burro , granulosa cell , immunosterilant , theca cell
Clinical relevance
• Quantification of SPACA3 in domestic and free- roaming mares provides insight into a potential future immunos- terilant for horses.
• Environmental and physiological differences may influ- ence the expression values of SPACA3.
• The reproductive role of SPACA3 can also be explored in other mammals such as humans, dogs, cats, and mice, allow- ing for a better understanding of reproduction systems.
aaep.org/wiley-redirect Equine Vet Educ. 2024;36:42.
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