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Type of publication: Journal Article
Type of document: Full Paper

Year: 2016

Authors: Merkl, M; Ertl, R; Handschuh, S; Aurich, C; Schäfer-Somi, S

Title: The cholesterol transporter ABCA1 is expressed in stallion spermatozoa and reproductive tract tissues.

Source: Theriogenology. 2016; 85(6):1080-1089



Authors Vetmeduni Vienna:

Aurich Christine
Ertl Reinhard
Handschuh Stephan
Merkl Maximiliane
Schäfer-Somi Sabine

Vetmed Research Units
University Clinic for Small Animals, Clinical Unit of Obstetrics, Gynaecology and Andrology
Insemination and Embryotransfer Platform
VetCore


Project(s): Untersuchungen zum Cholesteroltransport in der Membran von Hengstspermatozoen-Einfluss osmotischer und Temperaturänderungen sowie der Zusammensetzung des Seminalplasmas


Abstract:
In the present study, we assessed the presence of the ATP-binding-cassette (ABC) transporter molecules ABCA1 in spermatozoa of adult stallions and in testicular and epididymal tissue of prepubertal and adult stallions. For this purpose, semen samples from six fertile Shetland pony stallions aged 4 to 19 years were collected. Semen was collected from each stallion on three consecutive days. Ejaculates were analyzed immediately after collection, and only ejaculates meeting minimal requirements for fertile stallions were further evaluated. ABCA1 immunosignal was localized after staining of semen smears with different antibodies and counterstaining with Fluorescein isothiocyanate (FITC)-peanut agglutinin (PNA) and 4',6-Diamidin-2-phenylindol (DAPI). In a total of three samples, capacitation and acrosome reaction were induced by means of capacitation medium and progesterone substitution, respectively. Testicular and epididymal tissues were obtained from five prepubertal stallions aged 8 to 12 months and five adult stallions aged 4 to 9 years. For quantitative RT-PCR (qPCR), testicular and epididymal tissue of another seven adult (aged 1.5-14.5 years) and five prepupertal stallions (6-8 months) was used. For immunohistochemistry, sections from the caput, corpus, and cauda of the testes and epididymes were stained with the same specific antibodies as for immunocytochemistry. In stallion spermatozoa, strong immunosignal for ABCA1 was detected in the acrosomal area, the equatorial zone, and the principle piece of the flagellum but not in the caudal part of the head and the midpiece. In damaged or acrosome-reacted spermatozoa the FITC-PNA signal vanished together with the ABCA1 signal in most spermatozoa. In testicular tissue, strong immunostaining for ABCA1 was mainly visible in the heads and flagella of round spermatids and weaker signals in late spermatids and released spermatozoa. No staining was assessed in the Sertoli cells and spermatogonia of adult stallions, whereas strong signals in Leydig cells were present in prepubertal stallions. In prepubertal stallions, the ABCA1 messenger RNA level in testicular tissue was significantly higher than in adult stallions. We conclude that the ABCA1 transport molecule is present in adult and prepubertal stallion spermatozoa as well as testicular and epididymal tissue. ABCA1 is supposed to contribute to cholesterol transport and to support capacitation; however, this remains to be proven by functional studies. Species-specific differences concerning the localization inside the spermatozoa membrane are alike.Copyright © 2016 Elsevier Inc. All rights reserved.

Keywords Pubmed: ATP Binding Cassette Transporter 1metabolism
Acrosome Reaction
Animals
Biological Transport
Cholesterolmetabolism
Epididymismetabolism
Horsesmetabolism
Immunohistochemistryveterinary
Male
Semen Analysisveterinary
Species Specificity
Spermatozoametabolism
Testismetabolism

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