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Vertebrate reproductive science and technology
RESEARCH ARTICLE (Open Access)

Transit along the vas deferens results in a high percentage of filiform spermatozoa with compacted chromatin in the rooster (Gallus domesticus)

B. Bernal https://orcid.org/0000-0001-9144-1554 A * , A. Behnamifar B , C. Álvarez-Rodríguez C , A. Toledano-Díaz https://orcid.org/0000-0001-6679-485X D , C. Castaño D , R. Velázquez D , M. G. Gil A , A. Gutiérrez-Adán https://orcid.org/0000-0001-9893-9179 D , Henri Woelders E , E. Blesbois F and J. Santiago-Moreno https://orcid.org/0000-0001-5551-8120 D
+ Author Affiliations
- Author Affiliations

A Department of Animal Breeding, INIA-CSIC, Madrid, Spain.

B Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran.

C Department of Reproductive Biology, FES-Iztacala, UNAM, Iztacala, Mexico.

D Department of Animal Reproduction, INIA-CSIC, Madrid, Spain.

E Department of Animal Breeding and Genomics, Wageningen University and Research, Wageningen, Netherlands.

F PRC, CNRS, IFCE, INRAE, Université de Tours, Nouzilly, France.


Handling Editor: Marc Yeste

Reproduction, Fertility and Development 34(10) 699-712 https://doi.org/10.1071/RD21209
Published online: 4 April 2022

© 2022 The Author(s) (or their employer(s)). Published by CSIRO Publishing. This is an open access article distributed under the Creative Commons Attribution 4.0 International License (CC BY)

Abstract

The present work aimed to evaluate the chromatin compaction of rooster spermatozoa along the male reproductive tract, and to study the vas deferens lining cells, potentially involved in sperm maturation. Chromomycin A3 (CMA3) was used to determine the chromatin compaction of spermatozoa from testis (T), proximal (including epididymis, V1), intermediate (V2) and distal (V3) vas deferens, and ejaculate (E). Six Birchen Leonesa roosters were used. E was obtained in vivo by dorso-ventral massage. V1, V2 and V3 sperm were obtained post mortem (six pairs of vasa deferentia), by flushing. T was obtained by washing the testes, cut in halves. The fixed cells were stained with CMA3 and propidium iodide for flow cytometry assessment. Results showed higher (P < 0.01) median fluorescence intensity (lower chromatin compaction) of T (693.8 ± 30.2) than V1 (546.3 ± 17.7), V2 (515.1 ± 12.1), V3 (517.6 ± 12.3) and E (491.4 ± 16.1). Regarding the percentage of intensely stained cells, T differs (P < 0.05) from V2, V3 and E, V1 differs (P < 0.05) from V3 and E, while V2, V3 and E do not differ. The histological analysis revealed secretory capacity of the vas deferens. Our findings specified that the transit though the vas deferens results in high percentage of compacted chromatin spermatozoa in E.

Keywords: chromatin condensation, flow cytometry, histology, protamines, rooster sperm chromatin, rooster sperm decondensation, sperm maturation process, vas deferens.


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