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Characterization of in vivo autophagy during avian spermatogenesis1.

Abstract

Spermatogenesis is a complex cellular process that includes many subcellular events that are essential for the production of healthy spermatozoa. Autophagy is a physiological process that plays a significant role in the process of spermatogenesis; however, autophagy during avian spermatogenesis has not yet been reported. In the current study, we characterized in vivo autophagy throughout the process of domestic fowl spermatogenesis. Autophagy-specific markers, including microtubule-associated protein light chain 3 (LC3), sequestosome 1 (p62), and autophagy-related 7 (Atg7), were used to confirm the occurrence of autophagy in testicular germ cells. The protein expression of Atg7, LC3, and p62 in domestic fowl testes was confirmed by Western blotting. The immunohistochemical staining indicated a strong localization of LC3 and Atg7 within spermiogenic cells (intermediate and late spermatids) and primary spermatocytes. However, poorly expressed in cells (spermatogonia) that were located near the basement membrane. The immunofluorescence staining results showed the opposite tendency for LC3 and p62. LC3 was more strongly localized within the elongated spermatids, while p62 was strongly localized within the early spermatids. Moreover, the ultrastructural components of autophagy were revealed by transmission electron microscopy. Well-developed autophagosomes and multivesicular bodies were found to be prominent in primary spermatocytes (zygotene and pachytene) and spermiogenic cells. Furthermore, other vesicular structures, such as early endosomes and amphisomes, were also observed during spermatogenesis. The above findings collectively suggest that autophagy is active during spermatogenesis and that the level of autophagy increases from the basal to the luminal regions of the seminiferous tubules of domestic fowl testes. We propose that autophagic pathways may be involved in multiple functions to sustain spermatogenesis.

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  • Authors+Show Affiliations

    ,

    Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Jiangsu Province 210095, China. Faculty of Veterinary and Animal Sciences, University of Poonch Rawalakot, Azad Kashmir 12350, Pakistan.

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    Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Jiangsu Province 210095, China.

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    Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Jiangsu Province 210095, China.

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    Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Jiangsu Province 210095, China.

    ,

    Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Jiangsu Province 210095, China.

    ,

    Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Jiangsu Province 210095, China.

    ,

    Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Jiangsu Province 210095, China.

    ,

    Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Jiangsu Province 210095, China.

    ,

    Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Jiangsu Province 210095, China.

    Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Jiangsu Province 210095, China.

    Source

    Poultry science : 2019 Jun 14 pg

    Pub Type(s)

    Journal Article

    Language

    eng

    PubMed ID

    31198935

    Citation

    Haseeb, A, et al. "Characterization of in Vivo Autophagy During Avian Spermatogenesis1." Poultry Science, 2019.
    Haseeb A, Bai X, Vistro WA, et al. Characterization of in vivo autophagy during avian spermatogenesis1. Poult Sci. 2019.
    Haseeb, A., Bai, X., Vistro, W. A., Tarique, I., Chen, H., Yang, P., ... Chen, Q. (2019). Characterization of in vivo autophagy during avian spermatogenesis1. Poultry Science, doi:10.3382/ps/pez320.
    Haseeb A, et al. Characterization of in Vivo Autophagy During Avian Spermatogenesis1. Poult Sci. 2019 Jun 14; PubMed PMID: 31198935.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - Characterization of in vivo autophagy during avian spermatogenesis1. AU - Haseeb,A, AU - Bai,X, AU - Vistro,W A, AU - Tarique,I, AU - Chen,H, AU - Yang,P, AU - Gandahi,N S, AU - Iqbal,A, AU - Huang,Y, AU - Chen,Q, Y1 - 2019/06/14/ PY - 2019/01/09/received PY - 2019/05/25/accepted PY - 2019/6/15/entrez KW - Atg7 KW - LC3 KW - MVB KW - autophagosomes KW - autophagy KW - domestic fowl KW - p62 KW - spermatogenesis JF - Poultry science JO - Poult. Sci. N2 - Spermatogenesis is a complex cellular process that includes many subcellular events that are essential for the production of healthy spermatozoa. Autophagy is a physiological process that plays a significant role in the process of spermatogenesis; however, autophagy during avian spermatogenesis has not yet been reported. In the current study, we characterized in vivo autophagy throughout the process of domestic fowl spermatogenesis. Autophagy-specific markers, including microtubule-associated protein light chain 3 (LC3), sequestosome 1 (p62), and autophagy-related 7 (Atg7), were used to confirm the occurrence of autophagy in testicular germ cells. The protein expression of Atg7, LC3, and p62 in domestic fowl testes was confirmed by Western blotting. The immunohistochemical staining indicated a strong localization of LC3 and Atg7 within spermiogenic cells (intermediate and late spermatids) and primary spermatocytes. However, poorly expressed in cells (spermatogonia) that were located near the basement membrane. The immunofluorescence staining results showed the opposite tendency for LC3 and p62. LC3 was more strongly localized within the elongated spermatids, while p62 was strongly localized within the early spermatids. Moreover, the ultrastructural components of autophagy were revealed by transmission electron microscopy. Well-developed autophagosomes and multivesicular bodies were found to be prominent in primary spermatocytes (zygotene and pachytene) and spermiogenic cells. Furthermore, other vesicular structures, such as early endosomes and amphisomes, were also observed during spermatogenesis. The above findings collectively suggest that autophagy is active during spermatogenesis and that the level of autophagy increases from the basal to the luminal regions of the seminiferous tubules of domestic fowl testes. We propose that autophagic pathways may be involved in multiple functions to sustain spermatogenesis. SN - 1525-3171 UR - https://www.unboundmedicine.com/medline/citation/31198935/Characterization_of_in_vivo_autophagy_during_avian_spermatogenesis1 L2 - https://academic.oup.com/ps/article-lookup/doi/10.3382/ps/pez320 DB - PRIME DP - Unbound Medicine ER -