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ABSTRACT
The ectoplasmic specialization (ES) is essential for Sertoli-germ cell communication to support all phases of germ cell development and maturity. Its formation and remodeling requires rapid reorganization of the cytoskeleton. However, the molecular mechanism underlying the regulation of ES assembly is still largely unknown. Here, we show that Sertoli cell-specific disruption of autophagy influenced male mouse fertility due to the resulting disorganized seminiferous tubules and spermatozoa with malformed heads. In autophagy-deficient mouse testes, cytoskeleton structures were disordered and ES assembly was disrupted. The disorganization of the cytoskeleton structures might be caused by the accumulation of a negative cytoskeleton organization regulator, PDLIM1, and these defects could be partially rescued by Pdlim1 knockdown in autophagy-deficient Sertoli cells. Altogether, our works reveal that the degradation of PDLIM1 by autophagy in Sertoli cells is important for the proper assembly of the ES, and these findings define a novel role for autophagy in Sertoli cell-germ cell communication.
Abbreviations
| aES | = | apical ectoplasmic specialization |
| APES | = | 3-aminopropyl-triethoxysilane |
| bES | = | basal ectoplasmic specialization |
| BTB | = | blood–testis barrier |
| CHX | = | cycloheximide |
| CQ | = | chloroquine |
| DAB | = | 3, 3′-diaminobenzidine |
| DAPI | = | 4′, 6-diamidino-2- phenylindole |
| ER | = | endoplasmic reticulum |
| ES | = | ectoplasmic specialization |
| H&E | = | hematoxylin and eosin staining |
| HRP | = | horseradish peroxidase |
| IF | = | immunofluorescent staining |
| IHC | = | immunohistochemistry |
| IP | = | immunoprecipitation |
| MEF | = | mouse embryonic fibroblast |
| MS | = | mass spectrometry |
| NH4Cl | = | ammonium chloride |
| OCT | = | optimum cutting temperature compound |
| PBS | = | phosphate-buffered saline |
| PFA | = | paraformaldehyde |
| PI | = | propidium iodide |
| SSCs, | = | spermatogonial stem cells |
| STRING | = | Search Tool for the Retrieval of Interacting Genes/Proteins |
| TBC | = | tubulobulbar complex |
| TEM | = | transmission electron microscopy |
| TUNEL | = | terminal deoxynucleotidyl transferase dUTP nick end-labeling |
| VAP | = | average path velocity |
| VCL | = | curvilinear velocity |
| VSL | = | straight-line velocity |
Disclosure of potential conflicts of interest
The authors declare that they have no conflict of interest.
Acknowledgments
We thank Noboru Misushima and Masaaki Komatsufor providing the Atg5 and Atg7 floxed mice. We thank Hong Zhang for critical reading of the manuscript.
Funding
This work was supported by the National Natural Science Foundation of China (Grant No. 31471277, 31171374), the Major Basic Research Program (Grant No. 2012CB944404) and the Knowledge Innovation Program of Chinese Academy of Science (Grant No. KSCX2-YW-N-071).