Pattern of protein expression in the epididymis of Oligoryzomys nigripes (Cricetidae, Sigmodontinae)
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In the epididymis, epithelial cells work in a concerted manner to create a luminal environment for sperm maturation, transport, and storage. However, the cell functions may be affected by anthropogenic factors, causing negative impacts on male fertility. In our study, we describe the pattern of protein expression in the epithelium and luminal fluid from epididymis of Oligoryzomys nigripes, a South American sigmodontine rodent whose reproductive biology has been little studied. Nine animals were captured from a preserved area of Atlantic Forest, where the exposure to anthropogenic influences is minimal. Epididymides were processed for histological analysis under light and epifluorescence microscopy, in which we used cell-specific markers aquaporin 9 (AQP9), vacuolar H+-ATPase (V-ATPase), and cytokeratin 5 (KRT5). Other samples were assessed for protein expression using shotgun proteomics. Similar to laboratory rodents, principal cells expressed AQP9 in their stereocilia. Basal cells, identified by KRT5 labeling, presented lateral body projections and a few axiopodia going toward the lumen. Clear cells expressed V-ATPase in their sub-apical vesicles and microplicae, and showed different shapes along the duct. Shotgun proteomics detected 51 proteins from epididymal supernatant. Most of them have been previously described in other species, indicating that they are well conserved. Twenty-three proteins detected in O. nigripes have not been described in epididymis from other South American sigmodontine rodents, confirming that the secretion pattern is species-specific. Our findings in O. nigripes from a protected area may help to create a baseline for studies investigating the effects of anthropogenic factors on functionality of the epididymal epithelium.
KeywordsEpithelial cells Epididymal fluid Proteome Morphometry Reproductive biology
The authors thank Ana Carolina Torre Morais, Maytê Koch Balarini, Kyvia Lugate, Nayara Magalhães, Rafael Reis and Mário José de Oliveira for their support during the collection of the specimens, and Mariana M Castro for revising the manuscript. This research was supported by Dr. Arlindo A A Moura and Dr. Sylvie Breton along with their research teams. Tatiana P Menezes has been granted a scholarship of Brazilian Federal Agency for Support and Evaluation of Graduate School (CAPES). Mariana Machado-Neves participated as a visiting professor in the Program of Membrane Biology at Massachusetts General Hospital (MGH) with a fellowship from Fundação de Amparo a Pesquisa de Minas Gerais (FAPEMIG; process number ECT00054-1).
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Conflict of interest
None of the authors have any conflict of interest to declare.
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