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AMPA receptor expression in mouse testis and spermatogonial GC‐1 cells: A study on its regulation by excitatory amino acids
Author(s) -
Santillo Alessandra,
Falvo Sara,
Di Fiore Maria M.,
Di Giacomo Russo Federica,
Chieffi Paolo,
Usiello Alessandro,
Pinelli Claudia,
Baccari Gabriella Chieffi
Publication year - 2019
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.28382
Subject(s) - ampa receptor , glutamate receptor , nmda receptor , biology , incubation , receptor , microbiology and biotechnology , mapk/erk pathway , extracellular , protein kinase b , biochemistry , kinase , signal transduction
Excitatory amino acids (EAAs) are found present in the nervous and reproductive systems of animals. Numerous studies have demonstrated a regulatory role for Glutamate (Glu), d ‐aspartate ( d ‐Asp) and N‐methyl‐ d ‐aspartate (NMDA) in the control of spermatogenesis. EAAs are able to stimulate the Glutamate receptors, including the α‐amino‐3‐hydroxy‐5‐methyl‐4‐isoxazolepropionic acid receptor (AMPAR). Here in, we assess expression of the main AMPAR subunits, GluA1 and GluA2/3, in the mouse testis and in spermatogonial GC‐1 cells. The results showed that both GluA1 and GluA2/3 were localized in mouse testis prevalently in spermatogonia. The subunit GluA2/3 was more highly expressed compared with GluA1 in both the testis and the GC‐1 cells. Subsequently, GC‐1 cells were incubated with medium containing l ‐Glu, d ‐Glu, d ‐Asp or NMDA to determine GluA1 and GluA2/3 expressions. At 30 minutes and 2 hours of incubation, EAA‐treated GC‐1 cells showed significantly higher expression levels of both GluA1 and GluA2/3. Furthermore, p‐extracellular signal‐regulated kinase (ERK), p‐Akt, proliferating cell nuclear antigen (PCNA), and Aurora B expressions were assayed in l ‐Glu‐, d ‐Glu‐, and NMDA‐treated GC‐1 cells. At 30 minutes and 2 hours of incubation, treated GC‐1 cells showed significantly higher expression levels of p‐ERK and p‐Akt. A consequent increase of PCNA and Aurora B expressions was induced by l ‐Glu and NMDA, but not by d ‐Glu. Our study demonstrates a direct effect of the EAAs on spermatogonial activity. In addition, the increased protein expression levels of GluA1 and GluA2/3 in EAA‐treated GC‐1 cells suggest that EAAs could activate ERK and Akt pathways through the AMPAR. Finally, the increased PCNA and Aurora B levels may imply an enhanced proliferative activity.