Related Experiment Video
Updated: Dec 9, 2025
Examination of Anatomical Features of Retinal Ganglion Cells Under N-methyl-D-aspartic Acid (NMDA)-induced Excitotoxicity
Published on: September 19, 2025
Agonist and antagonist NMDA receptor effect on cell fate during germ cell differentiation and regulate apoptotic
Ghazaleh Modirshanechi1, Mohammad Amin Eslampour1, Zohreh Abdolmaleki2
1Department of Clinical Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Abstract:
In this work, agonist and antagonist N-methyl-D-aspartate (NMDA) receptor activation effect on cell fate during germ cell differentiation and regulate apoptotic process in 3D organ culture were studied. Afterwards, the effect of D-serine, retinoic acid (RA) and MK801 on spermatogenesis development was investigated. The animals were injected a single dose (40 mg/kg, intraperitoneal) of busulfan. After confirming the model, ten 5-day-old NMRI mice were used as spermatogonial stem cells (SSCs) transplantation donors. The SSCs were confirmed by detecting the promyelocytic leukaemia zinc finger (PLZF) protein. Then, tissue culture of the azoospermia model which had received SSCs was performed in various conditions (seven groups). The apoptosis markers levels of cells were significantly decreased in differentiation media containing RA and serine. In contrast, the expression of apoptotic markers including caspase 3, caspase 9 and Bax was increased in the presence of MK801. In conclusion, a new in vitro system capable of producing mature spermatozoa was developed that would be useful for investigating the medicinal effects of agents on the male reproductive system. Also, a comparison of spermatogenesis development in different media revealed that the presence of D-serine and RA (retinoic acid) in the culture medium has a positive effect on spermatogenesis.
Insights
This study explored how N-methyl-D-aspartate (NMDA) receptor modulators affect germ cell development and apoptosis. D-serine and retinoic acid (RA) promoted spermatogenesis, while MK801 increased apoptosis markers.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Cell Biology
Background:
- N-methyl-D-aspartate (NMDA) receptor activity influences cell fate and apoptosis.
- Understanding germ cell differentiation is crucial for male reproductive health.
Purpose of the Study:
- To investigate the impact of NMDA receptor agonists and antagonists on germ cell differentiation and apoptosis in a 3D organ culture system.
- To evaluate the effects of D-serine, retinoic acid (RA), and MK801 on spermatogenesis.
Main Methods:
- Established an azoospermia model in mice using busulfan injection.
- Utilized spermatogonial stem cells (SSCs) for transplantation and confirmed their identity via PLZF protein detection.
- Cultured SSCs in a 3D organ culture system under various conditions, including the presence of D-serine, RA, and MK801.
Main Results:
- Differentiation media supplemented with RA and D-serine significantly reduced apoptosis markers.
- MK801 treatment led to increased expression of apoptosis markers (caspase 3, caspase 9, Bax).
- The presence of D-serine and RA positively influenced spermatogenesis development in vitro.
Conclusions:
- A novel in vitro system for producing mature spermatozoa was developed, offering a platform for studying male reproductive system pharmacology.
- D-serine and RA enhance spermatogenesis, while NMDA receptor antagonists like MK801 may impede it by promoting apoptosis.
