## 4 Discussion
### Effects on CG and Microvilli Distribution  
CGs, essential for preventing polyspermy during fertilization, are important markers of oocyte quality and cytoplasmic maturation and are used to assess oocyte maturity and organelle organization [87]. Our data, although not statistically significant, suggest that Lindane exposure prompted the dose-related reduction and misdistribution of CGs. This could impair the oocyte's ability to prevent polyspermy, leading to potential fertilization issues and infertility [88; 89; 90]. These results align with previous research showing that organochlorines can cause polyspermy by altering the zona pellucida. Additionally, the observed changes in CG distribution indicate that Lindane may alter the cytoskeleton, which regulates CG movement [87].  
Moreover, results from this work showed irregular and reduced microvilli distribution after Lindane exposure, which plays a key role in sperm-oocyte interactions [91; 92; 93], during fertilization. Alteration in the peripheral oolemma may result in diminished oocyte quality and an increased risk of fertilization failure.  
Hypothesis on Role of Cumulus Cells in Lindane-Induced Oocyte Damage, Study Limitations, and Future Perspectives  
In our recent in vitro study on mouse oocytes exposed to the endocrine disruptor Mancozeb, we observed significant ultrastructural changes only at the highest concentration (1 \(\upmu\)g/mL), suggesting that cumulus cells may have a protective role toward the oocytes [43]. In contrast, this current work revealed ultrastructural alterations in mouse oocytes exposed to Lindane across all experimental groups, with more severe damage observed in the L100 group.  
It is well-established that cumulus cells surround oocytes, forming COCs, and are essential in oocyte maturation and fertilization through bidirectional communication and metabolic interdependence [94; 95]. Acting as a biological barrier, cumulus cells rely on gap junctions formed by connexin proteins, such as Cx43, to facilitate the exchange of small molecules. This connection supports the transfer of metabolites and regulatory molecules, enhancing cytoplasmic maturation and aiding early embryo development through coordinated cellular changes [96; 97].  
Considering the Lindane-induced inhibition of gap junctional intercellular communication, as previously demonstrated in numerous studies [34; 98], the changes here observed in mouse oocytes may be associated with its activity as a gap junction blocker, thus explaining its possible entry into the oocyte to exert its cytotoxic effect. Results from this study evidenced the action of Lindane in affecting the bidirectional communication between cumulus cells and oocytes by abolishing gap junctions and interrupting the functional and protective role of cumulus cells toward the oocyte. However, due to the limited research on Lindane, further studies are needed to fully explore its reproductive toxicity and comprehensively examine the specific mechanisms underlying its effects on reproductive health and related pathologies. The limited availability of comparable ultrastructural data on Lindane exposure, particularly in mammalian oocytes, also restricts broader comparative analysis.  
Recent applications of artificial intelligence and recurrent neural networks in monitoring and diagnostic fields, such as the study by [PERSON] et al. (2025), which integrates LSTM and U-Net models for anomaly detection in electrical systems, underscore the potential of deep learning techniques for identifying complex patterns. In the future, a similar approach could be applied to the automated analysis of ultrastructural images, aiming to improve the early detection of toxic effects induced by environmental pollutants on oocytes [99].