A 3D analysis revealed complexe mitochondria morphologies in porcine cumulus cells

In the ovarian follicle, a bilateral cell-to-cell communication exists between the female germ cell and the cumulus cells which surround the oocyte. This communication allows the transit of small size molecules known to impact oocyte developmental competence. Pyruvate derivatives produced by mitochondria, are one of these transferred molecules. Interestingly, mitochondria may adopt a variety of morphologies to regulate their functions. In this study, we described mitochondrial morphologies in porcine cumulus cells. Active mitochondria were stained with TMRM (Tetramethylrhodamine, Methyl Ester, Perchlorate) and observed with 2D confocal microscopy showing mitochondria of different morphologies such as short, intermediate, long, and very long. The number of mitochondria of each phenotype was quantified in cells and the results showed that most cells contained elongated mitochondria. Scanning electron microscopy (SEM) analysis confirmed at nanoscale resolution the different mitochondrial morphologies including round, short, intermediate, and long. Interestingly, 3D visualisation by focused ion-beam scanning electron microscopy (FIB-SEM) revealed different complex mitochondrial morphologies including connected clusters of different sizes, branched mitochondria, as well as individual mitochondria. Since mitochondrial dynamics is a key regulator of function, the description of the mitochondrial network organisation will allow to further study mitochondrial dynamics in cumulus cells in response to various conditions such as in vitro maturation.

Automated summary

Bilateral cell-to-cell communication exists in the ovarian follicle between the female germ cell and the cumulus cells. This communication allows the transfer of small molecules that impact oocyte developmental competence, including pyruvate derivatives produced by mitochondria. In this study, mitochondrial morphologies in porcine cumulus cells were described using various microscopy techniques. The results showed different morphologies of mitochondria, with elongated mitochondria being the most common. Further analysis revealed complex mitochondrial morphologies, including connected clusters and branched mitochondria. Understanding the organization of the mitochondrial network in cumulus cells will contribute to the study of mitochondrial dynamics in response to different conditions.