Dysmorphic patterns are associated with cytoskeletal alterations in human oocytes.
Are specific morphological anomalies in human mature oocytes, as revealed by transmitted light microscopy, associated with intrinsic damage to the meiotic spindle and actin cytoskeleton?



Aggregates of smooth endoplasmic reticulum (SER) and domains of centrally localized granular cytoplasm (GC) reflect intrinsic damage to the oocyte cytoskeleton, namely alterations in spindle size, chromosome misalignment and cortical actin disorganization.



In preparation for ICSI, oocytes are often selected for use in treatment by morphological criteria, but the rationale and implications of this practice are controversial. Very little information is available on the relationship between oocyte morphology and intrinsic cellular characteristics, such as the actin cytoskeleton, meiotic spindle and chromosome alignment.



A total of 170 metaphase II (MII) oocytes were donated by consenting IVF patients and analysed; 62 were classified as morphologically normal (control), 54 had SER clusters and 54 had centrally localized GC.



Supernumerary oocytes were fixed within 3 h from recovery and stained for tubulin, chromatin and actin. Spindles were analysed for 1D and 2D characteristics by high-performance confocal microscopy. Chromosomes were classified as scattered or aligned and the conformation and intensity of cortical actin was evaluated.



In comparison with control oocytes, both SER and GC oocytes showed greater spindle length (P = 0.033 and 0.003, respectively) and GC oocytes also showed greater spindle width (P= 0.049) and area (P= 0.036). Control and SER oocytes had statistically comparable rates of chromosome displacement from the metaphase plate, unlike GC oocytes where chromosome displacement occurred at higher rate (P = 0.013). In situations where a complete Z-stack was reconstructed from a polar angle, chromosome disposition was classified as being normal when two sets of concentric arrays were visible. Based on these parameters, the proportions of oocytes with normal chromosomal arrangement or partial/total disarrangement was not statistically different between control and SER oocytes. Conversely, in GC oocytes, chromosome disarrangement was higher (P = 0.002). All control oocytes displayed a continuous meshwork of suboolemmal actin, which appeared as an uninterrupted ring in thin optical sections. In contrast, in SER and  GC groups, integrity of suboolemmal actin was observed in only 66.7 and 42.9% of oocytes, respectively (P = 0.0001).



N/A.



Only two of several known oocyte dysmorphisms were investigated, while oocyte quality was assessed only by cytoskeletal criteria.



This study represents a significant step toward a more objective assessment of oocyte morphology, offering information that can assist embryologists to make a more aware and rationally founded decision on whether, and with what possible implications, oocytes with certain dysmorphic characters should be used for treatment or discarded. More generally, it also demonstrates that morphometric parameters of the cytoskeleton and chromosome organization can be used as biomarkers of oocyte quality.



This study was funded by Biogenesi Reproductive Medicine Centre (Monza, Italy). All authors declare no conflict of interests.
Biogenesi Reproductive Medicine Centre, Istituti Clinici Zucchi, Via Zucchi, Monza,Italy.
Mariabeatrice
M
Dal Canto
Biogenesi Reproductive Medicine Centre, Istituti Clinici Zucchi, Via Zucchi, Monza,Italy.
Maria Cristina
MC
Guglielmo
Biogenesi Reproductive Medicine Centre, Istituti Clinici Zucchi, Via Zucchi, Monza,Italy.
Mario
M
Mignini Renzini
Biogenesi Reproductive Medicine Centre, Istituti Clinici Zucchi, Via Zucchi, Monza,Italy.
Rubens
R
Fadini
Biogenesi Reproductive Medicine Centre, Istituti Clinici Zucchi, Via Zucchi, Monza,Italy.
Clarissa
C
Moutier
Biogenesi Reproductive Medicine Centre, Istituti Clinici Zucchi, Via Zucchi, Monza,Italy.
Maria
M
Merola
Department of Medical Physics, ASST Monza, Via Pergolesi, Monza, Italy.
Elena
E
De Ponti
Biogenesi Reproductive Medicine Centre, Istituti Clinici Zucchi, Via Zucchi, Monza,Italy.
Giovanni
G
Coticchio
D015415:Biomarkers; D002875:Chromosomes / Q000648:ultrastructure; D003593:Cytoplasm / Q000648:ultrastructure; D003599:Cytoskeleton / Q000648:ultrastructure*; D018871:Endoplasmic Reticulum, Smooth / Q000648:ultrastructure; D005260:Female; D006801:Humans; D059471:In Vitro Oocyte Maturation Techniques; D008677:Metaphase; D009865:Oocytes / Q000166:cytology / Q000648:ultrastructure*; D008941:Spindle Apparatus / Q000648:ultrastructure*
D015415:Biomarkers
IVF; actin; chromosomes; confocal microscopy; cytoskeleton; morphology; oocytes; spindle