Phenotype Classes
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Bold names designate phenotype classes. Associated defect categories and their scoring criteria are listed underneath.
Aberrant Cytoplasmic Structures
Areas Devoid of Yolk Granules
Aberrant cytoplasmic structures excluding yolk granules
Asymmetry of Division
P0 Pronuclear Meeting
Position near the cortex or centrally (WT: near the centre of the posterior half of the embryo)
P0 Spindle Rocking
No or excessive spindle rocking
P0 Spindle Positioning-Asymmetry
Spindle positioning at telophase other than along the longitudinal axis with the posterior pole shifted posteriorly approx. 10-15%
P0 Spindle Poles
Irregular shape, in particular lack of flattening of posterior pole in telophase
P1 Nuclear Migration/Rotation
Lack of migration of P1 nucleus towards posterior cortex, lack of rotation of P1 spindle before nuclear envelope breakdown
AB Spindle Orientation
Rotation of AB spindle (WT: AB spindle keeps orientation,whereas P1 spindle rotates)
P1/AB Asynchrony of Divisions
Delay between AB and P1 cleavage furrow initiation is either shorter than 2 min or exceeds 5 min (WT: approx. 2-3 min)
Four Cell Stage Configuration
Par-like configurations of blastomeres
Centrosome Attachment
P0 Pronuclear Migration (male)
No migration of male PN from posterior cortex towards centre of the embryo
P0 Pronuclear Meeting
Position near the cortex or centrally (WT: near the centre of the posterior half of the embryo)
P0 Spindle Integrity
Irregular length (WT: 25-30% the length of the embryo during metaphase), or Thickness (WT: 20-25% the width of the embryo at metaphase), or lack of Rigidity
P1/AB Nuclei-Number
Numbers of nuclei in daughter cells below or above 1
Chromosome Segregation (karyomeres)
P1/AB Nuclei-Number
Numbers of nuclei in daughter cells below or above 1
Four Cell Stage Nuclei-Size/Shape
Irregular size and/or shape of nuclei in daughter cells
Complex Phenotype
Cortical Dynamics
P0 Pseudo-Cleavage Furrow
No or excessive furrowing (WT: Covering 20-30% the width of the embryo)
P0 Cortical Ruffling
Excessive or no early cortical ruffling (WT: wave of membrane ruffling from posterior to anterior resulting in Pseudo Cleavage Furrow)
P1/AB Cortical Activity
Excessive membrane ruffling and blebbing
Cytokinesis
P0 Cytokinesis-Furrow Ingression
Little or no ingression, or uneven ingression from the two sites
P0 Cytokinesis- Completion/Stability
Regression of the furrow
P1/AB Cytokinesis
Aberrant furrow initiation or ingression or completion
Tetrapolar Spindle
4 poles visible by exclusion of yolk granules
Unclear-Multinucleate
Aberrant numbers of nuclei
Polar Bodies
Aberrant number (fewer or more than two) or size (matches or exceeds size of early pronuclei) or internalization of polar bodies
Egg Size
Egg Shape/Size
Small(<2/3 of WT), large (>4/3 of WT), or irregularly shaped eggs
Entry Into Interphase
P0 Pseudo-Cleavage Furrow
No or excessive furrowing (WT: Covering 20-30% the width of the embryo)
P0 Pronuclei-Size/Shape
Size >30% smaller or larger than WT (WT: diameter approx. 25% the width of the embryo at onset of PN migration), or irregular shape of pronuclei
General Pace of Development
Overall Pace of Events
Timing; scored were events where delay between pronuclear meeting and initiation of AB cleavage furrow exceeded 30 min (WT: 18-22min)
Integrity of Membranous Organelles
Yolk Granules-Size
Aberrant size of individual or all granules
Yolk Granules-Density
Reduction >30%
Nuclear Appearance
P1/AB Nuclei-Size/Shape
Size >30% smaller or larger than WT (diameter approx. 25% of AB blastomere), or irregular shape of nuclei
Four Cell Stage Nuclei-Size/Shape
Irregular size and/or shape of nuclei in daughter cells
P1/AB Asynchrony of Divisions
Delay between AB and P1 cleavage furrow initiation is either shorter than 2 min or exceeds 5 min (WT: approx. 2-3 min)
Osmotic Integrity
Osmotic Sensitivity
Swelling of the embryo to fill the egg shell and/or irregular granule movements
Unclear-Multinucleate
Aberrant numbers of nuclei
Pace of P-Lineage
P1/AB Asynchrony of Divisions
Delay between AB and P1 cleavage furrow initiation is either shorter than 2 min or exceeds 5 min (WT: approx. 2-3 min)
P1/AB Nuclei-Size/Shape
Size >30% smaller or larger than WT (diameter approx. 25% of AB blastomere), or irregular shape of nuclei
Passage Through Meiosis
P0 Pronuclei-Number
Numbers of pronuclei above or below 2 (WT: one female, one male PN)
Osmotic Sensitivity
Swelling of the embryo to fill the egg shell and/or irregular granule movements
Meiotic Arrest
No visible pronuclei, little or no cytoplasmic events, embryo often fills egg shell
Polar Body Extrusion
P0 Pronuclei-Number
Numbers of pronuclei above or below 2 (WT: one female, one male PN)
P0 Pronuclei-Size/Shape
Size >30% smaller or larger than WT (WT: diameter approx. 25% the width of the embryo at onset of PN migration), or irregular shape of pronuclei
Polar Bodies
Aberrant number (fewer or more than two) or size (matches or exceeds size of early pronuclei) or internalization of polar bodies
P1/AB Nuclei-Number
Numbers of nuclei in daughter cells below or above 1
Pronuclear Migration
P0 Pronuclei-Number
Numbers of pronuclei above or below 2 (WT: one female, one male PN)
P0 Pronuclear Rotation
Rotation takes place after PN envelope breakdown (WT: approx. 2-3min before PNEBD)
P0 Pronuclear Migration (male)
No migration of male PN from posterior cortex towards centre of the embryo
P0 Pronuclear Meeting
Position near the cortex or centrally (WT: near the centre of the posterior half of the embryo)
P0 Pronuclear Centration
Position off centre
P0 Spindle Assembly-Bipolarity
Lack of 2 visible poles, i.e. two regularly shaped sites of yolk granule exclusion
P0 Cytokinesis-Furrow Specification
Fewer or more than 2 sites of furrowing and/or aberrant positioning (WT: two sites, intersecting the long axis by a ratio of approx. 3:2)
P0 Cytokinesis- Completion/Stability
Regression of the furrow
Pronuclear/Nuclear Appearance
P0 Pronuclei-Size/Shape
Size >30% smaller or larger than WT (WT: diameter approx. 25% the width of the embryo at onset of PN migration), or irregular shape of pronuclei
P1/AB Nuclei-Size/Shape
Size >30% smaller or larger than WT (diameter approx. 25% of AB blastomere), or irregular shape of nuclei
P1/AB Asynchrony of Divisions
Delay between AB and P1 cleavage furrow initiation is either shorter than 2 min or exceeds 5 min (WT: approx. 2-3 min)
Four Cell Stage Nuclei-Size/Shape
Irregular size and/or shape of nuclei in daughter cells
Severe Pleiotropic Defects
Osmotic Sensitivity
Swelling of the embryo to fill the egg shell and/or irregular granule movements
P0 Pronuclei-Number
Numbers of pronuclei above or below 2 (WT: one female, one male PN)
Polar Bodies
Aberrant number (fewer or more than two) or size (matches or exceeds size of early pronuclei) or internalization of polar bodies
P0 Spindle Integrity
Irregular length (WT: 25-30% the length of the embryo during metaphase), or Thickness (WT: 20-25% the width of the embryo at metaphase), or lack of Rigidity
P1/AB Nuclei-Number
Numbers of nuclei in daughter cells below or above 1
Four Cell Stage Nuclei-Size/Shape
Irregular size and/or shape of nuclei in daughter cells
Areas Devoid of Yolk Granules
Aberrant cytoplasmic structures excluding yolk granules
Overall Pace of Events
Timing; scored were events where delay between pronuclear meeting and initiation of AB cleavage furrow exceeded 30 min (WT: 18-22min)
Sister Chromatid Separation (Cross-eyed)
P1/AB Nuclei-Number
Numbers of nuclei in daughter cells below or above 1
P1/AB Nuclei-Size/Shape
Size >30% smaller or larger than WT (diameter approx. 25% of AB blastomere), or irregular shape of nuclei
Four Cell Stage Cross-Eyed
Re-forming daughter nuclei stay closely attached to the central cortex
Four Cell Stage Nuclei-Size/Shape
Irregular size and/or shape of nuclei in daughter cells
P1/AB Asynchrony of Divisions
Delay between AB and P1 cleavage furrow initiation is either shorter than 2 min or exceeds 5 min (WT: approx. 2-3 min)
P1/AB Cytokinesis
Aberrant furrow initiation or ingression or completion
Spindle Assembly
P0 Spindle Assembly-Bipolarity
Lack of 2 visible poles, i.e. two regularly shaped sites of yolk granule exclusion
P0 Cytokinesis-Furrow Specification
Fewer or more than 2 sites of furrowing and/or aberrant positioning (WT: two sites, intersecting the long axis by a ratio of approx. 3:2)
P1/AB Cortical Activity
Excessive membrane ruffling and blebbing
P1/AB Spindle Assembly
Aberrant bipolarity or length or thickness
P1/AB Cytokinesis
Aberrant furrow initiation or ingression or completion
Spindle Elongation/Integrity
P0 Spindle Integrity
Irregular length (WT: 25-30% the length of the embryo during metaphase), or Thickness (WT: 20-25% the width of the embryo at metaphase), or lack of Rigidity
P0 Spindle Elongation
Spindle covers less or more than 50-60% of the length of the embryo at telophase
Sterility/Impaired Fertility in F0
Number/Age of Embryos
Absence or reduction of one and two cell stage embryos, suggesting reduced fertility of the parent worm
Osmotic Sensitivity
Swelling of the embryo to fill the egg shell and/or irregular granule movements