GO:0032154 cleavage furrow: Components, Assembly and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0032154 cleavage furrow is the actomyosin-based invagination of the plasma membrane that physically separates a dividing cell into two daughter cells.
• Furrow positioning is determined by the mitotic spindle and central spindle through the centralspindlin complex and RhoA signaling.
• Actin and myosin II are the core machinery, but recent work shows that furrow formation can occur without F-actin in some organisms, indicating alternative mechanisms.
• Calcium spikes and chloride intracellular channel proteins such as CLIC4 regulate cortical cytoskeleton stability during furrow ingression.
• Defects in cleavage furrow formation lead to cytokinesis failure, polyploidy, and aneuploidy, which are hallmarks of cancer and other proliferative disorders.
• CRISPR-based knockout, knock-in, and overexpression models are essential to dissect the molecular players of cleavage furrow assembly and function.
Description
The cleavage furrow is a specialized actomyosin-rich structure that forms at the equatorial cortex of a dividing cell during late mitosis and physically separates the two daughter cells. This process, known as cytokinesis, is essential for maintaining genome stability and tissue homeostasis. The cleavage furrow is a dynamic and highly regulated structure whose positioning and ingression are controlled by signals from the mitotic spindle and central spindle. Understanding how the cleavage furrow forms and functions is fundamental to cell biology and has direct implications for cancer, developmental disorders, and regenerative medicine. Researchers study the cleavage furrow using a combination of live-cell imaging, genetic perturbation, and biochemical assays to identify the molecular components and signaling pathways that drive its assembly and contraction.
cleavage furrow At A Glance
| GO ID | GO:0032154 |
|---|---|
| GO term | cleavage furrow |
| Ontology | biological_process |
| Synonym | None |
| Major function | Actomyosin-based invagination of the plasma membrane that separates daughter cells during cytokinesis |
| Related cellular component | Actomyosin contractile ring, midbody, central spindle |
| Key regulators | RhoA, centralspindlin, anillin, ECT2, MgcRacGAP |
| Disease relevance | Cytokinesis failure leads to polyploidy and aneuploidy, which are associated with cancer and megakaryocyte disorders |
| Model organisms | Dictyostelium, Chlamydomonas, fission yeast, mammalian cells |
What Is GO:0032154?
The cleavage furrow (GO:0032154) is the actomyosin-based invagination of the plasma membrane that occurs during cytokinesis in animal cells and some other eukaryotes. It is the physical structure that pinches the cell into two daughter cells. The furrow is enriched in filamentous actin and myosin II, which generate the contractile force for ingression. Its position is determined by the mitotic spindle and the central spindle through the centralspindlin complex and RhoA signaling. While actin and myosin are canonical components, recent studies have shown that furrow formation can occur without F-actin in certain organisms, highlighting the diversity of cytokinetic mechanisms.
Why Is cleavage furrow Important in Cell Biology?
The cleavage furrow is essential for cytokinesis, the final step of cell division that ensures each daughter cell receives a complete copy of the genome. Failure of cleavage furrow formation or ingression results in binucleation or multinucleation, which can lead to genomic instability and cancer. In addition, proper furrow positioning is critical for asymmetric cell division and tissue morphogenesis. Studying the cleavage furrow provides insights into fundamental mechanisms of cell division and identifies potential therapeutic targets for diseases characterized by aberrant cell proliferation.
• Ensures accurate chromosome segregation and genome stability by physically separating daughter cells.
• Defects in cleavage furrow formation cause cytokinesis failure, leading to polyploidy and aneuploidy, hallmarks of cancer.
• Furrow positioning is critical for asymmetric cell division and cell fate determination during development.
• The cleavage furrow is a target for anti-cancer drugs that inhibit cytokinesis.
• Understanding furrow formation in model organisms like Dictyostelium and Chlamydomonas reveals evolutionary diversity.
• Calcium signaling and CLIC4 regulate furrow ingression and cortical stability, linking ion channels to cytokinesis.
• Centralspindlin and RhoA are key regulators that coordinate spindle position with furrow formation.
• Research on cleavage furrow informs regenerative medicine and tissue engineering by controlling cell division.
What Happens During cleavage furrow?
Furrow Positioning and Specification
In simple terms: The cell decides where to pinch based on signals from the mitotic spindle.
The position of the cleavage furrow is determined by the mitotic spindle and the central spindle. The centralspindlin complex, composed of MKLP1 and MgcRacGAP, localizes to the central spindle and recruits the RhoGEF ECT2, which activates RhoA at the equatorial cortex. This signaling cascade ensures that the furrow forms precisely between the two sets of segregated chromosomes. In Dictyostelium, furrow positioning is also influenced by the spindle and involves similar molecular players.
Actomyosin Ring Assembly and Contraction
In simple terms: A ring of actin and myosin assembles at the equator and squeezes the cell like a drawstring.
Once RhoA is activated, it promotes the assembly of a contractile ring composed of filamentous actin and myosin II. This ring generates the force that drives membrane invagination. Myosin II uses ATP to slide actin filaments, constricting the ring and deepening the furrow. In Chlamydomonas, however, cleavage furrow formation can occur without F-actin, indicating that alternative mechanisms exist.
Membrane Remodeling and Ingression
In simple terms: The membrane is pulled inward and reshaped as the furrow deepens.
As the actomyosin ring contracts, the plasma membrane must be remodeled to accommodate the increasing curvature. CLIC4, a chloride intracellular channel protein, localizes to the cleavage furrow and regulates cortical cytoskeleton stability during ingression. Calcium spikes accompany furrow ingression and cell separation in fission yeast, suggesting that calcium signaling is important for membrane dynamics.
Abscission and Cell Separation
In simple terms: The final cut that separates the two daughter cells.
After the furrow has ingressed sufficiently, the midbody forms and abscission occurs, severing the remaining connection between the daughter cells. This step requires precise coordination of membrane trafficking and cytoskeletal disassembly. Defects in abscission can lead to binucleation and genomic instability.
Key Genes Involved in GO:0032154 cleavage furrow
The following genes and proteins are key players in cleavage furrow formation and function, as supported by the verified literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| RhoA | Small GTPase that activates actomyosin ring assembly at the equatorial cortex | Central regulator of furrow formation; knockout causes cytokinesis failure |
| ECT2 | RhoGEF that activates RhoA at the central spindle | Essential for furrow positioning; knockdown leads to furrow defects |
| MgcRacGAP | Component of centralspindlin; regulates RhoA and Rac | Required for central spindle formation and furrow ingression |
| MKLP1 | Kinesin motor in centralspindlin; bundles microtubules | Critical for central spindle assembly and furrow specification |
| Anillin | Actin-binding protein that links the contractile ring to the membrane | Scaffold for ring assembly; depletion causes furrow instability |
| Myosin II | Motor protein that generates contractile force | Core component of the contractile ring; inhibition blocks furrow ingression |
| Actin | Filamentous polymer that forms the contractile ring | Main structural component; depolymerization prevents furrow formation |
| CLIC4 | Chloride intracellular channel protein; regulates cortical cytoskeleton stability | Modulates furrow ingression; knockdown affects cortical actin |
| Calcium channels | Mediate calcium spikes during furrow ingression | Regulate furrow progression in fission yeast |
| F-actin | Filamentous actin; canonical component of the contractile ring | In Chlamydomonas, furrow can form without F-actin, indicating alternative mechanisms |
| Centralspindlin | Complex of MKLP1 and MgcRacGAP; key furrow regulator | Central to Rappaport's cleavage signaling |
| Rappaport's cleavage signaling | Signaling from the central spindle to the cortex | Model for furrow positioning |
| Polyploidization regulators | Genes controlling megakaryocyte polyploidization | Relevant to platelet production and leukemia |
| Cytokinesis regulators | Various proteins involved in abscission | Targets for cancer therapy |
How Is cleavage furrow Regulated?
Cleavage furrow formation is regulated by multiple signaling pathways. The centralspindlin complex and RhoA are central regulators that link spindle position to furrow formation. Calcium signaling also plays a role, with calcium spikes accompanying furrow ingression in fission yeast. CLIC4 regulates cortical cytoskeleton stability during furrow ingression. Additionally, phosphorylation events and small GTPases modulate the timing and location of furrow formation.
cleavage furrow and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| RhoA | Cancer, cytokinesis failure | Knockout in cancer cell lines; rescue with point mutants |
| ECT2 | Cancer, aneuploidy | Knockdown or knockout in HeLa cells; live-cell imaging |
| MgcRacGAP | Cancer, cytokinesis defects | Knockout in mouse models; conditional alleles |
| CLIC4 | Cancer, cortical cytoskeleton instability | Knockout in mammalian cells; overexpression studies |
| Myosin II | Cancer, contractile ring defects | Point mutations in myosin II; inhibitor studies |
Cancer and Genomic Instability
Failure of cleavage furrow formation leads to cytokinesis failure, resulting in tetraploidy or aneuploidy, which are common features of cancer cells. Many cancer cells have defects in cytokinesis regulators such as RhoA, ECT2, or MgcRacGAP, contributing to genomic instability and tumor progression.
Megakaryocyte Polyploidization
Megakaryocytes undergo polyploidization through repeated rounds of DNA replication without cell division, a process that involves abortive cytokinesis and cleavage furrow formation. Defects in this process can lead to thrombocytopenia or leukemia.
Developmental Disorders
Proper cleavage furrow positioning is essential for asymmetric cell division during development. Mutations in genes regulating furrow formation can cause developmental abnormalities and tissue morphogenesis defects.
From cleavage furrow-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does gene X regulate furrow positioning? | Knockout cell lines with live-cell imaging of furrow markers |
| What is the role of a specific phosphorylation site in furrow ingression? | Point mutation knock-in of the phospho-mutant |
| How does a disease-associated mutation affect furrow formation? | Knock-in of the patient mutation; phenotypic analysis |
| Where does protein X localize during cytokinesis? | Tagged knock-in with fluorescent protein; live imaging |
| Does overexpression of gene Y cause furrow defects? | Overexpression cell lines; time-lapse microscopy |
| What is the transcriptional profile during furrow ingression? | RNA-seq of synchronized cells; bioinformatics analysis |
How to Study the cleavage furrow Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Live-cell imaging | Dynamics of furrow formation and ingression | Visualizing actin/myosin ring contraction |
| CRISPR knockout | Loss-of-function phenotype | Identifying essential furrow genes |
| RNA-seq | Transcriptional changes during cytokinesis | Discovering new furrow regulators |
| Proteomics | Protein composition of the furrow | Identifying novel furrow components |
| Calcium imaging | Calcium spikes during furrow ingression | Studying calcium signaling in cytokinesis |
| FRAP | Protein turnover at the furrow | Measuring dynamics of ring components |
| Electron microscopy | Ultrastructure of the furrow | Visualizing membrane invagination |
Live-Cell Imaging
Live-cell imaging with fluorescently tagged actin, myosin II, or RhoA allows real-time visualization of cleavage furrow formation and ingression. This method is essential for studying the dynamics and positioning of the furrow.
Genetic Perturbation
Knockout, knockdown, or overexpression of candidate genes followed by phenotypic analysis of furrow formation can identify essential regulators. CRISPR-Cas9 is commonly used for generating knockout cell lines.
Biochemical Assays
Biochemical assays such as pull-downs and co-immunoprecipitation can identify protein-protein interactions within the cleavage furrow. For example, the centralspindlin complex was identified through such methods.
Calcium Imaging
Calcium indicators can be used to monitor calcium spikes during furrow ingression, as demonstrated in fission yeast.
How CRISPR Can Be Used to Study GO:0032154 cleavage furrow
Knockout
CRISPR knockout of genes such as RhoA, ECT2, or CLIC4 can reveal their essential roles in cleavage furrow formation. Knockout cell lines often exhibit cytokinesis failure, binucleation, or furrow defects.
Point Mutation
Point mutation knock-in can be used to study the function of specific phosphorylation sites or disease-associated mutations in furrow regulators. For example, mutating the catalytic residue of ECT2 can abolish its RhoGEF activity.
Knock-in
Knock-in of fluorescent tags (e.g., GFP) allows real-time visualization of protein localization during furrow formation. Tagged knock-in of myosin II or actin has been instrumental in understanding ring dynamics.
Overexpression
Overexpression of furrow regulators can cause dominant-negative effects or hyperactivation, leading to furrow defects. For instance, overexpression of constitutively active RhoA can induce ectopic furrows.
How EDITGENE Supports cleavage furrow Research
Researchers studying cleavage furrow-related genes often need to determine whether a candidate gene is causally involved in furrow formation and ingression. This requires precise genetic manipulation, which can be achieved through CRISPR-based knockout, point mutation, knock-in, or overexpression models. EDITGENE provides comprehensive services to generate these models and analyze their phenotypes.
Contact EDITGENE today to design your custom CRISPR model for cleavage furrow research.
Frequently Asked Questions About cleavage furrow
What is the cleavage furrow?
The cleavage furrow is the actomyosin-based invagination of the plasma membrane that separates a dividing cell into two daughter cells during cytokinesis.
What genes are involved in cleavage furrow formation?
Key genes include RhoA, ECT2, MgcRacGAP, MKLP1, anillin, myosin II, actin, and CLIC4.
How is the cleavage furrow positioned?
The furrow is positioned by signals from the central spindle, involving the centralspindlin complex and RhoA activation.
What is the role of actin in the cleavage furrow?
Actin filaments form the contractile ring that generates the force for membrane invagination.
Can cleavage furrow form without actin?
Yes, in Chlamydomonas, cleavage furrow formation can occur without F-actin, indicating alternative mechanisms.
What happens if the cleavage furrow fails?
Failure leads to cytokinesis defects, resulting in binucleation, polyploidy, and genomic instability, which are associated with cancer.
How do calcium signals regulate the cleavage furrow?
Calcium spikes accompany furrow ingression and cell separation in fission yeast, suggesting a role in membrane dynamics.
What is CLIC4 and its role in cytokinesis?
CLIC4 is a chloride intracellular channel protein that localizes to the cleavage furrow and regulates cortical cytoskeleton stability.
What model organisms are used to study the cleavage furrow?
Common models include Dictyostelium, Chlamydomonas, fission yeast, and mammalian cells.
How can CRISPR help study cleavage furrow genes?
CRISPR knockout, knock-in, point mutation, and overexpression models allow precise functional analysis of furrow regulators.
Conclusion
The cleavage furrow (GO:0032154) is a fundamental structure in cell division, essential for cytokinesis and genome stability. Its formation and ingression are driven by a complex interplay of cytoskeletal, signaling, and membrane remodeling proteins. Defects in furrow formation are linked to cancer and developmental disorders, making it a critical area of research. Advances in CRISPR-based models and imaging techniques continue to uncover new regulators and mechanisms, offering potential therapeutic targets.
References
- 1. Onishi M et al.. 2020. Cleavage-furrow formation without F-actin in Chlamydomonas.. Proc Natl Acad Sci U S A 117(31):18511-18520 PMID: 32690698
- 2. Okada A et al.. 2023. Cleavage furrow positioning in dividing Dictyostelium cells.. Cytoskeleton (Hoboken) 80(11-12):448-460 PMID: 37650534
- 3. D'Avino PP et al.. 2005. Cleavage furrow formation and ingression during animal cytokinesis: a microtubule legacy.. J Cell Sci 118(Pt 8):1549-58 PMID: 15811947
- 4. Burgess DR et al.. 2005. Site selection for the cleavage furrow at cytokinesis.. Trends Cell Biol 15(3):156-62 PMID: 15752979
- 5. Mazzi S et al.. 2018. Megakaryocyte and polyploidization.. Exp Hematol 57:1-13 PMID: 29111429
- 6. Mishima M. 2016. Centralspindlin in Rappaport's cleavage signaling.. Semin Cell Dev Biol 53:45-56 PMID: 26964770
- 7. Poddar A et al.. 2021. Calcium spikes accompany cleavage furrow ingression and cell separation during fission yeast cytokinesis.. Mol Biol Cell 32(1):15-27 PMID: 33175606
- 8. Peterman E et al.. 2020. CLIC4 is a cytokinetic cleavage furrow protein that regulates cortical cytoskeleton stability during cell division.. J Cell Sci 133(9) PMID: 32184265
