Here, we take advantage of the low chromosome number (2N=6) and distinctively large kinetochores of female Indian muntjac cells to investigate the molecular mechanism underlying k-fiber maturation. We describe steps for monitoring kinetochore-microtubule dynamics over time. Specifically, we detail the combination of live-cell super-resolution CH-STED microscopy of microtubule growth events within individual k-fibers and a laser-mediated k-fiber injury/repair assay. These tools provide a direct assessment of microtubule amplification mechanisms within k-fibers in metazoans. For complete details on the use and execution of this protocol, please refer to Almeida et al. (2022)..
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9846000 | PMC |
| http://dx.doi.org/10.1016/j.xpro.2022.102011 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
An evolutionary perspective on the relationship between kinetochore size and CENP-E dependence for chromosome alignment.
J Cell Sci
December 2024
i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen 208, 400-135 Porto, Portugal.
Chromosome alignment during mitosis can occur as a consequence of bi-orientation or is assisted by the CENP-E (kinesin-7) motor at kinetochores. We previously found that Indian muntjac chromosomes with larger kinetochores bi-orient more efficiently and are biased to align in a CENP-E-independent manner, suggesting that CENP-E dependence for chromosome alignment negatively correlates with kinetochore size. Here, we used targeted phylogenetic profiling of CENP-E in monocentric (localized centromeres) and holocentric (centromeres spanning the entire chromosome length) clades to test this hypothesis at an evolutionary scale.
View Article and Find Full Text PDFMicrotubule search-and-capture model evaluates the effect of chromosomal volume conservation on spindle assembly during mitosis.
Phys Rev E
September 2023
School of Mathematical and Computational Sciences, Indian Association for the Cultivation of Science, Kolkata 700032, India.
Variation in the chromosome numbers can arise from the erroneous mitosis or fusion and fission of chromosomes. While the mitotic errors lead to an increase or decrease in the overall chromosomal substance in the daughter cells, fission and fusion keep this conserved. Variations in chromosome numbers are assumed to be a crucial driver of speciation.
View Article and Find Full Text PDFA Camera-Trap Survey of Mammals in Thung Yai Naresuan (East) Wildlife Sanctuary in Western Thailand.
Animals (Basel)
April 2023
Department of Environmental Conservation, University of Massachusetts, Amherst, MA 01003, USA.
The Thung Yai Naresuan (East) Wildlife Sanctuary (TYNE), in the core area of the Western Forest Complex of Thailand, harbors a diverse assemblage of wildlife, and the region has become globally significant for mammal conservation. From April 2010 to January 2012, 106 camera traps were set, and, in 1817 trap-nights, registered 1821 independent records of 32 mammal species. Of the 17 IUCN-listed (from Near Threatened to Critically Endangered) mammal species recorded, 5 species listed as endangered or critically endangered included the Asiatic elephant (), tiger (), Malayan tapir (), dhole (), and Sunda pangolin ().
View Article and Find Full Text PDFOptimized protocol for live-cell analysis of kinetochore fiber maturation in Indian muntjac cells.
STAR Protoc
March 2023
Instituto de Investigação e Inovação em Saúde (i3S), Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal; Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal; Cell Division Group, Department of Biomedicine, Faculdade de Medicina, Universidade do Porto, Alameda Professor Hernâni Monteiro, 4200-319 Porto, Portugal. Electronic address:
Here, we take advantage of the low chromosome number (2N=6) and distinctively large kinetochores of female Indian muntjac cells to investigate the molecular mechanism underlying k-fiber maturation. We describe steps for monitoring kinetochore-microtubule dynamics over time. Specifically, we detail the combination of live-cell super-resolution CH-STED microscopy of microtubule growth events within individual k-fibers and a laser-mediated k-fiber injury/repair assay.
View Article and Find Full Text PDFAugmin-dependent microtubule self-organization drives kinetochore fiber maturation in mammals.
Cell Rep
April 2022
Instituto de Investigação e Inovação em Saúde (i3S), Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal; Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal; Cell Division Group, Department of Biomedicine, Faculdade de Medicina, Universidade do Porto, Alameda Professor Hernâni Monteiro, 4200-319 Porto, Portugal. Electronic address:
Chromosome segregation in mammals relies on the maturation of a thick bundle of kinetochore-attached microtubules known as k-fiber. How k-fibers mature from initial kinetochore microtubule attachments remains a fundamental question. By combining molecular perturbations and phenotypic analyses in Indian muntjac fibroblasts containing the lowest known diploid chromosome number in mammals (2N = 6) and distinctively large kinetochores, with fixed/live-cell super-resolution coherent-hybrid stimulated emission depletion (CH-STED) nanoscopy and laser microsurgery, we demonstrate a key role for augmin in kinetochore microtubule self-organization and maturation, regardless of pioneer centrosomal microtubules.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!