Actin, myosin, cytokeratins and spectrin are components of the guinea pig sperm nuclear matrix

Ocampo, Juan; Mondragón, Ricardo; Roa-Espitia, Ana Lilia; Chiquete-Félix, Natalia; Salgado, Zaira O.; Mújica, Adela

doi:10.1016/j.tice.2005.03.003

Cited by 27 publications

(25 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…As to one promising candidate of this group, there is now sufficient evidence that actin is implicated in several nuclear activities together with a growing number of actin-binding proteins (34). Actin, myosin, cytokeratins, and spectrin have all been identified in NM filaments by immunodetection (35). Indeed, in this study, many actin-binding proteins were enriched in the NM fraction (e.g.…”

Section: Nuclear Matrix Proteomics Of Colorectal Cancermentioning

confidence: 56%

Subnuclear Proteomics in Colorectal Cancer

Albrethsen

Knol

Piersma

et al. 2010

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

Abnormalities in nuclear phenotype and chromosome structure are key features of cancer cells. Investigation of the protein determinants of nuclear subfractions in cancer may yield molecular insights into aberrant chromosome function and chromatin organization and in addition may yield biomarkers for early cancer detection. Here we evaluate a proteomics work flow for profiling protein constituents in subnuclear domains in colorectal cancer tissues and apply this work flow to a comparative analysis of the nuclear matrix fraction in colorectal adenoma and carcinoma tissue samples. First, we established the reproducibility of the entire work flow. In a reproducibility analysis of three nuclear matrix fractions independently isolated from the same colon tumor homogenate, 889 of 1,047 proteins (85%) were reproducibly identified at high confidence (minimally two peptides per protein at 99% confidence interval at the protein level) with an average coefficient of variance for the number of normalized spectral counts per protein of 30%. This indicates a good reproducibility of the entire work flow from biochemical isolation to nano-LC-MS/MS analysis. Second, using spectral counting combined with statistics, we identified proteins that are significantly enriched in the nuclear matrix fraction relative to two earlier fractions (the chromatin-binding and intermediate filament fractions) isolated from six colorectal tissue samples. The total data set contained 2,059 non-redundant proteins. Gene ontology mining and protein network analysis of nuclear matrixenriched proteins revealed enrichment for proteins implicated in "RNA processing" and "mRNA metabolic process." Finally, an explorative comparison of the nuclear matrix proteome in colorectal adenoma and carcinoma tissues revealed many proteins previously implicated in oncogenesis as well as new candidates. A subset of these differentially expressed proteins also exhibited a corresponding change at the mRNA level. Together, the results show that subnuclear proteomics of tumor tissue is feasible and a promising avenue for exploring oncogenesis. Molecular & Cellular Proteomics 9:988 -1005, 2010.

show abstract

Section: Nuclear Matrix Proteomics Of Colorectal Cancermentioning

confidence: 56%

Subnuclear Proteomics in Colorectal Cancer

Albrethsen

Knol

Piersma

et al. 2010

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

show abstract

“…For nuclei isolation, a previously described technique was followed (Ocampo et al 2005) with slight modifications. NaCl-washed spermatozoa were resuspended in buffer A (50 mM Tris-HCl (pH 7.4) plus a commercial mixture of protease inhibitors, Complete, according to the manufacturer's instructions), and treated with 25 mM DTT for 20 min with constant agitation at 4 8C.…”

Section: Sperm Nuclei Isolationmentioning

confidence: 99%

In spermatozoa, Stat1 is activated during capacitation and the acrosomal reaction

Bastián

Zepeda‐Bastida²,

Uribe³

et al. 2007

Reproduction

View full text Add to dashboard Cite

A role for sperm-specific proteins during the early embryonic development has been suggested by a number of recent studies. However, little is known about the participation of transcription factors in that stage. Here, we show that the signal transducer and activator of transcription 1 (Stat1), but not Stat4, was phosphorylated in response to capacitation and the acrosomal reaction (AR). Moreover, Stat1 phosphorylation correlated with changes in its localization: during capacitation, Stat1 moved from the cytoplasm to the theca/flagellum fraction. During AR, Stat1 phosphorylation increased again. In addition, blocking protein kinase A (PKA) and PKC during capacitation abolished both phosphorylation and migration of Stat1. Our results show tight spatio-temporal rearrangements of Stat1, suggesting that after fertilization Stat1 participates in the first rounds of transcription within the male pronucleus.

show abstract

“…The general appearance of the cytoskeletal framework is similar to that described previously (Jamil, 1984). The present method is mild in that it extracts the soluble proteins leaving the general cytoarchitechture of the sperm intact and includes cytokeratins and spectrin in addition to actin and myosin (Ocampo et al, 2005). Further, in sperm, the chromatin and RNA are densely packed by the nucleoproteins in the sperm head at spermiogenesis with only a miniscule amount of cytoplasm unlike in other cells giving the microtrabecular lattice appearance seen post detergent extraction (Fey et al, 1986) to the sperm head.…”

Section: Conventional Electron Microscopy Of Thementioning

confidence: 52%

“…The actinomyosin complex and the microtubule assembly in addition to being a basic engine for locomotion, are also involved in physiological functions. The contractile actinomyosin complex is reported to play a critical role during sperm capacitation and acrosome reaction (Breitbart et al, 2005;Brener et al, 2003) and chromatin decondensation (Ocampo et al, 2005). Reports indicate that…”

mentioning

confidence: 99%

Spatial Distribution of Actin and Tubulin in Human Sperm Nuclear Matrix‐intermediate Filament Whole Mounts—A New Paradigm

Kadam¹,

D’Souza²,

Natraj³

2007

Microscopy Res & Technique

View full text Add to dashboard Cite

Sperm is a highly differentiated cell streamlined for fertilization. The function is thus heavily dependent on the cytoskeletal organization. Conventional methods limit the appreciation and correlation of this intricate cytoskeletal filament network in the context of an entire sperm. Our recent successful localization of nonmuscle myosin IIA on sperm nuclear matrix-intermediate filament (NM-IF) preparations from fertile men by embedment-free electron microscopy (EF-EM), prompted us to investigate the antigenic distribution of two major cytoskeletal proteins-actin and tubulin. The NM-IF preparations were subjected to a cocktail of buffered paraformaldehyde (2%) with a low concentration of glutaraldehyde (0.05%). These proteins were localized by indirect immunogold technique using EF-EM on sperm NM-IF whole mounts. Ultrastructure analysis revealed well preserved centrioles, outer dense fibers, axonemal filaments, and submitochondrial reticulum in the sperm NM-IF. Immunoreactive actin was localized along the length of the sperm whereas beta-tubulin was present in the axoneme alone. The spatial distribution of actin and tubulin in normal human sperm NM-IF reported here together with that of myosin on whole mount offers a powerful technique to understand sperm cytoskeletal supramolecular structure.

show abstract

Actin, myosin, cytokeratins and spectrin are components of the guinea pig sperm nuclear matrix

Cited by 27 publications

References 53 publications

Subnuclear Proteomics in Colorectal Cancer

Subnuclear Proteomics in Colorectal Cancer

In spermatozoa, Stat1 is activated during capacitation and the acrosomal reaction

Spatial Distribution of Actin and Tubulin in Human Sperm Nuclear Matrix‐intermediate Filament Whole Mounts—A New Paradigm

Contact Info

Product

Resources

About