Nuclear lamina and matrices were prepared from sperm pronuclei assembled in Xenopus egg extracts using a fractionation and extraction procedure. Indirect immunofluorescence revealed that while chromatin was efficiently removed from nuclei during the extraction procedure, the distribution of lamins was unaffected. Consistent with this data, the amount of lamin B3, determined by immunoblotting, was not affected through the extraction procedure. Nuclear matrices were visualised in DGD sections by TEM. Within these sections filaments were observed both at the boundary of the nucleus (the lamina) and within the body of the nucleus (internal nuclear matrix filaments). To improve resolution, nuclear matrices were also prepared as whole mounts and viewed using field emission in lens scanning electron microscopy (FEISEM). This technique revealed two distinct networks of filaments. Filaments lying at the surface of nuclear matrices interconnected nuclear pores. These filaments were readily labelled with monoclonal anti-lamin B3 antibodies. Filaments lying within the body of the nuclear matrix were highly branched but were not readily labelled with antilamin B3 antibodies. Nuclear matrices were also prepared from sperm pronuclei assembled in lamin B3 depleted extracts. Using FEISEM, filaments were also detected in these preparations. However, these filaments were poorly organised and often appeared to aggregate. To confirm these results nuclear matrices were also observed as whole mounts using TEM. Nuclear matrices prepared from control nuclei contained a dense array of interconnected filaments. Many (but not all) of these filaments were labelled with anti-lamin B3 antibodies. In contrast, nuclear matrices prepared from “lamin depleted nuclei' contained poorly organised or aggregated filaments which were not specifically labelled with anti-lamin B3 antibodies.
Nuclear lamina and nuclear matrix organization in sperm pronuclei assembled in Xenopus egg extract
C. Zhang, H. Jenkins, M.W. Goldberg, T.D. Allen, C.J. Hutchison; Nuclear lamina and nuclear matrix organization in sperm pronuclei assembled in Xenopus egg extract. J Cell Sci 1 September 1996; 109 (9): 2275–2286. doi: https://doi.org/10.1242/jcs.109.9.2275
Download citation file:
Advertisement
Cited by
JCS Journal Meeting 2023: Imaging Cell Dynamics

Our 2023 Journal Meeting on ‘Imaging Cell Dynamics’ will be held from 14-17 May 2023 in Lisbon, Portugal. Due to popular demand, we can currently only accept applications for online attendance. Apply now to attend this meeting virtually. Registration deadline: 31 March.
Call for papers: Cell and Tissue Polarity
-PolarityCFP.png?versionId=4696)
We are welcoming submissions for our next special issue, which will focus on ‘Cell and tissue polarity’ and will be guest edited by David Bryant. Submission deadline: 15 July.
Editorial: Publishing where it matters
Editor-in-Chief Michael Way outlines Journal of Cell Science’s plans for the upcoming year and introduces Seema Grewal as our new Executive Editor.
preLights 5th Birthday webinar

preLights, our preprint highlighting service, is celebrating its 5th birthday this year. To mark the occasion, join us online on 14 March 2023 at 16:00 GMT for a discussion, led by four preLights alumni, on how to identify and navigate the challenges and opportunities while shaping your career as an early-career researcher.
Cell Scientists to Watch

As a community-focused journal, Journal of Cell Science is keen to support the next generation of cell biologists. Check out Cell Scientists to Watch, our interview series featuring talented researchers who have recently set up their own labs.