Research Methodologies for the Investigation of Cell Nucleus

Bustamante, J. O.

doi:10.1007/0-387-27747-1_13

Cited by 1 publication

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“…The indirect hourglass technique has a handicap that, as a consequence of the multivariate mathematical formula used, it has a large intrinsic error or uncertainty (see Appendix B). Furthermore, the technique is based on a series of assumptions that do not appear to be completely justified (Bustamante 2002b(Bustamante , 2005. The developers of the technique interpret their leakage current as resulting from 8 channels peripheral to the well-established (central) channel of the NPC Enss et al 2003).…”

Section: Indirect Techniques Used To Estimate Npc Ion Conductancementioning

confidence: 99%

Current concepts in nuclear pore electrophysiologyThis paper is one of a selection of papers published in this Special Issue, entitled The Nucleus: A Cell Within A Cell.

Bustamante

2006

Can. J. Physiol. Pharmacol.

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Over 4 decades ago, microelectrode studies of in situ nuclei showed that, under certain conditions, the nuclear envelope (NE) behaves as a barrier opposing the nucleocytoplasmic flow of physiological ions. As the nuclear pore complexes (NPCs) of the NE are the only pathways for direct nucleocytoplasmic flow, those experiments implied that the NPCs are capable of restricting ion flow. These early studies validated electrophysiology as a useful approach to quantify some of the mechanisms by which NPCs mediate gene activity and expression. Since electron microscopy (EM) and other non-electrophysiological investigations, showed that the NPC lumen is a nanochannel, the opinion prevailed that the NPC could not oppose the flow of ions and, therefore, that electrophysiological observations resulted from technical artifacts. Consequently, the initial enthusiasm with nuclear electrophysiology faded out in less than a decade. In 1990, nuclear electrophysiology was revisited with patch-clamp, the most powerful electrophysiological technique to date. Patch-clamp has consistently demonstrated that the NE has intrinsic ion channel activity. Direct demonstrations of the NPC on-off ion channel gating behavior were published for artificial conditions in 1995 and for intact living nuclei in 2002. This on-off switching/gating behavior can be interpreted in terms of a metastable energy barrier. In the hope of advancing nuclear electrophysiology, and to complement the other papers contained in this special issue of the journal, here I review some of the main technical, experimental, and theoretical issues of the field, with special focus on NPCs.

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Section: Indirect Techniques Used To Estimate Npc Ion Conductancementioning

confidence: 99%

Current concepts in nuclear pore electrophysiologyThis paper is one of a selection of papers published in this Special Issue, entitled The Nucleus: A Cell Within A Cell.

Bustamante

2006

Can. J. Physiol. Pharmacol.

View full text Add to dashboard Cite

show abstract

Research Methodologies for the Investigation of Cell Nucleus

Cited by 1 publication

References 63 publications

Current concepts in nuclear pore electrophysiologyThis paper is one of a selection of papers published in this Special Issue, entitled The Nucleus: A Cell Within A Cell.

Current concepts in nuclear pore electrophysiologyThis paper is one of a selection of papers published in this Special Issue, entitled The Nucleus: A Cell Within A Cell.

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