Presence of tubular and reticular structures in the nucleus of human vascular smooth muscle cells

Abstract

In recent decades, studies addressing nuclear calcium (Ca(2+)) homeostasis and signaling contributed to redefining the role of the nucleus. Yet many aspects of nuclear Ca(2+) signaling and homeostasis are only modestly understood. The present study aimed at investigating the presence of nuclear structures which could contribute to the regulation of nuclear Ca(2+) homeostasis. Using real 3D confocal microscopy, coupled to utilization of appropriate organelle probes and specific antibodies, we identified two entities in the nuclei of intact human vascular smooth muscle cells (hVSMCs) as well as in isolated hVSMCs nuclei. Our results demonstrate the presence of an ER-like nuclear reticular structure in nuclei of intact hVSMCs and in isolated nuclei. Similar to the ER/SR, this structure possesses thapsigargin binding sites, IP(3)Rs and RyRs, thus it was named nucleoplasmic reticulum (NR). Furthermore, nuclear tubular structures were also detected. The latter, similar to the nuclear envelope membranes, possess nuclear pores, thapsigargin binding sites, Angiotensin II receptor AT(2), and are associated with Lamin A/C. However, unlike the NR and the nuclear envelope membranes, these tubular structures disappeared when the nuclei were isolated from the cells. The nuclear tubular structures were called Nuclear T-Tubules (NTTs). Our calcium studies in isolated nuclei utilizing IP(3) and Ryanodine suggest that the NR may participate in nuclear Ca(2+) signaling. On the other hand, presence of nuclear pores on the NTTs suggests that these structures can play a role in cytosol-nucleus exchange. In conclusion, two distinct structures are present in the nucleus of hVSMCs and might play an important role in nuclear Ca(2+) homeostasis.