Loss of polyubiquitin gene Ubb leads to metabolic and sleep abnormalities in mice.

AIMS: Ubiquitin performs essential roles in a myriad of signalling pathways required for cellular function and survival. Recently, we reported that disruption of the stress-inducible ubiquitin-encoding gene Ubb reduces ubiquitin content in the hypothalamus and leads to adult-onset obesity coupled with a loss of arcuate nucleus neurones and disrupted energy homeostasis in mice. Neuropeptides expressed in the hypothalamus control both metabolic and sleep behaviours. In order to demonstrate that the loss of Ubb results in broad hypothalamic abnormalities, we attempted to determine whether metabolic and sleep behaviours were altered in Ubb knockout mice. METHODS: Metabolic rate and energy expenditure were measured in a metabolic chamber, and sleep stage was monitored via electroencephalographic/electromyographic recording. The presence of neurodegeneration and increased reactive gliosis in the hypothalamus were also evaluated. RESULTS: We found that Ubb disruption leads to early-onset reduced activity and metabolic rate. Additionally, we have demonstrated that sleep behaviour is altered and sleep homeostasis is disrupted in Ubb knockout mice. These early metabolic and sleep abnormalities are accompanied by persistent reactive gliosis and the loss of arcuate nucleus neurones, but are independent of neurodegeneration in the lateral hypothalamus. CONCLUSIONS: Ubb knockout mice exhibit phenotypes consistent with hypothalamic dysfunction. Our data also indicate that Ubb is essential for the maintenance of the ubiquitin levels required for proper regulation of metabolic and sleep behaviours in mice.

Preoperative uterine artery embolization and evacuation in the management of cervical pregnancy: report of two cases.

Preoperative uterine artery embolization and cervical evacuation as conservative management of cervical pregnancy has been tried in recent years. However, cervical suturing, vasoconstrictor injection, or cervical ballooning was frequently used as an ancillary measures in those procedures in most of the previous studies. We report two cases of cervical pregnancy that were successfully treated with preoperative uterine artery embolization and removal of gestational material without ancillary procedures. Our therapeutic modality seems to be safe and effective for conservative management of cervical pregnancy.

Development of reverse transcription-competitive polymerase chain reaction method to quantitate the expression levels of human sodium iodide symporter.

The sodium iodide symporter (NIS) is the plasma membrane protein that mediates active iodide uptake into thyroid follicular cells. To investigate whether human NIS (hNIS) mRNA levels in papillary thyroid carcinomas (PCs) correlate with the ability of tumors to concentrate radioiodide, we developed a reverse transcription-competitive polymerase chain reaction (RT-cPCR) method to quantify the hNIS mRNA levels in thyroid tissues. We studied 7 normal thyroid tissues, 8 PCs, and 1 follicular adenoma. hNIS mRNA levels in PCs were generally lower than those found in normal thyroid tissues. The reduced radioiodide concentrating activity of PCs is due, at least in part, to the reduced expression and/or the decreased stability of hNIS mRNA.

An immunohistochemical study of Na+/I- symporter in human thyroid tissues and salivary gland tissues.

The human Na+/I- symporter (hNIS) is the plasma membrane protein that mediates active iodide uptake into several tissues, such as the thyroid and salivary glands. To study the distribution and cellular localization of the hNIS protein, we have generated a polyclonal antibody that could detect the hNIS protein by immunohistochemical staining on tissue sections. In normal thyroids, hNIS expression is heterogeneous, and it is only detected in sporadic thyrocytes of a given follicle. The hNIS protein was not detected in thyroid carcinomas, yet it was detected in the majority of thyrocytes in Graves' thyroids. In salivary glands, hNIS protein was not detected in acinar cells, but it was detected in ductal cells. The hNIS proteins are clustered in the basal and lateral membranes in cells stained positive for hNIS.

Promoter characterization of the human Na+/I- symporter.

The Na+/I- symporter (NIS) is the plasma membrane protein that mediates active iodide uptake into thyroid follicular cells. To understand the molecular mechanisms of human NIS (hNIS) gene regulation, the 2-kb immediate 5'-flanking region of hNIS was characterized. The tentative hNIS transcriptional start site was mapped to -375 nucleotide relative to the ATG site. Various 5'-genomic DNA fragments were tested for promoter activity in thyroid and nonthyroid cells. Our results indicate that the DNA regulatory elements in the 2-kb immediate 5'-flanking region were not sufficient to confer thyroid-selective transcription of the hNIS gene. In addition, hNIS minimal promoter was localized to a region of 144 bp that contains a 90-bp stretch of a highly conserved DNA fragment between hNIS and rat NIS.

Promoter characterization of the rat Na+/I- symporter gene.

The Na+/I- symporter is the molecule that mediates active iodide uptake in the thyroid gland. A 16.4 kb genomic DNA fragment of the rat Na+/I- symporter gene (rNIS) was isolated, restriction mapped and a 2 kb region immediate 5' to the ATG site was sequenced. The transcription start site for rNIS was mapped to -98 nucleotide (nt) relative to the ATG translation initiation site. A series of 5' genomic DNA fragments ranging from 233 bp to 8 kb were tested for promoter activity in both thyroid and non-thyroid cells. Our results indicate that the DNA regulatory elements within 8 kb of the 5' flanking region of rNIS are not sufficient to confer thyroid-selective transcription. However, consistent with the clinical observation that NIS expression is reduced in thyroid tumors, the rNIS promoter activity is suppressed in ret/PTC1 transformed NIH3T3 cells, compared to non-transformed NIH3T3 cells.

Expression, exon-intron organization, and chromosome mapping of the human sodium iodide symporter.

The active iodide uptake of the thyroid gland in humans is mediated by the human sodium iodide symporter (hNIS). In this report, we show that hNIS expression was detected primarily in thyroid tissue, but also in breast, colon, and ovary tissues. Expression of hNIS is greatly reduced in thyroid tumors compared to normal thyroid tissue. Among tumor tissues, hNIS expression appears to be variable, consistent with the variable response to radioiodide treatment observed for thyroid carcinomas. The coding region of hNIS is interrupted by 14 introns, and the nucleotide sequence of each exon-intron junction is reported. Using this information, an alternatively spliced form of hNIS was identified. Finally, the chromosome location of the hNIS gene was mapped to chromosome 19p.