Long non-coding RNAs and small nucleolar RNA host gene 6 contribute to the depression-like behavior of hypothyroid mice by promoting methylation of the brain-derived neuropathic factor promoter that is mediated by DNA methyltransferase 1.

Long non-coding RNAs (lncRNAs) and small nucleolar RNA host gene 6 (SNHG6) have attracted extensive attention due to their involvement in various pathological processes. However, the functional role of lncRNA SNHG6 in depression-like behavior induced by hypothyroidism is still largely unknown. Our study was designed to explore the biological role of lncRNA SNHG6 in depression-like behavior induced by hypothyroidism and the underlying mechanisms. First, the depression-like behavior of hypothyroid mice was investigated after lncRNA SNHG6 knockdown. Subsequently, the regulation of the methylation levels of the brain-derived neurotrophic factor (BDNF) promoters by lncRNA SNHG6 was evaluated. To reveal the underlying mechanisms of lncRNA SNHG6 in the methylation of the BDNF promoters, RNA pull-down assays, RNA immunoprecipitation, and co-immunoprecipitation were performed. Further experiments were also conducted to investigate the roles of DNA (cytosine-5)-methyltransferase 1 (DNMT1) in the depression-like behavior induced by hypothyroidism. In this study, elevated levels of lncRNA SNHG6 were noted in the hippocampus in hypothyroid mice. Function assays proved that lncRNA SNHG6 knockdown alleviated the depression-like behavior induced by hypothyroidism. Furthermore, a mechanistic investigation validated that lncRNA SNHG6 stabilized DNMT1 by blocking UHRF1-mediated DNMT1 ubiquitination, which increased the methylation levels of the BDNF promoters. Moreover, DNMT1 was found to be involved in depression-like behavior in hypothyroid mice. Our findings revealed a novel mechanism by which lncRNA SNHG6 promotes the methylation levels of the BDNF promoters by stabilizing DNMT1 and sheds light on potential therapeutic strategies for depression-like behavior induced by hypothyroidism.

A comparative study of stress-related gene expression under single stress and intercross stress in rice.

Plant hormones play important roles in the crosstalk between biotic and abiotic stresses in rice throughout its entire growth period. However, these interactions are not completely understood. In this study, the physiological performance of rice seedlings under a single stress and a sequential combination of various stresses (intercross stress) was determined. We found that catalase, superoxide dismutase, and peroxidase activities and malondialdehyde were highly regulated by intercross stresses. Furthermore, the expression levels of pathogenesis-related genes and drought stress-related genes under various treatments were analyzed. We demonstrated that under drought-disease intercross stress, the expression levels of the PR4, PAL, and Cht-1 genes were significantly upregulated, while under salt-disease intercross stress, the expression levels of the PR1a, PBZ1, Gns1, and Cht-1 genes underwent significant changes. Regardless of the type of intercross stress, the expression of LOX-RLL was significantly affected. We also showed that the expression of drought stress-related genes OsSKIPa, OsNADPH1, JRC0594, and OsGL1-2 was significantly regulated, suggesting that these genes play important roles in the interaction between biotic and abiotic stresses. We, therefore, conclude that the interactions between various types of biotic and abiotic stresses vary in a complex pattern and would require further in-depth investigation.

Effect of thyroxine on SNARE complex and synaptotagmin-1 expression in the prefrontal cortex of rats with adult-onset hypothyroidism.

Thyroid hormone insufficiency in adulthood causes a wide range of brain impairments, including altered synaptic proteins in the prefrontal cortex (PFC). The present study investigated whether adult-onset hypothyroidism altered the expression of soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes and synaptotagmin-1 (syt-1) in the PFC of rats. Sprague-Dawley rats were randomly divided into 4 groups: control, hypothyroid, and hypothyroid treated with T(4) [5 or 20 µg/100 g body weight (BW)]. Adult-onset hypothyroidism was induced in rats with the antithyroid drug 6-n-propyl-2-thiouracil (ip injection). PFC levels of synaptosomal-associated protein of 25 kDa (SNAP-25), syntaxin-1, vesicle-associated membrane protein 2 (VAMP-2) and syt-1 were determined by immunohistochemistry and western blot analyses. The results showed that syntaxin-1 and syt-1 were expressed at significantly lower levels in hypothyroid rats, VAMP-2 levels were not altered, and SNAP-25 levels were much higher compared to controls. A 2-week treatment with 5 µg T(4)/100 g BW partially normalized levels of SNARE complex and syt-1, and 20 µg T(4)/100 g BW restored these proteins closer to normal levels. Our findings indicate that dysregulation of SNARE complex and syt-1 in PFC of adult-onset hypothyroidism can be restored by T(4) treatment.

Effect of thyroxine on synaptotagmin 1 and SNAP-25 expression in dorsal hippocampus of adult-onset hypothyroid rats.

Adult-onset hypothyroidism causes cognitive dysfunctions of learning and memory, in which many synaptic proteins in hippocampus are involved. In our work, we studied the effect of adult-onset hypothyroidism on the expression of synaptotagmin 1 (syt 1) and SNAP-25 in dorsal hippocampus as well as its recovery by levothyroxine (L-T(4)) replacement therapy. Rats were divided into 4 groups: control, hypothyroidism, and hypothyroid rats treated with 5 µg T(4)/100 g body weight (BW) and 20 µg L-T(4)/100 g BW, respectively. Protein levels of syt 1 and SNAP-25 in dorsal hippocampus were determined by Western blot and immunohistochemistry. The immunoblot analysis indicated that syt 1 was expressed at a significantly lower level in hypothyroid rats, while the level of SNAP-25 was much higher compared to controls. Furthermore, using immunostaining, we found that on the one hand, expression of syt 1 was significantly down-regulated in the examined layers of CA1 and CA3 subregions but not dentate gyrus (DG); however, on the other hand, expression of SNAP-25 was up-regulated in the layers of CA1, CA3, and DG. Two-week treatment with 20 µg LT(4)/ 100 g BW fully restored the levels of syt 1 and SNAP-25 to the normal level, which was more effective than 5 µg LT(4)/ 100 g BW that partially restored the levels of both proteins. These results suggest that adult-onset hypothyroidism caused down-regulation of syt 1 and up-regulation of SNAP- 25 level in dorsal hippocampus, which could be restored by L-T(4) treatment, and the recovery degree is related to the LT(4) dosage.

Direct interactions between immunocytes and neurons after axotomy in Aplysia.

Axon growth during development and after injury has processes in common, but also differs in that regeneration requires the participation of cells of the immune system. To investigate how neuron-immunocyte interactions might influence regeneration, we developed an in vitro model whereby neurons and hemocytes from Aplysia californica were cocultured. The hemocytes, which behave like vertebrate macrophages, migrated randomly throughout the dish. When a neuron was encountered, some hemocytes exhibited an avoidance response, whereas others formed stable contacts. Hemocytes did not distinguish between neurons from different animals. Stable contacts occurred on neurites and growth cones, but not the cell soma, and were benign in that the hemocytes did not impede neurite growth. When hemocytes attached to the cell body, it presaged the destruction of the neuron. Destruction was a dynamic process that was initiated when groups of one to three hemocytes adhered to various regions of the cell soma. Each group was then joined by other hemocytes. They did not contact the neuron, but interconnected the initial groups, forming a network around the neuron. The network then contracted to dismember the cell. Once a neuron was destroyed, hemocytes removed the debris by phagocytosis. Both damaged neurons and those without apparent damage were targets for destruction. Severing neurites with a needle resulted in the destruction of only one of six cells. Our studies suggest that hemocytes, and by extrapolation, vertebrate macrophages, exhibit highly complex interactions with neurons that can exert a variety of influences on the course of nerve regeneration.

Inflammation causes a long-term hyperexcitability in the nociceptive sensory neurons of Aplysia.

Nerve injury, tissue damage, and inflammation all cause hyperalgesia. A factor contributing to this increased sensitivity is a long-term (>24 hr) hyperexcitability (LTH) in the sensory neurons that mediate the responses. Using the cluster of nociceptive sensory neurons in Aplysia californica as a model, we are examining how inflammation induces LTH. A general inflammatory response was induced by inserting a gauze pad into the animal Within 4 days, the gauze is enmeshed in an amorphous material that contains hemocytes, which comprise a cellular immune system. Concurrently, LTH appears in both ipsilateral and contralateral sensory neurons. The LTH is manifest as increased action potential discharge to a normalized stimulus. Immunocytochemistry revealed that hemocytes have antigens recognized by antibodies to TGFbeta1, IL-6, and 5HT. When a localized inflammation was elicited on a nerve, hemocytes containing the TGFbeta1 antigen were present near axons within the nerve and those containing the IL-6 were on the surface. Western blots of hemocytes, or of gauze that had induced a foreign body response, contained a 28-kD polypeptide recognized by the anti-TGFbeta1 antibody. Exposure of the nervous system to recombinant human TGFbeta1 elicited increased firing of the nociceptive neurons and a decrease in threshold. The TGFbeta1 also caused an activation of protein kinase C (PKC) in axons but did not affect a kinase that is activated in axons after injury. Our findings, in conjunction with previous results, indicate that a TGFbeta1-homolog can modulate the activity of neurons that respond to noxious stimuli. This system could also contribute to interactions between the immune and nervous systems via regulation of PKC.

Transforming growth factor-alpha and rhinitis.

OBJECTIVES: Transforming growth factor-alpha (TGF-alpha) has been implicated in diverse physiologic and pathophysiologic functions including immunological, inflammatory, and neoplastic processes. TGF-alpha has been localized in the hyperproliferative, inflammatory environment of chronic otitis media, cholesteatoma, and asthmatic airways. TGF-beta1, which must be present with TGF-alpha to transform fibroblasts, has been found in rhinitic mucosa and in asthma in prior studies. The authors sought to identify whether TGF-alpha also played a role in the inflammatory cascade and fibrosis of rhinitis. STUDY DESIGN: A nonrandomized, prospective study was carried out in which samples of inferior turbinate and nasal polyps from rhinitic and nonrhinitic patients were subjected to immunohistochemistry and Western blotting to determine the presence of TGF-alpha. METHODS: Twenty-seven subjects undergoing surgery for rhinitis, obstructive sleep apnea, nasal fracture, and rhinoplasty were recruited for this study, the latter three groups acting as controls. Immunohistochemical and Western blotting techniques were employed to identify the presence of TGF-alpha in inferior-turbinate and nasal-polyp samples of rhinitic subjects. RESULTS: Immunohistochemistry demonstrated the selective staining of TGF-alpha in the basement membrane and extracellular matrix, including lymphatic, vascular, and glandular structures, in most turbinate samples and the absence of staining in corresponding controls. Further, TGF-alpha was isolated to a discrete 30-kD band in both inferior turbinate and polyp tissues by Western blotting without staining in the corresponding controls. CONCLUSIONS: These results suggest that TGF-alpha may play a role in the inflammatory derangement of rhinitis.