CircRNA-WNK2 acts as a ceRNA for miR-328a-3p to promote AANAT expression in the male rat pineal gland

Noncoding RNAs (ncRNAs), including miRNAs and circRNAs, which are expressed with a daily rhythm in the rat pineal gland, are associated with the regulation of melatonin secretion and other biological functions. However, the mechanisms of these molecules in the rat pineal gland are not yet fully understood. In this study, we found circR-WNK2 was highly expressed at night, which may be involved in the regulation of melatonin secretion through the ceRNA mechanism. By dual luciferase reporter, RNA pulldown, and FISH assays, we found that miR-328a-3p can target circR-WNK2 and the Aa-nat mRNA 3’UTR. Transfection experiments indicated that circR-WNK2 could competitively bind to miR-328a-3p, reduce miR-328a-3p expression, and promote Aa-nat gene expression and melatonin secretion. And by constructing an SCGx rat model, we found that ncRNAs expressed in the pineal gland was regulated by signals from the SCN. This finding supports the hypothesis that these noncoding RNAs may interact to shape the circadian rhythm through transcriptional processing in melatonin synthesis.

FEATURES OF MORPHOLOGICAL REARRANGEMENTS OF STRUCTURAL COMPONENTS IN THE RAT PINEAL GLAND UNDER THE IMPACT OF HEAVY METAL SALTS

The development of the pathology of individual organs and systems undoubtedly depends on adverse environmental factors. Particular attention of researchers attracts heavy metal salts. Epiphyseal hormones play a key role in regulating and maintaining basic body functions. The article presents the results of the study on morphological and morphometric rearrangements of structural changes in the rat pineal gland influence for 30 days of heavy metal salts (sulfates of zinc, copper, iron, manganese, plumbers’ nitrite and chrome oxide). The selected concentration of salts in the mixture was due to the presence of similar concentrations of these salts in the soil and drinking water in some regions of Ukraine according to literature sources. General morphological and statistical research methods were used (histological, morphometric and variational statistics methods). The 30-days impact of the heavy metal salts combination on the rat body caused in the pineal gland morphological changes of nonspecific polymorphic nature, which were expressed in the increased vascular area, active glial response, hypertrophy of pinealocyte nuclei and increase of their optical density. Cytological signs in most pinealocytes indicated the predominance of indolamine synthesis over polypeptides. The main pathogenetic mechanisms of the influence of heavy metal salts on the organ have been established: change in the vascular lumen area, blood rheological properties impairment, tissue hypoxia, nucleus hypertrophy and change in their optical density. Morphological changes in the gland comply with the stress phase of the general adaptation syndrome. The above morphological changes negatively affected the processes of hormones evacuation into the blood, the course of the general adaptation syndrome and the homeostasis restoration in the organ.

The Circadian Rhythms of STAT3 in the Rat Pineal Gland and Its Involvement in Arylalkylamine-N-Acetyltransferase Regulation

In rodents, the melatonin production by the pineal gland is controlled through adrenergic signaling from the suprachiasmatic nuclei and regulation of the principal enzyme in its synthesis, arylalkylamine-N-acetyltransferase (AANAT). In the present study, we identified increased isoprenaline-induced aa-nat expression and nocturnal AANAT activity in the pineal glands in response to the silencing of the signal transducer and activator of transcription 3 (STAT3) with siRNA or STAT3 inhibitors WP1066 and AZD1480. This AANAT activity enhancement in vivo did not interfere with light-induced AANAT suppression. Systemic or in vitro lipopolysaccharide (LPS) administration markedly increased Stat3 expression and STAT3 phosphorylation, but it did not significantly affect AANAT expression or activity. Simultaneous LPS administration and Stat3 silencing enhanced the aa-nat transcription and AANAT activity to a similar extent as Stat3 inhibition without LPS co-administration. Furthermore, we describe the circadian rhythmicity in Stat3 expression and the phosphorylated form of STAT3 protein in the rat pineal gland. Our data suggest that the higher nocturnal endogenous level of STAT3 in the pineal gland decelerates or hampers the process of NA-induced AANAT activation or affects the AANAT enzyme stability.

Morphological structure of rat epiphysis exposed to electromagnetic radiation from communication devices

Pineal gland is one of the most important components of homeostasis - the supporting system of the body. It participates in the launch of stress responses, restriction of their development, prevention of adverse effects on the body. There was proved an impact of electromagnetic radiation on the epiphysis. However, morphological changes in the epiphysis under exposure to electromagnetic radiation of modern communication devices are studied not sufficiently. For the time present the population is daily exposed to electromagnetic radiation, including local irradiation on the brain. These date determined the task of this research - the study of the structure of rat pineal gland under the exposure to electromagnetic radiation from personal computers and mobile phones. These date determined the task of this research - the study of the structure of rat pineal gland under the exposure to electromagnetic radiation from personal computers and mobile phones. Performed transmission electron microscopy revealed signs of degeneration of dark and light pinealocytes. These signs were manifested in the development of a complex of general and specific morphological changes. There was revealed the appearance of signs of aging and depletion transmission electron microscopy both in light and dark pinealocytes. These signs were manifested in the accumulation of lipofuscin granules and electron-dense "brain sand", the disappearance of nucleoli, cytoplasm vacuolization and mitochondrial cristae enlightenment.