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Vol 54, No 3 (2023)
Original study articles
Analysis of the Ovarian Marker Genes Expression Revealed the Antagonistic Effects of Serotonin and Androstenedione on the Functional State of Mouse Granulosa Cells in Primary Culture
Abstract
Primary culture of granulosa cells is a prerequisite for a complete study of the normal functioning of the ovary and its pathologies. In this work, we selected the optimal protocol for obtaining a primary culture of mouse granulosa cells in the most functionally active state and revealed the effects of androstenedione and serotonin on the expression of ovarian markers that reflect the functional status of granulosa cells. The morpho-functional analysis of the ovary after PMSG stimulation revealed that 48 hours after PMSG stimulation is the optimal time for obtaining granulosa cells in the most active functional state. Using the set of 14 ovarian functional state marker genes we reveal that androstenedione inhibits cumulus and immature granulosa markers but stimulates genes characteristic of the mature state of granulosa. At the same time, granulosa cells express serotonergic receptors and transporter SERT. The ovarian marker genes expression analysis revealed that serotonin affects the expression of genes characterizing the differentiation of granulosa cells towards cumulus cells. Summarizing, we can conclude that serotonin and androstenedione have an antagonistic effect on the functional state of mouse granulosa cells in primary culture in vitro.
183-195
Expression of the engrailed Homologue in Larvae and Juveniles of the Annelid Alitta virens Characterizes the Formation of Segments from the Growth Zone
Abstract
The evolutionary origin of segmentation remains to be a mystery. In arthropods, the engrailed gene is recognized as one of the most important and conservative members of the segmentation developmental program. Orthologues of this gene have been identified in annelids, but their role is interpreted ambiguously, because in some species their expression precedes subdivision of the body into segments, but in others it does not. Here we studied the expression of engrailed in the nereid polychaete Alitta virens during metamorphosis and development of the first postlarval segments. Our data support the possible involvement of this gene in the process of segment formation from the growth zone in A. virens. At the larval stages, engrailed is expressed in neuroectodermal cells, in the growth zone, as well as in metameric epidermal cell rows at the anterior boundary of each segment. Upon transition from the metatrochophore to the nectochaete stage, the circular expression domain in the growth zone expands and then resolves into two serial domains. Over time, the distance between these circular domains increases indicating the growth of the first postlarval segment anlage. Formation of subsequent postlarval segments occurs in a similar way. Analyzing our results and literature data, we compared engrailed expression patterns in annelids and arthropods. Our work indicates an absence of conservation in patterning of sequentially developing segments from the growth zone in protostomes. We suggest that in A. virens the anteroposterior axis elongation occurs simultaneously with the specification of a new segment. These features differ from the known models of the growth zone and indicate the possibility that nereids have a specific mechanism of segmentation.
196-204
Sharp Increase in Expression of Subunit Genes and Activity of Proteasomes in Laringeal Ontogeny Is Associated with the Development of Squamous Cell Carcinoma
Abstract
Proteasomes, the most important participants in protein catabolism, maintain proteostasis and regulate cellular processes in ontogeny. Deviations in the functioning of proteasomes are associated with the development of various pathologies, including a number of oncological diseases. In this work, we studied changes in subunit gene expression and proteasome activity in laryngeal cancer tissue and epithelium of patients with chronic hyperplastic diseases of the larynx, which are considered obligate precancer. The activity of circulating proteasomes was also studied in the same groups of patients. The level of gene expression was assessed using quantitative reverse transcriptase PCR in real time. A method for assessing chymotrypsin-like (CTL) and caspase-like (CL) activities of proteasomes has been modified for the analysis of small volumes of biopsy samples. An increase in the level of expression of the proteasome genes (PSMB6, PSMB7, PSMB5, and PSMB4) in the tissues of squamous cell carcinoma of the larynx was shown compared to pre-tumor samples. An increase in CTL and CL activities of intracellular proteasomes in the malignant epithelium of the larynx was also found in comparison with the conditionally normal tissue and with the epithelium of patients with chronic hyperplastic diseases of the larynx. An increase in chymotrypsin-like activity was observed in circulating proteasomes. ROC-analysis (Receiver operating characteristic) revealed the relationship between PSMB5 mRNA expression and CTP activity of tissue proteasomes with the development of laryngeal cancer in patients with chronic hyperplastic diseases of the larynx. In the future, it is possible to use these indicators to develop a method for predicting the transition of precancer of the larynx to cancer.
205-213
Influence of Temperature on the Life Cycle of Invasive Species Gmelinoides fasciatus (Crustacea: Amphipoda) in Lake Onego
Abstract
The invasive species Gmelinoides fasciatus (Stebbing 1899) (Crustacea: Amphipoda) is able to successfully form stable populations in the recipient reservoir Lake Onego. In conditions of latitudes 62° N (Petrozavodsk Bay)—63° N (Kumsa area of the Povenets Bay) of Lake Onego, the temperature factor does not limit the development of the amphipod population. Successful naturalization of G. fasciatus under new conditions on the northern border of the range of the European part of Russia (Petrozavodsk Bay and Povenets Bay of Lake Onego) is explained by a sufficient number of degree-days (2269–2626 for the period end of May–beginning of October), which determine the one-year life cycle with generations previous year and current year.
214-225
Short communications
Structural Organization of GABA- and Nitroxidergic Systems of Subfornical Organ in Wistar Rats Postnatal Development
Abstract
The subfornical organ (SFO) is one of the circumventricular organs (CVOs) of the mammalian nervous system responsible for maintaining the energy and water and sodium balance. Despite notable interest in the SFO and its physiological functions, the organization of individual populations of SFO cells, as well as their interactions remain to be clearly established. In this study we examined GABA and nitroxidergic systems of SFO using immunohistochemical (IHC) methods. The brain of male Wistar rats at different stages of postnatal development: postnatal day 7 (P7), 14 (P14) and adult (4–6 months), was examined. The data obtained allowed us to characterize changes in the activity of the GABA- and nitroxidergic systems of the SFO during development. In adult rats, three subpopulations of nitroxidergic cells, differing in the intensity of the reaction and tissue localization, can be distinguished. The revealed morphological heterogeneity of nitroxidergic cells in SFO may reflect their diverse functional status.
226-231
Generation of a Null Mutation of the toothrin Gene by Targeted Homologous Recombination in Drosophila melanogaster
Abstract
Drosophila toothrin gene is related to the conserved d4 family of genes that encode specific transcription factors. In early works, we investigated the effect of tth gene overexpression and characterized the specific pattern of its expression in the nervous system, but mutations in the tth gene have not yet been found. Null mutants are essential for analyzing gene function and screening of partner genes. Here we report the generation of the first tth gene knockout in Drosophila using a homologous recombination technique.
232-238