Detection of Trace Processes in the Networks of Neurons Cultured on Microelectrode Arrays

Sukhanova, Anna L.; Mineyeva, Olga A.; Kiselev, I. I.; Burtsev, Mikhail; Anokhin, Konstantin V.

doi:10.1007/s10517-012-1774-9

Cited by 2 publications

(3 citation statements)

References 10 publications

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“…Stained cultures were digitized and analyzed (see Section 4.2). The number of c-Fos-positive nuclei was expressed as a percentage of the total number of nuclei [22]; the arithmetic mean was calculated for 10 fields of view in each culture.…”

Section: Immunocytochemical Detection Of Proteinsmentioning

confidence: 99%

“…In cell cultures, the expression of immediate early genes can be induced by diverse stimuli. For instance, electrical stimulation significantly increases c-Fos expression in the neuronal culture [22] and in rat hippocampus [23], while KCl application was followed by the appearance of c-Fos protein in cultured hippocampal slices [24]. A possible relationship between Lf and c-Fos induction at the level of individual cells is still not explored.…”

Section: Introductionmentioning

confidence: 99%

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Lactoferrin Modulates Induction of Transcription Factor c-Fos in Neuronal Cultures

Kopaeva

Азиева

Cherepov

et al. 2023

IJMS

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Lactoferrin (Lf) is a multifunctional protein from the transferrin family. Of particular interest is the ability of Lf to affect a wide range of neuronal processes by modulating the expression of genes involved in long-term neuroplasticity. The expression of the immediate early gene c-fos that is rapidly activated in response to external influences, and its product, transcription factor c-Fos, is widely used as a marker of long-term neuronal plasticity. The present study aims to examine the effect of human Lf on the induction of transcription factor c-Fos in the primary mouse neuronal cultures after stimulation and to determine the cellular localization of human Lf and its colocalization with induced c-Fos protein. Primary dissociated cultures of hippocampal cells were obtained from the brains of newborn C57BL/6 mice (P0-P1). On day 7 of culturing, human Lf was added to the medium. After 24 h (day 8 in culture), c-Fos protein was induced in cells by triple application of 50 mM KCl. c-Fos content was analyzed using the immunofluorescent method 2 h after stimulation. Stimulation promoted exogenous Lf translocation into the nuclei of cultured neuronal cells, which correlated with increased induction of transcription factor c-Fos and was accompanied by nuclear colocalization of these proteins. These results attest to the potential of Lf as a modulator of neuronal processes and open up new prospects in studying the mechanisms of the regulatory effects of lactoferrin on cell function.

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Section: Immunocytochemical Detection Of Proteinsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Lactoferrin Modulates Induction of Transcription Factor c-Fos in Neuronal Cultures

Kopaeva

Азиева

Cherepov

et al. 2023

IJMS

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“…В культуре экспрессия немедленных ранних генов может быть индуцирована различными стимулами. Ранее было показано, что электрическая стимуляция вызывает значительное увеличение экспрессии c-Fos в нейрональной культуре [6], а аппликация KCl -появление белка c-Fos в культурах срезов гиппокампа [7]. В наших предыдущих исследованиях методами иммуногистохимии были выявлены высокоспецифичные места связывания лактоферрина человека (чЛф) в ядрах нейронов в мозге мыши [8].…”

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Human lactoferrin enhances the expression of transcription factor c-Fos in neuronal cultures under stimulated conditions

Копаева¹,

Азиева²,

Cherepov³

et al. 2021

Nauchno-Prakticheskii Zhurnal «Patogenez»

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Целью настоящей работы стало исследование влияния лактоферрина (Лф) человека на экспрессию транскрипционного фактора c-Fos в первичных нейрональных культурах после физиологической стимуляции, определение клеточной локализации Лф человека и возможной колокализации экзогенного белка с индуцированной экспрессией c-Fos. Методы. Первичные диссоциированные клеточные культуры получали из гиппокампа головного мозга новорожденных мышей (Р0-Р1) линии С57Вl/6. Индукцию экспрессии белка c-Fos в клетках осуществляли путем трехкратного добавления 50 мМ KСl в культуральную среду на 8-й день культивирования in vitro. Анализ содержания c-Fos проводили иммунофлюоресцентным методом через 2 часа после стимуляции. Результаты. Лф детектировался как в цитоплазме, так и в ядрах отдельных клеток культуры после стимуляции KСl. В ядрах некоторых клеток была выявлена колокализация включения Лф и экспрессии c-Fos. Было обнаружено, что предварительное введение Лф в культуральную среду увеличивало количество клеток, экспрессирующих c-Fos после добавления 50 мМ KСl. The aims of this research were 1) to study the effect of human lactoferrin (Lf) on the expression of the c-Fos transcription factor in primary neuronal cultures after physiological stimulation; 2) to determine the cellular localization of human Lf and possible colocalization of an exogenous protein with induced c-Fos expression. Methods. Primary dissociated cell cultures were obtained from the hippocampus of newborn C57Bl/6 mice (P0-P1). The expression of c-Fos was induced by addition of 50 mM KCl to the culture medium at 8 day in vitro. c-Fos content was analyzed by immunofluorescence 2 hrs after stimulation. Results. Lf was detected in cytoplasm and in nuclei after stimulation KCl. Lf inclusion and c-Fos expression were colocalized in the nuclei of some cells. Thus, results showed that pretreatment with Lf led to increase in the number of cells expressing c-Fos after exposure to 50 mM KCl.

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Detection of Trace Processes in the Networks of Neurons Cultured on Microelectrode Arrays

Cited by 2 publications

References 10 publications

Lactoferrin Modulates Induction of Transcription Factor c-Fos in Neuronal Cultures

Lactoferrin Modulates Induction of Transcription Factor c-Fos in Neuronal Cultures

Human lactoferrin enhances the expression of transcription factor c-Fos in neuronal cultures under stimulated conditions

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