Selection of suitable reference genes is the first aim of any research dedicated to the investigation of gene expression, because the interpretation of obtained results largely depends on selection of appropriate reference genes. Nowadays, there are many mathematical approaches allowing to select not only single reference gene but also a group of the most stably expressed reference genes. The use of mathematical models which take into account multiple reference genes will allow to obtain more accurate data on the expression of target genes.

Despite abundant research studies on the physiological and biochemical nature of embryonic neurotransmitter function, little is known about the molecular genetic mechanisms involved. The expression of the main components of the serotonergic system during early Xenopus embryogenesis was investigated using RT-PCR, real time PCR and in situ hybridization. Transcripts encoding the serotonin receptors HTR2C and HTR7, as well as the vesicular monoamine transporter VMAT2, the serotonin transporter (SERT) and the serotonin synthesis enzymes tryptophan hydroxylase (TPH2) and aromatic amino acid decarboxylase (AAAD) were found to be expressed during the cleavage division stages, whereas the degradation enzyme monoamine oxidase A (MAOA) was absent. The main components of the serotonergic system were found to be expressed during the earliest stages of embryonic development. The embryonic transmitter mechanism, its complexity, and its variability among various species are discussed.

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Diterpenoid plant hormone gibberellic acid (GA) plays an important role in regulation of plant growth and development and is commonly used in agriculture for activation of plant growth and food production. It is known that many plant-derived compounds have miscellaneous biological effects on animals and humans, influencing specific cellular functions and metabolic pathways. However, the effect of GA on animal and human cells remains controversial. We investigated the effect of GA on cultured human cell lines of epidermoid origin—immortalized non-tumorigenic keratinocytes HaCaT and carcinoma A431 cells. We found that at a non-toxic dose, GA upregulated the expression of genes associated with the ER stress response—CHOP, sXBP1, GRP87 in both cell lines, and ATF4 predominantly in A431 cells. We also showed that GA was more effective in upregulating the production of ER stress marker GRP78, autophagy marker LC3B-II, and differentiation markers involucrin and filaggrin in A431 cells than in HaCaT. We conclude that GA induces mild ER stress in both cell lines, followed by the activation of differentiation via upregulation of autophagy. However, in comparison with immortalized keratinocytes HaCaT, GA is more effective in inducing differentiation of carcinoma A431 cells, probably due to the inherently lower differentiation status of A431 cells. The activation of differentiation in poorly differentiated and highly malignant A431 cells by GA may lower the level of malignancy of these cells and decrease their tumorigenic potential.

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