Novel Pituitary Actions of TAC4 Gene Products in Teleost

Tachykinin 4 (TAC4) is the latest member of the tachykinin family involved in several physiological functions in mammals. However, little information is available about TAC4 in teleost. In the present study, we firstly isolated TAC4 and six neurokinin receptors (NKRs) from grass carp brain and pituitary. Sequence analysis showed that grass carp TAC4 could encode two mature peptides (namely hemokinin 1 (HK1) and hemokinin 2 (HK2)), in which HK2 retained the typical FXGLM motif in C-terminal of tachyinin, while HK1 contained a mutant VFGLM motif. The ligand-receptor selectivity showed that HK2 could activate all 6 NKRs but with the highest activity for the neurokinin receptor 2 (NK2R). Interestingly, HK1 displayed a very weak activation for each NKR isoform. In grass carp pituitary cells, HK2 could induce prolactin (PRL), somatolactin α (SLα), urotensin 1 (UTS1), neuromedin-B 1 (NMB1), cocaine- and amphetamine-regulated transcript 2 (CART2) mRNA expression mediated by NK2R and neurokinin receptor 3 (NK3R) via activation cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA), phospholipase C (PLC)/inositol 1,4,5-triphosphate (IP3)/protein kinase C (PKC) and calcium2+ (Ca2+)/calmodulin (CaM)/calmodulin kinase-II (CaMK II) cascades. However, the corresponding stimulatory effects triggered by HK1 were found to be notably weaker. Furthermore, based on the structural base for HK1, our data suggested that a phenylalanine (F) to valine (V) substitution in the signature motif of HK1 might have contributed to its weak agonistic actions on NKRs and pituitary genes regulation.

Inflammatory markers in the nasal mucosa and polyp-modified tissues in chronic rhinosinusitis with nasal polyps

Introduction. To date, chronic rhinosinusitis with nasal polyps (CRSwNP) has not yet been extensively studied: the molecular factors and mechanisms involved in the initiation of polypous transformations in nasal mucosa (NM) and sustaining their recurrence probability are still to be determined. Simultaneously, it is necessary to understand the molecular rearrangement in NM tissues to make clinical prognosis and choose an adequate therapeutic or surgical strategy for CRSwNP treatment. The aim of the study was to identify the features of how inflammatory markers localize and are distributed in the NM and polyps in various morphological CRSwNP types. Materials and methods. We studied morphological and chemical structure of nasal polyps and mucosa of the inferior turbinates. The material was obtained during surgical management of patients diagnosed with CRSwNP. The comparison group involved the patients with a deviated septum who underwent septorhi-noplasty and had neither polyposis nor concomitant inflammatory/allergic pathology. The NM removed in surgeries was used to compare morphological and chemical changes. Immunohistochemistry was applied to determine the localization and distribution of SP, NK1, nNOS, iNOS, and IL1b in the tissues. Results. The formation of nasal polyps was found to be accompanied by morphological and chemical altera-tions in the mucous membrane of the inferior turbinates. In polyps of different morphological types, the changes in the activity of inflammatory markers were specific. Conclusion. The data obtained indicate that changes in the NM of the inferior turbinates, which accompany polyposis development, give certain pathological causes that induce and maintain the pathological process. We have revealed the features of the specific signaling microenvironment in the nasal cavity, which provide special conditions for the formation of polyps of various types. The specificity of the activity and distribu-tion of inflammatory markers in the polyps of different morphological types may serve as a prerequisite for the development of personalized therapy for the disease. Keywords: chronic rhinosinusitis with nasal polyps, inflammation, neurokinin receptors, substance P, nitric oxide

The Neurokinins: Peptidomimetic Ligand Design and Therapeutic Applications

The neurokinins are indisputably essential neurotransmitters in numerous pathoand physiological events. Being widely distributed in the Central Nervous System (CNS) and peripheral tissues, their discovery rapidly promoted them to drugs targets. As a necessity for molecular tools to understand the biological role of this class, endogenous peptides and their receptors prompted the scientific community to design ligands displaying either agonist and antagonist activity at the three main neurokinin receptors, called NK1, NK2 and NK3. Several strategies were implemented for this purpose. With a preference to small non-peptidic ligands, many research groups invested efforts in synthesizing and evaluating a wide range of scaffolds, but only the NK1 antagonist Aprepitant (EMENDT) and its prodrug Fosaprepitant (IVEMENDT) have been approved by the Food Drug Administration (FDA) for the treatment of Chemotherapy-Induced and Post-Operative Nausea and Vomiting (CINV and PONV, respectively). While non-peptidic drugs showed limitations, especially in side effect control, peptidic and pseudopeptidic compounds progressively regained attention. Various strategies were implemented to modulate affinity, selectivity and activity of the newly designed ligands. Replacement of canonical amino acids, incorporation of conformational constraints, and fusion with non-peptidic moieties gave rise to families of ligands displaying individual or dual NK1, NK2 and NK3 antagonism, that ultimately were combined with non-neurokinin ligands (such as opioids) to target enhanced biological impact.

Characterisation, localisation and expression of porcine TACR1, TACR2 and TACR3 genes

Substance P is involved in many physiological and pathophysiological processes. This functional diversity is mediated by three neurokinin receptor subtypes (NK1R, NK2R and NK3R) encoded by the TACR1, TACR2 and TACR3 genes, respectively. Despite the increasing interest in using pigs (Sus scrofa) to study human disease mechanisms, the sequences of these receptors are still unconfirmed or in the case of the NK1 receptor, not yet even unpredicted. We employed in silico analysis to define the localisation of the porcine tachykinin receptor genes, and to predict the structures and amino acid sequences of the respective proteins. A reverse transcription polymerase chain reaction (RT-PCR) assay was performed to analyse the expression of tachykinin receptor genes in different porcine tissues. The data show that the TACR1 gene is located on chromosome 3, TACR2 on chromosome 14 and TACR3 on chromosome 8. All three genes encode proteins with structures that incorporate features of G-protein-coupled receptors with sizes of 407, 381 and 464 amino acids, respectively. The receptors display a high degree of similarity to other mammalian neurokinin receptors. The NK1R subtype is expressed in both the central nervous system and peripheral tissues, while NK2R expression seems to be localised mostly to peripheral tissues. The expression of NK3R is found mainly in the central nervous system. This report provides for the first time the results of a comprehensive analysis of the structure and distribution of porcine NK1R, as well as other porcine neurokinin receptors and their genes. We hope that our data may offer an invaluable foundation for the future studies on the function of diverse tachykinin peptides in the central nervous system and peripheral tissues.