Alternative splicing of CRH‐R1 receptors in human and mouse skin: identification of new variants and their differential expression

Pisarchik, Alexander; Słomiński, Andrzej

doi:10.1096/fj.01-0487fje

Cited by 170 publications

(224 citation statements)

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“…All samples were standardized for analysis by the amplification of housekeeping gene GAPDH as described previously [25]. Mouse TPH was amplified by a single PCR, while serotonin AANAT was amplified by nested PCR.…”

Section: Rna Extraction and Cdna Preparationmentioning

confidence: 99%

Characterization of the serotoninergic system in the C57BL/6 mouse skin

Slominski

Pisarchik

Semak

et al. 2003

European Journal of Biochemistry

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127

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We showed expression of the tryptophan hydroxylase gene and of tryptophan hydroxylase protein immunoreactivity in mouse skin and skin cells. Extracts from skin and melanocyte samples acetylated serotonin to N-acetylserotonin and tryptamine to N-acetyltryptamine. A different enzyme from arylalkylamine N-acetyltransferase mediated this reaction, as this gene was defective in the C57BL6 mouse, coding predominantly for a protein without enzymatic activity. Serotonin (but not tryptamine) acetylation varied according to hair cycle phase and anatomic location. Serotonin was also metabolized to 5-hydroxytryptophol and 5-hydroxyindole acetic acid, probably through stepwise transformation catalyzed by monoamine oxidase, aldehyde dehydrogenase and aldehyde reductase. Activity of the melatonin-forming enzyme hydroxyindole-O-methyltransferase was notably below detectable levels in all samples of mouse corporal skin, although it was detectable at low levels in the ears and in Cloudman melanoma (derived from the DBA/2 J mouse strain). In conclusion, mouse skin has the molecular and biochemical apparatus necessary to produce and metabolize serotonin and N-acetylserotonin, and its activity is determined by topography, physiological status of the skin, cell type and mouse strain.Keywords: mouse skin; serotonin acetylation; arylalkylamine N-acetyltransferase; tryptophan hydroxylase; hair cycle.The skin is the largest body organ and functions as a metabolically active biological barrier regulating internal homeostasis and separating the internal milieu from noxious environmental factors [1]. These functions are mediated by the skin immune, pigmentary, neuroendocrine, adnexal and vascular systems [1][2][3][4][5][6][7]. Most recently we have uncovered local serotoninergic and melatoninergic systems as novel elements of the cutaneous neuroendocrine components of human and hamster skin [8][9][10][11][12]. Serotonin is the product of a multistep metabolic pathway that starts with the hydroxylation of the aromatic aminoacid L-tryptophan by tryptophan hydroxylase (TPH) [13,14]. Serotonin can be acetylated by arylalkylamine N-acetyltransferase (AANAT) to N-acetylserotonin (NAS), which is further transformed to melatonin by hydroxyindole-O-methyltransferase (HIOMT) [13,15].Serotonin can act as a neurohormone, regulator of vascular tone, immunomodulator and growth factor, while melatonin can act as a hormone, neurotransmitter, cytokine or biological modifier [2,[15][16][17]. Some of these functions may be pertinent to skin physiology, which exhibits basic differences among the mammalian species. In rodents (mostly nocturnal animals) the skin is shielded from the damaging effect of solar radiation by fur [18], and the morphology of the entire mouse skin changes in close coordination with the cyclic activity of the hair follicle [19]. Mouse hair follicle cycling is characterized by a precisely regulated, time frame-restricted and differential pattern in the expression and activity of melanogenesis related proteins, PH, pterins and thioredoxin ...

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Section: Rna Extraction and Cdna Preparationmentioning

confidence: 99%

Characterization of the serotoninergic system in the C57BL/6 mouse skin

Slominski

Pisarchik

Semak

et al. 2003

European Journal of Biochemistry

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127

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“…Thus, it has been reported that activation of protein kinases (PKA and PKC) and environmental insults (UV irradiation) can lead to modulation of CRH-R1 mRNA expression in keratinocytes and melanoma cells [34]. In addition, physiological changes, such as the onset of labour, can alter the relative expression of CRH-R1 variants in the uterus [102].…”

Section: Mechanisms Of Alternative Splicing Of Gpcrsmentioning

confidence: 99%

“…A fascinating example of the importance of splicing in GPCR biology and hormone actions is the CRH-R1 receptor, which orchestrates mammalian responses to stressful stimuli [33]. In humans the CRH-R1 gene spans over 50 kb and contains 14 exons, and at least 14 alternatively spliced mRNAs for CRH-R1 variants have been detected to date [33,34]. The complete gene product incorporating all 14 exons encodes a 444-amino acid GPCR, termed CRH-R1β, which displays weak agonist binding and impaired signalling properties [93][94][95].…”

Section: Introductionmentioning

confidence: 99%

“…Deletion of exon 3 results in CRH-R1c, which lacks 40 amino acids within the N-terminus and no longer binds CRH [96]. Various exon deletions or insertions of "cryptic" exons (exons that are absent in the normal form, but occur in an alternative form) result in frame-shifts and the potential introduction of a premature termination codon, leading to the expression of soluble or severely truncated membrane proteins (termed CRH-R1e-h) [34,97]. Another splice variant, termed CRH-R1d, contains a 42 base-pair exon deletion resulting in a 14 amino acid deletion from TM7 [87].…”

Section: Introductionmentioning

confidence: 99%

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Alternative splicing of G protein-coupled receptors: physiology and pathophysiology

Markovic

Challiss

2009

Cell. Mol. Life Sci.

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Abstract. The G protein-coupled receptors (GPCRs) are a superfamily of transmembrane receptors that have a broad distribution and can collectively recognize a diverse array of ligands. Activation or inhibition of GPCR signalling can affect many (patho)physiological processes and consequently they are a major target for existing and emerging drug therapies.A common observation has been that the pharmacological, signalling and regulatory properties of GPCRs can vary in a cell-and tissue-specific manner. Such "phenotypic" diversity might be attributable to post-translational modifications and/or association of GPCRs with accessory proteins, however, post-transcriptional mechanisms are also likely to contribute. Although, approximately 50% of GPCR genes are intronless, those that possess introns can undergo alternative splicing, generating GPCR subtype isoforms that may differ in their pharmacological, signalling and regulatory properties. In this Review we shall highlight recent research into GPCR splice-variation, and discuss the potential consequences this might have for GPCR function in health and disease.

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“…The CRH-R1 gene expresses multiple subtypes (Ross et al, 1994) (Grammatopoulos et al, 1999) (Pisarchik and Slominski, 2001), which are produced by differential exon splicing. Similar exon rearrangements have been described for the calcitonin and parathyroid hormone/parathyroid hormone-related peptide receptors.…”

Section: Crh Receptorsmentioning

confidence: 99%