Molecular Cloning and Pharmacological Characterization of Two Novel GnRH Receptors in the Lamprey (Petromyzon marinus)

Center for Molecular and Comparative Endocrinology and Department of Molecular, Cellular and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire 03824, USA.
Endocrinology (Impact Factor: 4.5). 05/2012; 153(7):3345-56. DOI: 10.1210/en.2012-1217
Source: PubMed


This paper reports the identification, expression, binding kinetics, and functional studies of two novel type III lamprey GnRH receptors (lGnRH-R-2 and lGnRH-R-3) in the sea lamprey, a basal vertebrate. These novel GnRH receptors share the structural features and amino acid motifs common to other known gnathostome GnRH receptors. The ligand specificity and activation of intracellular signaling studies showed ligands lGnRH-II and -III induced an inositol phosphate (IP) response at lGnRH-R-2 and lGnRH-R-3, whereas the ligand lGnRH-I did not stimulate an IP response. lGnRH-II was a more potent activator of lGnRH-R-3 than lGnRH-III. Stimulation of lGnRH-R-2 and lGnRH-R-3 testing all three lGnRH ligands did not elicit a cAMP response. lGnRH-R-2 has a higher binding affinity in response to lGnRH-III than lGnRH-II, whereas lGnRH-R-3 has a higher binding affinity in response to lGnRH-II than IGnRH-III. lGnRH-R-2 precursor transcript was detected in a wide variety of tissues including the pituitary whereas lGnRH-R-3 precursor transcript was not as widely expressed and primarily expressed in the brain and eye of male and female lampreys. From our phylogenetic analysis, we propose that lGnRH-R-1 evolved from a common ancestor of all vertebrate GnRH receptors and lGnRH-R-2 and lGnRH-R-3 likely occurred due to a gene duplication within the lamprey lineage. In summary, we propose from our findings of receptor subtypes in the sea lamprey that the evolutionary recruitment of specific pituitary GnRH receptor subtypes for particular physiological functions seen in later evolved vertebrates was an ancestral character that first arose in a basal vertebrate.

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    • "The only specificity of interaction was found for lGnRH-I which was only capable of stimulating signal transduction via the lGnRH-R 1 (24, 35). The distribution of expression of lamprey GnRH receptors as reported from RT-PCR and in situ experiments (36, 37) includes the pituitary for all isoforms but also a much wider diversity of brain areas, including neuronal populations in the telencephalon, epithalamus, thalamus, preoptic area, and in the midbrain. Some sexual dimorphic features of this distribution was also noted (37). "
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    ABSTRACT: The gonadotropin-releasing hormone (GnRH) system is well known as the main regulator of reproductive physiology in vertebrates. It is also part of a network of brain structures and pathways that integrate information from the internal and external milieu and coordinate the adaptive behavioral and physiological responses to social and reproductive survival needs. In this paper we review the state of knowledge of the GnRH system in relation to the behavior, external, and internal factors that control reproduction in one of the oldest lineage of vertebrates, the lampreys.
    Frontiers in Endocrinology 10/2013; 4:151. DOI:10.3389/fendo.2013.00151
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    • "Multiple forms of GnRH and GnRH-Rs have been identified and recently reviewed (Millar, 2005; Tsai and Zhang, 2008). Three distinct paralogous forms of GnRH (Morgan and Millar, 2004) have been identified in the vertebrate lineage, with most vertebrates encoding two GnRH forms and the presence of all three forms is found only in a few species of teleost fish (Joseph et al., 2012; Kah et al., 2007). Three GnRH-R subtypes exist in vertebrates (Millar et al., 2004) with most vertebrate species possessing two GnRH-R subtypes. "
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    ABSTRACT: Diverse external and internal environmental factors are integrated in the hypothalamus to regulate the reproductive system. This is mediated through the pulsatile secretion of GnRH into the portal system to stimulate pituitary gonadotrophin secretion, which in turn regulates gonadal function. A single subpopulation of neurons termed 'KNDy neurones' located in the hypothalamic arcuate nucleus co-localise kisspeptin (Kiss), neurokinin B (NKB) and dynorphin (Dyn) and are responsive to negative feedback effects of sex steroids. The co-ordinated secretion from KNDy neurones appears to modulate the pulsatile release of GnRH, acting as a proximate pacemaker. This review briefly describes the neuropeptidergic control of reproduction in the avian class, highlighting the status of reproductive neuropeptide signalling systems homologous to those found in mammalian genomes. Genes encoding the GnRH system are complete in the chicken with similar roles to the mammalian counterparts, whereas genes encoding Kiss signalling components appear missing in the avian lineage, indicating a differing set of hypothalamic signals controlling avian reproduction. Gene sequences encoding both NKB and Dyn signalling components are present in the chicken genome, but expression analysis and functional studies remain to be completed. The focus of this article is to describe the avian complement of neuropeptidergic reproductive hormones and provide insights into the putative mechanisms that regulate reproduction in birds. These postulations highlight differences in reproductive strategies of birds in terms of gonadal steroid feedback systems, integration of metabolic signals and seasonality. Also included are propositions of KNDy neuropeptide gene silencing and plasticity in utilisation of these neuropeptides during avian evolution.
    General and Comparative Endocrinology 06/2013; 190. DOI:10.1016/j.ygcen.2013.05.018 · 2.47 Impact Factor
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    • "First strand cDNA was synthesized in a 10 μl final reaction volume containing 1 μg of the total RNA, 100 U of Superscript III reverse transcriptase (Invitrogen, Carlsbad, California) and 200 ng of NotI-(dT)18 primer according to Daukss et al. (2012). These cDNA syntheses were tested by PCR amplification of Elongation Factor 1α (Joseph et al., 2012). "
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    ABSTRACT: Two recently cloned gonadotropin-releasing hormone (GnRH) receptors (lamprey GnRH-R-2 and lamprey GnRH-R-3) along with lamprey (l) GnRH-R-1 were shown to share similar structural features and amino acid motifs common to other vertebrate receptors. Here we report on our findings of RNA expression of these three GnRH receptors in the three major life stages (larval, parasitic, and adult phases) of the sea lamprey, Petromyzon marinus, a basal vertebrate. For each stage, we examined the expression of messenger RNA encoding the receptors in the brain, pituitary, gonad, heart, muscle, liver, eye, intestine, kidney, skin, thyroid, gill, and endostyle by RT-PCR. In adult lampreys, the spatial expression of the three receptors in the brain and pituitary was investigated by in situ hybridization. In general, the receptors were more widely expressed in adult tissues as compared to parasitic-phase tissues and least widely expressed in the larval tissues. There were noted differences in male and female lampreys in the adult and parasitic phases for all three receptors. The data showed the presence of all three receptor transcripts in brain tissues for adult and parasitic phases and all three receptor transcripts were expressed in the adult pituitaries, but not in the parasitic pituitaries. However, in the larval phase, only lGnRH-R-1 was expressed in the larval brain and pituitary. In situ hybridization revealed that lGnRH-R-2 and -3 were expressed in the pineal tissue of adult female lampreys while lGnRH-R-1 was expressed in the pineal in adult male lampreys, all restricted to the pineal pellucida. In summary, these data provide an initial comparative analysis of expression of three lamprey GnRH receptors suggesting differential regulation within males and females at three different life/reproductive stages.
    Frontiers in Neuroscience 05/2013; 7(7):88. DOI:10.3389/fnins.2013.00088 · 3.66 Impact Factor
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