Identification and three-dimensional reconstruction of oxytocin receptor expressing astrocytes in the rat and mouse brain

Right here, we current a step-by-step protocol for three-dimensional reconstruction of astrocyte morphology, utilized to the central amygdala oxytocin receptor-expressing astrocytes. This contains RNAse-free perfusion, mixture of RNAscope and immunohistochemistry, and confocal imaging. This protocol supplies detailed details about tissue dealing with and a complete description of the RNAScope approach to label rat and mouse oxytocin receptor mRNA. We additionally describe three-dimensional reconstruction that enables the evaluation of greater than 70 totally different mobile parameters, highly effective for finding out astrocyte morphology and astrocyte-astrocyte interactions. For full particulars on the use and execution of this protocol, please seek advice from Wahis et al. (2021) and Althammer et al. (2020).

Oxytocin as a possible pharmacological device to fight weight problems

The neuropeptide oxytocin (OT) has emerged as an necessary anorexigen within the regulation of meals consumption and power stability. It has been proven that the discharge of OT and activation of hypothalamic OT neurons coincide with meals ingestion. Its results on feeding have largely been attributed to limiting meal dimension via interactions in key regulatory mind areas governing the homeostatic management of meals consumption such because the hypothalamus and hindbrain along with key feeding reward areas such because the nucleus accumbens and ventral tegmental space.
  • Moreover, the magnitude of an anorexigenic response to OT and feeding-related activation of the mind OT circuit are modified by the composition and taste of a food plan, in addition to by a social context by which a meal is consumed.
  • OT is especially efficient in lowering consumption of carbohydrates and candy tastants. Pharmacologic, genetic, and pair-feeding research point out that OT-elicited weight reduction can’t be totally defined by reductions of meals consumption and that the general influence of OT on power stability is additionally partly a results of OT-elicited modifications in lipolysis, power expenditure, and glucose regulation.
  • Peripheral administration of OT mimics lots of its results when it’s given into the central nervous system, elevating the questions of whether or not and to what extent circulating OT acts via peripheral OT receptors to control power stability.
  • Though OT has been discovered to elicit weight reduction in feminine mice, current research have indicated that intercourse and estrous cycle could influence oxytocinergic modulation of meals consumption.
  • Regardless of the general promising primary analysis knowledge, makes an attempt to make use of OT within the medical setting to fight weight problems and overeating have generated considerably combined outcomes.
  • The main focus of this mini-review is to briefly summarize the position of OT in feeding and metabolism, tackle gaps and inconsistencies in our data, and talk about among the limitations to the potential use of power OT that ought to assist information future analysis on OT as a tailored anti-obesity therapeutic.

Do generally used herbicides (atrazine and glyphosate) have the potential to impair the contractions, prostaglandin releasing and conducting of oxytocin sign on the bovine cervix in vitro?

Glyphosate (Gly) and atrazine (Atr) are among the many mostly used herbicides in international agriculture. It was beforehand proven that each Atr and Gly impair the ovarian and uterine secretion of regulators of myometrial motility (oxytocin (OT) or prostaglandins (PGs)) in cows, and Atr may also lower the power of contractions in strips from the uterine horn. Therefore, the purpose of this examine was to match the consequences of Atr and Gly on the motor and secretory perform of the bovine cervix in vitro in addition to receptivity and sign transduction in cervical cell cultures. Cervical strips or cells obtained from cows earlier than ovulation have been handled with environmental doses of Atr or Gly (0.1-10 ng/ml) since these herbicides exerted no cytotoxic impact at a dose of 100 ng/ml.
Solely Atr elevated the power of cervical contractions, whereas each Atr and Gly decreased the secretion of prostaglandins (PGs) with out disturbing their synthesis. Furthermore, Atr decreased the mRNA expression and protein stage of oxytocin receptor (OTR), whereas Gly elevated OTR protein ranges.
Each Atr and Gly decreased the contents of hole junction proteins (GAPs), Atr decreased the contents of second messengers (diacylglycerol – DAG, inositol-tris-phosphate – IP3), and Gly decreased the extent of myosin gentle chain kinase (MLCK) however elevated DAG ranges. Atr straight enhanced the cervical strips contractions. Each herbicides disturbed mobile signalling and inhibited PGs secretion. It counsel that Atr and Gly have the potential to impair the exercise of cervical cells in vitro, which may be adopted by failure of upkeep with gestation.

Modified Sawhorse Waveform for the Voltammetric Detection of Oxytocin

Carbon fiber microelectrodes (CFMEs) have been used to detect neurotransmitters and different biomolecules utilizing fast-scan cyclic voltammetry (FSCV) for the previous few many years. This system measures neurotransmitters equivalent to dopamine and, extra lately, physiologically related neuropeptides. Oxytocin, a pleiotropic peptide hormone, is physiologically necessary for adaptation, growth, replica, and social habits. This neuropeptide capabilities as a stress-coping molecule, an anti-inflammatory agent, and serves as an antioxidant with protecting results particularly throughout adversity or trauma.
Right here, we measure tyrosine utilizing the Modified Sawhorse Waveform (MSW), enabling enhanced electrode sensitivity for the amino acid and oxytocin peptide. Making use of the MSW, decreased floor fouling and enabled codetection with different monoamines. As oxytocin comprises tyrosine, the MSW was additionally used to detect oxytocin. The sensitivity of oxytocin detection was discovered to be 3.99 ± 0.49 nA/μM, (n=5). Moreover, we display that making use of the MSW on CFMEs permits for actual time measurements of exogenously utilized oxytocin on rat mind slices. These research could function novel assays for oxytocin detection in a quick, sub-second timescale with attainable implications for in vivo measurements and additional understanding of the physiological position of oxytocin.

The Evolution of Oxytocin and Vasotocin Receptor Genes in Jawed Vertebrates: A Clear Case for Gene Duplications Via Ancestral Complete-Genome Duplications

The neuronal and neuroendocrine peptides oxytocin (OT) and vasotocin (VT), together with vasopressins, have six cognate receptors encoded by six receptor subtype genes in jawed vertebrates. The peptides elicit a broad vary of responses which might be particularly mediated by the receptor subtypes together with neuronal capabilities regulating habits and hormonal actions on replica and water/electrolyte stability.
Beforehand, we’ve got demonstrated that these six receptor subtype genes, which we designated VTR1AVTR1BOTRVTR2AVTR2B and VTR2C, arose from a syntenic ancestral gene pair, one VTR1/OTR ancestor and one VTR2 ancestor, via the early vertebrate whole-genome duplications (WGD) referred to as 1R and 2R. This was supported by each phylogenetic and chromosomal conserved synteny knowledge. Extra lately, different research have targeted on confounding components, such because the OTR/VTR orthologs in cyclostomes, to query this situation for the origin of the OTR/VTR gene household; proposing as a substitute much less parsimonious interpretations involving just one WGD adopted by advanced sequence of chromosomal or segmental duplications.

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Right here, we’ve got up to date the phylogeny of the OTR/VTR gene household, together with a bigger variety of vertebrate species, and revisited seven consultant neighboring gene households from our earlier conserved synteny analyses, including chromosomal info from newer high-coverage genome assemblies from species that occupy key phylogenetic positions: the polypteriform fish reedfish (Erpetoichthys calabaricus), the cartilaginous fish thorny skate (Amblyraja radiata) and a more moderen high-quality meeting of the Western clawed frog (Xenopus tropicalis) genome.
Our analyses as soon as once more add sturdy help for four-fold symmetry, i.e., chromosome quadruplication in the identical time window because the WGD occasions early in vertebrate evolution, previous to the jawed vertebrate radiation. Thus, the evolution of the OTR/VTR gene household might be most parsimoniously defined by two WGD occasions giving rise to the six ancestral genes, adopted by differential gene losses of VTR2 genes in several lineages. We additionally argue for extra coherence and readability within the nomenclature of OT/VT receptors, primarily based on probably the most parsimonious situation.