Publications by authors named "Francesca Tatini"

17 Publications

  • Page 1 of 1

Photobiomodulation of Human Fibroblasts and Keratinocytes with Blue Light: Implications in Wound Healing.

Biomedicines 2021 Jan 5;9(1). Epub 2021 Jan 5.

Istituto di Fisica Applicata "Nello Carrara", Consiglio Nazionale delle Ricerche (CNR-IFAC), 50019 Florence, Italy.

In recent years, photobiomodulation (PBM) has been recognized as a physical therapy in wound management. Despite several published research papers, the mechanism underlying photobiomodulation is still not completely understood. The investigation about application of blue light to improve wound healing is a relatively new research area. Tests in selected patients evidenced a stimulation of the healing process in superficial and chronic wounds treated with a blue LED light emitting at 420 nm; a study in animal model pointed out a faster healing process in superficial wound, with an important role of fibroblasts and myofibroblasts. Here, we present a study aiming at evidencing the effects of blue light on the proliferation and metabolism in fibroblasts from healthy skin and keratinocytes. Different light doses (3.43, 6.87, 13.7, 20.6, 30.9 and 41.2 J/cm2) were used to treat the cells, evidencing inhibitory and stimulatory effects following a biphasic dose behavior. Electrophysiology was used to investigate the effects on membrane currents: healthy fibroblasts and keratinocytes showed no significant differences between treated and not treated cells. Raman spectroscopy revealed the mitochondrial Cytochrome C (Cyt C) oxidase dependence on blue light irradiation: a significant decrease in peak intensity of healthy fibroblast was evidenced, while it is less pronounced in keratinocytes. In conclusion, we observed that the blue LED light can be used to modulate metabolism and proliferation of human fibroblasts, and the effects in wound healing are particularly evident when studying the fibroblasts and keratinocytes co-cultures.
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http://dx.doi.org/10.3390/biomedicines9010041DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7824830PMC
January 2021

Experimental Study on Blue Light Interaction with Human Keloid-Derived Fibroblasts.

Biomedicines 2020 Dec 6;8(12). Epub 2020 Dec 6.

Istituto di Fisica Applicata "Nello Carrara", Consiglio Nazionale delle Ricerche (CNR-IFAC), 50019 Florence, Italy.

Keloids are an exuberant response to wound healing, characterized by an exaggerated synthesis of collagen, probably due to the increase of fibroblasts activity and to the reduction of their apoptosis rate: currently no standard treatments or pharmacological therapies are able to prevent keloid recurrence. To reach this goal, in recent years some physical treatments have been proposed, and among them the PhotoBioModulation therapy (PBM). This work analyses the effects of a blue LED light irradiation (410-430 nm, 0.69 W/cm power density) on human fibroblasts, isolated from both keloids and perilesional tissues. Different light doses (3.43-6.87-13.7-20.6-30.9 and 41.2 J/cm) were tested. Biochemical assays and specific staining were used to assess cell metabolism, proliferation and viability. Micro-Raman spectroscopy was used to explore direct effects of the blue LED light on the Cytochrome C (Cyt C) oxidase. We also investigated the effects of the irradiation on ionic membrane currents by patch-clamp recordings. Our results showed that the blue LED light can modulate cell metabolism and proliferation, with a dose-dependent behavior and that these effects persist at least till 48 h after treatment. Furthermore, we demonstrated that the highest fluence value can reduce cell viability 24 h after irradiation in keloid-derived fibroblasts, while the same effect is observed 48 h after treatment in perilesional fibroblasts. Electrophysiological recordings showed that the medium dose (20.6 J/cm) of blue LED light induces an enhancement of voltage-dependent outward currents elicited by a depolarizing ramp protocol. Overall, these data demonstrate the potentials that PBM shows as an innovative and minimally-invasive approach in the management of hypertrophic scars and keloids, in association with current treatments.
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http://dx.doi.org/10.3390/biomedicines8120573DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7762279PMC
December 2020

Blue LED light modulates inflammatory infiltrate and improves the healing of superficial wounds.

Photodermatol Photoimmunol Photomed 2020 03 10;36(2):166-168. Epub 2019 Dec 10.

Institute of Applied Physics, National Research Council (IFAC-CNR), Sesto Fiorentino, Italy.

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http://dx.doi.org/10.1111/phpp.12527DOI Listing
March 2020

Stable, Monodisperse, and Highly Cell-Permeating Nanocochleates from Natural Soy Lecithin Liposomes.

Pharmaceutics 2019 Jan 16;11(1). Epub 2019 Jan 16.

Department of Chemistry, University of Florence, Via U. Schiff 6, 50019 Sesto Fiorentino, Florence, Italy.

(1) Background: Andrographolide (AN), the main diterpenoid constituent of , has a wide spectrum of biological activities. The aim of this study was the development of nanocochleates (NCs) loaded with AN and based on phosphatidylserine (PS) or phosphatidylcholine (PC), cholesterol and calcium ions in order to overcome AN low water solubility, its instability under alkaline conditions and its rapid metabolism in the intestine. (2) Methods: The AN-loaded NCs (AN⁻NCs) were physically and chemically characterised. The in vitro gastrointestinal stability and biocompatibility of AN⁻NCs in J77A.1 macrophage and 3T3 fibroblasts cell lines were also investigated. Finally, the uptake of nanocarriers in macrophage cells was studied. (3) Results: AN⁻NCs obtained from PC nanoliposomes were suitable nanocarriers in terms of size and homogeneity. They had an extraordinary stability after lyophilisation without the use of lyoprotectants and after storage at room temperature. The encapsulation efficiency was 71%, while approximately 95% of AN was released in PBS after 24 h, with kinetics according to the Hixson⁻Crowell model. The in vitro gastrointestinal stability and safety of NCs, both in macrophages and 3T3 fibroblasts, were also assessed. Additionally, NCs had extraordinary uptake properties in macrophages. (4) Conclusions: NCs developed in this study could be suitable for both AN oral and parental administration, amplifying its therapeutic value.
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http://dx.doi.org/10.3390/pharmaceutics11010034DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6359122PMC
January 2019

Three-dimensional mapping of the orientation of collagen corneal lamellae in healthy and keratoconic human corneas using SHG microscopy.

J Biophotonics 2017 01 29;10(1):75-83. Epub 2016 Jul 29.

National Institute of Optics, National Research Council (INO-CNR), Via Nello Carrara 1, 50019, Sesto Fiorentino, Italy.

Keratoconus is an eye disorder that causes the cornea to take an abnormal conical shape, thus impairing its refractive functions and causing blindness. The late diagnosis of keratoconus is among the principal reasons for corneal surgical transplantation. This pathology is characterized by a reduced corneal stiffness in the region immediately below Bowman's membrane, probably due to a different lamellar organization, as suggested by previous studies. Here, the lamellar organization in this corneal region is characterized in three dimensions by means of second-harmonic generation (SHG) microscopy. In particular, a method based on a three-dimensional correlation analysis allows to probe the orientation of sutural lamellae close to the Bowman's membrane, finding statistical differences between healthy and keratoconic samples. This method is demonstrated also in combination with an epi-detection scheme, paving the way for a potential clinical ophthalmic application of SHG microscopy for the early diagnosis of keratoconus. SHG image acquired with sagittal optical sectioning (A) of a healthy cornea and (B) of a keratoconic cornea. Scale bars: 30 μm.
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http://dx.doi.org/10.1002/jbio.201600122DOI Listing
January 2017

Preparation and Photoacoustic Analysis of Cellular Vehicles Containing Gold Nanorods.

J Vis Exp 2016 05 2(111). Epub 2016 May 2.

Institute of Applied Physics, Italian National Research Council.

Gold nanorods are attractive for a range of biomedical applications, such as the photothermal ablation and the photoacoustic imaging of cancer, thanks to their intense optical absorbance in the near-infrared window, low cytotoxicity and potential to home into tumors. However, their delivery to tumors still remains an issue. An innovative approach consists of the exploitation of the tropism of tumor-associated macrophages that may be loaded with gold nanorods in vitro. Here, we describe the preparation and the photoacoustic inspection of cellular vehicles containing gold nanorods. PEGylated gold nanorods are modified with quaternary ammonium compounds, in order to achieve a cationic profile. On contact with murine macrophages in ordinary Petri dishes, these particles are found to undergo massive uptake into endocytic vesicles. Then these cells are embedded in biopolymeric hydrogels, which are used to verify that the stability of photoacoustic conversion of the particles is retained in their inclusion into cellular vehicles. We are confident that these results may provide new inspiration for the development of novel strategies to deliver plasmonic particles to tumors.
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http://dx.doi.org/10.3791/53328DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4942024PMC
May 2016

Observation of an improved healing process in superficial skin wounds after irradiation with a blue-LED haemostatic device.

J Biophotonics 2016 06 12;9(6):645-55. Epub 2016 Jan 12.

National Institute of Optics, National Research Council (INO-CNR), Florence, 50125, Italy.

The healing process of superficial skin wounds treated with a blue-LED haemostatic device is studied. Four mechanical abrasions are produced on the back of 10 Sprague Dawley rats: two are treated with the blue-LED device, while the other two are left to naturally recover. Visual observations, non-linear microscopic imaging, as well as histology and immunofluorescence analyses are performed 8 days after the treatment, demonstrating no adverse reactions neither thermal damages in both abraded areas and surrounding tissue. A faster healing process and a better-recovered skin morphology are observed: the treated wounds show a reduced inflammatory response and a higher collagen content. Blue LED induced photothermal effect on superficial abrasions.
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http://dx.doi.org/10.1002/jbio.201500191DOI Listing
June 2016

Mutations of Profilin-1 Associated with Amyotrophic Lateral Sclerosis Promote Aggregation Due to Structural Changes of Its Native State.

ACS Chem Biol 2015 Nov 27;10(11):2553-63. Epub 2015 Aug 27.

Department of Biomedical Experimental and Clinical Sciences, Section of Biochemistry, University of Florence , Viale Morgagni 50, I-50134, Florence, Italy.

The PFN1 gene, coding for profilin-1, has recently been associated with familial amyotrophic lateral sclerosis (fALS), as three mutations, namely C71G, M114T, and G118V, have been found in patients with familial forms of the disease and another, E117G, has been proposed to be a moderate risk factor for disease onset. In this work, we have purified the four profilin-1 variants along with the wild-type protein. The resulting aggregates appear to be fibrillar, to have a weak binding to ThT, and to possess a significant amount of intermolecular β-sheet structure. Using ThT fluorescence assays, far-UV circular dichroism, and dynamic light scattering, we found that all four variants have an aggregation propensity higher than that of the wild-type counterpart. In particular, the C71G mutation was found to induce the most dramatic change in aggregation, followed by the G118V and M114T substitutions and then the E117G mutation. Such a propensity was found not to strictly correlate with the conformational stability in this group of profilin-1 variants, determined using both urea-induced denaturation at equilibrium and folding/unfolding kinetics. However, it correlated with structural changes of the folded states, as monitored with far-UV circular dichroism, intrinsic fluorescence spectroscopy, ANS binding, acrylamide quenching, and dynamic light scattering. Overall, the results suggest that all four mutations increase the tendency of profilin-1 to aggregate and that such aggregation behavior is largely determined by the mutation-induced structural changes occurring in the folded state of the protein.
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http://dx.doi.org/10.1021/acschembio.5b00598DOI Listing
November 2015

Nonexocytotic serotonin release tonically suppresses serotonergic neuron activity.

J Gen Physiol 2015 Mar;145(3):225-51

Department of Neuroscience, Psychology, Drug Research and Child Health, University of Florence, 50121 Florence, Italy.

The firing activity of serotonergic neurons in raphe nuclei is regulated by negative feedback exerted by extracellular serotonin (5-HT)o acting through somatodendritic 5-HT1A autoreceptors. The steady-state [5-HT]o, sensed by 5-HT1A autoreceptors, is determined by the balance between the rates of 5-HT release and reuptake. Although it is well established that reuptake of 5-HTo is mediated by 5-HT transporters (SERT), the release mechanism has remained unclear. It is also unclear how selective 5-HT reuptake inhibitor (SSRI) antidepressants increase the [5-HT]o in raphe nuclei and suppress serotonergic neuron activity, thereby potentially diminishing their own therapeutic effect. Using an electrophysiological approach in a slice preparation, we show that, in the dorsal raphe nucleus (DRN), continuous nonexocytotic 5-HT release is responsible for suppression of phenylephrine-facilitated serotonergic neuron firing under basal conditions as well as for autoinhibition induced by SSRI application. By using 5-HT1A autoreceptor-activated G protein-gated inwardly rectifying potassium channels of patched serotonergic neurons as 5-HTo sensors, we show substantial nonexocytotic 5-HT release under conditions of abolished firing activity, Ca(2+) influx, vesicular monoamine transporter 2-mediated vesicular accumulation of 5-HT, and SERT-mediated 5-HT transport. Our results reveal a cytosolic origin of 5-HTo in the DRN and suggest that 5-HTo may be supplied by simple diffusion across the plasma membrane, primarily from the dense network of neurites of serotonergic neurons surrounding the cell bodies. These findings indicate that the serotonergic system does not function as a sum of independently acting neurons but as a highly interdependent neuronal network, characterized by a shared neurotransmitter pool and the regulation of firing activity by an interneuronal, yet activity-independent, nonexocytotic mechanism.
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http://dx.doi.org/10.1085/jgp.201411330DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4338157PMC
March 2015

In vitro assessment of antibody-conjugated gold nanorods for systemic injections.

J Nanobiotechnology 2014 Dec 5;12:55. Epub 2014 Dec 5.

Istituto di Fisica Applicata 'Nello Carrara', Consiglio Nazionale delle Ricerche, Via Madonna del Piano 10, 50019, Sesto Fiorentino, Italy.

Background: The interest for gold nanorods in biomedical optics is driven by their intense absorbance of near infrared light, their biocompatibility and their potential to reach tumors after systemic administration. Examples of applications include the photoacoustic imaging and the photothermal ablation of cancer. In spite of great current efforts, the selective delivery of gold nanorods to tumors through the bloodstream remains a formidable challenge. Their bio-conjugation with targeting units, and in particular with antibodies, is perceived as a hopeful solution, but the complexity of living organisms complicates the identification of possible obstacles along the way to tumors.

Results: Here, we present a new model of gold nanorods conjugated with anti-cancer antigen 125 (CA125) antibodies, which exhibit high specificity for ovarian cancer cells. We implement a battery of tests in vitro, in order to simulate major nuisances and predict the feasibility of these particles for intravenous injections. We show that parameters like the competition of free CA125 in the bloodstream, which could saturate the probe before arriving at the tumors, the matrix effect and the interference with erythrocytes and phagocytes are uncritical.

Conclusions: Although some deterioration is detectable, anti-CA125-conjugated gold nanorods retain their functional features after interaction with blood tissue and so represent a powerful candidate to hit ovarian cancer cells.
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http://dx.doi.org/10.1186/s12951-014-0055-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4266900PMC
December 2014

Size dependent biological profiles of PEGylated gold nanorods.

J Mater Chem B 2014 Sep 7;2(36):6072-6080. Epub 2014 Aug 7.

Institute of Applied Physics "Nello Carrara", National Research Council, Via Madonna del Piano 10, Sesto Fiorentino, 50019 Italy.

The perspective of introducing plasmonic particles for applications in biomedical optics is receiving much interest. However, their translation into clinical practices is delayed by various factors, which include a poor definition of their biological interactions. Here, we describe the preparation and the biological profiles of gold nanorods belonging to five different size classes with average effective radii between ∼5 and 20 nm and coated with polyethylene glycol (PEG). All these particles exhibit decent stability in the presence of representative proteins, low cytotoxicity and satisfactory compatibility with intravenous administration, in terms of their interference with blood tissue. However, the suspension begins to become unstable after a few days of exposure to blood proteins. Moreover, the cytotoxicity is a little worse for smaller particles, probably because their purification is more critical, while undesirable interactions with the mononuclear phagocyte system are minimal in the intermediate size range. Overall, these findings hold implications of practical relevance and suggest that PEGylated gold nanorods may be a versatile platform for a variety of biomedical applications.
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http://dx.doi.org/10.1039/c4tb00991fDOI Listing
September 2014

Graphene as a photothermal switch for controlled drug release.

Nanoscale 2014 Jul;6(14):7947-53

Institute of Applied Physics "Nello Carrara", National Research Council, via Madonna del Piano 10, I-50019 Sesto Fiorentino, Italy.

Graphene has recently emerged as a novel material in the biomedical field owing to its optical properties, biocompatibility, large specific surface area and low cost. In this paper, we provide the first demonstration of the possibility of using light to remotely trigger the release of drugs from graphene in a highly controlled manner. Different drugs including chemotherapeutics and proteins are firmly adsorbed onto reduced graphene oxide (rGO) nanosheets dispersed in a biopolymer film and then released by individual millisecond-long light pulses generated by a near infrared (NIR) laser. Here graphene plays the dual role of a versatile substrate for temporary storage of drugs and an effective transducer of NIR-light into heat. Drug release appears to be narrowly confined within the size of the laser spot under noninvasive conditions and can be precisely dosed depending on the number of pulses. The approach proposed paves the way for tailor-made pharmacological treatments of chronic diseases, including cancer, anaemia and diabetes.
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http://dx.doi.org/10.1039/c4nr01622jDOI Listing
July 2014

Photothermally activated hybrid films for quantitative confined release of chemical species.

Angew Chem Int Ed Engl 2013 Jun 9;52(23):5956-60. Epub 2013 May 9.

Institute of Applied Physics Nello Carrara, National Research Council via Madonna del Piano 10, 50019, Italy.

Illuminating films of a porous chitosan matrix containing gold nanorods and thermosensitive micelles loaded with a chemical stimulates local photothermal conversion of the gold nanorods. The heat produced activates the ejection of the chemical from the micelles (see scheme), and causes the transient permeabilization of adjacent cell membranes, resulting in a selective cellular uptake of the released chemical with control over spatiotemporal parameters and dosage.
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http://dx.doi.org/10.1002/anie.201207986DOI Listing
June 2013

Extracellular chaperones prevent Aβ42-induced toxicity in rat brains.

Biochim Biophys Acta 2013 Aug 16;1832(8):1217-26. Epub 2013 Apr 16.

Department of Biomedical Experimental and Clinical Sciences, University of Florence, V.le GB Morgagni 50, 50134, Italy.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterised by cognitive decline, formation of the extracellular amyloid β (Aβ42) plaques, neuronal and synapse loss, and activated microglia and astrocytes. Extracellular chaperones, which are known to inhibit amyloid fibril formation and promote clearance of misfolded aggregates, have recently been shown to reduce efficiently the toxicity of HypF-N misfolded oligomers to immortalised cell lines, by binding and clustering them into large species. However, the role of extracellular chaperones on Aβ oligomer toxicity remains unclear, with reports often appearing contradictory. In this study we microinjected into the hippocampus of rat brains Aβ42 oligomers pre-incubated for 1h with two extracellular chaperones, namely clusterin and α2-macroglobulin. The chaperones were found to prevent Aβ42-induced learning and memory impairments, as assessed by the Morris Water Maze test, and reduce Aβ42-induced glia inflammation and neuronal degeneration in rat brains, as probed by fluorescent immunohistochemical analyses. Moreover, the chaperones were able to prevent Aβ42 colocalisation with PSD-95 at post-synaptic terminals of rat primary neurons, suppressing oligomer cytotoxicity. All such effects were not effective by adding pre-formed oligomers and chaperones without preincubation. Molecular chaperones have therefore the potential to prevent the early symptoms of AD, not just by inhibiting Aβ42 aggregation, as previously demonstrated, but also by suppressing the toxicity of Aβ42 oligomers after they are formed. These findings elect them as novel neuroprotectors against amyloid-induced injury and excellent candidates for the design of therapeutic strategies against AD.
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http://dx.doi.org/10.1016/j.bbadis.2013.04.012DOI Listing
August 2013

Amyloid-β oligomer synaptotoxicity is mimicked by oligomers of the model protein HypF-N.

Neurobiol Aging 2013 Sep 18;34(9):2100-9. Epub 2013 Apr 18.

Department of Biomedical, Experimental and Clinical Sciences, University of Florence, Florence, Italy.

Protein misfolded oligomers are thought to be the primary pathogenic species in many protein deposition diseases. Oligomers by the amyloid-β peptide play a central role in Alzheimer's disease pathogenesis, being implicated in synaptic dysfunction. Here we show that the oligomers formed by a protein that has no link with human disease, namely the N-terminal domain of HypF from Escherichia coli (HypF-N), are also synaptotoxic. HypF-N oligomers were found to (i) colocalize with post-synaptic densities in primary rat hippocampal neurons; (ii) induce impairment of long-term potentiation in rat hippocampal slices; and (iii) impair spatial learning of rats in the Morris Water Maze test. By contrast, the native protein and control nontoxic oligomers had none of such effects. These results raise the importance of using HypF-N oligomers as a valid tool to investigate the pathogenesis of Alzheimer's disease, with advantages over other systems for their stability, reproducibility, and costs. The results also suggest that, in the context of a compromised protein homeostasis resulting from aggregation of the amyloid β peptide, a number of oligomeric species sharing common synaptotoxic activity can arise and cooperate in the pathogenesis of the disease.
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http://dx.doi.org/10.1016/j.neurobiolaging.2013.03.020DOI Listing
September 2013

Light-responsive nanocomposite sponges for on demand chemical release with high spatial and dosage control.

J Mater Chem B 2013 Feb 7;1(8):1096-1100. Epub 2013 Jan 7.

Institute of Applied Physics "Nello Carrara", National Research Council, via Madonna del Piano 10, 50019, Sesto Fiorentino, Italy.

We present a biocompatible device for on demand chemical release in the form of a light-activated sponge-like nanocomposite scaffold, which ensures excellent control over the principal parameters of chemical release and dosage in order to sustain effective therapeutic action. The sponge consists of a porous biopolymer scaffold containing a dispersion of gold nanorods, which acts as an absorber of the incoming laser light, and of thermosensitive micelles, which serve as a reservoir for the drug molecules to be released. The photothermal response of the nanoparticles contained inside the sponge triggers a contraction in proximal micelles, thus promoting the expulsion of the drug that in turn is released from the sponge to the external environment. The peculiar physiochemical and structural properties of the nanocomposite sponges impart a number of interesting features to the proposed drug release system, including the possibility of spatially confining the therapeutic treatment as well as precise control of the amount of released drug as a function of duration and power of the excitation light.
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http://dx.doi.org/10.1039/c2tb00310dDOI Listing
February 2013

Differential autoinhibition of 5-hydroxytryptamine neurons by 5-hydroxytryptamine in the dorsal raphe nucleus.

Neuroreport 2005 Aug;16(12):1351-5

Department of Preclinical and Clinical Pharmacology Mario Aiazzi-Mancini, University of Florence, Viale G. Pieraccini 6, 50139 Florence, Italy.

5-Hydroxytryptamine neurons in the dorsal raphe nucleus are under autoinhibitory control by endogenous 5-hydroxytryptamine. Tonic activation of 5-hydroxytryptamine 1A autoreceptors was demonstrated in awake animals, but was inconsistently observed in anaesthetized animals and slice preparations, leading to questioning of its physiological significance. We re-evaluated autoinhibition in single-unit recordings from deeply seated 5-hydroxytryptamine neurons in slices in which endogenous 5-hydroxytryptamine bioavailability was restored by supplementing its precursor L-tryptophan. In these conditions, the application of the neutral 5-hydroxytryptamine 1A receptor antagonist WAY-100635 markedly increased 5-hydroxytryptamine neuron firing. Responses to WAY-100635 in single experiments ranged from a lack of effect to a several-fold increase in firing rate, suggesting that 5-hydroxytryptamine neurons in the dorsal raphe nucleus represent a heterogeneous population regarding their susceptibility to autoinhibition by endogenous 5-hydroxytryptamine.
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http://dx.doi.org/10.1097/01.wnr.0000175249.25535.bfDOI Listing
August 2005