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1.
Micromachines (Basel) ; 14(11)2023 Oct 27.
Article in English | MEDLINE | ID: mdl-38004856

ABSTRACT

The electric stimulation (ES) of the cornea is a novel therapeutic approach to the treatment of degenerative visual diseases. Currently, ES is delivered by placing a mono-element electrode on the surface of the cornea that uniformly stimulates the eye along the electrode site. It has been reported that a certain degree of correlation exists between the location of the stimulated retinal area and the position of the electrode. Therefore, in this study, we present the development of a sectioned surface electrode for selective electric stimulation of the human cornea. The proposed device consists of 16 independent microelectrodes, a reference electrode, and 18 contact pads. The microelectrodes have a size of 200 µm × 200 µm, are arranged in a 4 × 4 matrix, and cover a total stimulation area of 16 mm2. The proposed fabrication process, based on surface micromachining technology and flexible electronics, uses only three materials: polyimide, aluminum, and titanium, which allow us to obtain a simplified, ergonomic, and reproducible fabrication process. The fabricated prototype was validated to laboratory level by electrical and electrochemical tests, showing a relatively high electrical conductivity and average impedance from 712 kΩ to 1.4 MΩ at the clinically relevant frequency range (from 11 Hz to 30 Hz). Additionally, the biocompatibility of the electrode prototype was demonstrated by performing in vivo tests and by analyzing the polyimide films using Fourier transform infrared spectroscopy (FTIR). The resulting electrode prototype is robust, mechanically flexible, and biocompatible, with a high potential to be used for selective ES of the cornea.

2.
Behav Brain Res ; 445: 114377, 2023 05 08.
Article in English | MEDLINE | ID: mdl-36868364

ABSTRACT

Human use of marijuana at an early age has been reported to lead to cognitive impairment. However, researchers have not yet clearly determined whether this impairment is due to marijuana-induced alterations in the developing nervous system and whether this deficit persists into adulthood after marijuana use has ceased. We administered anandamide to developing rats to assess the effect of cannabinoids on development. We subsequently evaluated learning and performance on a temporal bisection task in adulthood and assessed the expression of genes encoding principal subunits of NMDA receptors (Grin1, Grin2A, and Grin2B) in the hippocampus and prefrontal cortex. Rats in two age groups, namely, 21-day-old and 150-day-old rats, received intraperitoneal injections of anandamide or the vehicle for 14 days. Both groups performed a temporal bisection test, which included listening to tones of different durations and classifying them as short or long. The expression of the Grin1, Grin2A and Grin2B mRNAs was evaluated using quantitative PCR in both age groups after extracting mRNA from the hippocampus and prefrontal cortex. We observed a learning impairment in the temporal bisection task (p < 0.05) and changes in the response latency (p < 0.05) in rats that received anandamide. Furthermore, these rats exhibited decreased expression of Grin2b (p = 0.001) compared to those that received the vehicle. In human subjects, the use of cannabinoids during development induces a long-term deficit, but this deficit is not observed in subjects who use cannabinoids in adulthood. Rats treated with anandamide earlier in development took longer to learn the task, suggesting that anandamide exerts a harmful effect on cognition in developing rats. Administration of anandamide during early stages of development induced deficits in learning and other cognitive processes that depend on an adequate estimation of time. The cognitive demands of the environment must be considered when evaluating the cognitive effects of cannabinoids on developing or mature brains. High cognitive demands might induce differential expression of NMDA receptors that improves cognitive capacity, overcoming altered glutamatergic function.


Subject(s)
Cannabinoids , Hallucinogens , Time Perception , Humans , Rats , Animals , Infant, Newborn , Receptors, N-Methyl-D-Aspartate , Learning , Hippocampus/physiology
3.
BMC Womens Health ; 22(1): 356, 2022 08 26.
Article in English | MEDLINE | ID: mdl-36028805

ABSTRACT

BACKGROUND: Endometriosis is an estrogen-dependent and chronic inflammatory disease affecting up to 10% of women. It is the result of a combined interaction of genetic, epigenetic, environmental, lifestyle, reproductive and local inflammatory factors. In this study, we investigated whether single nucleotide polymorphisms (SNPs) mapping to TNF-alpha (TNF, rs1800629) and IL-1beta (IL1B, rs1143634) and variable number tandem repeat polymorphism mapping to IL1-Ra (IL1RN intron 2, rs2234663) genetic loci are associated with risk for endometriosis in a Mexican mestizo population. METHODS: This study included 183 women with confirmed endometriosis (ENDO) diagnosed after surgical laparoscopy and 186 women with satisfied parity and without endometriosis as controls (CTR). PCR/RFLP technique was used for genotyping SNPs (rs1800629 and rs1143634); PCR for genotyping rs2234663. RESULTS: We found no statistical differences in age between groups nor among stages of endometriosis and the CTR group. We observed no difference in genotype and allele frequencies, nor carriage rate between groups in none of the three studied polymorphisms. The prevalence of TNF*2-allele heterozygotes (p = 0.025; OR 3.8), TNF*2-allele (p = 0.029; OR 3.4), IL1B*2-allele heterozygotes (p = 0.044; OR 2.69) and its carriage rate (p = 0.041; OR 2.64) in endometriosis stage IV was higher than the CTR group. Surprisingly, the carriage rate of IL1RN*2-allele (ENDO: p = 0.0004; OR 0.4; stage I: p = 0.002, OR 0.38; stage II: p = 0.002, OR 0.35; stage III: p = 0.003, OR 0.33), as well as the IL1RN*2-allele frequencies (ENDO: p = 0.0008, OR 0.55; I: p = 0.037, OR 0.60; II: p = 0.002, OR 0.41; III: p = 0.003, OR 0.38) were lower than the CTR group. Women with endometriosis stage IV (severe) had frequencies more alike to the CTR group in the IL1RN*2 allele frequency (31.2% vs. 27.2%) and carriage rate (37.5% vs. 41.9%). CONCLUSION: Although these polymorphisms are not associated with the risk of endometriosis, Mexican mestizo women with severe stage of endometriosis have higher frequencies of TNF*2-, IL1B*2- and IL1RN*2-alleles, which may explain a possible correlation with disease severity rather than predisposition or risk.


Subject(s)
Endometriosis , Interleukin 1 Receptor Antagonist Protein , Interleukin-1beta , Tumor Necrosis Factor-alpha , Case-Control Studies , Endometriosis/genetics , Female , Gene Frequency , Genetic Predisposition to Disease , Genotype , Humans , Interleukin 1 Receptor Antagonist Protein/genetics , Interleukin-1beta/genetics , Mexico , Polymorphism, Single Nucleotide , Tumor Necrosis Factor-alpha/genetics
4.
Int J Mol Sci ; 22(21)2021 Oct 25.
Article in English | MEDLINE | ID: mdl-34768919

ABSTRACT

The generation of new neurons in the adult brain is a currently accepted phenomenon. Over the past few decades, the subventricular zone and the hippocampal dentate gyrus have been described as the two main neurogenic niches. Neurogenic niches generate new neurons through an asymmetric division process involving several developmental steps. This process occurs throughout life in several species, including humans. These new neurons possess unique properties that contribute to the local circuitry. Despite several efforts, no other neurogenic zones have been observed in many years; the lack of observation is probably due to technical issues. However, in recent years, more brain niches have been described, once again breaking the current paradigms. Currently, a debate in the scientific community about new neurogenic areas of the brain, namely, human adult neurogenesis, is ongoing. Thus, several open questions regarding new neurogenic niches, as well as this phenomenon in adult humans, their functional relevance, and their mechanisms, remain to be answered. In this review, we discuss the literature and provide a compressive overview of the known neurogenic zones, traditional zones, and newly described zones. Additionally, we will review the regulatory roles of some molecular mechanisms, such as miRNAs, neurotrophic factors, and neurotrophins. We also join the debate on human adult neurogenesis, and we will identify similarities and differences in the literature and summarize the knowledge regarding these interesting topics.


Subject(s)
Dentate Gyrus/cytology , Lateral Ventricles/cytology , Neurogenesis/physiology , Neurons/cytology , Ventral Striatum/cytology , Adult , Animals , Hippocampus/cytology , Humans , Mice , MicroRNAs/genetics , Neural Stem Cells/cytology , Neurogenesis/genetics , Rats
5.
Int J Mol Sci ; 22(14)2021 Jul 12.
Article in English | MEDLINE | ID: mdl-34299069

ABSTRACT

The endocannabinoid system (ECS) is a crucial modulatory system in which interest has been increasing, particularly regarding the regulation of behavior and neuroplasticity. The adolescent-young adulthood phase of development comprises a critical period in the maturation of the nervous system and the ECS. Neurogenesis occurs in discrete regions of the adult brain, and this process is linked to the modulation of some behaviors. Since marijuana (cannabis) is the most consumed illegal drug globally and the highest consumption rate is observed during adolescence, it is of particular importance to understand the effects of ECS modulation in these early stages of adulthood. Thus, in this article, we sought to summarize recent evidence demonstrating the role of the ECS and exogenous cannabinoid consumption in the adolescent-young adulthood period; elucidate the effects of exogenous cannabinoid consumption on adult neurogenesis; and describe some essential and adaptive behaviors, such as stress, anxiety, learning, and memory. The data summarized in this work highlight the relevance of maintaining balance in the endocannabinoid modulatory system in the early and adult stages of life. Any ECS disturbance may induce significant modifications in the genesis of new neurons and may consequently modify behavioral outcomes.


Subject(s)
Brain/drug effects , Cannabinoids/pharmacology , Neurodegenerative Diseases/physiopathology , Neurogenesis , Social Behavior , Stress, Psychological , Adolescent , Adult , Brain/metabolism , Humans , Neurodegenerative Diseases/chemically induced , Receptors, Cannabinoid/metabolism , Young Adult
6.
eNeuro ; 8(3)2021.
Article in English | MEDLINE | ID: mdl-33962969

ABSTRACT

The lateral habenula (LHb) is a phylogenetically primitive brain structure that plays a key role in learning to inhibit distinct responses to specific stimuli. This structure is activated by primary aversive stimuli, cues predicting an imminent aversive event, unexpected reward omissions, and cues associated with the omission of an expected reward. The most widely described physiological effect of LHb activation is acutely suppressing midbrain dopaminergic signaling. However, recent studies have identified multiple means by which the LHb promotes this effect as well as other mechanisms of action. These findings reveal the complex nature of LHb circuitry. The present paper reviews the role of this structure in learning from reward omission. We approach this topic from the perspective of computational models of behavioral change that account for inhibitory learning to frame key findings. Such findings are drawn from recent behavioral neuroscience studies that use novel brain imaging, stimulation, ablation, and reversible inactivation techniques. Further research and conceptual work are needed to clarify the nature of the mechanisms related to updating motivated behavior in which the LHb is involved. As yet, there is little understanding of whether such mechanisms are parallel or complementary to the well-known modulatory function of the more recently evolved prefrontal cortex.


Subject(s)
Habenula , Cues , Dopamine , Mesencephalon , Reward
7.
J Vis Exp ; (163)2020 09 09.
Article in English | MEDLINE | ID: mdl-32986037

ABSTRACT

One of the most important things in the field of adult hippocampal neurogenesis (AHN) is the identification of the newly generated cells. The immunodetection of thymidine analogs (such as 5-Bromo-2'-deoxyuridine (BrdU)) is a standard technique used for visualizing these newly generated cells. Therefore, BrdU is usually injected in small animals intraperitoneally, so the thymidine analog gets incorporated into dividing cells during DNA synthesis. Detection is performed by immunohistochemical analysis of brain slices. Every research group that has been using this technique can appreciate that it requires special attention to minute details to achieve a successful stain. For instance, an important step is DNA denaturation with HCl, which allows it to reach the cell nucleus to stain it. However, the existing scientific reports describe very few of such steps in detail. Therefore, standardizing the technique is challenging for new laboratories as it can take several months to yield positive and successful outcomes. The purpose of this work is to describe and elaborate the steps to obtain positive and successful outcomes of the immunostaining technique in detail when working with the thymidine analog BrdU. The protocol includes the reagent preparation and setup, administration of thymidine analog in a rodent, transcardial perfusion, tissue preparation, peroxidase immunohistochemical reaction, use of avidin-biotin complex, immunofluorescence, counterstaining, microscopy imaging, and cell analysis.


Subject(s)
Bromodeoxyuridine/metabolism , Immunohistochemistry/methods , Neurogenesis , Thymidine/analogs & derivatives , Animals , Antigens/metabolism , Cell Nucleus Shape , Cell Proliferation , Dentate Gyrus/cytology , Dissection , Fluorescent Antibody Technique , Image Processing, Computer-Assisted , Imaging, Three-Dimensional , Male , Rats, Wistar , Tissue Fixation
8.
J Vis Exp ; (162)2020 08 20.
Article in English | MEDLINE | ID: mdl-32894266

ABSTRACT

Brainwaves amplitude obtained from electroencephalography (EEG) has been well-recognized as a basis for cognitive capacity, memory, and learning on animals and humans. Adult neurogenesis mechanism is also linked to memory and learning improvement. Traditionally, researchers used to assess learning and memory parameters in rodent models by behavioral tasks. Therefore, the simultaneous monitoring of behavioral changes and EEG is particularly interesting in correlating data between brain activity and task-related behaviors. However, most of the equipment required to perform both studies are either complex, expensive, or uses a wired setup network that hinders the natural animals' movement. In this study, EEG was recorded with a wireless electrophysiology device along with the execution of a novel object recognition task (NORT). The animal's behavior was monitored simultaneously by a video tracking system. Both recordings were analyzed offline by their timestamps which were synched to link EEG signals with the animal's actions. Subjects consist of adult Wistar rats after medium-term environmental enrichment treatment. Six skull screw electrodes were fixed in pairs on both hemispheres over frontal, central, and parietal regions and were referenced to an electrode located posterior of the nasal bone. NORT protocol consists of exposing the animal to two identical objects for 10 min. After 2 h and 24 h, one of the objects was replaced with a novel one. Exploration time for each object was monitored by a behavioral tracking software (BTS) and EEG data recording. The analysis of the EEG synced with behavioral data consists of estimations of alpha and beta relative band power and comparisons between novel object recognition versus familiar object exploration, between three experimental stages. In this manuscript, we have discussed electrodes manufacturing process, epidural electrodes implantation surgery, environmental enrichment protocol, NORT protocol, BTS setup, EEG - BTS coupling for simultaneous monitoring in real-time, and EEG data analysis based on automatic events detection.


Subject(s)
Behavior, Animal/physiology , Electroencephalography/methods , Hippocampus/physiology , Memory/physiology , Wireless Technology , Animals , Brain Waves/physiology , Electrodes, Implanted , Electroencephalography/instrumentation , Hippocampus/cytology , Hippocampus/growth & development , Male , Neurogenesis , Rats , Rats, Wistar
9.
J Vis Exp ; (145)2019 03 17.
Article in English | MEDLINE | ID: mdl-30933076

ABSTRACT

The present article provides a guide for the conduction and analysis of three conditioning-based protocols to evaluate impulsivity in rats. Impulsivity is a meaningful concept because it is associated with psychiatric conditions in humans and with maladaptive behavior in non-human animals. It is believed that impulsivity is composed of separate factors. There are laboratory protocols devised to assess each of these factors using standardized automated equipment. Delay discounting is associated with the incapacity to be motivated by delayed outcomes. This factor is evaluated through intertemporal choice protocols, which consist of presenting the individual with a choice situation involving an immediate reward and a larger but delayed reward. Response inhibition deficit is associated with the incapacity to withhold prepotent responses. Differential reinforcement of low rates (DLR) and feature-negative discrimination protocols assess the response inhibition deficit factor of impulsivity. The former imposes a condition to a motivated individual in which most wait a minimum period of time for a response to be rewarded. The latter evaluates the capacity of individuals to refrain from food seeking responses when a signal of the absence of food is presented. The purpose of these protocols is to construct an objective quantitative measure of impulsivity, which serves to make cross-species comparisons, allowing the possibility of translational research. The advantages of these particular protocols include their easy set-up and application, which stems from the relatively small amount of equipment needed and the automated nature of these protocols.


Subject(s)
Impulsive Behavior/physiology , Laboratories , Animals , Choice Behavior , Delay Discounting/physiology , Discrimination, Psychological , Food , Haloperidol/pharmacology , Male , Rats, Inbred SHR , Rats, Wistar , Reinforcement, Psychology
10.
J Neurosci Methods ; 320: 9-15, 2019 05 15.
Article in English | MEDLINE | ID: mdl-30836105

ABSTRACT

BACKGROUND: In behavioral neuroscience studies, it is essential to accurately and easily record rodent sexual behavior. However, current systems are complex, expensive and often require a computer that hinders their use in situ. NEW METHOD: SEXRAT MALE is new software for the real-time annotation and process of live sexual behavior in male rodents. RESULTS: SEXRAT MALE works on smartphones and tablets Android™ operating system, and soon on iOS™. This software uses a simple algorithm based on chronometry and the counting of the behavioral states that the user taps on the screen. It can annotate, count and time a complex sequence of behavioral states by a human observer, such as genital exploration, mounting, intrusion, ejaculation, and up to nine wildcard events established by the user, such as rearing and female receptivity events. SEXRAT MALE reports time, order, duration, intervals, and the simple statistics of all events in the session through data files, which are easily importable from any statistical software. SEXRAT MALE showed greater precision among both expert and naïve observers analyzing sexual interaction, decreasing errors and providing greater accuracy of timing data. Additionally, it records and processes more information than the traditional recording, in terms of factors such as order, occurrence time, duration, events intervals, and other processed data. COMPARISON(S) WITH EXISTING METHOD: Compared to the traditional, manual method, SEXRAT MALE is more accurate, reliable, and simplifies the process of recording and processing. CONCLUSIONS: SEXRAT MALE provides a novel tool for accurately recording live male rodent sexual behavior.


Subject(s)
Behavior, Animal/physiology , Computers, Handheld , Mobile Applications , Neurosciences/instrumentation , Sexual Behavior/physiology , Animals , Male , Neurosciences/methods , Rats , Rodentia , Smartphone
11.
Genes Nutr ; 13: 1, 2018.
Article in English | MEDLINE | ID: mdl-29339975

ABSTRACT

BACKGROUND: Genetic polymorphisms of antioxidant enzymes CAT, GPX, and SOD are involved in the etiology of obesity and its principal comorbidities. The aim of the present study was to analyze the effect of aforementioned SNPs over the output of several variables in people with obesity after a nutritional intervention. The study included 92 Mexican women, which received a dietary intervention by 3 months. Participants were genotyped and stratified into two groups: (1) carriers; mutated homozygous plus heterozygous (CR) and (2) homozygous wild type (WT). A comparison between CR and WT was done in clinical (CV), biochemical (BV), and anthropometric variables (AV), at the beginning and at the end of the intervention. RESULTS: Participants (n = 92) showed statistically significant differences (p < 0.05) at the end of the nutritional intervention in several CV, BV, and AV. However, two kinds of responses were observed after genotyping participants: (A) CR and WT showed statistically significant differences (p < 0.05) in several CV, BV, and AV for the SNPs 599C>T GPX1 (rs1050450), - 251A>G SOD1 (rs2070424), and - 262C>T CAT (rs1001179). (B) Only CR showed statistically changes (p < 0.05) in several CV, BV, and AV for the SNPs - 21A>T CAT (rs7943316) and 47C>T SOD2 (rs4880). The dietary intervention effect was statistically significantly between the polymorphisms of 47C>T SOD2 and BMI, SBP, TBARS, total cholesterol, and C-LCL (p < 0.05) and between the polymorphisms of - 21A>T CAT (rs7943316) and SBP, DBP, total cholesterol, and atherogenic index (p < 0.05). CONCLUSION: People with obesity display different response in several CV, BV, and AV after a nutritional intervention, depending on the antioxidant genetic background of SOD and CAT enzymes.

12.
Naunyn Schmiedebergs Arch Pharmacol ; 391(3): 247-254, 2018 03.
Article in English | MEDLINE | ID: mdl-29349706

ABSTRACT

Cathinones, such as mephedrone (Meph), are often co-abused with alcoholic drinks. In the present study, we investigated the combined effects of Meph plus ethanol (EtOH) on neurotransmitter release in the nucleus accumbens (NAc) and the medial prefrontal cortex (mPFC). A guide canula was stereotaxically implanted into either the NAc or the mPFC of male Sprague-Dawley rats. Seven days after surgery, a microdialysis probe was inserted and rats were administered saline, EtOH (1 g/kg, i.p.), Meph (25 mg/kg, s.c.), or their combination, and dialysates were collected. Serotonin (5-HT), dopamine (DA), and their metabolites (5-HIAA, DOPAC and HVA) were determined through high-pressure liquid chromatography coupled to mass spectrometry. 5-HT and DA peaked 40 min after Meph administration (with or without EtOH co-treatment) in both areas. EtOH combined with Meph increased the 5-HT release compared with the rats receiving Meph alone (85% in NAc, 65% in mPFC), although the overall change in the area under the curve only reached statistical significance in the NAc. In mPFC, the increased release of 5-HT lasted longer in the combination than that in the Meph group. Moreover, EtOH potentiated the psychostimulant effect of Meph measured as a locomotor activity. Given that both 5-HT and DA are also related with reward and impulsivity, the observed effects point to an increased risk of abuse liability when combining Meph with EtOH compared with consuming these drugs alone.


Subject(s)
Dopamine/metabolism , Ethanol/pharmacology , Illicit Drugs/pharmacology , Methamphetamine/analogs & derivatives , Nucleus Accumbens/drug effects , Prefrontal Cortex/drug effects , Serotonin/metabolism , Animals , Drug Interactions , Locomotion/drug effects , Male , Methamphetamine/pharmacology , Nucleus Accumbens/metabolism , Prefrontal Cortex/metabolism , Rats, Sprague-Dawley
13.
J Psychopharmacol ; 30(7): 707-12, 2016 07.
Article in English | MEDLINE | ID: mdl-27147595

ABSTRACT

Methylenedioxypyrovalerone (MDPV) is a synthetic cathinone which has recently emerged as a designer drug of abuse. The objective of this study was to investigate the locomotor sensitization induced by MDPV in adolescent mice, and associated neuroplastic changes in the nucleus accumbens and striatum through deltaFosB and CREB expression. Behavioural testing consisted of three phases: Phase I: conditioning regimen with MDPV (0.3 mg/kg/day for five days) or saline; Phase II: resting (11 days); Phase III: challenged with MDPV (0.3 mg/kg), cocaine (10 mg/kg) or saline on day 16 for both groups. Mice repeatedly exposed to MDPV increased locomotor activity by 165-200% following acute MDPV or cocaine administration after an 11-day resting period, showing a MDPV-induced sensitization to itself and to cocaine. An explanation for this phenomenon could be the common mechanism of action between these two psychostimulants. Furthermore, the MDPV challenge resulted in higher levels of phospho-CREB in MDPV-conditioned mice compared with MDPV-naive mice, probably due to an up-regulation of the cAMP pathway. Likewise, MDPV exposure induced a persistent increase in the striatal expression of deltaFosB; the priming dose of MDPV also produced a significant increase in the accumbal expression of this transcription factor. This study constitutes the first evidence that an exposure to a low dose of MDPV during adolescence induces behavioural sensitization and provides a neurobiological basis for a relationship between MDPV and cocaine. We hypothesize that, similar to cocaine, both CREB and deltaFosB play a role in the induction of this behavioural sensitization.


Subject(s)
Behavior, Animal/drug effects , Benzodioxoles/pharmacology , Cyclic AMP Response Element-Binding Protein/metabolism , Proto-Oncogene Proteins c-fos/metabolism , Pyrrolidines/pharmacology , Alkaloids/pharmacology , Animals , Central Nervous System Stimulants/pharmacology , Cocaine/pharmacology , Corpus Striatum/drug effects , Designer Drugs/pharmacology , Male , Mice , Motor Activity/drug effects , Nucleus Accumbens/drug effects , Nucleus Accumbens/metabolism , Transcription Factors/metabolism , Up-Regulation/drug effects , Synthetic Cathinone
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