bluuebunny/crossref_metadata_2025 huggingface dataset:
Modulated electro hyperthermia inhibits tumor progression in a triple negative mouse breast cancer model
Authors: Hamar Péter , Danics Lea , Schvarcz Csaba , Zolcsák Zita , Benyó Zoltán , Kaucsár Tamás
Published: June 2021

Introduction

The effective therapy of triple‐negative breast cancer (TNBC) has not yet been achieved. Modulated electro‐hyperthermia (mEHT) is a novel adjuvant antitumor therapy, based on the highly selective heating of the tumor tissue by a 13.56 MHz radiofrequency current induced electric field.

Aims

Our aim was to investigate the effects of repeated mEHT treatment in a triple‐negative mammary carcinoma bearing mouse model.

Method

4T07 cells were inoculated orthotopically in female BALB/c mice. Tumor growth was monitored in vivo by digital caliper and ultrasound (Phillips Sonos 5500). The mEHT (n=8) or sham (n=9) treatments started 7 days after inoculation and were repeated 5 times, on every other day. Mice were euthanized 1 day after the fifth treatment and the tumors were dissected, weighed and processed for histology and molecular biology techniques. The ratio of the damaged area compared to the whole tumor area (Tissue Destruction Ratio, TDR) was evaluated on H&E and cleaved caspase‐3 stained sections, while HSP70, a common damage‐associated molecular signal, Ki67, a proliferation marker and p21, a tumor suppressor protein expression were analyzed on immunohistochemical staining with the HistoQuant module of the CaseViewer Software (3DHistech).

Results

There was a significant decrease in tumor growth (sham: 5.7x, mEHT: 2.4x relative to pre‐treatment (day 6) size, p<0.0001) and weight (sham: 288.3±58.1 mg vs mEHT: 85.3±21.3 mg, p<0.05) in the mEHT treated group, compared to the sham group. The HSP70 stained area in the non‐destructed tumor tissue was 5.2 fold higher in the mEHT treated group, compared to the sham group (p<0.05). Moreover, the Ki67 positive nucleus / mm2 count was significantly lower (sham: 2823.4±211.9 pcs/mm2 vs mEHT: 1736.7±315.3 pcs/mm2, p<0.05) and the p21 positive nucleus / mm2 count showed increasing tendency (sham: 127.0±25.3 pcs/mm2 vs mEHT: 242.2±78.2 pcs/mm2, p = 0,073) in the mEHT treated group, compared to the sham group.

Conclusion

Our findings suggest, that repeated mEHT could lower tumor cell proliferation by promoting cell cycle arrest in vivo. Thus, mEHT could be a possible alternative adjuvant therapeutic strategy for TNBC cancer patients. We plan next generation sequencing to elucidate the biological mechanism behind the effects of mEHT.

NVKP_16‐1‐2016‐0042

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anti‐angiogenic activity of memantine, a dual α7‐nAChR/ NMDAR antagonist, in human squamous cell lung cancer
Authors: Dasgupta Piyali , Robateau Zachary R , Nolan Nicholas A , Brown Kathleen C , Chen Yi Charlie , Tirona Maria R , Egleton Richard D
Published: June 2021

Squamous cell lung cancer (SCC‐L) bears a strong etiological association with smoking habits. Nicotine, the addictive component of cigarettes, promotes angiogenesis in lung cancer via the α7‐nicotinic acetylcholine receptor (α7‐nAChR) subunit on human lung endothelial cells. Our published data shows that nicotine upregulates the expression of α7‐nAChR in SCC‐L cell lines. Therefore, we conjectured that α7‐nAChR‐antagonists should display potent anti‐angiogenic and anti‐tumor activity in SCC‐Ls. Memantine is a dual α7‐nAChR/NMDAR antagonist, which is used in the clinic for the treatment of patients suffering from mild‐to‐moderate Alzheimer's disease. Receptor binding assays have shown that the affinity of Memantine for α7‐nAChR is greater than NMDAR. Tumor‐associated endothelial cells are biochemically and morphologically distinct from normal pulmonary endothelial cells. We observed that SCC‐L associated endothelial cells (STACE) and human microvascular endothelial cells of the lung (HMEC‐Ls) expressed robust levels of α7‐nAChR and NMDAR‐1 and NMDAR‐1. Our results indicate that memantine attenuated nicotine‐induced angiogenesis in HMEC‐L. Most interestingly, the levels of α7‐nAChR in tumor‐associated endothelial cells was greater than normal lung endothelial cells. The anti‐angiogeneic activity of memantine was mediated by the α7‐nAChR (and not by NMDARs) on lung endothelial cells. Furthermore, the α7‐nAChR antagonist memantine displayed potent anti‐angiogenic activity in chicken chorioallantoic membrane (CAM) model. Our studies show that α7‐nAChR antagonists may be useful anti‐tumor agents relevant for the treatment of human lung cancer.

Support or Funding Information

Funding for our study was supported by an NIH R15‐AREA Grant (2R15CA161491‐02), Edwards Cancer Center Pilot grant, Cabell Huntington Hospital, Marshall University. This work was supported in part by the West Virginia IDeA Network of Biomedical Research Excellence (WV‐INBRE) grant GM103434(PI: Dr. G. Rankin). A NASA Undergraduate Fellowship was also given to NAN.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
How the calcium influx and homeostasis might be disrupted in the Purkinje cell degeneration and cerebellar dysfunction?
Authors: Aslam Naveed , Alvi Farah
Published: June 2021

In cerebellar circuitry the Purkinje cells (PCs) are the most prominent neuronal element and are characterized by thick and highly branched dendritic tree. The signaling molecule PKCγ is critically linked to Purkinje cell function and has a strong negative impact on the dendritic growth and development of PCs. Recent evidence links enhanced PKCγ to long‐term alteration in calcium equilibrium in PCs. Observations also show that blocking the voltage gated calcium channels could partially or completely protect Purkinje cell dendrites from stunted growth thought to be associated with enhanced PKCγ activity. Additional evidence from moon walker mutant mice also link the increased calcium influx through TRPC3 channels to dampened dendritic morphology. In this study we investigate that how the calcium influx and homoeostasis is regulated in PCs? Here, we propose an integrated model of calcium equilibrium in PCs, by converging the mGluR1 induced membrane generation of DAG and calcium influx through TRPC3 channels into an integrated signaling cascade. The model proposed here link the DAG generation after membrane depolarization to PKCγ activation as well as the activation of TRPC3 through direct binding of DAG. This proposed signaling cascade suggests that DAG signaling in Purkinje cells is tightly regulated through a double‐negative feed‐back loop. The results show that DAG‐ PKCγ‐DGKγ axis forms the first negative feedback loop whereas, the DAG‐TRPC3‐PKCγ‐DGKγ axis forms the second negative feedback loop where, both PKCγ and DGKγ molecules in second negative feedback loop are contributing due to Ca+2 influx through TRPC3 channel, which in turn, stimulates the cytosol‐to‐membrane translocation of these molecules in PCs. Our results suggest that for wild‐type case the double negative feedback on DAG and subsequently on TRPC3 channel is exquisite and specific enough to maintain the calcium homeostasis in PCs, thus ensuring their viability. However, for PKCγ with enhanced activity the second double negative feedback loop is not able to tightly regulate the gating characteristics of TRPC3 channels thus disrupting the calcium homeostasis and leading to degeneration and dysfunction of PCs.

Support or Funding Information

Private Funding

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dissecting the Contribution of Specific Hindbrain Cell Populations in the Phenotypic Characterization of a Mouse Model of Central Congenital Hypoventilation Syndrome
Authors: Alzate‐Correa Diego , Liu Jillian , Jones Mikayla , Alves Michele Joana , Silva Talita Melo , Baer Lisa , Stanford Kristin , Czeisler Catherine , Otero Jose
Published: June 2021

Hindbrain respiratory neuron networks undergo critical developmental maturation during the late embryonic, perinatal and post‐natal periods in mammals. Thus, abnormalities in neuron network development may contribute to autonomic respiratory dysfunction. Central Congenital Hypoventilation Syndrome (CCHS) is a rare genetic disorder characterized by an inability to sense and respond to altered levels of CO2 or O2 (Chemosensation). CCHS patients present with a spectrum of central respiratory control pathophysiologies, ranging from deficiency in chemosensation, to most severe manifestations of neonatal apnea that requires life‐sustaining mechanical ventilation. Heterozygous Mutations of the transcription factor PHOX2B, classified as either poly‐alanine repeat (PARM) and non‐poly‐alanine repeat (NPARM), have been linked to the development of CCHS. In order to explore CCHS pathophysiology, a novel transgenic mouse expressing an inducible NPARM Phox2b mutation (Phox2bΔ8) was developed and characterized by Nobuta and collaborators (Nabuta et al. 2015). Following this study and aiming to reveal the contribution of specific hindbrain populations to CCHS, we have generated two additional mouse models in which of Phox2bΔ8 expression is restricted to cell populations derived from either NKX2.2+ or OLIG3+ progenitors. Expression of Phox2bΔ8 on NKX2.2+ derived cells resulted in a strong apneic phenotype on newborn pups. Detail examination of this mouse model hindbrains showed deficiencies in a spectrum of rhombomeric nuclei not developmentally derived from NKX2.2+‐expressing progenitors, indicating that the developmental neuropathologies identified result through non‐cell‐autonomous mechanisms. Additionally, we find that these pathologies result from early rhombomeric patterning defects in embryonic ventral neural progenitor zones, which showed reduced endogenous PHOX2B expression. On the other hand, preliminary results indicate that expression of Phox2bΔ8 on OLIG3+ derived cells result in a subtle deficit in chemosensation, but we have been unable to find any neuropathological change in the hindbrain. Together, our results help to dissect the contributions of different cell populations in the hindbrain and to further clarify the different manifestations and severity of chemosensation and respiratory deficits seen in human CCHS patients

Support or Funding Information

NIH R01HL132355

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterizing αSyn Pre‐Formed Fibril Animal Models and Exploring Endpoints Beyond αSyn for Parkinson Disease
Authors: Gocal Wiktoria , Gentzel Renee , Marcus Jacob
Published: June 2021

The second most common neurodegenerative disease, Parkinson disease (PD), has been characterized by conformational mutations in the synaptic protein, alpha‐synuclein (αSyn). In the disease state, the protein becomes increasingly insoluble and aggregates, forming Lewy bodies and impairing cellular function, as seen in PD. To further study this synucleinopathy in animals, a model has been established in which αSyn Lewy bodies are induced through the addition of exogenous αSyn preformed fibrils (PFFs) (Luk et al., 2009). These PFFs allow for the “seeding” of endogenous αSyn, producing the Lewy body neuropathology typical of PD. Our group used this seeding model in‐vivo, by injecting 4 uL and 8 uL doses of 2 ug/ul PFFs directly into the striatum of wild‐type rats, and histologically examining the spread of the pathology 30 days later. Pathological αSyn is correlated with phosphorylation of the protein at serine129 (pSer129), and thus all samples were stained for pSer129 using immunohistochemistry methods at the 30 day time point.

Additional endpoints were also explored to better understand the biology of PD. Aside from αSyn accumulation, PD is also characterized by neurodegeneration that could be due to an inflammatory or immune response to the aggregated protein. To explore this relationship, immunohistochemistry staining was performed for glial marker Iba‐1 to visualize microglia and GFAP to visualize astrocytes. Staining was also performed for lymphocyte‐activation gene 3 (LAG3) and major histocompatibility complex II (MHCII) as markers for a potential immune response. These results will enrich understanding of additional inflammation and immune components of this rodent model of PD.

Support or Funding Information

Merck Research Laboratories

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Fat Emboli in the Rat Brain in a Model of Fat Embolism Syndrome
Authors: Colson Jordan Dane , Siddiqi Ahsan , Van Dillen Michael , Fotouhi Ariana , Al‐Husseinawi Ethar , Al‐Allaf Jwan , Wells Brandon , Monaghan Paula , Poisner Alan , Molteni Agostino
Published: June 2021

Introduction

Fat embolism (FE) resulting from intravenous triolein (T) injection in an experimental rat model has been shown to result in severe pulmonary damage, septal and arterial inflammation, and eventually fibrosis. This tissue response appears to be biphasic in nature with a peak at 48 hours. Fat droplets have previously been reported in the brains of patients suffering severe fat embolism syndrome (FES). The current study was carried out to determine if FE in our model was also associated with fat droplets in the brain at 48 hours.

Materials and Methods

26 Sprague‐Dawley rats (250–330g) were divided into two groups (N=13 each) receiving either 0.2 mg intravenous T or normal saline. All animals were euthanized under isoflurane anesthesia 48 hours later. Lungs and brains were isolated, snap‐frozen, and processed for cryosectioning. Sections were stained for oil red o and hematoxylin and eosin. Sections were imaged at 400x magnification and counted by two pathologists blinded to section identity. These counts were decoded by an independent observer and statistically compared utilizing a student's T‐test.

Results

The brains of the control group exhibited fat droplets in 3 of the 12 brains considered. These droplets were small in size and primarily localized to the cerebral cortex with minimal associated parenchymal reaction. 10 of the 12 brains observed in the experimental group demonstrated fat droplets which were localized to the cerebral cortex, cortical arteries, and especially the meningeal arteries. These droplets were much larger and were associated with an appreciable vascular thickening within affected vessels. Lungs of the control group featured a small amount of small scattered fat droplets in the same 3 subjects while the lungs of the experimental group all featured large fat droplets with the expected septal and associated arterial damage pattern previously published.

Conclusions

While fat deposition is a non‐specific finding, the difference in tissue response demonstrated in rat lungs and brains is evident. While FE appears to provoke a marked inflammatory response within the lungs even at 48 hours the brain does not appear to reciprocate this change. While the droplets noted within the experimental group are larger and more associated with a mild vessel response the parenchyma of the brain appears much less affected compared to the lungs within the same animal. As the tissue reaction within the lungs is biphasic and the observed changes within the brain differ significantly further studies are necessary to ascertain if said changes might not evolve over a longer period of time.

Support or Funding Information

Supported by the Catherine T Geldmacher foundation, Saint Louis, MO.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
High scores in the embedded figures test correlate with tissue diagnostic speeded
Authors: ACERO EDWARD JAVIER
Published: June 2021

INTRODUCTION

In educational scenarios of tissue diagnostics, cognitive skills with high GEFT scores are valued because they are associated with more rapid diagnoses, but perceptive or confidence errors can be presented. The objective of this work was to show that students with high GEFT scores do not observe all the tissue diagnostic patterns of Fibrous Histiocytoma stained with H&E.

MATERIALS AND METHODS

Individually 63 medical students were challenge to diagnostic miscroscopic with digital image of Fibrous Histiocytoma stained with H&E, and while an eye track was made, diagnostic times of begin to finish was recorded. On other site and moment, too individually a Group Embedded Figures Test (GEFT) was made

RESULTS

High scores (Between 25 and 20 of GEFT) n= 35 students) had less time to diagnostic Fibrous Histiocytoma (X 115.1 s, SD = 13,2); and correlation (r = −0.84 P = 0.0041). and low scores less than 7 of GEFT) n= 14 students) nedded more time to diagnostic Fibrous Histiocytoma (X 132,6 s, SD = 18,5); and correlation (r = −43,1 P = 0.037). intermedium scores (Between 19 and 9 of GEFT) n= 11 students) had correlation non linear with shape "U" inverted with the more diagnostic longer times (X 157,2 s, SD = 26,8). Interesting find with eye tracking show how high scores of GEFT, not see clasical cells pattenrs of Fibrous Histiocytoma, but watch only collagen bundles; and those with intermedium and lower scores GEFT watch collagen and fibroblast bundles

DISCUSSION

As the laws of perceptual organization state that contrast cognitive loadis used for the recognition of GEFT, in students with high scores to GEFT a faster cognitive recognition processes to diagnostic Fibrous Histyocitoma is suggested with similar mechanism, achieving by visual search only to collagen bundles stained with H&E.

CONCLUSIONS

High scores on GEFT correlate with tissue diagnostic of Fibrous Histiocytoma stained with H&E more speeded, using patterns perceptive only to collagen bundles

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
RAGE Implications during DNA Double Strand Breaks in Trophoblast Cells
Authors: Tsai K YF , Hirschi K M , Knowlton M N , Mejia J F , Hall P , Davis T , Reynolds P R , Arroyo Juan A
Published: June 2021

Intrauterine growth restriction (IUGR) is an obstetric pathology characterized by an increase in trophoblast apoptosis and decreased cell invasion. Trophoblasts are critical for a successful pregnancy and they mediate important steps such as implantation, immune protection of the fetus, maternal blood flow to the placenta, and delivery. Previous mouse studies in our laboratory showed increased trophoblast apoptosis, decreased cell invasion and increased placental receptors for advanced glycation end‐products (RAGE) during second hand smoke (SHS) treatment. More recently, a role for nuclear RAGE (nRAGE) has been implicated in DNA double strand break (DNA‐DSB) repair. We tested the hypothesis that RAGE is involved during DNA‐DSB repair in trophoblast cells. DSBs were induced in human trophoblasts (Bewo and Sw71) with cigarette smoke extract (CSE) or bleomycin (BLM). Assessment of nRAGE and g‐H2AX (involved in DNA‐DSB repair) was done by immunostaining. Invasion was measured in trophoblast cells after CSE and BLM treatment. Immunoblot was used to quantify nRAGE and g‐H2AX in cells. DNA degradation was used to determine DNA integrity.

Increased Co‐expression of RAGE and y‐H2AX was observed in treated trophoblasts when compared to controls. Increases protein expression of RAGE and g‐H2AX was quantified by immunoblot in control and treated trophoblast cells . Trophoblast invasion was decreased 50% with 20ug/ml BLM treatment (p<0.0001) and 80% with 30ug/ml as compared to controls (p<0.0001). Invasion of trophoblast cells was decreased 92% (p<0.002) when treated with 0.5% CSE. Trophoblast DNA degradation was significantly detected following treatment with BLM or CSE. We conclude that CSE and BLM causes DNA‐DSBs in trophoblast cells and repair may plausibly be regulated by nRAGE and g‐H2AX. Confirmatory studies in the absence of RAGE are necessary in order to validate a possible link. DNA‐DSBs could be a factor in inhibited trophoblast invasion observed in compromised placenta. These studies provide a critical initial step in dissecting tobacco‐mediated IUGR progression.

Support or Funding Information

This work was supported by a grant from the Flight Attendant's Medical Research Institute (FAMRI, PRR and JAA) and a BYU Mentoring Environment Grant (JAA and PRR).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sex Differences in Anti‐allodynic Tolerance to Δ9‐THC in Desensitization‐resistant S426A/S430A and Wild‐type Mice
Authors: Henderson‐Redmond Angela N , Sepulveda Diana E , Hale David E , Morgan Daniel J
Published: June 2021

Tolerance to the anti‐allodynic effects of cannabinoids has the potential to limit their therapeutic potential. A growing body of clinical and preclinical evidence suggests that sex influences a variety of cannabinoid‐related outcomes, including cannabinoid‐induced anti‐allodynia. Male and female mice expressing a desensitization‐resistant form of the cannabinoid type‐1 (CB1) receptor (S426A/S430A) show delayed tolerance to Δ9‐tetrahydrocannabinol (Δ9‐THC) in the tail‐flick assay. Likewise, using the tail‐flick test, male wild‐type mice are more sensitive than females to the anti‐nociceptive effects of Δ9‐THC across the full dose response curve. While the tail‐flick is an acute model of thermal pain, it remains unknown whether these sex‐ and/or genotype differences extend to a chronic model of neuropathic pain. The purpose of the current study was to determine whether the S426A/S430A mutation also alters tolerance to anti‐allodynic effects of Δ9‐THC in a sex‐specific manner. Male and female wild‐type and S426A/S430A mice were assessed for tolerance to the anti‐allodynic effects of 6 and 10 mg/kg of Δ9‐THC in a cisplatin‐evoked model of neuropathic pain. Female mice were less sensitive to the anti‐allodynic effects of 6 mg/kg Δ9‐THC than males. Treatment with 6 mg/kg Δ9‐THC was able to completely reverse allodynia in male, but not female, mice. Female mice showed a complete reversal of allodynia following treatment with 10 mg/kg Δ9‐THC. Interestingly, the S426A/S430A mutation did not alter sensitivity or tolerance development to the anti‐allodynic effects of Δ9‐THC compared to wild‐type mice of either sex, suggesting that the action of Δ9‐ THC at the mutant form of CB1 may be masked by its substantial anti‐allodynic effects through CB2. This study highlights the therapeutic potential of Δ9‐THC in a model of neuropathic pain and suggests that these effects might be mediated in a sex‐specific manner that may have clinical implications for the use of cannabinoids in chronic pain management.

Support or Funding Information

National Institute on Drug Abuse grant DA037355 (DJM); Penn State University College of Medicine Department of Anesthesiology & Perioperative Medicine; Pennsylvania Department of Health using CURE Tobacco Settlement Funds (DJM)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Relative Contribution of Dopamine, Norepinephrine, and Serotonin to the Discriminative Stimulus Effects of MDPV
Authors: Seaman Robert W , Doyle Michelle R , Sulima Agnieszka , Rice Kenner C , Collins Gregory T
Published: June 2021

The abuse of synthetic cathinones (“bath salts”) has increased worldwide over the past decade. Bath salt preparations often contain multiple synthetic cathinones in addition to other stimulants such as cocaine or caffeine. Synthetic cathinones are known to interact with dopamine, norepinephrine, and serotonin transporters (DAT, NET, and SERT, respectively). Although recent studies suggest that interactions with DAT are crucial for the reinforcing effects of synthetic cathinones, actions at SERT were found to negatively modulate reinforcing effectiveness. The goal of the current study was to characterize the relative contributions of dopamine, norepinephrine, and serotonin to the discriminative stimulus effects of MDPV by evaluating the substitution profiles of 1) a series of structurally related cathinones (MDPBP, MDPPP, α‐PPP, and α‐PVP) and cocaine, and 2) direct‐acting dopamine (quinpirole [D2/3], SKF 82958 [D1/5], and apomorphine [D1‐5]), and norepinephrine (phenylephrine [α1], and clonidine [α2]) receptor agonists in male Sprague‐Dawley rats (n=8) trained to discriminate 1.0 mg/kg MDPV from saline under a fixed ratio 10 schedule of sucrose pellet delivery. The mu‐opioid receptor agonist fentanyl served as a negative control. As expected, each of the cathinones and cocaine produced dose‐dependent increases in MDPV‐appropriate responding, whereas fentanyl failed to increase MDPV‐appropriate responding at doses smaller than those that suppressed responding. The rank order potency of the synthetic cathinones to increase MDPV‐appropriate responding is positively correlated with their potency to inhibit DAT and NET, but not SERT, and is consistent with the rank order to maintain self‐administration. The D2/3 receptor‐preferring agonist quinpirole dose‐dependently increased MDPV‐appropriate responding whilst SKF 82958 [D1/5], apomorphine [D1‐5], phenylephrine [α1], and clonidine [α2] failed to increase MDPV‐appropriate responding at doses smaller than those that suppressed responding. Together, these data provide evidence that the discriminative stimulus effects of MDPV are primarily mediated by its capacity to inhibit DAT, and the subsequent activation of dopamine D2 and/or D3 receptors. Moreover, these studies suggest do not support a role for serotonergic or adrenergic systems in mediating/modulating the discriminative stimulus effects of MDPV. Future studies will utilize antagonists of dopamine, adrenergic, and serotoninergic receptors to better characterize their relative contributions to the discriminative stimulus effects of MDPV.

Support or Funding Information

This work was supported by NIH/NIDA grant (R01 DA039146; GTC) and NIDA‐ and NIAAA‐IRPs (KCR)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effects of Eating a High Fat Diet on Methamphetamine‐Induced Conditioned Place Preference
Authors: Serna Paloma , Galindo Kayla I , Serafine Katherine M
Published: June 2021

Eating a high fat diet can cause several negative health consequences, including dysfunction to dopamine systems. For example, eating a high fat diet enhances sensitivity of rats to methamphetamine‐induced locomotion. However, it is not known if sensitivity to the rewarding effects of methamphetamine are similarly enhanced in rats eating a high fat diet. Females are more sensitive than males to the behavioral effects of stimulants in general, and therefore might also be particularly vulnerabile to high fat diet‐induced enhanced sensitivity to stimulant drugs. To test the hypothesis that eating high fat chow enhances sensitivity of rats to the rewarding effects of methamphetamine, female Sprague‐Dawley rats were fed standard laboratory chow (17% kcal from fat) or high fat chow (60% kcal from fat) for 4 weeks prior to conditioned place preference (CPP) training, using a biased design. Rats were trained on alternating days rats with saline or methamphetamine (0.32 or 1.0 mg/kg, i.p.) and were restricted to one of two sides (wire mesh vs rod flooring) of a CPP apparatus. After 8 alternating training days, rats had free access to the entire apparatus, and time spent in the drug‐paired side versus the saline‐paired side was examined. Methamphetmaine induced a significant CPP in all rats, regardless of diet or dose of methamphetamine; however, there were no differences in magnitude of CPP between rats eating high fat chow and rats eating standard chow. Previous literature suggests that females might be more sensitive to the rewarding effects of methamphetamine, necessitating the use of smaller doses. As such, future studies will examine a wider range of doses of methamphetamine, as well as other conditioned behavior (i.e., self‐administration) and will include male subjects to study potential sex differences.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A selective phosphodiesterase 4D inhibitor BPN14770 reverses beta amyloid‐induced memory impairment in humanized PDE4D mice
Authors: Xu Ying , Yang Mingxin , Zhang Han‐Ting , Gurney Mark , O'Donnell James
Published: June 2021

Beta amyloid peptides (Aβ) are known risk factors involved in cognitive impairment in Alzheimer's disease (AD). Type 4 phosphodiesterase (PDE4) inhibitors such as rolipram could upregulate intracellular cAMP level, which further activate downstream target cAMP‐responsive‐ element‐binding protein (CREB), brain‐derived neurotrophic factor (BDNF) and nerve growth factor (VGF) signaling. Although conventional PDE4 inhibitors have benefits on memory enhancement in preclinical and clinical studies, they were prevented to be developed as a novel class of memory‐enhancing drugs due to their adverse effects such as GI irritation. BPN14770, a novel allosteric inhibitor of PDE4D, exhibits high selectivity on PDE4D based on its close interaction with a single amino acid present at primate PDE4D. The present study used the humanized PDE4D (hPDE4D) mice, which provide a unique and powerful genetic tool for assessing the PDE4D target engagement, to investigate the effects of BPN14770 on memory impairment induced by Ab1‐42. Results suggested that BPN14770‐treated mice exhibited significant learning and memory enhancement, as shown in Morris water maze and Y maze tests. The morphological analyses suggested that both the number of dendrites and the total length were significantly increased after treatment hPDE4D mice with BPN14770 for two weeks. The subsequent neurochemical assay indicated that neuroprotective proteins such as pCREB, BDNF and VGF in Ab‐treated hPDE4D mice were upregulated after treatment with BPN14770. These findings provide evidence that BPN14770 may be a potential therapeutic drug for treating cognitive deficits in Alzheimer's disease and its underlying mechanism may be involved in the cAMP‐pCREB‐BDNF/VGF signaling.

Support or Funding Information

This project was supported by R44 MH091791 to Drs. Mark Gurney and Ying Xu.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Overexpression of TIPE2, a Negative Regulator of Innate and Adaptive Immunity, Attenuates Cognitive Deficits in APP/PS1 Mice
Authors: Zhang Fang , Miao Yong‐zhen , Xu Zi‐han , Ju Chuan‐xia , Wang Lei , Wang Ke‐wei
Published: June 2021

Neuroinflammation plays an early and prominent role in the pathology of Alzheimer's disease (AD). Tumor necrosis factor‐a‐induced protein 8‐like 2 (TIPE2) has been identified as a negative regulator of innate and adaptive immunity. However, whether TIPE2 affects cognitive functions in AD‐like mouse models remains unknown. In this study, we compared the gene and protein expressions of TIPE2 and β‐amyloid in the brain between the APP/PS1 mice and the age‐matched wild type (WT) mice at different stages of developments using assays of immunofluorescence, western blot and qPCR. Deposition of Aβ plaques appeared in the hippocampus from the 6‐month‐old APP/PS1 mice, and the hippocampal expression of the TIPE2 mRNA and protein in APP/PS1 mice were significantly higher than that of the WT mice starting from 6 months to 10 months. However, the difference of the TIPE2 expression between the APP/PS1 mice and the WT mice declined in a time‐dependent manner. The spatial learning and memory deficit from 8‐month‐old APP/PS1 mice was observed in the Y‐maze test and fear conditioning task. Interestingly, overexpression of TIPE2 by intra‐hippocampal injection of AAV‐TIPE2 into the 7‐month old APP/PS1 mice resulted in an improvement of learning and memory. Taken together, our findings show that the TIPE2 expression level was negatively correlated with the Alzheimer's disease, suggesting TIPE2 is a potential target for pharmacological intervention and improvement of cognitive deficits.

Support or Funding Information

This work was supported by research grants to Fang ZHANG from the Natural Science Foundation of Shandong Province of China, No. ZR2016 HM 46; Post doctoral application foundation of Qingdao Municipal Bureau of Social Sciences, No. 2016062; Qingdao science and technology bureau, No 18‐6‐1‐75‐nsh; Clinical medicine +X Project of Qingdao University Medical Department; Innovation and Entrepreneurship Training Program of undergraduate student in Qingdao University. No. 201711065152.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular and behavioral mechanisms mediating paclitaxel‐induced changes in affect‐like behavior in mice
Authors: Meade Julie , Toma Wisam , Alkhlaif Yasmin , Bagdas Deniz , Obeng Samuel , Selley Dana E. , Damaj M. Imad
Published: June 2021

The antineoplastic paclitaxel is associated with negative affective outcomes, such as depression, anxiety, and decreased quality of life during treatment and convalescence. With the Baby Boomer population approaching peak cancer age, it is dire that the mechanisms behind paclitaxel‐induced changes in mood are uncovered. Cancer‐free male and female C57BL/6J mice were treated with one set of four injections of vehicle or paclitaxel (32mg/kg cumulative), or two sets of four injections of vehicle or paclitaxel (64mg/kg cumulative), and periodically assessed for depression‐like behaviors. Paclitaxel caused significant, time‐dependent deficits in sucrose preference and operant responding for palatable food. Because there is growing evidence to support the role of kappa opioid receptors (KORs) in stress‐mediated depression and reward dysfunction, we investigated KOR signaling as a putative mechanism of paclitaxel‐induced depression‐like behaviors. The selective KOR antagonist norbinaltorphimine (norBNI) reversed paclitaxel‐induced attenuation of sucrose preference. At the molecular level, paclitaxel time‐dependently induced an increase in the expression of Prodynorphin mRNA, the precursor for endogenous KOR agonists, in the nucleus accumbens (NAc). Using the [35S]GTPγS assay, we discovered that a history of paclitaxel time‐dependently attenuated activation of dopamine D2 receptors (D2R) and KORs in the NAc but not caudate putamen. These data suggest that paclitaxel‐induced changes in affect‐like behavior may be due to time‐ and region‐dependent dysregulation of KOR and D2R signaling. These observations help to establish the roles of KOR and D2R systems in paclitaxel‐induced disruption of behavioral reward, thus revealing potential neurochemical targets for therapeutic intervention in cancer survivors with treatment‐resistant depression.

Support or Funding Information

Supported by: Ruth L. Kirschstein National Research Service Award Individual Predoctoral Fellowship to Promote Diversity in Health‐Related Research 1F31CA224930‐01A1; NIH grant 1R01‐CA206028‐01; NIHNCI Cancer Center Support Grant P30‐CA016059.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lower Pain Sensitivity in Ovariectomized Mice Using HIV Gp120 and Chemotherapy‐induced Chronic Pain Models
Authors: Rodríguez Ismael Rodríguez Lugo , García Dianise Rodríguez , Brelsfoard Jennifer , Lilley Jennifer , Blanton Henry , Guindon Josee
Published: June 2021

Pain is an uncomfortable sensation linked with a vast majority of health complications. It is a result of complex interactions involving the peripheral and central nervous system, which vary between different types of pain. It is important to improve the analgesic effects of drugs for different types of pain while maintaining patient well‐being. For this reason, our laboratory focused on understanding the behavioral antinociceptive effects of cisplatin (a chemotherapeutic drug) and HIV gp120 on both male and female mice. We also tested the analgesic effects of CP55, 940 (a cannabinoid CB1/CB2 receptor agonist) in an HIV‐gp120 mouse model after neuropathic effects were documented. Cannabinoid receptor agonists have been shown to be effective in pain animal models by inhibiting acute nociceptive responses and reversing the behavioral hypersensitivity. Because of this, we hypothesize that neuropathic pain on this HIV‐gp120 mouse model will also be reversed. All of this was experimented in ovariectomized mice in order to improve our understanding of pro‐ or anti‐ nociceptive processes and the potential role of sex hormones. We tested the effect of cisplatin (0.5 mg/kg intraperitoneal injection once a week for 4 weeks) or HIV‐gp120 (200 ng) treatment on mechanical (von Frey) and cold (acetone) allodynia in control (male and female) and ovariectomized mice. We also conducted daily vaginal lavages in the female mice. Our results suggest that both, cisplatin and HIV‐gp120 mouse models, induce mechanical and cold pain sensitivity in both male and female. Only HIV‐gp120 female mice show an increase in mechanical and cold pain sensitivity relative to male. Both, ovariectomized mice treated with cisplatin and ovariectomized HIV‐gp120 mice show a decrease in mechanical and cold pain sensitivity relative to control female mice. As for treatment with Cp55, 940 analgesic effects were clearly present at 10mg/kg for all mice. However, 1mg/kg had no analgesic effects. We also observed disturbance in the estrous cycle of the HIV‐gp120 mice group. As a conclusion, we confirm that hormonal changes are influencing pain sensitivity and therefore demonstrate the importance of sex‐hormones in pain transmission. Further studies are needed to better understand the mechanisms of action of these findings.

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This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Age‐dependent differences in pain‐related depression of nesting in mice
Authors: Miller Laurence , McPherson Sarah
Published: June 2021

Pain‐related inference with behavior is a key clinical diagnostic indicator and target in the treatment of pain. Increasingly, preclinical studies on the expression, mechanisms, and treatment of pain have been aimed at improving understanding of pain‐related interference with behavior. Although there are well‐known changes to sensory, motor, and cognitive functions across the lifespan, no studies have examined the expression, mechanisms, and treatment of pain‐related depression of behavior from an aging perspective. The present studies compare the effects of a noxious pain stimulus and a clinically relevant analgesic on nesting behavior in 12‐, 18‐, and 48‐ week old mice. Nesting was operationally defined as the clearance of pieces of cotton nesting material from six zones within the home cage. This is the first step of consolidating the nesting material into a single zone for nest construction. Nesting was observed over the course of a 60‐minute session. First, the rate of nesting was compared across the age groups. Early in the nesting session, 12‐week old mice cleared more zones of nesting material than 18‐ and 48‐week old mice, but there were no differences in the number of zones cleared after 60 minutes. Next, the potency of intraperitoneal injection of lactic acid to depress nesting was determined in each age group. Lactic acid was more potent in the 48‐week old mice than the 18‐week old mice, but no other age related differences were observed. Finally, the effects of the nonsteroidal anti‐inflammatory drug (NSAID), ketoprofen, on lactic acid‐induced depression of nesting were determined. There were no age‐related differences in the potency or efficacy of ketoprofen. Together, these findings support the validity of assays of pain‐related depression of behavior to examine age‐related changes in the expression and treatment of functional impairment caused by pain. Moreover, these findings suggest that aging may result in increased susceptibility to pain‐related functional impairment, and that aging does not affect the ability of NSAIDs to treat pain‐related functional impairment.

Support or Funding Information

Support was provided by the Augusta University Center for Undergraduate Research and Scholarship

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular Mechanism of Rap1A‐Dependent Activation of PLCɛ
Authors: Sieng Monita , Garland‐Kuntz Elisabeth E. , Selvia Arielle F. , Chakravarthy Srinivas , Lyon Angeline M.
Published: June 2021

Phospholipase C (PLC) enzymes hydrolyze phosphatidylinositol lipids to produce diacylglycerol (DAG) and inositol phosphates, leading to the activation of protein kinase C (PKC) and downstream signaling pathways, including cell growth and survival. The PLCɛ subfamily is a key player in cardiovascular function, where it contributes to maximum contractility. However, prolonged activation of PLCɛ results in cardiac hypertrophy and heart failure through its ability to regulate the expression of hypertrophic genes. This process is regulated by the small GTPase Rap1A, which is activated downstream of β‐adrenergic receptors. Rap1A binds to the C‐terminal Ras association (RA) domain of PLCɛ, simultaneously translocating the complex to the perinuclear region and activating PLCɛ. PLCɛ also contains an N‐terminal CDC25 domain, which has guanine nucleotide exchange factor (GEF) activity for Rap1A resulting in a feed forward activation loop and sustained lipid hydrolysis. However, the molecular mechanism of this process is not known. In this work, we seek to characterize the interactions between Rap1A and PLCɛ using structural and functional studies to map the Rap1A binding site on PLCɛ and determine whether activation results in conformational changes that release autoinhibition and/or increase membrane association. These studies provide the first molecular details of the Rap1A‐dependent activation of PLCɛ and open the door to the development of new therapeutic strategies for treating cardiac hypertrophy.

Support or Funding Information

This research is supported by the American Heart Association Predoctoral Fellowship 18PRE33990057 to M.S., American Heart Association Grant Scientist Development Grant 16SDG29920017 to A.M.L. and the NIH NLHBI 1R01HL 141076‐01 to A.M.L. SAXS data was collected and analyzed with the assistance of S. Chakravarthy at Argonne National Laboratory.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Building a Sound Wall in the Cochlea: Development of the Stria Vascularis
Authors: Basch Martin Leandro
Published: June 2021

The stria vascularis is a specialized epithelial structure of the mammalian cochlea that produces endolymph, the potassium‐rich fluid responsible for the positive endocochlear potential of the cochlear lumen. This positive extracellular potential is the major driving force for proper signal transduction by sensory cells in the ear, and thus normal hearing. The stria vascularis is the “battery” of the auditory organ: it is localized in the lateral wall of the cochlea, and consists of a multilayered highly vascularized epithelium. It is composed of three main cell types with distinct embryonic origins: the marginal cells derived from the otic epithelium , the intermediate cells which are specialized melanocyte‐like cells derived from migratory neural crest cells, and the basal cells that are derived from otic mesenchyme. Mesoderm‐derived blood vessels are interspersed between these cells throughout the stria. Defects in the development of the stria vascularis are part of a number of inheritable deafness syndromes in humans, including Hirschsprung disease and Waardenburg, Jervell and Lange‐Nielsen, Bartter and SeSAME syndromes. Strial degeneration is estimated to cause more than 30% of age related hearing loss or presbycusis, which affects between 25–40% of the population over 65 years and over 60% of the population over 80 years old. The causes of strial atrophy remain poorly understood. Over the past decade, a great effort has been made to understand the development of the sensory cells in the mammalian cochlea with the aim of applying their developmental principles to regenerative therapies. By contrast, very little is known about the developmental potential of the stria vascularis, or even the mechanisms of strial degeneration due to aging. Our focus is to uncover the normal development of the stria, the cellular interactions that lead to its formation, and the potential for regeneration in cases of congenital strial deafness.

Support or Funding Information

Hearing Health Foundation

The Hartwell Foundation

NIDCD R01DC015785

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
mGluR5 regulates ZBTB16 pathway of autophagy in Alzheimer's disease in a sex‐specific manner
Authors: Abd‐Elrahman Khaled S , Hamilton Alison , Ferguson Stephen S. G.
Published: June 2021

Alzheimer's disease (AD) is characterized by neurotoxicity mediated by the accumulation of beta amyloid (Aβ) oligomers, causing neuronal loss and progressive cognitive decline. We have previously showed that genetic deletion or chronic pharmacological inhibition of mGluR5 by the negative allosteric modulator CTEP, rescued cognitive function and reduced Aβ aggregation in male APPswe/PS1ΔE9 (APPswe) mouse model of AD 1,2. We have recently reported that the novel ZBTB16‐medaited mechanism of autophagy downstream of mGluR5 plays a key role in Huntington's disease (HD) pathology in zQ715 mice 3. In a follow‐up study, we showed that the favorable outcomes of mGluR5 silencing either genetically and pharmacologically in male APPswe were associated with a reduction in ZBTB16 similar to zQ175 mice 4. Here, we show that, unlike male mice, chronic (12 week) treatment with CTEP of female APPswe mice does not improve cognitive function or AD pathology. CTEP treatment did not mitigate memory deficits detected in female APPswe mice during novel object recognition and Morris water maze tasks. We then tested brain samples from male and female APPswe mice following 12‐week treatment with either vehicle or CTEP (2mg/kg) for changes in the autophagic clearance of Aβ oligomers via ZBTB16 pathway. When quantified using ELISA, both male and female APPswe mice exhibited higher level of Aβ oligomers at 6 months of age compared to control. To our surprise, CTEP significantly reduced Aβ burden in male mice yet, it exacerbated Aβ oligomers accumulation in female APPswe mice. mGluR5 blockade in male APPswe mice caused a GSK3β‐dependent degradation of ZBTB16 and stabilization of the autophagy adaptor ATG14. This was paralleled by a reduction in the autophagy marker p62. Interestingly, we did not detect any change in GSK3β phosphorylation, ZBTB16, ATG14 or p62 levels in female APPswe mice following CTEP treatment compared to control. It is worth noting that female APPswe brain also exhibited lower expression of mGluR5 compared to age‐matched APPswe male mice. This study supports the inhibitory role of mGluR5 on autophagy via GSK3β/ZBTB16 pathway that result in impaired clearance of Aβ burden in male APPswe only. We also provide evidence, and for the first time, that some aspects of mGluR5 signaling are not conserved between males and females and there are sex‐specific differences that must be considered when embracing mGluR5 as a potential drug target for neurodegenerative disease.

Support or Funding Information

Alberta Innovates Health Solutions and Canadian Institutes for Health Research

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.