Abstract List for 2021 meeting

#651: Parsing thirst and hunger circuits in brain using vasopressin-deficient Brattleboro rats

Brakey, Destiny J ¹; Schatz, Kelcie C ¹; Paul, Matthew J ¹; Daniels, Derek ²;

¹Department of Psychology, SUNY University at Buffalo, College of Arts and Sciences; ²Center for Ingestive Behavior Research and Department of Psychology, SUNY University at Buffalo;

Fluid imbalance is a pervasive and costly problem. Fluid intake is controlled, in part, by the central actions of glucagon-like peptide-1 (GLP-1); however, it is not yet known where or how GLP-1 is acting in the brain to control fluid intake. One problem in furthering our understanding of GLP-1’s role in the control of fluid intake is that GLP-1 affects both drinking and feeding and these behaviors are intertwined. In order to dissociate these normally intertwined behaviors, our lab has begun using a rat with a genetic mutation that causes extreme polydipsia without affecting food intake (Brattleboro; BB). We found that these rats were hypersensitive to the fluid intake suppression by a GLP-1 receptor agonist, Exendin-4 (Ex4), without any differences from wild-type (WT) rats in food intake suppression by the same drug. Additionally, we found different patterns of Fos expression in areas commonly associated with food and fluid intakes after injection of Ex4. In the nucleus tractus solitarius and area postrema, for instance, Ex4 treatment was associated with a greater number of Fos-positive cells. In the organum vasculosum of the lamina terminalis and median preoptic nucleus, however, we found no treatment-related differences in WT rats, but in BB rats we found that Ex4 was associated with fewer Fos-positive cells. Lastly, in the paraventricular nucleus, subfornical organ, and arcuate nucleus we found evidence for Ex4-induced increases in Fos in WT rats, but BB rats appear to have Ex4-induced inhibition of Fos expression. These experiments provide important data that can be used to guide future studies that will test for eating and drinking specific effects of GLP-1.

#622: Neurodevelopmental Risk Factors ADNP and POGZ Demonstrate Striking Functional Convergence

Conrow-Graham, Megan E ;

Department of Physiology and Biophysics, SUNY University at Buffalo, School of Medicine and Biomedical Sciences;

ADNP and POGZ are two top-ranking high-confidence risk factors for autism spectrum disorder and intellectual disability, but how they are linked to these neurodevelopmental disorders (NDD) is largely unknown. Both ADNP and POGZ are chromatin remodelers, which could profoundly affect gene transcription and cellular function in the brain. Using postmortem tissue from patients with autism, we found diminished protein levels of ADNP and POGZ in the prefrontal cortex (PFC), a region highly implicated in NDD. Thus, to understand the functional role of these important risk factors, we investigated how Adnp or Pogz reduction in mouse PFC affects behavioral, transcriptomic, and synaptic function. We used viral-based gene transfer to knock down Adnp and Pogz in PFC of wild-type mice during the critical period of synaptic regulation. Behavioral testing revealed impaired NDD-linked cognitive task performance with Adnp- or Pogz-deficiency. Further, patch-clamp recording of PFC pyramidal neurons showed impaired glutamatergic signaling in Adnp- or Pogz-deficient mice. RNA-sequencing demonstrated transcriptional upregulation induced by Adnp- or Pogz-deficiency, with significant overlap in differentially expressed genes. Gene ontology and HUB gene analyses indicated that inflammation is the most upregulated functional category, representing nearly half of the commonly upregulated genes. This finding parallels previous human cortical findings of increased pro-inflammatory and microglial signaling in NDD. Taken together, these findings suggest that a reduction in ADNP or POGZ leads to the elevation of inflammatory genes in the PFC, causing increased microglial activation and impaired synaptic transmission. This results in impairment of PFC-mediated cognitive behaviors. This study has provided novel insights into the convergent actions of two top risk factors for NDD and potential intervention targets.

#646: Huntingtin (HTT) in axon injury: HTT-Rab7 co-migrate on a retrogradely moving signaling endolysosome

Krzystek, Thomas J ; White, Joseph ; Thurston, Layne ; Hoffmar-Glennon, Hayley ; Li, Yichen ; Gunawardena, Shermali ;

Biology Department, SUNY University at Buffalo;

Huntingtin (HTT), the Huntington’s disease (HD) protein, moves bi-directionally within axons and loss/reduction of HTT leads to axonal transport defects. Although studies show that HTT is important for vesicular transport within axons, the types of cargo that HTT transports remain elusive. Previously, we found that HTT and Rab7 co-migrated within axons on a retrogradely moving single vesicle complex; however, the biological relevance of a moving HTT-Rab7 vesicle remains ambiguous. Here, we couple Drosophila genetics, in vivo microscopy, and biochemistry of mammalian vesicle membranes to unravel a unique retrogradely moving HTT-Rab7 vesicle complex aided by adaptor proteins Hip1 and Rilp. We found that the HTT-Rab7 vesicle is an axonal endolysosome that exhibits impaired motility upon disrupting autophagosome-endolysosome fusion with defective Syntaxin17, or by pharmacological inhibition of endolysosomal acidification and fusion. The HTT-Rab7 axonal endolysosome co-migrated with Phosphatidylinositol 3-phosphate (PI3P) and the BMP-signaling pathway receptor, Thickvein (Tkv), indicating that the HTT-Rab7 endolysosome is perhaps a signaling endolysosome. To test this proposal, we used a Drosophila nerve crush model to evaluate axon injury in vivo and found that HTT-Rab7 endolysosomes moved retrogradely away from the axon injury site, in contrast to synaptic vesicles which were immobile. Further, pharmacological inhibition of endolysosomal acidification and fusion impaired the motility of HTT-Rab7 endolysosomes, disrupting retrograde injury signaling as reported by the JNK-phosphatase, puckered. Together, our observations provide evidence for a retrogradely moving HTT-Rab7 signaling endolysosome, uncovering a novel role for HTT at the endolysosomal pathway in response to axon injury.

#637: Study of a novel neurosteroid as an acute neurotherapeutic following severe Traumatic Brain Injury

McGuire, Matthew J ; Gertz, Steven ; Wagner, Audrey ; Schwartz Byrne, Robin ; Newman, Michael ; Woodhouse, Colleen ; Poulsen, David ;

Department of Neurosurgery, SUNY University at Buffalo, School of Medicine and Biomedical Sciences;

Traumatic brain injury (TBI) is the leading cause of trauma related mortality and disability worldwide. Progesterone in many past preclinical studies is an effective treatment for models of brain injury, however, when attempted to translate these results to clinical trials it has failed. This was partially due to its poor solubility and short half-life, limiting progesterone’s ability to be properly delivered in emergent settings at effective levels of bioavailability. A novel neurosteroid, NTS-104, was developed to address these fundamental issues with the progesterone molecule. We tested the effectiveness of NTS-104 in adult male Wistar rats following severe TBI (sTBI). Animals underwent a T2 MRI scan 72 hours post injury to evaluate edema. We observed a significant reduction in cortical thickness and volume on the injured hemisphere when comparing the drug treated animals to vehicle (p=0.0186) (p<0.0001). 6 weeks post TBI we observed a drug dependent decrease in the size of thalamic calcifications detected by QSM (p=0.0525). At 5 weeks post TBI, cognitive assessments with the Morris water maze showed drug treated animals with either no difference from sham animals or significantly better performance from vehicle treated sTBI rats. Further, calcification volumes significantly negatively correlated with learning, (p=0.002), and memory (p=0.0018).

#632: Long-term effects of adolescent cannabis exposure: role of prenatal environment

Murlanova, Kateryna ; Jouroukhin, Yan ; Pletnikov, Mikhail ;

Department of Physiology and Biophysics, SUNY University at Buffalo;

The two-hit hypothesis of adult psychopathology predicts that a combination of prenatal and postnatal environmental insults increases risks for major psychiatric disorders, including psychotic disorders. Maternal immune activation (MIA) induced by in utero exposure to viral infection and adolescent cannabis use were associated with psychiatric conditions. Here, we hypothesized that MIA would interact with adolescent exposure to Δ⁹-tetrahydrocannabinol (THC), the major psychotropic ingredient of cannabis, to synergistically produce neurobehavioral abnormalities in adulthood, an environment x environment model (E x E). MIA was induced by administration of a viral mimetic, polyriboinosinic-polyribocytidilic acid [Poly (I:C), 5 mg/kg; IP] to gestating dams (CD1 mice, GD 12.5) and was validated by detecting increased levels of IFN-γ, IL-1β, IL-6 and TNF- α in maternal serum. Male and female offspring received THC (8 mg/kg; SC) or vehicle during adolescence (PND 30–51) and were evaluated in a series of behavioral tests as adults (PND 72–90). We found that E x E interaction increased exploratory behaviors in the Hole Board Test (increased head dipping), the Open Field (increased rearing), Elevated Plus Maze (increased open space exploration) and elevated sensitivity to amphetamine (2 mg/kg, IP) in female but not male mice. Our data indicate that E x E interaction synergistically produces neurobehavioral alterations in adult mice in a sex-dependent manner. On-going studies aim at identifying the cell type-specific molecular mechanisms underlying E x E interaction in this model.

#624: Life style change improves lupus life quality

Okamura, Kazuki K ; Phelps, Michael ; Jacob, Alexander A ; Alexander, Jessy A ;

Department of Biological Sciences, SUNY University at Buffalo, School of Medicine and Biomedical Sciences;

Systemic lupus erythematosus (lupus) is a global health problem where 20-80% patients display cognitive problems and central nervous system (CNS) dysfunction. Early diagnosis and treatment of lupus remains a clinical challenge. Exercise was shown to improve lupus nephritis but its effects in CNS lupus remains unknown. This study investigates the effects of controlled exercise (CE) that consisted of treadmill walking (5m/min for 10 min everyday) on experimental CNS lupus using the well-established mouse model, MRL/lpr mice. The MRL/lpr mice were subjected to CE from 8 weeks (preclinical) to 16 weeks (disease). Multiplex gene expression analysis revealed significant upregulation of genes involved in neurite growth, proliferation and synaptic plasticity, and a decrease in inflammatory genes including complement proteins, NFkB, chemokines and cytokines in exercised mice compared to the unmanipulated, age-matched controls. Exercise increased hippocampal area and reduced changes in cell morphology. The loss of blood-brain barrier integrity, astrogliosis and edema seen in MRL/lpr mice were reduced with exercise. Exercised mice performed better in behavioral assessments such as open field, nesting, and tail suspension test. For the first time our results show that a supervised, well regulated and controlled exercise regimen alleviates CNS lupus and could potentially serve as an intervention strategy to improve the quality of life. Exercise could also serve as an adjunct therapy for lupus and other neuroinflammatory diseases, thereby reducing the need for the current therapies with toxic side effects. The validity of the findings and a safe exercise regimen needs to be established by additional studies in patients. 

#642: Prognosticating Factors for Pediatric Concussion

Andrews, Mark ¹; Farooq, Osman ²;

¹Department of Medicine, ²Department of Pediatric Neurology, SUNY University at Buffalo, School of Medicine and Biomedical Sciences;

This study aims to identify prognostic indicators that are associated with either optimal or poor outcomes among pediatric concussion patients. Concussions are a common injury among children and adolescents. While most patients make full and uncomplicated recoveries, some experience persistent symptoms for weeks, months, and even years following injury. There is currently a paucity of data pertaining to prognostic indicators for severity of pediatric concussion. This study utilizes a database including premorbid, presenting, and longitudinal information for 235 pediatric concussion patients 1-19 years old. Chi square testing was used to measure association between outcomes and selected variables. The main outcomes tested are length of time for the patient to return to baseline following injury and whether the patient experienced school performance difficulty following concussion. Both outcomes are recorded as reported by the patient. Statistically significant associations were found between certain symptoms and patient outcomes. The primary recovery metric used was length of time for patient to return to baseline following injury. Patients who took longer than 14 days to recover were more likely to present with headaches (p=0.003), especially temporal (p=0.04) and occipital (p=0.01) headaches, than patients who recovered in 0-14 days. The longer recovery period was also associated with additional symptoms: photophobia (p=0.009), phonophobia (p=0.03), and vision disturbances (p=0.007). Difficulty in school was associated with frontal headaches (p=0.01).

#657: The presence of salivary proteins alters post-oral feedback

Ascencio Gutierrez, Verenice ¹; Martin, Laura E ²; James, Kimberly F ¹; Torregrossa, Ann-Marie ¹;

¹Department of Psychology, SUNY University at Buffalo; ²Department of Food Science and Technology, Oregon State University;

We have previously shown that salivary proteins (SPs) are upregulated in response to a quinine diet and SP expression is correlated with increases in meal size. As this suggested a role for SPs in post-oral feedback, we chose to examine the role of SPs in the gut using a licking suppression paradigm. Male, Long Evans rats, implanted with gastric catheters, were trained to lick a single bottle of Kool-Aid. As the animals licked they were simultaneously given a gastric infusion of 2mM quinine dissolved in water or donor saliva (with or without SPs). Donor saliva was collected from anesthetized rats, who were treated with injections of isoproterenol. All saliva was combined into a homogenous sample.  A subset was delivered to the catheterized rats with the SPs (5mg/ml). There was no difference in total licks to Kool-Aid when animals received either water or SPs alone in the gut (p = 0.17). Licking was suppressed within session (compared to the water control) when rats were infused with quinine (p = 0.02); however, when infused with quinine + SPs, licking increased to levels equivalent to the control group (p = 0.94). To determine whether the presence of protein itself rescued the lick response, the rats were infused with quinine + 5mg/ml bovine serum albumin (BSA). Licking to Kool-Aid decreased compared to the infusion of quinine + SPs (p = 0.03) and was equivalent to quinine + water (p = 0.52), suggesting the presence of protein alone does not reduce negative responding to quinine.

#659: Role of human alpha-synuclein(α-syn) & Leucine rich repeat kinase2(LRRK2) during neuronal cell death

Bajgain, Pratima ; Chakraborty, Piyali ; Gunawardena , Shermali ;

Department of Biological Sciences, SUNY University at Buffalo, College of Arts and Sciences;

Axonal transport is a complex process, which is very important for neuronal function and health. Any disruption to this process can result in axonal blockages seen in neurodegenerative diseases like Parkinson’s Disease (PD). α-syn, a PD causing protein, is the major constituent seen in accumulations known as Lewy bodies in both sporadic (sPD) and familial (fPD) forms of PD together with dopaminergic neuronal loss. LRRK2 is another PD protein that is mutated in fPD and is found localized to the periphery of Lewy bodies. Previous work in our lab showed that fPD LRRK2 mutations in the GTPase and WD40 domain rescued α-syn-mediated axonal blockages. Since axonal blockages can precede neuronal cell death, we tested the hypothesis that LRRK2 mutations in the GTPase and WD40 domain also rescues α-syn-mediated neuronal cell death. We observed neuronal cell death in Drosophila larval brains expressing wild type human LRRK2 (LRRK2-WT), indicating that excess LRRK2 is toxic to neurons. Larval brains expressing four fPD LRRK2 mutations: kinase domain mutants (LRRK2-G2019S and LRRK2-I2020T), WD40 domain mutant (LRRK2-G2385R) and GTPase domain mutant (LRRK2-Y1699C) also showed neuronal cell death, with enhanced amounts death seen in the WD40 domain mutant (LRRK2-G2385R), suggesting that perhaps this region is important for key functional associations. We next co-expressed α-syn-WT with LRRK2-WT and the fPD LRRK2 mutants. Surprisingly, co-expression of α-syn-WT and LRRK2 did not eliminate neuronal cell death. Together our observations suggest that while excess LRRK2 can induce neuronal cell death independent of α-syn, excess LRRK2 does not modulate α-syn-mediated cell death.

#621: Role of Complement activation in SARS-COV2 associated neuro-inflammation.

Bedi, Harneet ¹; Alexander, Jessy J ¹; Aalinkeel, Ravikumar ¹; Bruggemann, Liana ²; Schwartz, Stanley A ¹; Mahajan, Supriya D ¹;

¹Medicine, ²Biomedical Informatics, SUNY University at Buffalo, School of Medicine and Biomedical Sciences;

Neurological complications of SARS-CoV-2 infection are significant among patients with severe COVID-19. Neurovascular endothelial dysfunction, BBB breakdown, and CNS innate immune activation, potentially contributing to SARS-CoV-2-related neuropathology. Complement is a key innate immune pathway that rapidly responds to pathogens and cellular injury and amplifies a range of innate and adaptive immune processes. One of the key drivers of complement-mediated inflammation is the C5 cleavage product, C5a, which acts through two different seven-transmembrane receptors, namely C5aR1 and C5aR2 and mediates various biological activities. We have recently shown that SARS-COV-2 viral spike protein (Cat NRI-52307, BEI Resources Inc) via the ACE2 receptor on brain microvascular endothelial cell (BMVEC)  induces endothelial inflammation via hyper cytokinemia, specifically increased levels of IL-6, IL-8,IL-23, IL-33 and TNF-α resulting in altered BBB integrity which facilitates neuro-invasion of SARS-COV-2. We observed a significant increase in  C5aR1 and C5aR2 gene and protein expression in BMVEC treated with the SARS-CoV-2- recombinant spike protein as compared to the untreated control using QPCR, and western blot analysis. Additionally, we observed that the C5a receptor antagonist via targeting C5a significantly reduces inflammatory response. Further, studies using single cell gene expression analysis with Visium spatial technologies available from 10XGenomics are ongoing and will enable examining the effects of SARS-CoV-2- recombinant spike protein and /or C5a/C5aR antagonist in a single BMVEC that will facilitate a holistic understanding of the neuro-inflammatory response associated with SARS-CoV-2. We believe that targeting the C5a-C5aR1 signaling axis may have therapeutic benefit in SARS-COV-2 associated neuroinflammation.