RESUMO
The purpose of the 'First Regional Healthy Aging and Dementia Research Symposium' was to discuss the latest research in healthy aging and dementia research, public health trends related to neurodegenerative diseases of aging, and community-based programs and research studying health, nutrition, and cognition. This symposium was organized by the Garrison Institute on Aging (GIA) of the Texas Tech University Health Sciences Center (TTUHSC), and was held in Lubbock, Texas, October 24-25, 2018. The Symposium joined experts from educational and research institutions across the United States. The two-day Symposium included all GIA staff and researchers. Students, postdoctoral fellows, and faculty members involved in dementia research presented at the Symposium. Healthcare professionals, from geriatricians to social workers working with patients with neurodegenerative diseases, also presented. In addition, experts traveled from across the United States to participate. This event was comprised of multiple sessions, each with several oral presentations, followed by questions and answers, and discussion.
Assuntos
Pesquisa Biomédica/tendências , Congressos como Assunto/tendências , Demência/epidemiologia , Demência/psicologia , Envelhecimento Saudável/fisiologia , Envelhecimento Saudável/psicologia , Pesquisa Biomédica/métodos , Humanos , Texas/epidemiologiaRESUMO
Glycinergic synaptic inhibition is part of acoustic information processing in brain stem auditory pathways and contributes to the regulation of neuronal excitation. We found previously that unilateral cochlear ablation (UCA) in young adult guinea pigs decreased [3H]strychnine binding activity in several brain stem auditory nuclei. This study determined if the UCA-induced deficit could be regulated by protein kinase C (PKC), protein kinase A (PKA) or Ca2+/calmodulin-dependent protein kinase II (CaMKII). The specific binding of [3H]strychnine was measured in slices of the dorsal (DCN), posteroventral (PVCN) and anteroventral (AVCN) cochlear nucleus (CN), the lateral (LSO) and medial (MSO) superior olive, and the inferior colliculus (IC) 145 days after UCA. Tissues from age-matched unlesioned animals served as controls. UCA induced deficits in specific binding in the AVCN, PVCN, and LSO on the ablated side and in the MSO bilaterally. These deficits were reversed by 3 microM phorbol 1,2-dibutyrate, a PKC activator, or 0.2 mM dibutyryl-cAMP, a PKA activator. However, 50 nM Ro31-8220, a PKC inhibitor, and 2 microM H-89, a PKA inhibitor, had no effect in unlesioned controls and after UCA. In contrast, 4 microM KN-93, a CaMKII inhibitor, relieved or reversed the UCA-induced binding deficits and elevated binding in the IC. These findings suggest that a UCA-induced down-regulation of glycine receptor synthesis may have occurred via reduced phosphorylation of proteins that control receptor synthesis; this effect was reversed by diminishing CaMKII activity or increasing PKC and PKA activity.
Assuntos
Vias Auditivas/fisiologia , Tronco Encefálico/metabolismo , Cóclea/lesões , Lateralidade Funcional , Proteínas Quinases/fisiologia , Receptores de Glicina/metabolismo , Animais , Tronco Encefálico/anatomia & histologia , Tronco Encefálico/efeitos dos fármacos , Cóclea/fisiopatologia , Interações Medicamentosas , Ativação Enzimática/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Feminino , Cobaias , Masculino , Ligação Proteica/efeitos dos fármacos , Ligação Proteica/fisiologia , Proteínas Quinases/farmacologia , Trítio/metabolismoRESUMO
After left unilateral cochlear ablation (UCA) in young adult guinea pigs, the appearance of plasticities in auditory pathways suggested altered gene expression and modified phenotypic behaviors of auditory neurons. Because phosphorylated cyclic-AMP response element-binding protein (CREB-P) is a transcription factor that binds to certain genes to facilitate their expression, CREB-P levels were measured after UCA and correlated with postablation plasticities. After UCA, Western blotting was employed to quantify CREB-P levels and illustrate CREB levels in the anteroventral (AVCN), posteroventral (PVCN), and dorsal (DCN) cochlear nucleus; the lateral (LSO) and medial superior olive (MSO); the medial nucleus of the trapezoid body (MNTB); and the central nucleus of the inferior colliculus (ICc) for up to 145 days. We also quantified the levels of several protein synthesis regulators and synaptic markers in the AVCN at 60 days. Sucrose-based extraction buffer improved CREB-P recovery. CREB-P levels became depressed at 3 and 7 postablation days, except in the PVCN, where they were elevated at 7 days, and in the ICc, where they were elevated at both times. At 60 days, CREB-P levels in all the nuclei were elevated. In the AVCN, levels of the protein synthesis regulators and synaptic markers were also elevated at 60 days. By 145 days, CREB-P levels again declined, except in the AVCN, where elevations persisted and increased on the ablated side, and in the ICc, where CREB-P elevations remained. The changes in CREB-P levels coincided with several plasticities in glutamatergic and glycinergic transmitter release and receptor activities, and alterations in neurotrophic support, that developed after UCA. These findings suggest that UCA altered CREB-P levels, which in turn might have contributed to plasticities that appear after UCA.
Assuntos
Vias Auditivas/metabolismo , Tronco Encefálico/metabolismo , Cóclea/lesões , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Plasticidade Neuronal/fisiologia , Gânglio Espiral da Cóclea/lesões , Animais , Biomarcadores/metabolismo , Cóclea/fisiopatologia , Núcleo Coclear/metabolismo , Denervação , Modelos Animais de Doenças , Regulação da Expressão Gênica/fisiologia , Cobaias , Perda Auditiva Neurossensorial/metabolismo , Perda Auditiva Neurossensorial/fisiopatologia , Colículos Inferiores/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Núcleo Olivar/metabolismo , Terminações Pré-Sinápticas/metabolismo , Receptores de Glutamato/metabolismo , Receptores de Glicina/metabolismo , Gânglio Espiral da Cóclea/fisiopatologia , Sinapses/metabolismo , Transmissão Sináptica/fisiologia , Regulação para Cima/fisiologiaRESUMO
During attempts to quantify levels of phosphorylated cAMP response element binding protein (CREB-P) in guinea pig brain stem auditory nuclei by Western blotting, we compared the decay of CREB-P levels when tissues were homogenized in traditional Lysis buffer containing detergents or in 50 mM Tris-HCl buffer containing 0.32 M sucrose. The decay of CREB-P levels was retarded considerably in the Tris-Sucrose medium as compared to the Lysis buffer. Similarly, the levels of two other phospho-proteins, extracellular regulated kinases (ERK1/2-P) and stress activated protein/Jun-N-terminal kinase (SAP/JNK-P), were better preserved by the Tris-Sucrose medium. These findings imply that the detergents typically present in the Lysis buffer may disrupt organelles and increase the exposure of soluble phospho-proteins to hydrolyzing enzymes. In contrast, such exposure was probably minimized in the Tris-Sucrose medium, which is thought to preserve organelle integrity.
Assuntos
Química Encefálica , Tronco Encefálico/metabolismo , Técnicas de Laboratório Clínico , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/análise , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/isolamento & purificação , Animais , Western Blotting , Cobaias , Fosforilação , Proteínas/análise , Proteínas/isolamento & purificação , TrometaminaRESUMO
Injury to areas of the central nervous system can alter neurotrophin levels, which may influence postlesion neuronal survival and plasticity. To determine if sensorineural hearing loss induces such changes, we used an enzyme-linked immunosorbent assay (ELISA) to measure neurotrophin-3 (NT-3) and brain-derived neurotrophic factor (BDNF) levels in adult guinea pig brain stem auditory nuclei 3-60 days after a unilateral cochlear ablation (UCA). After UCA, which destroyed the cochlea and cochlear nerve on one side, NT-3 levels were usually depressed at 3 days by 22-44% but became elevated transiently at 7 days by 28-124%. BDNF levels were elevated transiently by 50% on the ablated side in the anteroventral (AVCN) and posteroventral (PVCN) cochlear nucleus at 3 days and may have signaled support for the survival of deafferented neurons. Coincident elevation at 3 and 7 days of BDNF or NT-3 and phosphorylated extracellular signal-regulated protein kinase 2 (ERK2-P) suggested a relationship to stimulated signal transduction activity. Elevated neurotrophin levels may have contributed to synaptogenesis in the AVCN and the superior olive and to changes in the synaptic biochemistry in the auditory nuclei after UCA. In contrast, deficiencies or failure to elevate neurotrophin levels within several days of the UCA correlated with upregulation of phosphorylated stress-activated protein kinase (SAPK-P), suggesting a relationship with stress-activated signal transduction and with the sparse degeneration of fibers observed in some of the auditory nuclei after UCA.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/metabolismo , Cóclea/lesões , Nervo Coclear/lesões , Núcleo Coclear/metabolismo , Perda Auditiva Neurossensorial/metabolismo , Neurotrofina 3/metabolismo , Animais , Sobrevivência Celular/fisiologia , Cóclea/fisiopatologia , Nervo Coclear/fisiopatologia , Denervação , Modelos Animais de Doenças , Feminino , Lateralidade Funcional/fisiologia , Cobaias , Perda Auditiva Neurossensorial/fisiopatologia , Masculino , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 8 Ativada por Mitógeno/metabolismo , Degeneração Neural/metabolismo , Degeneração Neural/fisiopatologia , Regeneração Nervosa/fisiologia , Plasticidade Neuronal/fisiologia , Transdução de Sinais/fisiologia , Regulação para Cima/fisiologiaRESUMO
Unilateral cochlear ablation (UCA) in adults deafferented one cochlear nucleus (CN) and induced several plasticities in central auditory pathways. To assess whether signal transduction could contribute to these changes, we determined if UCA induced activity in the extracellular signal-regulated kinase (ERK) and the stress-activated protein kinase (SAPK) signal transduction pathways. Using Western blots, we measured phosphorylated ERK1 (ERK1-P), ERK2 (ERK2-P), p46 and p54 SAPK (SAPK-P) and c-Jun (c-Jun-P) levels in the major subdivisions of the CN, the principal nuclei of the superior olivary complex (SOC) and the central nucleus of the inferior colliculus (ICc) for up to 145 days postablation. ERK1-P and ERK2-P were typically elevated at 7 and 145 days but depressed at 30 days, 60 days, or both. In addition, ERK1-P and ERK2-P were elevated at 3 days in the anteroventral (AVCN) and posteroventral CN (PVCN). Immunohistochemical labeling indicated that after 5 days, most ERK1/2-P in the CN was in neuronal nuclei. Only minor changes were evident in total ERK1 and ERK2 levels. Several correlations were evident between the postablation plasticities observed previously and altered ERK1-P and ERK2-P levels. Few changes were found in SAPK-P and c-Jun-P levels. Concomitant elevations of SAPK-P and c-Jun-P were not evident, except in the superficial dorsal CN (DCN) at postablation day 3, consistent with a cell-stress response. These findings suggest that signals induced as a consequence of UCA are transduced mainly through the neuronal ERK pathway. This activity probably influenced gene expression and cytoplasmic regulatory mechanisms that contributed to the plasticities induced by UCA.
Assuntos
Tronco Encefálico/enzimologia , Núcleo Coclear/enzimologia , Núcleo Coclear/lesões , Proteínas Quinases Ativadas por Mitógeno/biossíntese , Neurônios/enzimologia , Animais , Tronco Encefálico/química , Núcleo Coclear/química , Feminino , Cobaias , Sistema de Sinalização das MAP Quinases/fisiologia , Masculino , Proteínas Quinases Ativadas por Mitógeno/análise , Proteínas Quinases Ativadas por Mitógeno/genética , Neurônios/química , FosforilaçãoRESUMO
Tyrosine kinase B (TrkB) levels in the adult guinea pig cochlear nucleus (CN) were determined from Western blots for up to 60 days after unilateral cochlear ablation (UCA). Compared to TrkB levels on the intact side, those on the lesioned side were elevated in the anteroventral CN (AVCN) at 7 and 60 days and in the posteroventral CN (PVCN) at 30 days. TrkB levels were depressed in the AVCN and the dorsal CN (DCN) at 30 days. Elevations in the AVCN on the lesioned side at 7 days coincided with a period of synaptogenesis. Other changes were temporally related to up- or downregulations of transmitter release and synaptic receptor activities that were evident after UCA. The findings suggest that changes in signaling through TrkB may contribute to the plasticities in the CN that were evident after UCA.