Role of fibroblast growth factor 8 in neurite outgrowth from spiral ganglion neurons in vitro.

Many neurons degenerate after injuries resulting from overstimulation, drugs, genetic mutations, and aging. Although several growth factors and neurotrophins delay degeneration and promote regrowth of neural processes, the role of fibroblast growth factor 8 (FGF8) in mammalian spiral ganglion neurons (SGN) neurite outgrowth has not been examined. This study develops and uses SGN cell cultures suitable for experimental analysis, it investigates whether FGF8a and FGF8b isoforms affect the neurite outgrowth from SGN cultured in vitro.

Protein kinases regulate glycine receptor binding in brain stem auditory nuclei after unilateral cochlear ablation.

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).

Phosphorylated cAMP response element-binding protein levels in guinea pig brainstem auditory nuclei after unilateral cochlear ablation.

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.

Phospho-CREB and other phospho-proteins: improved recovery from brain tissue.

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.

Regulation of NT-3 and BDNF levels in guinea pig auditory brain stem nuclei after unilateral cochlear ablation.

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%.

ERK and SAPK signaling in auditory brainstem neurons after unilateral cochlear ablation.

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.

TrkB levels in the cochlear nucleus after unilateral cochlear ablation: correlations with post-lesion plasticity.

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.