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H Cline

Publications and source records attributed to H Cline.

11 recordsLinked to original sources

The molecular basis of CaMKII function in synaptic and behavioural memory.

Long-term potentiation (LTP) in the CA1 region of the hippocampus has been the primary model by which to study the cellular and molecular basis of memory. Calcium/calmodulin-dependent protein kinase II (CaMKII) is necessary for LTP induction, is persistently activated by stimuli that elicit LTP, and can, by itself, enhance the efficacy of synaptic transmission. The analysis of CaMKII autophosphorylation and dephosphorylation indicates that this kinase could serve as a molecular switch that is capable of long-term memory storage. Consistent with such a role, mutations that prevent persistent activation of CaMKII block LTP, experience-dependent plasticity and behavioural memory. These results make CaMKII a leading candidate in the search for the molecular basis of memory.

Animals↗

Nitric oxide is an essential negative regulator of cell proliferation in Xenopus brain.

Mechanisms controlling the transition of a neural precursor cell from proliferation to differentiation during brain development determine the distinct anatomical features of the brain. Nitric oxide (NO) may mediate such a transition, because it can suppress DNA synthesis and cell proliferation. We cloned the gene encoding the neuronal isoform of Xenopus NO synthase (XNOS) and found that in the developing brain of Xenopus tadpoles, a zone of XNOS-expressing cells lies adjacent to the zone of dividing neuronal precursors. Exogenous NO, supplied to the tadpole brain in vivo, decreased the number of proliferating cells and the total number of cells in the optic tectum. Conversely, inhibition of NOS activity in vivo increased the number of proliferating cells and the total number of cells in the optic tectum. NOS inhibition yielded larger brains with grossly perturbed organization. Our results indicate that NO is an essential negative regulator of neuronal precursor proliferation during vertebrate brain development.

Animals↗

The scaffold protein, Homer1b/c, regulates axon pathfinding in the central nervous system in vivo.

Homer proteins are a family of multidomain cytosolic proteins that have been postulated to serve as scaffold proteins that affect responses to extracellular signals by regulating protein-protein interactions. We tested whether Homer proteins are involved in axon pathfinding in vivo, by expressing both wild-type and mutant isoforms of Homer in Xenopus optic tectal neurons. Time-lapse imaging demonstrated that interfering with the ability of endogenous Homer to form protein-protein interactions resulted in axon pathfinding errors at stereotypical choice points. These data demonstrate a function for scaffold proteins such as Homer in axon guidance. Homer may facilitate signal transduction from cell-surface receptors to intracellular proteins that govern the establishment of axon trajectories.

Animals↗

Dendritic dynamics in vivo change during neuronal maturation.

In vivo imaging of optic tectal neurons in the intact Xenopus tadpole permits direct observation of the structural dynamics that occur during dendritic arbor formation. Based on images of single DiI-labeled neurons collected at daily intervals over a period of 6 d, we divided tectal cell development into three phases according to the total length of the dendritic arbor. During phase 1, the cell differentiates from a neuroepithelial cell type and extends an axon out of the tectum. The total dendritic branch length (TDBL) is <100 micrometers. During phase 2, when TDBL is 100-400 micrometers, the dendritic arbor grows rapidly. During phase 3, when TDBL is >400 micrometers, the dendritic arbor grows slowly and appears stable. Neurons at different positions along the rostrocaudal developmental axis of the tectum were imaged at 2 hr intervals over 6 hr and at 24 hr intervals over several days. Images collected at 2 hr intervals were analyzed to determine rates of branch additions and retractions. Morphologically complex, phase 3 neurons show half the rate of branch additions and retractions as phase 2 neurons. Therefore, rapidly growing neurons have dynamic dendritic arbors, and slower-growing neurons are structurally stable. The change in growth rate and dendritic arbor dynamics from phase 2 to phase 3 correlates with the developmental increase in synaptic strength in neurons located along the rostrocaudal tectal axis. The data are consistent with the idea that strong synaptic inputs stabilize dendritic arbor structures and that weaker synaptic inputs are permissive for a greater degree of dynamic rearrangements and a faster growth rate in the dendritic arbor.

Animals↗

Design and experimental verification of thin acoustic lenses for the coagulation of large tissue volumes.

Large focal volumes are desired in ultrasound surgery to reduce the total treatment time when large tumours are thermally coagulated. Phased arrays are capable of producing enlarged focal volumes in addition to providing the ability for on-line modification of focal shape and location. Although phased arrays have several advantages over their non-phased counterparts, the complexity of these arrays also presents some disadvantages regarding cost and complexity. One less costly alternative is the use of thin acoustic lenses to alter the field shape of a single-focus transducer. Four polystyrene lenses have been designed using the sector-vortex principle developed for phased arrays by Cain and Umemura. Measurements of the acoustic fields produced with the lenses are in good agreement with the simulated fields. The transmission measurements through each of the four lenses ranged from 76% to 84%, and over 52 W of total acoustic power has been delivered through each of the lenses during in vivo experiments without any damage to the lenses or the transducer. The in vivo results showed an increase in rate of necrosis to 10.1 +/- 1.4 cm3 h-1 using the mode 4 lens, or 5.2 +/- 0.7 times higher than the focused transducer alone.

Acoustic Stimulation↗

Computer-assisted three-dimensional planning in craniofacial surgery.

Three-dimensional surface reconstruction from computed tomographic (CT) data has been used to plan craniofacial operations. Cephalometric and anthropometric databases were integrated with three-dimensional CT reconstructions to quantitate the skeletal deformity and to assist in the design of the surgical procedure. Interactive techniques were developed to simulate osteotomies and skeletal movements in three dimensions on the computer-generated surface images. The ocular globes were referenced to position the orbital segments; i.e., the osteotomized segments were transposed into normal anatomic relationship with respect to the eyes. The measurements from the computer graphic simulation were used intraoperatively to establish the correct position of the skeletal segments.

Adolescent↗

Routine quantitative analysis of brain and cerebrospinal fluid spaces with MR imaging.

A computerized system for processing spin-echo magnetic resonance (MR) imaging data was implemented to estimate whole brain (gray and white matter) and cerebrospinal fluid volumes and to display three-dimensional surface reconstructions of specified tissue classes. The techniques were evaluated by assessing the radiometric variability of MR volume data and by comparing automated and manual procedures for measuring tissue volumes. Results showed (a) the homogeneity of the MR data and (b) that automated techniques were consistently superior to manual techniques. Both techniques, however, were affected by the complexity of the structure, with simpler structures (eg, the intracranial cavity) showing less variability and better spatial correlation of segmentation results between raters. Moreover, the automated techniques were completed for whole brain in a fraction of the time required to complete the equivalent segmentation manually. Additional evaluations included interrater reliability and an evaluation that included longitudinal measurement, in which one subject was imaged sequentially 24 times, with reliability computed from data collected by three raters over 1 year. Results showed good reliability for the automated segmentation procedures.

Body Fluid Compartments↗

No change in NMDA receptor-mediated response rise-time during development: evidence against transmitter spillover.

Glutamatergic transmission was examined in tadpole optic tectum to test the possibility that transmitter concentration reaching N-methyl-D-aspartate (NMDA) receptors increases over development. Pharmacologically isolated NMDA receptor-mediated transmission was monitored with whole-cell recordings. Synaptic responses were recorded from cells at different locations in the optic tectum, corresponding to different stages of development. Rise-times and decay-times of NMDA currents were analyzed. We found no significant correlation between rise-time and developmental stage. As NMDA rise-times can correlate with concentration for glutamate concentrations below 200 microM, these results argue that, if there is developmental variation in transmitter concentration, this occurs for values greater than 200 microM. Furthermore, we found a correlation between rise-times and decay-times, consistent with a model in which transmitter concentration is high and rise-time is controlled by channel closings. These results argue against synaptic models in which low concentrations of transmitter (as by spillover from nearby release sites) selectively activates NMDA receptors.

Animals↗