Biomedical subjects

S M Wu

Publications and source records attributed to S M Wu.

At least 145 records · Page 8Linked to original sources

PubMed1989

Effects of prolonged light exposure, GABA, and glycine on horizontal cell responses in tiger salamander retina.

1. The effects of prolonged light exposure, gamma-aminobutyric acid (GABA), and glycine on the horizontal cell (HC) light responses were studied in the superfused flat-mounted isolated retinas of the larval tiger salamander. 2. Under prolonged dark-adapted conditions, the time-to-peak of the HC light response was approximately 2-4 s, and after the termination of prolonged (6-8 min) light exposure, the time-to-peak became approximately 0.5-1 s. 3. This prolonged light-induced change in response rise time was not observed in either photoreceptors or bipolar cells, and thus the change in HC response rise time may occur postsynaptically in the HC membrane. 4. Application of 100 microM of GABA mimicked prolonged darkness and reversibly slowed down the HC response rise time, and application of 100 microM bicuculline mimicked prolonged light exposure and reversibly sped up the HC response rise time. 5. Glycine also slowed down the HC response rise course, but its effect was not observable until the concentration was raised to 1-3 mM. Strychnine did not exert any effect on HC responses when applied alone, but it could reverse the glycine actions. 6. The actions of glycine disappeared in the presence of bicuculline, indicating that the GABA and glycine pathways were probably not independent. Application of 5-10 mM glycine produced an increase of flow of preloaded 3H-GABA from the retina. 7. These results indicate that GABA may be the primary modulator that slows down the kinetics of the postsynaptic membrane proteins in the HCs. The extracellular concentration of GABA is probably high in prolonged darkness, and it is low after prolonged light exposure. Glycine, when applied at high dose, results in an increase of GABA release that slows down the HC response time course. 8. Prolonged darkness and light exposure appear to modulate the HC response in the time domain through GABA, and this change in HC response time course is probably responsible for shaping the bipolar cell responses and making the retinal signals more transient under light-adapted conditions.

Acclimatization↗

PubMed1989

Effects of background illumination on the horizontal cell responses in the tiger salamander retina.

Synaptic transmission between photoreceptors and horizontal cells (HCs) was studied in the flat-mounted isolated retinas of the tiger salamander. Background illumination expedited the rise time of the HC light response, and the HC response rise time (HCRRT) reached steady state about 2 sec after the onset of the background illumination. The change in HCRRT is probably responsible for the background-induced enhancement of the HC responses to short light stimuli. The amplitude of the HC responses to 100 msec light steps in the presence of background illumination was 2-5 times larger than that measured under dark-adapted conditions. Background illumination exerted little effect on the response rise time in cones and bipolar cells, and thus it caused no significant response enhancement in those cells. The background-induced change in HCRRT correlated closely with the rod voltage but not with the HC voltage. These results suggest that the background-induced change in HCRRT is probably mediated by postsynaptic events in HCs because no significant time course change is observed in photoreceptors and bipolar cells (which share the same synapses with the HCs). A suppressive rod action on the cone inputs in HCs may be responsible for modulating the HCRRT. By shortening the HCRRT, background illumination regulates the frequency response of the photoreceptor-HC synapse and alters the capacity of spatial resolution of retinal bipolar cells.

Ambystoma↗

PubMed1989

A preliminary report on the derivatization-gas chromatographic determination of nalidixic acid and 3,7-dicarboxynalidixic acid in urine.

A preliminary study on the gas chromatographic analysis of urine spiked with nalidixic acid and 3,7-dicarboxynalidixic acid is described. The method is based on the transfer of an ion-pair of NA or CNA with tetradecyldimethylbenzylammonium chloride (benzalkonium chloride) from alkaline aqueous solution into methyl isobutyl ketone (MIBK), the organic phase, where these salts are derivatized with pentafluorobenzyl bromide. The derivatives formed by this process are chromatographed on an analytical column packed with 1.5% OV-101 and detected with a flame-ionization detector. Several parameters affecting the transfer and/or derivatization of NA or CNA were investigated. These parameters include the phase transfer catalyst employed, the organic solvent used, the concentration of the acid and base added, the amount of derivatizing agent required, and reaction time and temperature.

Chemical Phenomena↗

PubMed1988

The coexistence of three neuroactive substances in amacrine cells of the chicken retina.

For the first time in the vertebrate retina, we have demonstrated that 3 neuroactive substances coexist in a single neuron. Using immunofluorescence, immunoperoxidase, and autoradiographic techniques, we found a subpopulation of amacrine cells in the chicken retina exhibited enkephalin- and neurotensin-like immunoreactivity and high-affinity uptake of glycine. Biochemical evidence showed that glycine release in this retina is inhibited by enkephalin and enhanced by neurotensin1.14. It is therefore possible that the two peptides form a push-pull system in self-regulating the glycine release from retinal amacrine cells.

Animals↗

PubMed1988

Melatonin enhances horizontal cell sensitivity in salamander retina.

Intracellular electrophysiological recording techniques were utilized to investigate the possible function of retinal melatonin in the larval tiger salamander. Endogenous retinal melatonin was present and appeared to bind a membrane-enriched fraction of the salamander retina, as determined by radioimmunoassay and receptor binding studies. Melatonin added through the perfusion bath to flat-mounted retinas resulted in a horizontal cell (HC) hyperpolarization of 10-20 mV. Additionally, the amplitude of HC responses to short test flashes increased in the presence of melatonin. Voltage-intensity plots revealed that application of 500 microM of melatonin caused an increase of the HC light sensitivity and this effect was reversible. These results suggest that melatonin synthesized and released during the dark period of the diurnal cycle may alter the sensitivity of second-order neurons at a time of day when photopic input is at its lowest level.

Animals↗

PubMed1988

Localization of tyrosine-hydroxylase-like-immunoreactive amacrine cells in the larval tiger salamander retina.

Immunocytochemistry was used to localize the populations of tyrosine-hydroxylase-like (TH)-immunoreactive cells in the tiger salamander retina. Ninety percent of these cells possessed somas that were situated in the innermost cell row of the inner nuclear layer and were classified as amacrine cells. Ten percent of TH-immunoreactive somas were located in the ganglion cell layer and were tentatively designated as those of displaced amacrine cells. The processes of TH-immunoreactive cells ramified most heavily in sublayer 1 of the inner plexiform layer, while a relatively small number of TH-labelled processes distributed in sublayers 3 and 5. Less than 1% of TH-immunoreactive cells in the amacrine cell layer exhibited a short process of somal origin that extended distally toward the outer plexiform layer. However, these processes did not cross the whole of the inner nuclear layer, and no immunolabelling was observed in the outer plexiform layer. An examination of retinal whole-mounts revealed that TH-immunoreactive amacrine and displaced amacrine cells were distributed throughout the center and periphery of the retina. The density of TH-immunolabelled amacrine cells was calculated to be 49 +/- 13 (mean +/- standard error) cells per mm2. The vast majority of TH-immunoreactive amacrine and displaced amacrine cells exhibited a stellate appearance and gave rise to three or more primary dendrites. A few TH-amacrine and displaced amacrine cells possessed two primary dendrites that emerged from opposite sides of their somas. The processes of TH-immunoreactive cells were generally poorly branched and varicose with terminal branches sometimes appearing thin and beaded. Because some TH-immunolabelled processes were very long, there was considerable overlap between the dendritic fields of neighboring TH-cells. Lastly, individual TH-immunoreactive amacrine and displaced amacrine cells were often observed in whole-mounts to provide processes that ramified at more than one level of the inner plexiform layer.

Ambystoma↗

PubMed1988

Quantitative studies of enkephalin's coexistence with gamma-aminobutyric acid, glycine and neurotensin in amacrine cells of the chicken retina.

Previous double-label studies demonstrate that enkephalin coexists with gamma-aminobutyric acid, glycine or neurotensin in amacrine cells of the chicken retina. The present study utilizes double- and triple-label paradigms to quantitatively analyze these coexisting relationships. Twenty-eight percent of enkephalin-like immunoreactive amacrine cells were found to exhibit high-affinity uptake of [3H]GABA, while 53% of enkephalin-amacrine cells specifically accumulate [3H]glycine. Moreover, the present study predicts that at least 26% of enkephalin-amacrine cells which accumulate [3H]glycine should also be immunoreactive for neurotensin.

Animals↗

PubMed1988

Localization of neurotensin-like immunoreactive amacrine cells in the larval tiger salamander retina.

Light microscopic immunocytochemistry was used to localize the populations of NT-like immunoreactive amacrine cells in the larval tiger salamander retina. Seventy-nine percent of NT-immunostained cells observed in transverse cryo-prepared sections were classified as Type 1 amacrine cells. Another 6% were classified as Type 2 amacrine cells, while 15% of the NT-cells had their cell bodies situated in the ganglion cell layer and were tentatively designated as displaced amacrine cells. Each type of NT-like immunoreactive cell was observed in the central and peripheral retina. NT-immunostained processes were observed to ramify in sublayers 3 and 5 of the inner plexiform layer. An examination of retinal whole mounts revealed that NT-amacrine cells were distributed throughout the center and periphery of the retina at a density of 82 +/- 24 cells/mm2. The dendritic fields of NT-immunostained amacrine and displayed amacrine cells were observed to be either symmetrically or asymmetrically distributed about their somas. Symmetrical dendritic fields were generally oval-shaped and ranged in diameter from 250 to 500 micron (major axis) by 150 to 250 micron (minor axis). Asymmetrical dendritic fields were observed to encompass one-half or less of an imaginary circle surrounding their soma of origin and were orientated in all directions. The processes forming asymmetrical dendritic fields ranged from 75 to 260 micron in length. Furthermore, partial overlap was often observed between the dendritic fields of adjacent NT-amacrine cells.

Animals↗

PubMed1988

The off-overshoot responses of photoreceptors and horizontal cells in the light-adapted retinas of the tiger salamander.

Depolarizing overshoot responses at the cessation of a test light step were observed in horizontal cells (HCs) and in a population of photoreceptors (rodCS) in light-adapted retinas of the tiger salamander. An anode break regenerative conductance may contribute to the overshoot responses in rodcS(o-wave). The overshoot responses in HCs consist of two components: a fast alpha-wave whose amplitude and time course follow those of the o-wave; and a slow beta-wave whose amplitude and time course vary with the HC membrane voltage. These results are consistent with the notion that the alpha-wave is a postsynaptic response to the voltage overshoots of the o-waves in rodCS and the beta-wave is mediated by voltage-dependent conductances in the HC membrane. A possible function of the HC overshoot responses is to reset the amplitude of the light-adapted HC responses during repetitive or rapidly changing light stimulation.

Action Potentials↗

PubMed1988

Neural interactions mediating the detection of motion in the retina of the tiger salamander.

The neural circuitry underlying movement detection was inferred from studies of amacrine cells under whole-cell patch clamp in retinal slices. Cells were identified by Lucifer yellow staining. Synaptic inputs were driven by "puffing" transmitter substances at the dendrites of presynaptic cells. Spatial sensitivity profiles for amacrine cells were measured by puffing transmitter substances along the lateral spread of their processes. Synaptic pathways were separated and identified with appropriate pre- and postsynaptic pharmacological blocking agents. Two distinct amacrine cell types were found: one with narrow spread of processes that received sustained excitatory synaptic current, the other with very wide spread of processes that received transient excitatory synaptic currents. The transient currents found only in the wide-field amacrine cell were formed presynaptically at GABAB receptors. They could be blocked with baclofen, a GABAB agonist, and their time course was extended by AVA, a GABAB antagonist. Baclofen and AVA had no direct affect upon the wide-field amacrine cell, but picrotoxin blocked a separate, direct GABA input to this cell. The narrow-field amacrine cell was shown to be GABAergic by counterstaining with anti-GABA antiserum after it was filled with Lucifer yellow. Its narrow, spatial profile and sustained synaptic input are properties that closely match those of the GABAergic antagonistic signal that forms transient activity (described above), suggesting that the narrow-field amacrine cell itself is the source of the GABAergic interaction mediating transient activity in the inner plexiform layer (IPL). Other work has shown a GABAB sensitivity at some bipolar terminals, suggesting a population of bipolars as the probable site of interaction mediating transient action. The results suggest that two local populations of amacrine cell types (sustained and transient) interact with the two populations of bipolar cell types (transient forming and nontransient forming). These interactions underlie the formation of the change-detecting subunits. We suggest that local populations of these subunits converge to form the receptive fields of movement-detecting ganglion cells.

Animals↗

PubMed1988

Electrical coupling between rods and cones in the tiger salamander retina.

Electrical coupling between rods and cones was studied in the salamander (Ambystoma tigrinum) retina by measuring the light responses and spectral sensitivities of rods and cones and by measuring the voltage responses from a rod to current pulses injected into a cone. A population of 10-20% of the photoreceptors exhibited a mixed-response waveform of the rod and the cone under dark-adapted conditions, and a response waveform closely resembled that of a cone in the presence of background illumination. Lucifer yellow injection revealed that these cells are morphologically identical to rods, and thus they are named rodcs. Dark-adapted rodcs exhibited a rod-like spectral sensitivity with a peak at approximately 520 nm that shifted to a cone-like spectral sensitivity with a peak at approximately 620 nm in response to background light (Purkinje shift). The voltage response of a rodc to a -1-nA current step injected into an adjacent cone is approximately 3.6 times larger than that of a rod to the same current step. These results indicate that there is a population of rods (rodcs) in the tiger salamander retina that is strongly coupled to the cones and that these cells allow significant mixture of rod and cone signals at the photoreceptor level.

Ambystoma↗

PubMed1988

Sequence duplication and internal deletion in the integrated human papillomavirus type 16 genome cloned from a cervical carcinoma.

Integrated human papillomavirus type 16 (HPV16) sequences were cloned from a cervical carcinoma and analyzed by restriction mapping and nucleotide sequencing. The viral integration sites were mapped within the E1 and E2 open reading frames (ORFs). The E4 and E5 ORFs were entirely deleted. An internal deletion of 376 base pairs (bp) was found disrupting the L1 and L2 ORFs. Sequencing analysis showed that an AGATGT/ACATCT inverted repeat marked the deletion junction with two flanking direct repeats 14 and 8 bp in length. A 1,330-bp sequence duplication containing the long control region (LCR) and the E6 and E7 ORFs was also found. The duplication junction was formed by two 24-bp direct repeats with 79% (19 of 24) homology located within the LCR and the E2 ORF of the prototype viral genome, respectively. This observation leads us to propose that the initial viral integration involved an HPV16 dimer in which the direct repeats in tandem units recombined, resulting in reiteration of only a portion of the original duplication. A guanosine insertion between nucleotides 1137 and 1138 created a continuous E1 ORF which was previously shown to be disrupted. Results from this study indicate that sequence reiteration and internal deletion in the integrated, and possibly in the episomal, HPV16 genome are influenced by specific nucleotide sequences in the viral genome. Moreover, reiteration of the LCR/E6/E7 sequences further supports the hypothesis that the E6/E7 ORFs may code for oncogenic proteins and that regulatory signals in the LCR may play a role in cellular transformation.

Base Sequence↗

PubMed1988

Synaptic transmission from rods to horizontal cells in dark-adapted tiger salamander retina.

Synaptic transmission between rods and horizontal cells (HCs) was studied by simultaneously recording from the pre- and postsynaptic cells in the living retinal slices of the larval tiger salamander. Voltage tails after the termination of a flash were observed in dark-adapted rods and HC somata, but not in cones. These simultaneously recorded voltage tails were used to isolate the rod input from the cone input to the HCs. The rod voltage versus the HC voltage in the response tails yielded the input-output relation of the rod-HC synapse. Two factors, a luminance-dependent signal delay between the rod and HC responses and a voltage-dependent depolarizing current in the HCs, complicate the analysis of the rod-HC synapse. A single input-output relation of the rod-HC synapse was obtained after these two factors had been corrected. The rod-HC synapse was approximately linear near the rod dark potential (-40 mV) with a synaptic gain around 12, but it became progressively nonlinear at more hyperpolarized potentials. The synaptic gain at Vrod = -50 mV was about 0.65.

Ambystoma↗

PubMed1987

Interactions between enkephalin and gamma-aminobutyric acid in the larval tiger salamander retina.

Both double-label and intracellular electrophysiological recording techniques were utilized to investigate the interactions between enkephalin and gamma-aminobutyric acid in the larval tiger salamander retina. Double-label studies revealed that the vast majority (greater than 96%) of enkephalin-immunostained amacrine cells also exhibit high affinity uptake of [3H]gamma-aminobutyric acid. Electrophysiological evidence demonstrated that morphine and gamma-aminobutyric acid exert opposite effects on a population of On-Off ganglion cells. gamma-Aminobutyric acid decreased the activity of these cells, while enkephalin increased their activity. These findings support the idea that opiate-mediated pathways inhibit GABAergic pathways in the vertebrate retina.

Animals↗

PubMed1987

Light-dependent synaptic delay between photoreceptors and horizontal cells in the tiger salamander retina.

A long delay between the time to peak of the photoreceptor and horizontal cell light responses was observed in dark-adapted tiger salamander retinas. The rise of the HC light responses follows a biphasic time-course which is probably mediated by the removal of the rod and cone transmitters, but not by voltage-dependent mechanisms in the HC membrane. A bleaching white light shortens the signal delay and results in a faster and monophasic rise of the HC light responses mediated predominantly by the cones.

Action Potentials↗

PubMed1987

Changes in response waveform of retinal horizontal cells during dark and light adaptation.

Changes in response waveform of retinal horizontal cells during dark and light adaptation were examined in the tiger salamander retina. In addition to sensitivity changes, the response waveform of HC was altered during visual adaptation: following bleaching or in the presence of background illumination, a transient depolarizing overshoot was observed at the cessation of the test flash. In contrast to the hyperpolarizing light responses, the depolarizing off-overshoots do not follow the standard threshold recovery time course during dark adaptation, neither do they obey Weber's law for increment thresholds during light adaptation. Simultaneous intracellular recordings from photoreceptors and HC revealed that the off-overshoot responses in light-adapted HC are probably originated from a population of cells in the photoreceptor layer which are strongly coupled with the cones.

Adaptation, Ocular↗

PubMed1987

Synaptic connections between neurons in living slices of the larval tiger salamander retina.

Synaptic connections between retinal cells were studied by recording simultaneously from pre- and postsynaptic cells in the retinal slice preparation. The time course and waveform of the pre- and postsynaptic light responses were monitored, and the postsynaptic voltage responses to presynaptic current injection were measured. Results obtained provide direct evidence showing that the rod-HBC and rod-HC synapses are sign-preserving, and the rod-DBC, HC-HBC and amacrine-DBC synapses are sign-inverting. Moreover, the synaptic delays between rods and bipolar cells are shorter than that between rods and horizontal cells. The methodology of making retinal slices and the subsequent recording and stimulation procedures are described. The advantages of the retinal slice preparation and its potential in retinal research are discussed.

Ambystoma↗

PubMed1986

Effect of vector type, host strains and transcription terminator on heterologous gene expression in yeast.

Using the surface antigen gene of the hepatitis B virus, and the promoter and terminator sequences of the yeast pho5 gene as a model system, a series of closely related expression plasmids were constructed to investigate the effect of vector type, genetic background of host strains and the presence of transcription terminator on the expression of heterologous gene in yeast. Plasmids carrying the replication origin of the 2 mu plasmids were found to be much more stable than those either independently or simultaneously carrying ars1 sequences. Gene expression was also higher with 2 micron-based plasmids. Yeast selection marker (trp1 or leu2) and therefore the host strains used did not have significant effects on gene expression. Addition of transcription terminator sequences downstream to the HBsAg gene also contributed only limited increases in gene expression levels.

Escherichia coli↗