Amacrine-signaled loss of intrinsic axon growth ability by retinal ganglion cells

The central nervous system (CNS) loses the ability to regenerate early during development, but it is not known why. The retina has long served as a simple model system for study of CNS regeneration. Here we show that amacrine cells signal neonatal rat retinal ganglion cells (RGCs) to undergo a profound and apparently irreversible loss of intrinsic axon growth ability. Concurrently, retinal maturation triggers RGCs to greatly increase their dendritic growth ability. These results suggest that adult CNS neurons fail to regenerate not only because of CNS glial inhibition but also because of a loss of intrinsic axon growth ability.     

Hyperacuity in cat retinal ganglion cells

Cat X retinal ganglion cells that can resolve sine gratings of only 2.5 cycles per degree can nevertheless respond reliably to displacements of a grating of approximately 1 minute of arc. This is a form of hyperacuity comparable in magnitude to that seen in human vision. A theoretical analysis of this form of hyperacuity reveals it to be a result of the high gain and low noise of ganglion cells. The hyperacuity expected for the best retinal ganglion cells is substantially better than that observed in behavioral experiments. Thus the brain, rather than improving on the retinal signal-to-noise ratio by pooling signals from many ganglion cells, is unable to make use of all the hyperacuity information present in single ganglion cell responses.     

Dendritic computation of direction selectivity by retinal ganglion cells

Direction-selective ganglion cells (DSGCs) in the retina respond strongly when stimulated by image motion in a preferred direction but are only weakly excited by image motion in the opposite null direction. Such coding represents an early manifestation of complex information processing in the visual system, but the cellular locus and the synaptic mechanisms have yet to be elucidated. We recorded the synaptic activity of DSGCs using strategies to observe the asymmetric inhibitory inputs that underlie the generation of direction selectivity. The critical nonlinear interactions between the excitatory and inhibitory inputs took place postsynaptically within the dendrites of the DSGCs.     

Synapses onto different morphological types of retinal ganglion cells

The percentage of bipolar and amacrine synapses onto ganglion cell dendrites of the ground squirrel has been determined by electron microscopy of cells impregnated by the Golgi method. One group of ganglion cells has mainly amacrine input (approximately 97 percent); the other group has an approximately equal bipolar and amacrine input. Morphologically distinct types of ganglion cells usually have a consistent synaptic input, but exceptions may exist.     

Responses of retinal ganglion cells to exponentially increasing light stimuli

Action potentials of ganglion cells were recorded extracellularly from opened cat eyes. It was found that inhibition, as judged by discharge frequency, may depend upon rate of change of light intensity. Apparently the balance between excitatory and inhibitory inputs at the ganglion cell level depends upon rate of change of illumination. Visual purple bleaching or sensory adaptation taking place during the stimulation does not explain the results.     

Phototransduction by retinal ganglion cells that set the circadian clock

Light synchronizes mammalian circadian rhythms with environmental time by modulating retinal input to the circadian pacemaker-the suprachiasmatic nucleus (SCN) of the hypothalamus. Such photic entrainment requires neither rods nor cones, the only known retinal photoreceptors. Here, we show that retinal ganglion cells innervating the SCN are intrinsically photosensitive. Unlike other ganglion cells, they depolarized in response to light even when all synaptic input from rods and cones was blocked. The sensitivity, spectral tuning, and slow kinetics of this light response matched those of the photic entrainment mechanism, suggesting that these ganglion cells may be the primary photoreceptors for this system.     

Spontaneous impulse activity of rat retinal ganglion cells in prenatal life

The existence of spontaneous neural activity in mammalian retinal ganglion cells during prenatal life has long been suspected. This activity could play a key role in the refinement of retinal projections during development. Recordings in vivo from the retinas of rat fetuses between embryonic day 17 and 21 found action potentials in spontaneously active ganglion cells at all the ages studied.     

Nicotinic antagonists enhance process outgrowth by rat retinal ganglion cells in culture

Functional nicotinic cholinergic receptors are found on mammalian retinal ganglion cell neurons in culture. The neurotransmitter acetylcholine (ACh) can be detected in the medium of many of these retinal cultures, after release presumably from the choline acetyltransferase-positive amacrine cells. The postsynaptic effect of endogenous or applied ACh on the ganglion cells can be blocked with specific nicotinic antagonists. Here it is shown that within 24 hours of producing such a pharmacologic blockade, the retinal ganglion cells begin to sprout or regenerate neuronal processes. Thus, the growth-enhancing effect of nicotinic antagonists may be due to the removal of inhibition to growth by tonic levels of ACh present in the culture medium. Since there is a spontaneous leak of ACh in the intact retina, the effects of nicotinic cholinergic drugs on process outgrowth in culture may reflect a normal control mechanism for growth or regeneration of retinal ganglion cell processes that is exerted by ACh in vivo.     

Monoclonal antibody to Thy-1 enhances regeneration of processes by rat retinal ganglion cells in culture

Ganglion cells were dissociated from postnatal rat retinas, identified by specific fluorescent labels, and maintained in culture on a variety of substrates. Regeneration of processes by retinal ganglion cells was enhanced when the cells were plated on glass coated with a monoclonal antibody against the Thy-1 determinant. Plain glass and glass coated with polylysine, collagen, fibronectin, or other monoclonal antibodies supported the growth of neural processes, but were less effective than antibody to Thy-1.     

Electrical responses of the retinal nerve and optic ganglion of the squid

Recordings were obtained from the retinal nerves and optic ganglia of intact squid, which were maintained in good condition by perfusing their mantles with sea water. Only "on" discharges were found in the nerves, whereas "on" and "off" discharges as well as spontaneous activity and tactile responses were obtained from the ganglia.     

New reflected-light microscope for viewing unstained brain and ganglion cells

We have designed and constructed a new type of reflected-light microscope to form images including only light reflected near the plane of the object. This selectivity of image formation is based on a mechanical flying-spot technique. Objects difficult or impossible to see with earlier microscopes, such as unstained cells and cell processes in brains of living salamanders and in excised dorsal root ganglia of frogs, have been observed routinely with this microscope.     

Generation of adrenergic and cholinergic potentials in sympathetic ganglion cells

Norepinephrine elicited a hyperpolarizing response, and acetylcholine (during nicotinic blockade) elicited a depolarizing one. Both responses showed no increase in membrane conductance. The norepinephrine response was suppressed by initial depolarization; the acetylcholine response (frog cells); by hyperpolarization. These neurotransmitters apparently can activate electrogenic mechanisms which do not involve movement of ions down their electrochemical gradients.     

Hematite: intrinsic and defect ferromagnetism

Both intrinsic (spin-canted) ferromagnetism and defect ferromagnetism occur in hematite. Because defect ferromagnetism is sensitive to structure, it is altered by stress or heating and could give spurious paleomagnetic information. Annealing experiments now suggest that the defect remanence of fine-grain hematites and red sediments, unlike that of single crystals, is magnetically softer than the spin-canted remanence and can be erased by partial demagnetization.     

Goldfish retina: sign of the rod input in opponent color ganglion cells

After light adaptation, all "on-center" ganglion cells in the dark became "red on-center," and all "off-center" cells turned into "red off-center" cells. On a chance basis, this similitude of effect between the rods and the red cones in opponent color cells was not expected. These findings indicate that in the goldfish there is some similarity between the connections of the rods and of the long-wavelength cones.     

永生化猪视网膜色素上皮细胞的建立及初步应用

【目的】建立永生化猪视网膜色素上皮细胞系,为病毒学研究提供一种新的细胞材料。【方法】合成猪腺病毒3型E1基因,将其克隆至真核表达质粒pCI-neo中,构建E1基因真核表达质粒pCI-neo-E1,进行PCR和测序鉴定。提取pCI-neo-E1质粒,转染原代猪视网膜色素上皮细胞,用新霉素G418筛选猪永生化视网膜色素上皮细胞系（RPECs）。检测RPECs的角蛋白18和19,通过细胞计数测定其生长曲线,流式细胞仪检测细胞周期,并对其进行染色体核型分析。分别将猪流行性腹泻病毒（PEDV）CV77毒株、猪圆环病毒2型（PCV2）DBN-SX07株和猪伪狂犬病毒（PRV）Bartha-K61株接种RPECs,同时将PEDV CV77毒株接种绿猴肾细胞（Vero细胞）,将PCV2 DBN-SX07株接种猪肾细胞（PK15细胞）,将PRV Bartha-K61株接种仓鼠肾成纤维细胞（BHK细胞）,观察细胞病变效应（CPE）,并采用间接免疫荧光法测定病毒的滴度。【结果】PCR和测序结果显示,真核表达质粒pCI-neo-E1构建成功。将pCI-neo-E1转染原代猪视网膜色素上皮细胞,经G418筛选得到永生化细胞株,该细胞株表达角蛋白18和19,传代50代仍保持上皮样细胞形态,增殖活性良好,细胞周期及染色体核型特征与正常的二倍体细胞一致。PEDV CV77毒株感染猪RPECs 24 h后,CPE明显,滴度为10^5.25TCID₅₀/mL,低于其在Vero细胞中的滴度（10^8.125 TCID₅₀/mL）。PCV2 DBN-SX07毒株感染猪RPECs后可产生明显CPE,病毒滴度为10^6.125TCID₅₀/mL,略高于其在PK15细胞上的滴度（10^6.0TCID₅₀/mL）。PRV Bartha-K61毒株感染猪RPECs后,CPE明显,病毒滴度为10^8.75 TCID₅₀/mL,略低于其在BHK上的滴度（10^8.875 TCID₅₀/mL）。【结论】成功构建了永生化猪视网膜细胞系,可用于猪源病毒的培养,尤其是培养PCV2可产生明显CPE,易于观察,为后续开展疫苗工艺的提升提供了一种新的细胞材料。     

Plasticity and differentiation of embryonic retinal cells after terminal mitosis

The relation between terminal mitosis and the events that determine the developmental fate of embryonic precursor cells is not well understood. This relation has now been investigated with [3H]thymidine autoradiography to determine the time of cell birth and with a culture system that allows the testing of the developmental potential of cells isolated from the chick embryo retina. Contrary to the situation in vivo, where neuronal differentiation always precedes photoreceptor differentiation, photoreceptor differentiation occurs prematurely and precedes neuronal differentiation when precursor cells are isolated from the retina at early embryonic stages. Thus, cells born by embryonic day 5 (ED-5) give rise predominantly to photoreceptors when isolated for culture on ED-6 but develop mainly as neurons when isolated on ED-8. This suggests that retinal precursor cells retain after terminal mitosis the capacity to develop either as neurons or as photoreceptors. Moreover, photoreceptor differentiation appears to represent a constitutive or "default" pathway that precursor cells follow in the absence of neuron-inducing signals.     

Technology characteristics, choice architecture, and farmer knowledge: the case of phytase

Phytase is an enzyme that frees the phosphorus bound in feed grains and thus reduces the amount of dicalcium phosphate supplementation required for non-ruminants, reducing phosphorous excretion and thus reducing water pollution. This innovation has been widely adopted by feed companies in the US due to decreased phytase production costs and increased dicalcium phosphate costs. The roles played by phytase characteristics and choice architecture in the widespread use of this win–win technology are examined. A recent survey has also revealed that Midwestern farmers are largely unaware of this technology even though they are using it. One implication is that further research on win–win technologies that will be adopted by industries, rather than being dependent on adoption by individuals, may be beneficial.     

Slow synaptic excitation in sympathetic ganglion cells: evidence for synaptic inactivation of potassium conductance

The slow excitatory postsynaptic potential (EPSP) was investigated in frog sympathetic ganglion cells. In contrast to the increased conductance associated with other known EPSP's, during the slow EPSP resting membrane conductance was decreased. Electrical depolarization of the membrane potentiated the slow EPSP, whereas progressive hyperpolarization decreased its size and then reversed it to a hyperpolarizing potential (the opposite of the effect of membrane polarization on other EPSP's). The reversal potential of the slow EPSP was close to the potassium equilibrium potential. We propose that the slow EPSP, in contrast to classical EPSP's, is generated by an inactivation of resting potassium conductance.     

Transient morphological features of identified ganglion cells in living fetal and neonatal retina

The function and morphology of retinal ganglion cells in the adult mammalian visual system has been well studied, but little is known about how the adult state is achieved. To address this question, the morphological changes that retinal ganglion cells undergo during development were studied. Ganglion cells were first identified by retrograde labeling with rhodamine latex microspheres deposited in retinorecipient targets in fetal and early postnatal cats. The structure of ganglion cells was then revealed by intracellular injection of Lucifer yellow in living retinas removed and maintained in vitro. As early as 2 weeks before birth, a morphologically diverse assortment of ganglion cells is present, some of which resemble the alpha, beta, and gamma classes found in the adult. However, in contrast to the adult, developing ganglion cells exhibit several transient features, including excessive axonal and dendritic branching and exuberant somatic and dendritic spines. These morphological features indicate that there is a transient network of connectivity that could play an important role in the final determination of retinal ganglion cell form and function.