scholarly journals NKCC1 Does Not Accumulate Chloride in Developing Retinal Neurons

2007 ◽  
Vol 98 (1) ◽  
pp. 266-277 ◽  
Author(s):  
Ling-Li Zhang ◽  
Eric Delpire ◽  
Noga Vardi
Keyword(s):  
Ganglion Cells ◽  
Chloride Concentration ◽  
Müller Cells ◽  
Amacrine Cells ◽  
Muller Cells ◽  
Retinal Neurons ◽  
High Concentration ◽  
Intracellular Chloride ◽  
Cellular Markers ◽  
Intracellular Chloride Concentration

GABA excites immature neurons due to their relatively high intracellular chloride concentration. This initial high concentration is commonly attributed to the ubiquitous chloride cotransporter NKCC1, which uses a sodium gradient to accumulate chloride. Here we tested this hypothesis in immature retinal amacrine and ganglion cells. Western blotting detected NKCC1 at birth and its expression first increased, then decreased to the adult level. Immunocytochemistry confirmed this early expression of NKCC1 and localized it to all nuclear layers. In the ganglion cell layer, staining peaked at P4 and then decreased with age, becoming undetectable in adult. In comparison, KCC2, the chloride extruder, steadily increased with age localizing primarily to the synaptic layers. For functional tests, we used calcium imaging with fura-2 and chloride imaging with 6-methoxy- N-ethylquinolinium iodide. If NKCC1 accumulates chloride in ganglion and amacrine cells, deleting or blocking it should abolish the GABA-evoked calcium rise. However, at P0-5 GABA consistently evoked a calcium rise that was not abolished in the NKCC1-null retinas, nor by applying high concentrations of bumetanide (NKCC blocker) for long periods. Furthermore, intracellular chloride concentration in amacrine and ganglion cells of the NKCC1-null retinas was ∼30 mM, same as in wild type at this age. This concentration was not lowered by applying bumetanide or by decreasing extracellular sodium concentration. Costaining for NKCC1 and cellular markers suggested that at P3, NKCC1 is restricted to Müller cells. We conclude that NKCC1 does not serve to accumulate chloride in immature retinal neurons, but it may enable Müller cells to buffer extracellular chloride.

Differential expression and distribution of Kir5.1 and Kir4.1 inwardly rectifying K+ channels in retina

2003 ◽  
Vol 285 (2) ◽  
pp. C260-C267 ◽  
Author(s):  
Masaru Ishii ◽  
Akikazu Fujita ◽  
Kaori Iwai ◽  
Shunji Kusaka ◽  
Kayoko Higashi ◽  
...  
Keyword(s):  
Cell Body ◽  
Müller Cells ◽  
Amacrine Cells ◽  
Distal Portion ◽  
Acid Decarboxylase ◽  
Muller Cells ◽  
Retinal Neurons ◽  
Functional Roles ◽  
Inwardly Rectifying ◽  
Specific Polyclonal Antibody

Kir5.1 is an inwardly rectifying K+ channel subunit whose functional role has not been fully elucidated. Expression and distribution of Kir5.1 in retina were examined with a specific polyclonal antibody. Kir5.1 immunoreactivity was detected in glial Müller cells and in some retinal neurons. In the Kir5.1-positive neurons the expression of glutamic acid decarboxylase (GAD65) was detected, suggesting that they may be GABAergic-amacrine cells. In Müller cells, spots of Kir5.1 immunoreactivity distributed diffusely at the cell body and in the distal portions, where Kir4.1 immunoreactivity largely overlapped. In addition, Kir4.1 immunoreactivity without Kir5.1 was strongly concentrated at the endfoot of Müller cells facing the vitreous surface or in the processes surrounding vessels. The immunoprecipitant obtained from retina with anti-Kir4.1 antibody contained Kir5.1. These results suggest that heterotetrameric Kir4.1/Kir5.1 channels may exist in the cell body and distal portion of Müller cells, whereas homomeric Kir4.1 channels are clustered in the endfeet and surrounding vessels. It is possible that homomeric Kir4.1 and heteromeric Kir4.1/Kir5.1 channels play different functional roles in the K+-buffering action of Müller cells.

scholarly journals Shift of Intracellular Chloride Concentration in Ganglion and Amacrine Cells of Developing Mouse Retina

2006 ◽  
Vol 95 (4) ◽  
pp. 2404-2416 ◽  
Author(s):  
Ling-Li Zhang ◽  
Hemal R. Pathak ◽  
Douglas A. Coulter ◽  
Michael A. Freed ◽  
Noga Vardi
Keyword(s):  
Gaba Receptors ◽  
Chloride Concentration ◽  
Reversal Potential ◽  
Amacrine Cells ◽  
Resting Potential ◽  
Equilibrium Potential ◽  
Mouse Retina ◽  
Intracellular Chloride ◽  
Excitatory Action ◽  
Intracellular Chloride Concentration

GABA and glycine provide excitatory action during early development: they depolarize neurons and increase intracellular calcium concentration. As neurons mature, GABA and glycine become inhibitory. This switch from excitation to inhibition is thought to result from a shift of intracellular chloride concentration ([Cl−]i) from high to low, but in retina, measurements of [Cl−]i or chloride equilibrium potential ( ECl) during development have not been made. Using the developing mouse retina, we systematically measured [Cl−]i in parallel with GABA's actions on calcium and chloride. In ganglion and amacrine cells, fura-2 imaging showed that before postnatal day (P) 6, exogenous GABA, acting via ionotropic GABA receptors, evoked calcium rise, which persisted in HCO3−- free buffer but was blocked with 0 extracellular calcium. After P6, GABA switched to inhibiting spontaneous calcium transients. Concomitant with this switch we observed the following: 6-methoxy- N-ethylquinolinium iodide (MEQ) chloride imaging showed that GABA caused an efflux of chloride before P6 and an influx afterward; gramicidin-perforated-patch recordings showed that the reversal potential for GABA decreased from −45 mV, near threshold for voltage-activated calcium channel, to −60 mV, near resting potential; MEQ imaging showed that [Cl−]i shifted steeply around P6 from 29 to 14 mM, corresponding to a decline of ECl from −39 to −58 mV. We also show that GABAergic amacrine cells became stratified by P4, potentially allowing GABA's excitatory action to shape circuit connectivity. Our results support the hypothesis that a shift from high [Cl−]i to low causes GABA to switch from excitatory to inhibitory.

Neurotransmitter-specific identification and characterization of neurons in the all-cone retina of Anolis carolinensis II: Glutamate and aspartate

1992 ◽  
Vol 9 (3-4) ◽  
pp. 313-323 ◽  
Author(s):  
David M. Sherry ◽  
Robert J. Ulshafer
Keyword(s):  
Ganglion Cell ◽  
Ganglion Cells ◽  
Müller Cells ◽  
Plexiform Layer ◽  
Amacrine Cells ◽  
Bipolar Cells ◽  
Anolis Carolinensis ◽  
Horizontal Cells ◽  
Inner Plexiform Layer ◽  
Muller Cells

AbstractImmunocytochemical and autoradiographic methods were used to identify neurons in the pure cone retina of the lizard (Anolis carolinensis) that are likely to employ glutamate (GLU) or aspartate (ASP) as a neurotransmitter.GLU immunocytochemistry demonstrated high levels of endogenous GLU in all cone types and numerous bipolar cells. Moderate GLU levels were found in horizontal and ganglion cells. Müller cells and most amacrine cells had very low GLU levels. GLU immunoreactivity (GLU-IR) in the cones was present from the inner segment to the synaptic pedicle. A large spherical cell type with moderate GLU-IR was identified in the proximal inner plexiform layer (IPL). These cells also contain ASP and have been tentatively identified as amacrine cells. Uptake of [3H]-L-GLU labeled all retinal layers. All cone types and Müller cells sequestered [3H]-D-ASP, a substrate specific for the GLU transporter.Anti-ASP labeling was observed in cones, horizontal cells, amacrine cells, and cells in the ganglion cell layer. ASP immunoreactivity (ASP-IR) in the cones was confined to the inner segment. One ASP-containing pyriform amacrine cell subtype ramifying in IPL sublamina b was identified.Analysis of GLU-IR, ASP-IR, and GABA-IR on serial sections indicated that there were two distinct populations of horizontal cells in the Anolis retina: one containing GABA-IR, GLU-IR, and ASP-IR; and another type containing only GLU-IR and ASP-IR. Light GLU-IR was frequently found in GABA-containing amacrine cells but ASP-IR was not.The distinct distributions of GLU and ASP may indicate distinctly different roles for these amino acids. GLU, not ASP, is probably the major neurotransmitter in the cone-biploar-ganglion cell pathway of the Anolis retina. Both GLU and ASP are present in horizontal cells and specific subpopulations of amacrine cells, but it is unclear if GLU or ASP have a neurotransmitter role in these cells.

Relationships between Müller cells and neurons in a primitive tetrapod, the Australian lungfish

1997 ◽  
Vol 14 (4) ◽  
pp. 795-800 ◽  
Author(s):  
Stephen R. Robinson
Keyword(s):  
Ganglion Cells ◽  
Horizontal Cell ◽  
Müller Cells ◽  
Amacrine Cells ◽  
Müller Cell ◽  
Muller Cells ◽  
Cone Photoreceptors ◽  
Rod Photoreceptors ◽  
Muller Cell ◽  
Australian Lungfish

AbstractWe recently proposed a model of cytogenesis which assumes that primitive ancestral mammals and premammalian vertebrates had a retinal composition that consisted of about seven neurons per Müller cell, comprising 1–2 cone photoreceptors, 1–2 rod photoreceptors, 2–3 bipolar cells, 1–2 amacrine cells, less than 1 ganglion cell, and less than 1 horizontal cell (Reichenbach & Robinson, 1995). The Australian lungfish (Neoceratodus forsten) closely resembles the lobe-finned ancestors of land vertebrates, and has an extremely plesiomorphic nervous system. The present study, therefore, has examined the relative frequencies of retinal neurons and Müller cells (identified by immunolabelling for glutamine synthetase) in the lungfish retina. It was found that for each Müller cell there is an average of 1.9 cone photoreceptors, 1.7 rod photoreceptors, 3.1 amacrine/bipolar/horizontal cells, and 0.6 ganglion cells; amounting to a ratio of 7.3 neurons per Müller cell. These results support our conjecture that the sequence of cytogenesis in mammals is constrained by a developmental program that predates the evolution of mammals. The study also provides the first detailed morphological descriptions of lungfish Müller cells and their relationship with adjacent neurons. It was found that individual Müller cells in lungfish have a volume (more than 12,000 μm3) that is an order of magnitude higher than in mammals, yet the proportion of total retinal volume occupied by these cells (20%) is very similar.

Development of the rabbit retina, III: Differential retinal growth, and density of projection neurons and interneurons

1993 ◽  
Vol 10 (3) ◽  
pp. 479-498 ◽  
Author(s):  
A. Reichenbach ◽  
J. Schnitzer ◽  
E. Reichelt ◽  
N. N. Osborne ◽  
B. Fritzsche ◽  
...  
Keyword(s):  
Ganglion Cell ◽  
Ganglion Cell Layer ◽  
Ganglion Cells ◽  
Cell Layer ◽  
Quantitative Description ◽  
Müller Cells ◽  
Amacrine Cells ◽  
Projection Neurons ◽  
Muller Cells ◽  
Single Plane

AbstractTo provide a quantitative description of postnatal retinal expansion in rabbits, a new procedure was developed to map the retinae, which cover the inner surface of hemispheres or parts of rotation ellipsoids, in situ, onto a single plane. This method, as well as the known distribution of Müller cells per unit retinal surface area, were used to estimate the redistribution of specific subpopulations of Müller cells within different topographic regions of the retinae. Müller cells are known to exist as a stable population of cells 1 week after birth and can therefore be used as “markers” for determining tissue expansion. Our results show that differential retinal expansion occurs during development. Peripheral retinal regions expand at least twice as much as the central ones. Furthermore, there is a greater vertical than horizontal expansion. This differential retinal expansion leads to a corresponding redistribution of 5-hydroxytryptamine (5-HT) accumulating amacrine cells. Differential retinal expansion, however, does not account for all of the changes in the centro-peripheral density gradient of cells in the ganglion cell layer (GCL) — mostly retinal ganglion cells — during postnatal development. The changes in the ganglion cell layer were evaluated in Nissl-stained wholemount retinal preparations. Additionally, the difference between expansion-related redistribution of cells in the GCL and Müller cells was confirmed in wholemount preparations where Müller cells (identified as vimentin positive) and cells in the GCL (identified by fluorescent supravital dyes) were simultaneously labeled. It is assumed that many of the ganglion cells within the retinal center are not translocated during retinal expansion, possibly because their axons are fixed. In contrast, 5-HT accumulating amacrine cells — which are interneurons without a retinofugal axon — display a passive redistribution together with the surrounding retinal tissue.

Long-Term Imaging Reveals a Circadian Rhythm of Intracellular Chloride in Neurons of the Suprachiasmatic Nucleus

2022 ◽  
pp. 074873042110597
Author(s):  
Nathan J. Klett ◽  
Olga Cravetchi ◽  
Charles N. Allen
Keyword(s):  
Suprachiasmatic Nucleus ◽  
Chloride Concentration ◽  
Intracellular Chloride ◽  
Voltage Gated Calcium Channels ◽  
Excitatory Gaba ◽  
Ratiometric Probe ◽  
Gaba Transmission ◽  
Circadian Physiology ◽  
Intracellular Chloride Concentration

Both inhibitory and excitatory GABA transmission exist in the mature suprachiasmatic nucleus (SCN), the master pacemaker of circadian physiology. Whether GABA is inhibitory or excitatory depends on the intracellular chloride concentration ([Cl−]i). Here, using the genetically encoded ratiometric probe Cl-Sensor, we investigated [Cl−]i in AVP and VIP-expressing SCN neurons for several days in culture. The chloride ratio (RCl) demonstrated circadian rhythmicity in AVP + neurons and VIP + neurons, but was not detected in GFAP + astrocytes. RCl peaked between ZT 7 and ZT 8 in both AVP + and VIP + neurons. RCl rhythmicity was not dependent on the activity of several transmembrane chloride carriers, action potential generation, or the L-type voltage-gated calcium channels, but was sensitive to GABA antagonists. We conclude that [Cl−]i is under circadian regulation in both AVP + and VIP + neurons.

scholarly journals GABA-mediated repulsive coupling between circadian clock neurons in the SCN encodes seasonal time

2015 ◽  
Vol 112 (29) ◽  
pp. E3920-E3929 ◽  
Author(s):  
Jihwan Myung ◽  
Sungho Hong ◽  
Daniel DeWoskin ◽  
Erik De Schutter ◽  
Daniel B. Forger ◽  
...  
Keyword(s):  
Circadian Clock ◽  
Synaptic Input ◽  
Chloride Concentration ◽  
Phase Relationship ◽  
Day Length ◽  
Intracellular Chloride ◽  
Circadian Oscillators ◽  
Length Changes ◽  
Excitatory Gaba ◽  
Intracellular Chloride Concentration

The mammalian suprachiasmatic nucleus (SCN) forms not only the master circadian clock but also a seasonal clock. This neural network of ∼10,000 circadian oscillators encodes season-dependent day-length changes through a largely unknown mechanism. We show that region-intrinsic changes in the SCN fine-tune the degree of network synchrony and reorganize the phase relationship among circadian oscillators to represent day length. We measure oscillations of the clock gene Bmal1, at single-cell and regional levels in cultured SCN explanted from animals raised under short or long days. Coupling estimation using the Kuramoto framework reveals that the network has couplings that can be both phase-attractive (synchronizing) and -repulsive (desynchronizing). The phase gap between the dorsal and ventral regions increases and the overall period of the SCN shortens with longer day length. We find that one of the underlying physiological mechanisms is the modulation of the intracellular chloride concentration, which can adjust the strength and polarity of the ionotropic GABAA-mediated synaptic input. We show that increasing day-length changes the pattern of chloride transporter expression, yielding more excitatory GABA synaptic input, and that blocking GABAA signaling or the chloride transporter disrupts the unique phase and period organization induced by the day length. We test the consequences of this tunable GABA coupling in the context of excitation–inhibition balance through detailed realistic modeling. These results indicate that the network encoding of seasonal time is controlled by modulation of intracellular chloride, which determines the phase relationship among and period difference between the dorsal and ventral SCN.

Influence of WNK3 on intracellular chloride concentration and volume regulation in HEK293 cells

2012 ◽  
Vol 464 (3) ◽  
pp. 317-330 ◽  
Author(s):  
Silvia Cruz-Rangel ◽  
Gerardo Gamba ◽  
Gerardo Ramos-Mandujano ◽  
Herminia Pasantes-Morales
Keyword(s):  
Volume Regulation ◽  
Chloride Concentration ◽  
Hek293 Cells ◽  
Intracellular Chloride ◽  
Intracellular Chloride Concentration

scholarly journals A dialogue between glia and neurons in the retina: modulation of neuronal excitability

2004 ◽  
Vol 1 (3) ◽  
pp. 245-252 ◽  
Author(s):  
ERIC A. NEWMAN
Keyword(s):  
Synaptic Transmission ◽  
Glial Cells ◽  
Neuronal Excitability ◽  
Ganglion Cells ◽  
Brain Slices ◽  
Müller Cells ◽  
Muller Cells ◽  
Signaling Mechanisms ◽  
Bidirectional Signaling ◽  
Stimulation Of

Bidirectional signaling between neurons and glial cells has been demonstrated in brain slices and is believed to mediate glial modulation of synaptic transmission in the CNS. Our laboratory has characterized similar neuron–glia signaling in the mammalian retina. We find that light-evoked neuronal activity elicits Ca2+ increases in Müller cells, which are specialized retinal glial cells. Neuron to glia signaling is likely mediated by the release of ATP from neurons and is potentiated by adenosine. Glia to neuron signaling has also been observed and is mediated by several mechanisms. Stimulation of glial cells can result in either facilitation or depression of synaptic transmission. Release of D-serine from Müller cells might also potentiate NMDA receptor transmission. Müller cells directly inhibit ganglion cells by releasing ATP, which, following hydrolysis to adenosine, activates neuronal A1 receptors. The existence of bidirectional signaling mechanisms indicates that glial cells participate in information processing in the retina.

Immunohistochemical analysis of the neurotrophins BDNF and NT-3 and their receptors trk B, trk C, and p75 in the developing chick retina

1997 ◽  
Vol 14 (5) ◽  
pp. 835-842 ◽  
Author(s):  
Indranil Das ◽  
Barbara L. Hempstead ◽  
Peter R. Macleish ◽  
Janet R. Sparrow
Keyword(s):  
Ganglion Cells ◽  
Plexiform Layer ◽  
Amacrine Cells ◽  
Immunohistochemical Analysis ◽  
Neurotrophin Receptor ◽  
Inner Plexiform Layer ◽  
P75 Neurotrophin Receptor ◽  
Retinal Neurons ◽  
Chick Retina ◽  
Trk B

AbstractThe neurotrophins are trophic and mitogenic factors critical for the development of specific classes of neurons in the central and peripheral nervous systems. In the retina, BDNF and NT-3 have been shown to promote the survival of differentiated ganglion cells (Rodriguez-Tebar et al., 1989; De La Rosa et al., 1994). NT-3 has also been demonstrated to support the survival of amacrine cells and facilitates the differentiation of retinal neurons in culture (De La Rosa et al., 1994). Here, we examine immunohistochemically the expression of BDNF and NT-3 proteins, their cognate receptors, trk B and trk C, respectively, and the p75 neurotrophin receptor in the developing chick retina. At E8, the earliest stage of retinal development examined, all of these proteins exhibit diffuse expression throughout the width of the retina, with the strongest reactivity in the innermost layers. A gradual restriction in expression to ganglion cells and amacrine cells, the staining of which is most prominent at E15, is followed by a downregulation of expression with the strongest immunoreactivity persisting in the ganglion cell layer. Overlapping patterns of expression throughout embryonic development indicate a colocalization of the neurotrophins and their receptors, although NT-3 and p75 alone are present in the inner plexiform layer and only p75 is observed in the outer plexiform layer. Although some of the immunoreactivity for BDNF, NT-3, and their receptors in retina may reflect trophic mechanisms operating in association with the optic tectum and isthmo-optic nucleus, the colocalization of ligands and receptors in retina strengthens the assertion that these neurotrophins function locally during development.

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