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1.
Front Physiol ; 12: 756542, 2021.
Article in English | MEDLINE | ID: mdl-34721079

ABSTRACT

A sedentary lifestyle is the top preventable cause of death and accounts for substantial socioeconomic costs to society. The rostral ventrolateral medulla regulates blood pressure under normal and pathophysiological states, and demonstrates inactivity-related structural and functional neuroplasticity, which is subregionally specific. The purpose of this study was to examine pro- and mature forms of brain-derived neurotrophic factor (BDNF) and their respective receptors in the male rat rostral ventrolateral medulla (RVLM) and its rostral extension following sedentary vs. active (running wheels) conditions (10-12weeks). We used subregionally specific Western blotting to determine that the mature form of BDNF and its ratio to its pro-form were lower in more caudal subregions of the rostral ventrolateral medulla of sedentary rats but higher in the rostral extension when both were compared to active rats. The full-length form of the tropomyosin receptor kinase B receptor and the non-glycosylated form of the 75 kilodalton neurotrophin receptor were lower in sedentary compared to active rats. The rostrocaudal patterns of expression of the mature form of BDNF and the full-length form of the tropomyosin receptor kinase B receptor were remarkably similar to the subregionally specific patterns of enhanced dendritic branching, neuronal activity, and glutamate-mediated increases in sympathetic nerve activity observed in previous studies performed in sedentary rats. Our studies suggest signaling pathways related to BDNF within subregions of both the rostral ventrolateral medulla and its rostral extension contribute to cardiovascular disease and premature death related to a sedentary lifestyle.

2.
Front Neural Circuits ; 15: 624563, 2021.
Article in English | MEDLINE | ID: mdl-33746717

ABSTRACT

Dopamine (DA) modulates the activity of nuclei within the ascending and descending auditory pathway. Previous studies have identified neurons and fibers in the inferior colliculus (IC) which are positively labeled for tyrosine hydroxylase (TH), a key enzyme in the synthesis of dopamine. However, the origins of the tyrosine hydroxylase positive projections to the inferior colliculus have not been fully explored. The lateral lemniscus (LL) provides a robust inhibitory projection to the inferior colliculus and plays a role in the temporal processing of sound. In the present study, immunoreactivity for tyrosine hydroxylase was examined in animals with and without 6-hydroxydopamine (6-OHDA) lesions. Lesioning, with 6-OHDA placed in the inferior colliculus, led to a significant reduction in tyrosine hydroxylase immuno-positive labeling in the lateral lemniscus and inferior colliculus. Immunolabeling for dopamine beta-hydroxylase (DBH) and phenylethanolamine N-methyltransferase (PNMT), enzymes responsible for the synthesis of norepinephrine (NE) and epinephrine (E), respectively, were evaluated. Very little immunoreactivity for DBH and no immunoreactivity for PNMT was found within the cell bodies of the dorsal, intermediate, or ventral nucleus of the lateral lemniscus. The results indicate that catecholaminergic neurons of the lateral lemniscus are likely dopaminergic and not noradrenergic or adrenergic. Next, high-pressure liquid chromatography (HPLC) analysis was used to confirm that dopamine is present in the inferior colliculus and nuclei that send projections to the inferior colliculus, including the cochlear nucleus (CN), superior olivary complex (SOC), lateral lemniscus, and auditory cortex (AC). Finally, fluorogold, a retrograde tracer, was injected into the inferior colliculus of adult rats. Each subdivision of the lateral lemniscus contained fluorogold within the somata, with the dorsal nucleus of the lateral lemniscus showing the most robust projections to the inferior colliculus. Fluorogold-tyrosine hydroxylase colocalization within the lateral lemniscus was assessed. The dorsal and intermediate nuclei neurons exhibiting similar degrees of colocalization, while neurons of the ventral nucleus had significantly fewer colocalized fluorogold-tyrosine hydroxylase labeled neurons. These results suggest that several auditory nuclei that project to the inferior colliculus contain dopamine, dopaminergic neurons in the lateral lemniscus project to the inferior colliculus and that dopaminergic neurotransmission is poised to play a pivotal role in the function of the inferior colliculus.


Subject(s)
Inferior Colliculi , Acoustics , Animals , Auditory Pathways , Dopamine , Olivary Nucleus , Pons , Rats
3.
J Comp Neurol ; 529(9): 2311-2331, 2021 06.
Article in English | MEDLINE | ID: mdl-33347606

ABSTRACT

The rostral ventrolateral medulla (RVLM) is a brain region involved in normal regulation of the cardiovascular system and heightened sympathoexcitatory states of cardiovascular disease (CVD). Among major risk factors for CVD, sedentary lifestyles contribute to higher mortality than other modifiable risk factors. Previous studies suggest excessive glutamatergic excitation of presympathetic neurons in the RVLM occurs in sedentary animals. Therefore, the purpose of this study was to examine neuroplasticity in the glutamatergic system in the RVLM of sedentary and physically active rats. We hypothesized that relative to active rats, sedentary rats would exhibit higher expression of glutamate N-methyl-d-aspartic acid receptor subunits (GluN), phosphoGluN1, and the excitatory scaffold protein postsynaptic density 95 (PSD95), while achieving higher glutamate levels. Male Sprague-Dawley rats (4 weeks old) were divided into sedentary and active (running wheel) conditions for 10-12 weeks. We used retrograde tracing/triple-labeling techniques, western blotting, and magnetic resonance spectroscopy. We report in sedentary versus physically active rats: 1) fewer bulbospinal non-C1 neurons positive for GluN1, 2) significantly higher expression of GluN1 and GluN2B but lower levels of phosphoGluN1 (pSer896) and PSD95, and 3) higher levels of glutamate in the RVLM. Higher GluN expression is consistent with enhanced sympathoexcitation in sedentary animals; however, a more complex neuroplasticity occurs within subregions of the ventrolateral medulla. Our results in rodents may also indicate that alterations in glutamatergic excitation of the RVLM contribute to the increased incidence of CVD in humans who lead sedentary lifestyles. Thus, there is a strong need to further pursue mechanisms of inactivity-related neuroplasticity in the RVLM.


Subject(s)
Medulla Oblongata/metabolism , Neuronal Plasticity/physiology , Physical Conditioning, Animal/physiology , Receptors, N-Methyl-D-Aspartate/biosynthesis , Sedentary Behavior , Animals , Male , Physical Conditioning, Animal/methods , Rats , Rats, Sprague-Dawley , Signal Transduction/physiology
4.
J Comp Neurol ; 528(6): 1053-1075, 2020 04.
Article in English | MEDLINE | ID: mdl-31642070

ABSTRACT

Neurons in the rostral ventrolateral medulla (RVLM) regulate blood pressure through direct projections to spinal sympathetic preganglionic neurons. Only some RVLM neurons are active under resting conditions due to significant, tonic inhibition by gamma-aminobutyric acid (GABA). Withdrawal of GABAA receptor-mediated inhibition of the RVLM increases sympathetic outflow and blood pressure substantially, providing a mechanism by which the RVLM could contribute chronically to cardiovascular disease (CVD). Here, we tested the hypothesis that sedentary conditions, a major risk factor for CVD, increase GABAA receptors in RVLM, including its rostral extension (RVLMRE ), both of which contain bulbospinal catecholamine (C1) and non-C1 neurons. We examined GABAA receptor subunits GABAAα1 and GABAAα2 in the RVLM/RVLMRE of sedentary or physically active (10-12 weeks of wheel running) rats. Western blot analyses indicated that sedentary rats had lower expression of GABAAα1 and GABAAα2 subunits in RVLM but only GABAAα2 was lower in the RVLMRE of sedentary rats. Sedentary rats had significantly reduced expression of the chloride transporter, KCC2, suggesting less effective GABA-mediated inhibition compared to active rats. Retrograde tracing plus triple-label immunofluorescence identified fewer bulbospinal non-C1 neurons immunoreactive for GABAAα1 but a higher percentage of bulbospinal C1 neurons immunoreactive for GABAAα1 in sedentary animals. Sedentary conditions did not significantly affect the number of bulbospinal C1 or non-C1 neurons immunoreactive for GABAAα2 . These results suggest a complex interplay between GABAA receptor expression by spinally projecting C1 and non-C1 neurons and sedentary versus physically active conditions. They also provide plausible mechanisms for both enhanced sympathoexcitatory and sympathoinhibitory responses following sedentary conditions.


Subject(s)
Medulla Oblongata/metabolism , Motor Activity/physiology , Neurons/metabolism , Receptors, GABA-A/metabolism , Animals , Male , Rats , Rats, Sprague-Dawley
5.
J Comp Neurol ; 524(10): 2059-79, 2016 07 01.
Article in English | MEDLINE | ID: mdl-26587737

ABSTRACT

In the retina, dopamine is a key molecule for daytime vision. Dopamine is released by retinal dopaminergic amacrine cells and transmits signaling either by conventional synaptic or by volume transmission. By means of volume transmission, dopamine modulates all layers of retinal neurons; however, it is not well understood how dopamine modulates visual signaling pathways in bipolar cells. Here we analyzed Drd1a-tdTomato BAC transgenic mice and found that the dopamine D1 receptor (D1R) is expressed in retinal bipolar cells in a type-dependent manner. Strong tdTomato fluorescence was detected in the inner nuclear layer and localized to type 1, 3b, and 4 OFF bipolar cells and type 5-2, XBC, 6, and 7 ON bipolar cells. In contrast, type 2, 3a, 5-1, 9, and rod bipolar cells did not express Drd1a-tdTomato. Other interneurons were also found to express tdTomato including horizontal cells and a subset (25%) of AII amacrine cells. Diverse visual processing pathways, such as color or motion-coded pathways, are thought to be initiated in retinal bipolar cells. Our results indicate that dopamine sculpts bipolar cell performance in a type-dependent manner to facilitate daytime vision. J. Comp. Neurol. 524:2059-2079, 2016. © 2015 Wiley Periodicals, Inc.


Subject(s)
Receptors, Dopamine D1/metabolism , Retina/cytology , Retinal Bipolar Cells/classification , Retinal Bipolar Cells/metabolism , Visual Pathways/physiology , Amacrine Cells/metabolism , Animals , Biotin/analogs & derivatives , Biotin/metabolism , Choline O-Acetyltransferase/metabolism , Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels/metabolism , Luminescent Proteins/genetics , Luminescent Proteins/metabolism , Mice , Mice, Inbred C57BL , Mice, Transgenic , Nerve Tissue Proteins/metabolism , Opsins/metabolism , Potassium Channels/metabolism , RNA, Messenger/metabolism , Receptors, Dopamine D1/genetics , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Retinal Bipolar Cells/cytology , Synaptotagmin II/metabolism , Transcription Factors/metabolism
6.
Front Neuroanat ; 9: 88, 2015.
Article in English | MEDLINE | ID: mdl-26257610

ABSTRACT

Dopamine (DA) modulates the effects of amino acid neurotransmitters (AANs), including GABA and glutamate, in motor, visual, olfactory, and reward systems (Hnasko et al., 2010; Stuber et al., 2010; Hnasko and Edwards, 2012). The results suggest that DA may play a similar modulatory role in the auditory pathways. Previous studies have shown that deafness results in decreased GABA release, changes in excitatory neurotransmitter levels, and increased spontaneous neuronal activity within brainstem regions related to auditory function. Modulation of the expression and localization of tyrosine hydroxylase (TH; the rate limiting enzyme in the production of DA) in the IC following cochlear trauma has been previously reported (Tong et al., 2005). In the current study the possibility of co-localization of TH with AANs was examined. Changes in the gene expression of TH were compared with changes in the gene expression of markers for AANs in the cochlear nucleus (CN) and inferior colliculus (IC) to determine whether those deafness related changes occur concurrently. The results indicate that bilateral cochlear ablation significantly reduced TH gene expression in the CN after 2 months while in the IC the reduction in TH was observed at both 3 days and 2 months following ablation. Furthermore, in the CN, glycine transporter 2 (GLYT2) and the GABA transporter (GABAtp) were also significantly reduced only after 2 months. However, in the IC, DA receptor 1 (DRDA1), vesicular glutamate transporters 2 and 3 (VGLUT2, VGLUT3), GABAtp and GAD67 were reduced in expression both at the 3 days and 2 months time points. A close relationship between the distribution of TH and several of the AANs was determined in both the CN and the IC. In addition, GLYT2 and VGLUT3 each co-localized with TH within IC somata and dendrites. Therefore, the results of the current study suggest that DA is spatially well positioned to influence the effects of AANs on auditory neurons.

7.
Biotechniques ; 57(5): 245-53, 2014 Nov.
Article in English | MEDLINE | ID: mdl-25391913

ABSTRACT

Green fluorescent protein (GFP) and its derivatives are broadly used in biomedical experiments for labeling particular cells or molecules. In the mouse retina, the light (~500 nm) used to excite GFP can also lead to photoreceptor bleaching (peak ~500 nm), which diminishes photoreceptor-mediated synaptic transmission in the retinal network. To overcome this problem, we investigated the use of infrared fluorescent protein (iRFP) as a marker since it is excited by light in the near-infrared range that would not damage the photoresponsiveness of the retina. Initially, we tested iRFP expression in human embryonic kidney 293 (HEK293) cells to confirm that conventional fluorescence microscopy can detect iRFP fluorescence. We next introduced the iRFP plasmid into adeno-associated virus 2 (AAV-2) and injected the resulting AAV-2 solution into the intraocular space. Retinal neurons were found to successfully express iRFP three weeks post-injection. Light-evoked responses in iRFP-marked cells were assessed using patch clamping, and light sensitivity was found to be similar in iRFP-expressing cells and non-iRFP-expressing cells, an indication that iRFP expression and detection do not affect retinal light responsiveness. Taken together, our results suggest iRFP can be a new tool for vision research, allowing for single-cell recordings from an iRFP marked neuron using conventional fluorescence microscopy.


Subject(s)
Bacterial Proteins/analysis , Green Fluorescent Proteins/analysis , Luminescent Proteins/analysis , Retina/chemistry , Retina/cytology , Animals , Bacterial Proteins/biosynthesis , Excitatory Postsynaptic Potentials/physiology , Green Fluorescent Proteins/biosynthesis , HEK293 Cells , Humans , Luminescent Proteins/biosynthesis , Mice , Mice, Inbred C57BL , Mice, Transgenic , Microscopy, Fluorescence/methods , Photic Stimulation/methods , Photochemical Processes , Retina/metabolism
8.
J Neurosci ; 34(26): 8761-71, 2014 Jun 25.
Article in English | MEDLINE | ID: mdl-24966376

ABSTRACT

In the visual system, diverse image processing starts with bipolar cells, which are the second-order neurons of the retina. Thirteen subtypes of bipolar cells have been identified, which are thought to encode different features of image signaling and to initiate distinct signal-processing streams. Although morphologically identified, the functional roles of each bipolar cell subtype in visual signal encoding are not fully understood. Here, we investigated how ON cone bipolar cells of the mouse retina encode diverse temporal image signaling. We recorded bipolar cell voltage changes in response to two different input functions: sinusoidal light and step light stimuli. Temporal tuning in ON cone bipolar cells was diverse and occurred in a subtype-dependent manner. Subtypes 5s and 8 exhibited low-pass filtering property in response to a sinusoidal light stimulus, and responded with sustained fashion to step-light stimulation. Conversely, subtypes 5f, 6, 7, and XBC exhibited bandpass filtering property in response to sinusoidal light stimuli, and responded transiently to step-light stimuli. In particular, subtypes 7 and XBC were high-temporal tuning cells. We recorded responses in different ways to further examine the underlying mechanisms of temporal tuning. Current injection evoked low-pass filtering, whereas light responses in voltage-clamp mode produced bandpass filtering in all ON bipolar cells. These findings suggest that cone photoreceptor inputs shape bandpass filtering in bipolar cells, whereas intrinsic properties of bipolar cells shape low-pass filtering. Together, our results demonstrate that ON bipolar cells encode diverse temporal image signaling in a subtype-dependent manner to initiate temporal visual information-processing pathways.


Subject(s)
Retina/physiology , Retinal Bipolar Cells/physiology , Retinal Cone Photoreceptor Cells/physiology , Visual Pathways/physiology , Animals , Male , Mice , Mice, Inbred C57BL , Photic Stimulation , Retina/cytology , Retinal Bipolar Cells/cytology , Retinal Cone Photoreceptor Cells/cytology , Synapses/physiology , Visual Pathways/cytology
9.
J Comp Neurol ; 501(6): 891-903, 2007 Apr 20.
Article in English | MEDLINE | ID: mdl-17311321

ABSTRACT

The hyperpolarization-activated and cyclic nucleotide-gated (HCN) channel isoforms HCN1, HCN2, and HCN4 were localized by immunofluorescence in the rat retina. Double labeling with the vesicular glutamate transporter (VGLUT1) was used to identify bipolar cell axon terminals in the inner retina. The HCN1 channel was localized to two cell types with differing intracellular distributions, insofar as staining was seen in the dendrites of a putative OFF-type cone bipolar cell and in the axon terminals of an ON-type bipolar that ramifies in stratum 3 (s3) of the inner plexiform layer (IPL). Staining for HCN4 was seen in two sets of bipolar axon terminals located in s2 and s3 and positioned between the two bands of choline acetyltransferase (ChAT) staining. The cells that ramify in s2 were identified as type 3 cone bipolar cells and the cells that ramify in s3 cells as a subclass of type 5 cone bipolars. The latter group, designated here as type 5b, exhibit diffuse axon terminals and can be distinguished from the narrowly stratifying type 5a cells. Double labeling showed that type 5b cone bipolar cells express both HCN1 and HCN4 as well as HCN2. Superposition of HCN channel labeling with VGLUT1 staining confirmed the presence of a cone bipolar cell whose terminals ramify in the same stratum of the IPL as type 5b cells but that do not express these HCN channels.


Subject(s)
Ion Channels/metabolism , Nerve Tissue Proteins/metabolism , Retina/metabolism , Retinal Bipolar Cells/metabolism , Retinal Cone Photoreceptor Cells/metabolism , Animals , Cyclic Nucleotide-Gated Cation Channels , Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels , Immunohistochemistry , Ion Channels/classification , Male , Potassium Channels , Protein Isoforms , Rats , Rats, Sprague-Dawley , Retina/ultrastructure , Retinal Bipolar Cells/ultrastructure , Retinal Cone Photoreceptor Cells/ultrastructure , Tissue Distribution , Vesicular Glutamate Transport Protein 1/metabolism , Visual Pathways/cytology , Visual Pathways/metabolism
10.
Vis Neurosci ; 21(4): 551-65, 2004.
Article in English | MEDLINE | ID: mdl-15579221

ABSTRACT

Retrieval of glutamate from extracellular sites in the retina involves at least five excitatory amino acid transporters. Immunocytochemical analysis of the cat retina indicates that each of these transporters exhibits a selective distribution which may reflect its specific function. The uptake of glutamate into Muller cells or astrocytes appears to depend upon GLAST and EAAT4, respectively. Staining for EAAT4 was also seen in the pigment epithelium. The remaining transporters are neuronal with GLT-1alpha localized to a number of cone bipolar, amacrine, and ganglion cells and GLT-1v in cone photoreceptors and several populations of bipolar cells. The EAAC1 transporter was found in horizontal, amacrine, and ganglion cells. Staining for EAAT5 was seen in the axon terminals of both rod and cone photoreceptors as well as in numerous amacrine and ganglion cells. Although some of the glutamate transporter molecules are positioned for presynaptic or postsynaptic uptake at glutamatergic synapses, others with localizations more distant from such contacts may serve in modulatory roles or provide protection against excitoxic or oxidative damage.


Subject(s)
Cats/metabolism , Monosaccharide Transport Proteins/metabolism , Retina/cytology , Retina/metabolism , Subcellular Fractions/metabolism , Amacrine Cells/metabolism , Animals , Astrocytes/metabolism , Glutamic Acid/metabolism , Immunohistochemistry/methods , Retinal Ganglion Cells/metabolism , Staining and Labeling , Tissue Distribution
11.
J Comp Neurol ; 475(4): 518-30, 2004 Aug 02.
Article in English | MEDLINE | ID: mdl-15236233

ABSTRACT

Vesicular transporters play an essential role in the packaging of glutamate for synaptic release and so are of particular importance in the retina, where glutamate serves as the neurotransmitter for photoreceptors, bipolar cells, and ganglion cells. In the present study, we have examined the distribution of the three known isoforms of vesicular glutamate transporter (VGLUT) in the cat retina. VGLUT1 was localized to all photoreceptor and bipolar cells, whereas VGLUT2 was found in ganglion cells. This basic pattern of complementary distribution for the two transporters among known populations of glutamatergic cells is similar to previous findings in the brain and spinal cord. However, the axon terminals of S-cone photoreceptors were found to express both VGLUT1 and VGLUT2 and some ganglion cells labeled for both VGLUT2 and VGLUT3. Such colocalizations suggest the existence of dual modes of regulation of vesicular glutamate transport in these neurons. Staining for VGLUT2 was also present in a small number of varicose processes, which were seen to ramify throughout the inner plexiform layer. These fibers may represent axon collaterals of ganglion cells. The most prominent site of VGLUT3 immunoreactivity was in a population of amacrine cells; the axon terminals of B-type horizontal cells were also labeled at their contacts with rod spherules. The presence of the VGLUT3 transporter at sites not otherwise implicated in glutamate release may indicate novel modes of glutamate signaling or additional roles for the transporter molecule.


Subject(s)
Amino Acid Transport System X-AG/physiology , Membrane Transport Proteins , Retina/anatomy & histology , Retina/physiology , Vesicular Transport Proteins , Amino Acid Transport System X-AG/ultrastructure , Amino Acid Transport Systems, Acidic/physiology , Amino Acid Transport Systems, Acidic/ultrastructure , Animals , Carrier Proteins/physiology , Carrier Proteins/ultrastructure , Cats , Immunohistochemistry , Microscopy, Electron , Neurons/cytology , Neurons/physiology , Neurons/ultrastructure , Retina/ultrastructure , Vesicular Glutamate Transport Protein 1 , Vesicular Glutamate Transport Protein 2
12.
J Comp Neurol ; 464(1): 104-13, 2003 Sep 08.
Article in English | MEDLINE | ID: mdl-12866131

ABSTRACT

It has been generally accepted that rod photoreceptor cells in the mammalian retina make synaptic contact with only a single population of rod bipolar cells, whereas cone photoreceptors contact a variety of cone bipolar cells. This assumption has been challenged in rodents by reports of a type of cone bipolar cell which receives input from both rods and cones. Questions remained as to whether similar pathways are present in other mammals. We have used an antiserum against the glutamate transporter GLT1-B to visualize a population of cone bipolar cells in the cat retina which make flat contacts with axon terminals of both rod and cone photoreceptor cells. These cells are identified as OFF-cone bipolar cells and correspond morphologically to type cb1 (CBa2) cone bipolar cells which are a major source of input to OFF-beta ganglion cells in the cat retina. The GLT1-B transporter was also localized to processes making flat contacts with photoreceptor terminals in rat and rabbit retinas. Examination of tissue processed for the GluR1 glutamate receptor subunit showed that cb1 cone bipolar cells, like their rodent counterparts, express this alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-selective receptor at their contacts with rod spherules. Thus, a direct excitatory pathway from rod photoreceptors to OFF-cone bipolar cells appears to be a common feature of mammalian retinas.


Subject(s)
Neurons/ultrastructure , Retina/cytology , Retinal Cone Photoreceptor Cells/ultrastructure , Retinal Rod Photoreceptor Cells/ultrastructure , Amino Acid Transport System X-AG/metabolism , Animals , Cats , Immunohistochemistry , Microscopy, Immunoelectron/instrumentation , Microscopy, Immunoelectron/methods , Neurons/classification , Neurons/metabolism , Rabbits , Rats , Receptors, AMPA/metabolism , Retinal Cone Photoreceptor Cells/metabolism , Retinal Rod Photoreceptor Cells/metabolism , Species Specificity
13.
J Comp Neurol ; 454(3): 341-9, 2002 Dec 16.
Article in English | MEDLINE | ID: mdl-12442324

ABSTRACT

Fast-acting excitatory neurotransmission in the retina is mediated primarily by glutamate, acting at alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) -selective and kainate-selective receptors. To localize these sites of action, cat retinas were stimulated with either AMPA or kainate and processed for histochemical visualization of cobalt uptake through calcium-permeable channels. Treatment with both agonists resulted in staining of A- and B-type horizontal cells and several types of OFF cone bipolar cells; there was no evidence for staining of ON cone bipolar cells or rod bipolar cells. The subpopulations of OFF cone bipolar cells differed in their responses with two distinct types that stained heavily with cobalt after exposure to AMPA and three different types that were preferentially labeled after exposure to kainate. Although many amacrine and ganglion cells appeared to respond to both agonists, AII amacrine cells were stained after stimulation by AMPA but not by kainate. The OFF cone bipolar cells that exhibit AMPA-stimulated cobalt uptake were found to have a high level of correspondence with cells that show immunocytochemical staining for the AMPA-selective glutamate receptor subunits GluR1 and GluR2/3. Similarly, the cone bipolar cells exhibiting kainate-stimulated cobalt uptake resemble those that are immunoreactive for the kainate subunit GluR5. The results indicate that, whereas many retinal neurons express both AMPA and kainate receptors, AII amacrine cells and subpopulations of OFF cone bipolar cells are limited to the expression of either AMPA or kainate receptors. This differential expression may contribute to the unique character of transmission by these cell types.


Subject(s)
Cobalt/metabolism , Excitatory Amino Acid Agonists/pharmacology , Neurons/metabolism , Receptors, AMPA/agonists , Receptors, AMPA/biosynthesis , Receptors, Kainic Acid/agonists , Receptors, Kainic Acid/biosynthesis , Retina/metabolism , Animals , Cats , Cobalt/analysis , Neurons/chemistry , Neurons/drug effects , Neurons/ultrastructure , Receptors, AMPA/analysis , Receptors, Kainic Acid/analysis , Retina/chemistry , Retina/drug effects , Retina/ultrastructure
14.
Vis Neurosci ; 19(5): 549-62, 2002.
Article in English | MEDLINE | ID: mdl-12507322

ABSTRACT

Immunocytochemical localization was carried out for five isoforms of protein kinase C (PKC) in the cat retina. In common with other mammalian species, PKCalpha was found in rod bipolar cells. Staining was also seen in a small population of cone bipolar cells with axon terminals ramifying near the middle of the inner plexiform layer (IPL). PKCbetaI was localized to rod bipolar cells, one class of cone bipolar cell, and numerous amacrine and displaced amacrine cells. Staining for PKCbetaI was seen in three types of cone bipolar cells as well as in amacrine and ganglion cells. Immunoreactivity for both PKCepsilon and PKCzeta was found in rod bipolar cells; PKCepsilon was also seen in a population of cone bipolar cells and a few amacrine and ganglion cells whereas PKCzeta was found in all ganglion cells. Double-label immunofluorescence studies showed that dendrites of the two PKCbetaII-positive OFF-cone bipolar cells exhibit immmunoreactivity for the kainate-selective glutamate receptor GluR5. The third PKCbetaII cone bipolar is an ON-type cell and did not stain for GluR5. The retinal distribution of these isoforms of PKC is consistent with a role in modulation of various aspects of neurotransmission including synaptic vesicle release and regulation of receptor molecules.


Subject(s)
Cats/metabolism , Protein Kinase C/metabolism , Retina/enzymology , Animals , Isoenzymes/metabolism , Microscopy, Electron , Microscopy, Fluorescence , Retina/ultrastructure , Tissue Distribution
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