Biomedical subjects

D M Lam

Publications and source records attributed to D M Lam.

At least 55 records · Page 3Linked to original sources

PubMed1986

Colocalization of immunoreactive substance P and neurotensin in amacrine cells of the goldfish retina.

Using an immunohistochemical double-label technique, neurotensin and substance P immunoreactivity were localized to amacrine cells in the goldfish retina. Both peptides were found in a single population of unistratified amacrine cells branching in sublamina 3 of the inner plexiform layer. A monostratified amacrine cell branching in sublamina 1 contained only substance P immunoreactivity and a bistratified cell branching in sublaminae 1 and 3 contained only neurotensin immunoreactivity.

Animals↗

PubMed1986

The light microscopic localization of substance P- and somatostatin-like immunoreactive cells in the larval tiger salamander retina.

Light microscopic immunocytochemistry was utilized to localize the populations of substance P (SP)- and somatostatin (SOM)-like immunoreactive cells in the larval tiger salamander retina. Of 104 SP-immunostained cells observed, 82% were Type 1 amacrine cells. Another 8% of the SP-cells were classified as Type 2 amacrine cells, while 10% of the SP-cells had their cell bodies located in the ganglion cell layer and were designated as displaced amacrine cells. Each type of SP-like immunoreactive cell was observed in the central and peripheral retina. SP-immunopositive processes were observed in the inner plexiform layer as a sparse plexus in sublamina 1 and as a denser network of fibers in sublamina 5. Seventy-eight percent of the 110 somatostatin-immunopositive cells observed were designated as Type 1 amacrine cells. Another 12% of SOM-cells were classified as displaced amacrine cells, while only two SOM-immunopositive Type 2 amacrine cells were observed. Nine percent of the SOM-cells were designated as interplexiform cells, based on their giving rise to processes distributing in the outer plexiform layer as well as processes ramifying in the inner plexiform layer. Each type of SOM-immunoreactive cell was observed in the central and peripheral retina, with the exception of the Type 2 amacrine cells, whose somas were only found in the central retina. Lastly, SOM-immunopositive processes in the inner plexiform layer appeared as a fine plexus in sublamina 1 and as a somewhat denser network of fibers in sublamina 5.

Ambystoma↗

PubMed1986

Enkephalin in the goldfish retina.

Enkephalin-like immunoreactive amacrine cells were visualized using the highly sensitive avidin-biotin method. The somas of these cells were situated in the inner nuclear and ganglion cell layers. Enkephalin-stained processes were observed in layers 1, 3, and 5 of the inner plexiform layer. The biosynthesis of sulfur-containing compounds in the goldfish retina was studied by means of a pulse-chase incubation with 35S-methionine. A 35S-labeled compound, which comigrated with authentic Met5-enkephalin on high-performance liquid chromatography (HPLC), was synthesized and was bound competitively by antibodies to enkephalin and by opiate receptors. This compound was tentatively identified as "Met5-enkephalin." The newly synthesized 35S-Met5-enkephalin was released upon depolarization of the retina with a high K+ concentration. This K+-stimulated release was greatly suppressed by 5 mM Co2+, suggesting that the release was Ca2+ dependent. Using a double-label technique, enkephalin immunoreactivity and gamma-aminobutyric acid (GABA) uptake were colocalized to some amacrine cells, whereas others labeled only for enkephalin or GABA. The possible significance of enkephalin-GABA interactions is also discussed.

Animals↗

PubMed1986

Specific labelling of ganglion cells in the cat retina by a monoclonal antibody.

The introduction of monoclonal antibody technology has made it possible to produce specific markers for various retinal cell types. We report here on a monoclonal antibody, AB5, which specifically labels the retinal ganglion cells of the cat. The labelled cells could be categorized as either alpha, beta or gamma subtype based on morphological criteria. This antibody will be useful for studies of the morphology, localization and synaptic connectivity of ganglion cells in the cat retina.

Animals↗

PubMed1986

The self-regulating synapse: a functional role for the co-existence of neuroactive substances.

Although the co-localizations of neuroactive substances, such as transmitters and peptides, in identified neurons is now a common histochemical phenomenon, the physiological roles and functional significance of such co-existence are largely unknown. Using the vertebrate retina as a model for the central nervous system, we have examined the relationship between co-existence and co-function. We propose here that the co-localization of neuroactive substances in a synaptic terminal provides the structural configuration to ensure the co-release of two or more predetermined substances into the same synaptic cleft, resulting in the capability of the presynaptic neuron to stringently regulate its own activities and output.

Animals↗

PubMed1986

Proton NMR spectroscopy of glucose consumption by cultured lens epithelial cells.

Proton NMR spectroscopy was performed on media collected from cultured lens epithelial cells of the rabbit eye incubated with Krebs-Ringer's solution containing 5.5mM 13C-glucose (labeled at the C-1 position). Comparing proton resonance intensities of the lactate-methyl group of the C-3 carbon (both 12C and 13C) enabled us to quantify the hexose monophosphate shunt (HMPS) activity. Our results showed that the epithelial cells remained stable in both lactate production and HMPS activity for at least 8 hours. In addition, although tBHP (1mM) resulted in an increase of glucose flow through the HMPS, the rate of lactate production was not affected. In contrast, KCN (2mM) caused a 72% increase of lactate production and a slight (6%) decrease of glucose consumption through the HMPS.

Animals↗

PubMed1986

Interactions between vasoactive intestinal peptide and dopamine in the rabbit retina: stimulation of a common adenylate cyclase.

Although 3,4-dihydroxyphenylethylamine (dopamine, DA) and vasoactive intestinal peptide (VIP) have been reported to stimulate adenylate cyclase activity in the rabbit retina, possible interactions between VIP-sensitive and DA-sensitive adenylate cyclase systems have not been previously investigated. To elucidate the interactions between these two putative transmitter-stimulated cyclase systems, the effects of VIP, DA, and VIP + DA on the conversion of [alpha-32P]ATP to [32P]cyclic AMP in rabbit retinal homogenates were measured. VIP stimulated adenylate cyclase activity in a biphasic manner, suggesting that two classes of VIP receptors may be involved in the induction of cyclic AMP formation. DA was less potent than VIP, and stimulated cyclase activity with a monophasic dose-response curve. When assayed together, these stimulations were partially nonadditive, implying the existence of a common adenylate cyclase pool that may be stimulated by both putative neurotransmitters. The dopaminergic antagonist (+)-butaclamol completely blocked dopaminergic stimulation, but had no significant effect on VIP-induced stimulation, indicating that VIP interacts with specific VIP receptor sites, which are distinct from the dopaminergic receptor sites. Furthermore, the specific D-2 dopaminergic receptor agonist LY141865 demonstrated no inhibitory effect on adenylate cyclase activity, suggesting that the interaction between the VIP- and DA-sensitive adenylate cyclase systems does not result from a D-2 receptor-mediated cyclase inhibition in the rabbit retina. Finally, at maximally effective concentrations, DA and VIP were less potent than fluoride or forskolin in the stimulation of cyclic AMP formation, suggesting that adenylate cyclase pools that are not sensitive to DA and VIP may also be present in this retina.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylyl Cyclases↗

PubMed1985

The presence of three neuroactive peptides in putative glycinergic amacrine cells of an avian retina.

Amacrine cells are retinal interneurons which serve to mediate transmission between bipolar and ganglion cells. To date, in addition to several classical neurotransmitters, a number of neuroactive peptides have been localized to these cells. We have previously demonstrated in the chicken that both peptide-transmitter and peptide-peptide colocalization exists in some amacrine cells. In this report, we show that 3 neuroactive peptides (enkephalin, neurotensin and somatostatin) are present in subpopulations of amacrine cells which also possess a high affinity uptake system for glycine. These observations suggest that the simultaneous visualization by autoradiography of [3H]glycine-uptake and immunocytochemistry of peptides may be useful for distinguishing between different types of putative glycinergic amacrine cells.

Animals↗

PubMed1985

Localization of enkephalin-like immunoreactive amacrine cells in the larval tiger salamander retina: a light and electron microscopic study.

Immunohistochemistry was utilized to examine the light and electron microscopic localization of enkephalin-like (enk) immunoreactive amacrine cells in the larval tiger salamander retina. The vast majority of enk-immunoreactive cells were typical amacrine cells whose round or oval cell bodies (14-16 microns) were situated in the innermost cell row of the inner nuclear layer. A relatively small number of enk-stained oval cell bodies (14-22 microns) were located in the ganglion cell layer and were designated as those of displaced amacrine cells. Enkephalin immunostaining was observed in the inner plexiform layer as a fine plexus in sublamina 1 and as a dense network of fibers in sublamina 5. In both the center and periphery of the retina the density of typical enk-amacrine cells was determined to be 250 +/- 16.36 cells per mm2 surface area of the retina. At the ultrastructural level typical enk-stained amacrine cells possessed a round, indented nuclear membrane. Enk-immunoreactive processes sometimes contained dense-core vesicles (60-115 nm) in addition to a rather homogeneous population of small, round, agranular synaptic vesicles (25-35 nm). In sublamina 1 the processes of enk-amacrine cells were presynaptic to amacrine and bipolar cells. They also contacted processes devoid of synaptic vesicles which possibly arise from ganglion cells. As the postsynaptic element in sublamina 1, they received synaptic input from amacrine cells. In sublamina 5 the processes of enk-amacrine cells were presynaptic to amacrine cells, bipolar cells, and the somas of cells situated in the ganglion cell layer.(ABSTRACT TRUNCATED AT 250 WORDS)

Ambystoma↗

PubMed1985

The coexistence of two neuroactive peptides in a subpopulation of retinal amacrine cells.

Amacrine cells are axonless interneurons of the vertebrate retina. Over the past few years many neuroactive peptides have been localized to these cells. We have previously demonstrated that a neuropeptide and a classical transmitter are colocalized in some amacrine cells. In this report, we show that two neuroactive peptides, which probably arise from different precursors, also coexist in some amacrine cells. Although some amacrine cells contain only enkephalin or neurotensin, others contain both of these neuroactive peptides.

Animals↗

PubMed1985

Localization of GABA immunoreactivity in the auditory brainstem of guinea pigs.

Using an antibody against GABA, neurons within the guinea pig hindbrain, midbrain and forebrain auditory nuclei were identified which demonstrate GABA-like immunoreactivity. GABA-positive cells were localized in the cochlear nucleus, superior olivary complex, lateral lemniscus, inferior colliculus, and medial geniculate body. GABA-positive terminals could be seen surrounding globular and spherical cells in ventral cochlear nucleus and principal cells in medial nucleus of the trapezoid body. In addition, numerous positive, punctate terminals appeared throughout the hindbrain auditory nuclei and, although fewer in number, in midbrain and forebrain auditory nuclei.

Animals↗

PubMed1985

A monoclonal antibody specific for retinal ganglion cells of mammals.

The development of specific markers for retinal ganglion cells is an area of great interest in retinal research. In this study we report on a monoclonal antibody (AB5) which specifically labels ganglion cells in rabbit, cat and monkey, as well as a variety of other mammalian species. Labelling of ganglion cells was also observed in isolated cell preparations of rabbit retina.

Animals↗

PubMed1985

Development of the subretinal space in the preterm human eye: ultrastructural and immunocytochemical studies.

To investigate the development of the subretinal space in the human infant, eyes were obtained from 12 live-born, anomaly-free, preterm infants from 20 to 32 weeks gestation and from one 3-month postterm infant. The retinas were studied by light microscopy, electron microscopy, and immunocytochemistry. The immunocytochemical studies utilized rabbit antiserum against purified bovine interstitial retinol binding protein (IRBP). A subretinal space containing IRBP was present in the central retina at 20 weeks and extended further into the periphery (expressed as a percentage of the distance from the optic disc to the ora serrata in the temporal hemisphere) as the retina developed. At 28 weeks, IRBP was absent only from the most peripheral 25% of the retina and reached the temporal ora serrata at 32 weeks. At 3 months postterm, IRBP immunofluorescence outlined fully developed photoreceptors, which were present from the optic disc to the ora serrata. The appearance of IRBP in the subretinal space correlated with the development of the first photoreceptor outer segment discs.

Fluorescent Antibody Technique↗

PubMed1985

The content and release of endogenous GABA in isolated horizontal cells of the goldfish retina.

The endogenous amino acid content of intact retinas and isolated horizontal cells was measured by high performance liquid chromatography (HPLC). In both of these preparations, the concentration of gamma-aminobutyric acid (GABA) was approximately 2 mM. The endogenous GABA of the isolated horizontal cells was specifically released upon depolarization of the cells by elevated external K+ concentrations. This K+ induced release of endogenous GABA has two components. There is a saturable, Ca2+-dependent portion which predominates with small depolarizations, and a Ca2+-independent fraction which is linearly dependent upon the membrane potential of the horizontal cells. The data presented suggest the possibility of a dual scheme for the release of GABA from goldfish horizontal cells. The first is a Ca2+-dependent, conventional synaptic release; and the second a Na+-driven, carrier mediated release, which is linearly dependent upon the cellular membrane potential and functions in the absence of extracellular Ca2+.

Amino Acids↗

PubMed1985

The signature hypothesis: co-localizations of neuroactive substances as anatomical probes for circuitry analyses.

The recent discoveries that a neuron in the vertebrate retina may contain more than one neuroactive substance (transmitter or neuropeptide) raise the possibility that within each class of neurons, every morphologically and physiologically distinct cell type may be uniquely identified and categorized by the neuroactive substances that it contains. This article examines the evidence to-date for such a conjecture and discusses some of its potential applications and implications.

Animals↗

PubMed1985

Opioid pathways in an avian retina. I. The content, biosynthesis, and release of Met5-enkephalin.

By means of an enzyme-linked immunosorbent assay, the concentration of enkephalin-immunoreactive substances was estimated to be about 25 nM in the chicken retina. The biosynthesis of 3H-Met5-enkephalin in this retina was studied by a pulse-chase incubation technique. Isolated retinas were incubated with 0.2 ml of oxygenated Ringer's solution containing 40 microCi of [3H]methionine and trasylol, a peptidase inhibitor, for 30 min at room temperature. The tissue was then rinsed three times in large volumes of Ringer's solution and incubated in the same solution containing unlabeled methionine (100 micrograms/ml) and trasylol for at least another hour. The products synthesized were extracted in acetic acid and assayed by high performance liquid chromatography (HPLC) and immunoassay. A peak of radioactivity that comigrated with Met5-enkephalin on HPLC and cross-reacted with antibodies against enkephalins was detected. The level of 3H-Met5-enkephalin radioactivity increased approximately 10-fold as the chase-incubation period increased from 0 to 120 min, suggesting that, as in other tissues, Met5-enkephalin may be synthesized as part of a larger precursor. The newly synthesized Met5-enkephalin could be released by depolarization of the retina with high extracellular K+ concentration. Furthermore, this K+-stimulated release was greatly suppressed by 5 mM Co2+ in the medium, suggesting that this release is Ca2+-dependent and may be synaptically mediated.

Animals↗

PubMed1985

Opioid pathways in an avian retina. II. Synaptic organization of enkephalin-immunoreactive amacrine cells.

Peroxidase-antiperoxidase immunocytochemistry was utilized to investigate the synaptic organization of enkephalin-like immunoreactive amacrine (Enk-amacrine) cells in the chicken retina. An initial light microscopic analysis revealed that immunostained cell bodies were situated in either the second or third tier of cells from the border of the inner nuclear and inner plexiform layers. The processes of such cells extended into the inner plexiform layer where they ramified as a fine plexus in sublaminae 1 and 3 to 5. At the ultrastructural level, enkephalin-positive somas exhibited a rather dense and evenly distributed peroxidase reaction product throughout their cytoplasm. Furthermore, the nucleus of Enk-amacrine cells was characterized by a round, unindented nuclear membrane. Immunoreactive processes of such cells were found to receive synaptic input from unstained amacrine and bipolar cells. Immunolabeled varicosities formed conventional synaptic contacts onto other vesicle-filled, nonimmunoreactive profiles tentatively suggested to originate from amacrine cells. Bipolar cell terminals did not receive synaptic input from stained varicosities of enkephalin-immunoreactive amacrine cells. Moreover, each of the above synaptic relationships were identified in each of sublaminae 1, and 3 to 5 of the inner plexiform layer. In addition, labeled varicosities of Enk-amacrine cells synapsed upon unstained processes which lacked synaptic vesicles and possibly arise from ganglion cells. Enkephalin-positive processes were also observed to synapse upon the vitreal pole of unstained somas situated in the innermost cell row of the inner nuclear layer. Lastly, large immunoreactive varicosities of Enk-amacrine cells were often characterized by the presence of large dense-core vesicles in addition to typical, small agranular synaptic vesicles.

Animals↗

PubMed1984

The identification and some functions of GABAergic neurons in the distal catfish retina.

Putative GABAergic neurons in the outer retina of the Texas channel catfish, Ictalurus punctatus, were studied using autoradiographic, biochemical and electrophysiological techniques. A red cone horizontal cell was found to accumulate exogenous GABA in the presence of red light. GABA could be released from these cells with high K+ Ringers solution. The release was only partially blocked by Co2+ and therefore may be only partially Ca2+ dependent. The red cone horizontal cells were found to contain significant activities of L-glutamic acid decarboxylase and GABA transaminase, the enzymes responsible for GABA synthesis and degradation respectively. These data suggest that catfish red cone horizontal cells are GABAergic. To substantiate this, recordings were made from photoreceptors and horizontal cells during the superfusion of the GABA blocking agents bicuculline methochloride or picrotoxin. These agents modified the cone responses in the manner specified if they were blocking the feedback pathway from horizontal cells to cones. Thus it is likely that the horizontal cells are using GABA as the transmitter in the feedback pathway. In addition, the GABA blocking agents were found to interfere with changes in horizontal cell responses which occur during light adaptation.

Adaptation, Ocular↗