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B A MacVicar

Publications and source records attributed to B A MacVicar.

67 records · Page 4Linked to original sources
PubMed1984

An incubation chamber for the simultaneous, on-slide treatment of brain sections with different reagents.

The design and use of a new microscope slide incubation chamber is described. The chief advantage of this device is that it allows the simultaneous incubation of tissue sections which are mounted on glass slides, either together with the same reagent, or individually with different reagents. Simple construction makes this chamber durable, reliable, and inexpensive. Advantages and disadvantages of this device, as well as, other published devices are discussed.

Animals↗
PubMed1983

Polyethylene glycol embedding: a technique compatible with immunocytochemistry, enzyme histochemistry, histofluorescence and intracellular staining.

A technique is described which permits rapid processing of neural tissue for light microscopic analysis of sections of 1-40 microns thickness. This technique was developed as an alternative to paraffin embedding. When compared to paraffin, polyethylene glycol (PEG) offers the following advantages: 10-15 degrees C lower embedding temperatures, net tissue shrinkage of less than 5% vs approximately 50% in paraffin, and approximately one-half the embedding time. Tissue orientation during embedding and sectioning is particularly easy to control, e.g. 500 microns brain slices can be routinely flat-embedded and sectioned at 5 microns to form excellent ribbons. Since PEG is water-soluble, tissue may be dehydrated with a series of aqueous PEG solutions; the embedding matrix is easily removed by washing with a variety of aqueous buffers. These procedures allow subsequent electron microscopic analysis of material with generally well preserved ultrastructure. However, PEG is hygroscopic, thus tissue blocks become soft and difficult to section in high (greater than 90%) relative humidity. PEG was found to be compatible with intracellular staining with Lucifer yellow, horseradish peroxidase enzyme histochemistry, aqueous histofluorescence and immunocytochemical demonstration of neuronal peptides and glial fibrillary acidic protein.

Animals↗
PubMed1982

Dye-coupling between pyramidal cells of rat hippocampus in vivo.

Lucifer Yellow was intracellularly injected into pyramidal cells of rat hippocampus in vivo. In 10 successful staining experiments there were 5 cases of dye-coupling, in which 2-4 cells were stained though only one was injected. These results indicate that dye-coupling and, therefore, presumably electrotonic coupling, occurs between neurons of the hippocampus in the whole animal and confirms previous findings in the hippocampal slice in vitro.

Animals↗
PubMed1982

Synaptic inputs and action potentials of magnocellular neuropeptidergic cells: intracellular recording and staining in slices of rat hypothalamus.

Excitatory postsynaptic potentials (EPSPs) and action potentials of magnocellular neuropeptidergic cells (MNCs) in the paraventricular (PVN) and supraoptic nuclei (SON) were studied with intracellular recording in coronal slices of rat hypothalamus. The fluorescent dye Lucifer Yellow (LY) was injected intracellularly and the cells were subsequently identified as magnocellular (somata greater than 15 x 15 micrometer). These cells generally had a large cytoplasm-to-nucleus ratio. In PVN it was frequently possible to trace filled dendrites to the ependyma of the third ventricle, and occasionally dendritic spines could be seen. Electrical stimuli in areas dorsolateral and ventrolateral to the fornix column evoked EPSPs in some anatomically identified MNCs of PVN, which indicates that presynaptic fibers innervating MNCs approach PVN from this region. Short-latency (less than 1 msec) spikes could be evoked in many MNCs of PVN by stimulation near SON, which is consistent with the known projection to the neurohypophysis of many MNCs. Action potentials in MNCs of PVN and SON had significantly longer durations at one-third spike height (mean +/- S.D. = 2.06 +/- 0.6 msec) than hippocampal CA1 pyramidal cells (1.17 +/- 0.29 msec). This suggests that neuroendocrine cells in mammals and some lower vertebrates and invertebrates are similar in this regard.

Action Potentials↗
PubMed1982

Intracellular electrophysiology of mammalian peptidergic neurons in rat hypothalamic slices.

The magnocellular neuropeptidergic cells (MNCs) of the paraventricular and supraoptic nuclei have been a model for biochemical and physiological studies of peptidergic neurons in the mammalian brain, but nearly all the electrophysiological studies of these vasopressinergic and oxytocinergic neuroendocrine cells are based on extracellular recordings. This paper reviews recent literature on electrophysiological properties of neurons in the magnocellular nuclei in which the rat in vitro slice preparation and intracellular recording were used. Spontaneously occurring action potentials and synaptic potentials (excitatory and inhibitory) have been observed in hypothalamic slices. The spike patterns have included slow and irregular firing, short rapid bursts of inactivating spikes, and slow phasic discharge with prolonged active and silent periods. Some studies have shown that increased osmolality causes neuronal firing, but this area is controversial. Intracellular injections of lucifer yellow have shown that some MNCs are dye-coupled and electron microscopic observations with the freeze-fracture technique have revealed occasional gap junctions, thus suggesting that some MNCs are electrotonically coupled. Both excitatory and inhibitory postsynaptic potentials have been evoked with extracellular stimulation. Therefore, action potentials, synaptic potentials, burst discharges, and probably electrotonic coupling have been found with intracellular recording in mammalian neuroendocrine cells. Future studies with intracellular recording and staining followed by immunohistochemical identification of cells should provide significant new information on the membrane physiology and synaptic pharmacology of vasopressinergic and oxytocinergic cells.

Action Potentials↗
PubMed1981

Electrotonic coupling between pyramidal cells: a direct demonstration in rat hippocampal slices.

Intracellular recordings from pairs of neurons in slices of rat hippocampus directly demonstrated electronic coupling between CA3 pyramidal cells. When two neurons were impaled simultaneously (as verified by subsequent double staining with horseradish peroxidase), current pulses injected into one cell caused voltage changes in other cells. These interactions were bidirectional. Fast prepotentials, historically thought to represent spike activity in dendrites, resulted from action potentials in other electronically coupled pyramidal cells. These data directly demonstrate electrotonic coupling between neurons in the mammalian brain and indicate that some fast prepotentials are coupling potentials. Coupling between pyramidal cells could mediate synchronization of normal rhythmic activity and of burst discharges during seizures.

Action Potentials↗
PubMed1981

Dye transfer through gap junctions between neuroendocrine cells of rat hypothalamus.

Most magnocellular neurosecretory cells that terminate in the posterior pituitary secrete either vasopressin, oxytocin, or enkephalin. Intracellular injection of the fluorescent dye Lucifer Yellow into single magnocellular neurons in slices of rat hypothalamus resulted in dye transfer between these cells. Freeze-fracture replicas of these cells occasionally revealed gap junctions, which presumably contain channels that mediate the dye coupling. These two independent techniques strongly suggest that some mammalian neuropeptidergic cells are electrotonically coupled, providing a possible means for recruitment and synchronization of their electrical activity.

Animals↗
PubMedDate not supplied

Membrane conductance oscillations in astrocytes induced by phorbol ester.

Glial cells in the central nervous systems (CNS) have complex functions which are difficult to decipher because of the intimate intertwining of glial cells with neurons. We have therefore developed an essentially neuron-free preparation of CNS astrocytes in the kainic acid lesioned hippocampal slice. With this preparation we have examined the effect of activating protein kinase C in astrocytes with a phorbol ester, TPA (12-O-tetradecanoyl-phorbol-13-acetate). In most cells, TPA induced rhythmic oscillations (0.1-3.0 Hz) of membrane potential which were typically 5-10 mV in amplitude and were associated with increases of up to eightfold in input resistance during the depolarizing phase. These large changes in membrane conductance are the first reported observations of endogenously generated conductance changes in astrocytes of the mammalian CNS and they could influence excitability of surrounding neurons, possibly by altering extracellular ion concentrations.

Animals↗