Glutamate microinjections in cerebellar cortex reproduce cerebrovascular effects of parallel fiber stimulation

1996 ◽  
Vol 271 (6) ◽  
pp. R1568-R1575 ◽  
Author(s):  
G. Yang ◽  
C. Iadecola
Keyword(s):  
Glutamate Receptors ◽  
Cerebellar Cortex ◽  
Glutamate Release ◽  
Molecular Layer ◽  
Parallel Fiber ◽  
Synaptic Activity ◽  
No Production ◽  
Doppler Flowmetry ◽  
Cranial Window ◽  
Fiber Stimulation

Electrical stimulation of cerebellar parallel fibers releases glutamate and increases local blood flow (BFcrb), an effect in part mediated by glutamate-induced nitric oxide (NO) production. We studied whether local microinjection of glutamate into the cerebellar cortex would produce increases in BFcrb comparable to those elicited by parallel fiber stimulation. In halothane-anesthetized rats equipped with a cranial window, glutamate was microinjected into the cerebellar molecular layer, and BFcrb was monitored by laser-Doppler flowmetry. Glutamate microinjections increased BFcrb dose dependently (2-200 pmol in 200 nl) (n = 9) and by 55 +/- 6% at 200 pmol (mean +/- SE). The magnitude and temporal profile of the increases in BFcrb compared favorably with the increase in flow produced by parallel fiber stimulation. The glutamate-induced BFcrb increase was attenuated by superfusion with the Na2+ channel blocker tetrodotoxin (10 microM; -50 +/- 10%; n = 5; P < 0.05; t-test) or by blocking synaptic activity by treatment of the cerebellar cortex with Ringer containing 20 mM Mg2+ and 0 mM Ca2+ (-80 +/- 4%; n = 6; P < 0.05). The glutamate-receptor antagonist kynurenate (10 mM) attenuated the increase in BFcrb by 59 +/- 6% (P < 0.05; n = 5). The relatively selective inhibitor of neuronal NO synthase 7-nitroindazole (100 mg/kg ip) reduced the flow response evoked by microinjection of glutamate (-46 +/- 7%; n = 5; P < 0.05) but not acetylcholine (10 microM; P > 0.05; n = 6). We conclude that glutamate microinjections increase local BFcrb via activation of glutamate receptors. The glutamate-induced vasodilation is mediated, in part, by neurally derived NO. The striking similarities between the vascular responses evoked by parallel fiber stimulation and that produced by microinjection of glutamate support the hypothesis that the increase in BFcrb produced by parallel fiber stimulation is mediated by glutamate release and activation of glutamate receptors. The data also strengthen the hypothesis that glutamate and NO are important mediators in the mechanisms linking synaptic activity to BFcrb in cerebellar cortex.

Local and Propagated Vascular Responses Evoked by Focal Synaptic Activity in Cerebellar Cortex

1997 ◽  
Vol 78 (2) ◽  
pp. 651-659 ◽  
Author(s):  
Costantino Iadecola ◽  
Guang Yang ◽  
Timothy J. Ebner ◽  
Gang Chen
Keyword(s):  
Cerebellar Cortex ◽  
Synaptic Activity ◽  
Field Potentials ◽  
Vascular Changes ◽  
Vasoactive Agents ◽  
Doppler Flowmetry ◽  
Vascular Responses ◽  
Cranial Window ◽  
Synaptic Release ◽  
Horizontal Beam

Iadecola, Costantino, Guang Yang, Timothy J. Ebner, and Gang Chen. Local and propagated vascular responses evoked by focal synaptic activity in cerebellar cortex. J. Neurophysiol. 78: 651–659, 1997. We investigated the local and remote vascular changes evoked by activation of the cerebellar parallel fibers (PFs). The PFs were stimulated (25–150 μA, 30 Hz) in halothane-anesthetized rats equipped with a cranial window. The changes in arteriolar and venular diameter produced by PF stimulation were measured with the use of a videomicroscopy system. Cerebellar blood flow (BFcrb) was monitored by laser Doppler flowmetry and the field potentials evoked by PF stimulation were recorded with the use of microelectrodes. PF stimulation increased the diameter of local arterioles (+26 ± 1%, mean ± SE) in the activated folium ( n = 10, P < 0.05). The vasodilation was greatest in smaller arterioles (16.5 ± 0.8 μm), was graded with the intensity of stimulation, and was less marked than the vasodilation produced by hypercapnia in comparably sized vessels (+58 ± 5%, CO2 pressure = 50–60 mmHg, n = 8). In addition, the vasodilation was greatest along the horizontal beam of activated PFs and was reduced in arterioles located away from the stimulated site in a rostrocaudal direction. The increases in vascular diameter were associated with increases in BFcrb in the activated area (+55 ± 4%, n = 5). PF stimulation increased vascular diameter (+10 ± 0.5%, n = 10) also in larger arterioles (30–40 μm) located in the folium adjacent to that in which the PFs were stimulated. Higher-order branches of these arterioles supplied the activated area. No field potentials were evoked by PF stimulation in the area where these upstream vessels were located. The data suggest that increased synaptic activity in the PF system produces a “local” hemodynamic response mediated by synaptic release of vasoactive agents and a “remote” response that is propagated to upstream arterioles from vessels residing in the activated folium. These propagated vascular responses are important in the coordination of segmental vascular resistance that is required to increase flow effectively during functional brain hyperemia.

Postsynaptic Current Mediated by Metabotropic Glutamate Receptors in Cerebellar Purkinje Cells

1998 ◽  
Vol 80 (2) ◽  
pp. 520-528 ◽  
Author(s):  
Filippo Tempia ◽  
Maria Concetta Miniaci ◽  
Davide Anchisi ◽  
Piergiorgio Strata
Keyword(s):  
Glutamate Receptors ◽  
Purkinje Cells ◽  
Metabotropic Glutamate Receptors ◽  
Parallel Fiber ◽  
High Concentration ◽  
Metabotropic Glutamate ◽  
Group I ◽  
Cerebellar Purkinje Cells ◽  
Mglur Antagonist ◽  
Fiber Stimulation

Tempia, Filippo, Maria Concetta Miniaci, Davide Anchisi, and Piergiorgio Strata. Postsynaptic current mediated by metabotropic glutamate receptors in cerebellar Purkinje cells. J. Neurophysiol. 80: 520–528, 1998. In rat cerebellar slices, repetitive parallel fiber stimulation evokes an inward, postsynaptic current in Purkinje cells with a fast component mediated by α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate receptors and a slower component mediated by metabotropic glutamate receptors (mGluR). The mGluR-mediated excitatory postsynaptic current (mGluR-EPSC) is evoked selectively by parallel fiber stimulation; climbing fiber stimulation is ineffective. The mGluR-EPSC is elicited most effectively with increasing frequencies of parallel fiber stimulation, from a threshold of 10 Hz to a maximum response at ∼100 Hz. The amplitude of the mGluR-EPSC is a linear function of the number of stimulus pulses without any apparent saturation, even with >10 pulses. Thus mGluRs at the parallel fiber-Purkinje cell synapse can function as linear detectors of the number of spikes in a burst of activity in parallel fibers. The mGluR-EPSC is present from postnatal day 15 and persists into adulthood. It is inhibited by the generic mGluR antagonist (RS)-a-methyl-4-carboxyphenylglycine and by the group I mGluR antagonist (RS)-1-aminoindan-1,5-dicarboxylic acid at a concentration selective for mGluR1. Although the intracellular transduction pathway involves a G protein, the putative mediators of mGluR1 (phospholipase C and protein kinase C) are not directly involved, indicating that the mGluR-EPSC studied here is mediated by a different and still unidentified second-messenger pathway. Heparin, a nonselective antagonist of inositol-trisphosphate (IP3) receptors, has no significant effect on the mGluR-EPSC, suggesting that also IP3 might be not required for the response. Buffering intracellular Ca2+ with a high concentration of bis-( o-aminophenoxy)- N,N,N′,N′-tetraacetic acid partially inhibits the mGluR-EPSC, indicating that Ca2+ is not directly responsible for the response but that resting Ca2+ levels exert a tonic potentiating effect on the mGluR-EPSC.

Nitric oxide contributes to functional hyperemia in cerebellar cortex

1995 ◽  
Vol 268 (5) ◽  
pp. R1153-R1162 ◽  
Author(s):  
C. Iadecola ◽  
J. Li ◽  
T. J. Ebner ◽  
X. Xu
Keyword(s):  
Nitric Oxide ◽  
Blood Flow ◽  
Catalytic Activity ◽  
Cerebellar Cortex ◽  
Ex Vivo ◽  
Molecular Layer ◽  
Nerve Fibers ◽  
Functional Hyperemia ◽  
Field Potentials ◽  
Doppler Flowmetry

We used the parallel fibers (PF) system of the cerebellar cortex as a model to investigate the role of nitric oxide (NO) in the increases in blood flow elicited by neural activation. Rats were anesthetized with halothane and ventilated. The vermis was exposed, and the site was superfused with Ringer (37 degrees C; pH 7.3-7.4). PF were stimulated electrically (100 muA; 30 Hz), and the associated changes in cerebellar cortex blood flow (BFcrb) were monitored by laser-Doppler flowmetry. The field potentials evoked by PF stimulation were recorded using microelectrodes. During Ringer superfusion (n = 7), PF stimulation increased BFcrb (+ 52 +/- 4%). Topical application of the NO synthase (NOS) inhibitor N omega-nitro-L-arginine (L-NNA; 0.1-1 mM) attenuated the increases in BFcrb dose dependently and by 50 +/- 4% at 1 mM (n = 9; P < 0.001; analysis of variance and Tukey's test). L-NNA (1 mM) inhibited NOS catalytic activity, assessed ex vivo using the citrulline assay, by 95 +/- 9% (P < 0.001). L-NNA did not influence the field potentials evoked by PF stimulation. D-NNA (1 mM; n = 6), the inactive stereoisomer of nitroarginine, did not attenuate the BFcrb response (P > 0.05). Methylene blue (1 mM; n = 7) reduced the response by 41 +/- 9% (P < 0.01) without affecting NOS catalytic activity (P < 0.05). The increases in BFcrb were not affected by lesioning the NOS-containing nerve fibers innervating cerebral vessels, indicating that these nerves are not the source of NO. Thus the increases in BFcrb elicited by activation of PF are, in part, mediated by NO produced in the molecular layer during neural activity. The results indicated that NO participates in the coupling of function activity to blood flow and support the hypothesis that NO is one of the mediators responsible for functional hyperemia in the central nervous system.

Laminar analysis of activity-dependent increases of CBF in rat cerebellar cortex: dependence on synaptic strength

1997 ◽  
Vol 273 (3) ◽  
pp. H1166-H1176 ◽  
Author(s):  
N. Akgoren ◽  
C. Mathiesen ◽  
I. Rubin ◽  
M. Lauritzen
Keyword(s):  
Electrical Stimulation ◽  
Purkinje Cells ◽  
Cerebellar Cortex ◽  
Synaptic Strength ◽  
Parallel Fiber ◽  
Climbing Fibers ◽  
Parallel Fibers ◽  
Doppler Flowmetry ◽  
Activity Dependent ◽  
Stimulation Of

The purpose of the present study was to examine mechanisms of activity-dependent changes of cerebral blood flow (CBF) in rat cerebellar cortex by laser-Doppler flowmetry, using two synaptic inputs that excite different regions of the same target cell and with different synaptic strength. The apical part of Purkinje cells was activated by electrical stimulation of parallel fibers, whereas the cell soma and the proximal part of the dendritic tree were activated by climbing fibers using harmaline (40 mg/kg ip) or electrical stimulation of the inferior olive. Glass microelectrodes were used for recordings of field potentials and single-unit activity of Purkinje cells. CBF increases evoked by parallel fibers were most pronounced in the upper cortical layers. In contrast, climbing fiber stimulation increased CBF in the entire cortex. Inhibition of nitric oxide (NO) synthase activity by NG-nitro-L-arginine (L-NNA) or guanylate cyclase activity by 1H-[1,2,4(oxadiazolo)4,3-a]quinoxaline-1-one did not affect basal or harmaline-induced Purkinje cell activity but attenuated harmaline- and parallel fiber-evoked CBF increases by approximately 40-50%. Application of 8-(p-sulfophenyl)theophylline and adenosine deaminase reduced the harmaline-evoked CBF increase without any effect on the parallel fiber-evoked CBF response. The results suggest that CBF increases elicited by activation of Purkinje cells are partially mediated by the NO-guanosine 3',5'-cyclic monophosphate system independent of the input function but that adenosine contributes as well when climbing fibers are activated. This is the first demonstration of variations of coupling as a function of postsynaptic activity in the same cell.

scholarly journals Integrative Versus Delay Line Characteristics of Cerebellar Cortex

1976 ◽  
Vol 3 (2) ◽  
pp. 85-97 ◽  
Author(s):  
W.A. MacKay ◽  
J.T. Murphy
Keyword(s):  
Cerebellar Cortex ◽  
Cell Activation ◽  
Delay Line ◽  
Granule Cells ◽  
Molecular Layer ◽  
Parallel Fiber ◽  
Response Latencies ◽  
Integrator Model ◽  
P Cells

SUMMARY:In order to determine which of two general models (“tapped delay line” or “integrator”) provides a more accurate description of mammalian Purkinje cell (P-cell) activation by natural stimulation, the spatial and temporal characteristics of a population of neurons in cerebellar cortex responsive to small controlled stretches of forelimb muscles were examined in awake, locally anesthetized cats. Stretch of a single wrist muscle excited P-cells over a distance of about 1 mm in the long axis of a folium, a span which is at most half the length of parallel fibers. Both granule cells and molecular layer interneurons were excited over a wider zone than P-cells.Furthermore, P-cells across a response zone all fired on the average at the same time, as determined by computing peristimulus cross-interval histograms from pairs of simultaneously recorded neurons. Consistent delays could only be demonstrated in the minimal response latencies as measured from peristimulus time histograms. These delays, however, were longer than could be ascribed to parallel fiber conduction velocity.No evidence, therefore, was found in cat cerebellum to support the “tapped delay line” model, which postulates the successive activation of P-cells as an excitatory volley travels along a parallel fiber beam. Instead, an integrative mode of operation seems to predominate: a relatively wide substratum of activated granule cells simultaneously activates a narrower focus of P-cells centrally situated with respect to the granule cell population. The role of inhibitory interneurons in promoting the “integrator” model is discussed.

Regional cerebral blood flow response to vibrissal stimulation in mice lacking type I NOS gene expression

1996 ◽  
Vol 270 (3) ◽  
pp. H1085-H1090 ◽  
Author(s):  
J. Ma ◽  
C. Ayata ◽  
P. L. Huang ◽  
M. C. Fishman ◽  
M. A. Moskowitz
Keyword(s):  
Blood Flow ◽  
Cerebral Blood Flow ◽  
Regional Cerebral Blood Flow ◽  
No Production ◽  
Type I ◽  
Regional Cerebral Blood ◽  
Doppler Flowmetry ◽  
Cranial Window ◽  
Whisker Stimulation ◽  
Neuronal Nos

The role of nitric oxide (NO) in cerebral blood flow-metabolism coupling was assessed in SV-129 wild-type (WT) and neuronal (type I) NO synthase (NOS) knockout mice (Kn). Regional cerebral blood flow (rCBF; laser-Doppler flowmetry) was measured over the contralateral cortical barrel field during unilateral mechanical vibrissal deflection (2-3 Hz, 60 s) under urethan anesthesia. The rCBF response was similar in WT and Kn and did not differ when recorded over the intact skull or closed cranial window preparations. Whisker stimulation increased rCBF by 41 +/- 8% (maximum) and 27 +/- 6% (mean) in WT (n = 6) and 41 +/- 7% (maximum) and 26 +/- 6% (mean) in Kn (n = 6) when recorded through a closed cranial window. After superfusion with topical N omega-nitro-L-arginine (L-NNA; 1 mM), the rCBF response was inhibited by approximately 45% in WT mice (P < 0.05), whereas there was no inhibition in Kn. Endothelium-dependent relaxation, assessed by pial vessel dilation in response to topical acetylcholine (100 microM) and inhibition by L-NNA (1 mM), was the same in both groups. Our results suggest that 1) endothelial NO production does not mediate the rCBF coupling to neuronal activity in Kn, 2) the inhibitory effect of L-NNA on the rCBF response to whisker stimulation in WT is a consequence of type I (neuronal) NOS inhibition, and 3) NO-independent mechanisms couple rCBF and metabolism during whisker stimulation in mice lacking expression of neuronal NOS.

Involvement of Kv1 Potassium Channels in Spreading Acidification and Depression in the Cerebellar Cortex

2005 ◽  
Vol 94 (2) ◽  
pp. 1287-1298 ◽  
Author(s):  
Gang Chen ◽  
Wangcai Gao ◽  
Kenneth C. Reinert ◽  
Laurentiu S. Popa ◽  
Claudia M. Hendrix ◽  
...  
Keyword(s):  
Potassium Channels ◽  
Cerebellar Cortex ◽  
Molecular Layer ◽  
Neutral Red ◽  
Parallel Fiber ◽  
Null Mouse ◽  
Gabaergic Neurotransmission ◽  
Multiple Cycles

Spreading acidification and depression (SAD) is a form of propagated activity in the cerebellar cortex characterized by acidification and a transient depression in excitability. This study investigated the role of Kv1 potassium channels in SAD using neutral red, flavoprotein autofluorescence, and voltage-sensitive dye optical imaging in the mouse cerebellar cortex, in vivo. The probability of evoking SAD was greatly increased by blocking Kv1.1 as well as Kv1.2 potassium channels by their specific blockers dendrotoxin K (DTX-K) and tityustoxin (TsTX), respectively. DTX-K not only greatly lowered the threshold for evoking SAD but also resulted in multiple cycles of spread and spontaneous SAD. The occurrence of spontaneous SAD originating from spontaneous parallel fiber-like beams of activity suggests that blocking Kv1 channels increased parallel fiber excitability. This was confirmed by the generation of parallel fiber-like beams with the microinjection of glutamate into the upper molecular layer in the presence of DTX-K. The dramatic effects of DTX-K suggest a possible connection between SAD and episodic ataxia type 1 (EA1), a Kv1.1 potassium channelopathy. The threshold for evoking SAD was significantly lowered in the Kv1.1 heterozygous knockout mouse compared with wild-type littermates. Carbamazepine and acetazolamide, both effective in the treatment of EA1, significantly decreased the likelihood of evoking SAD. Blocking GABAergic neurotransmission did not alter the effectiveness of DTX-K. The cyclin D2 null mouse, which lacks cerebellar stellate cells, also exhibited SAD. Therefore blocking Kv1 potassium channels establishes the conditions needed to generate SAD. Furthermore, the results are consistent with the hypothesis that SAD may underlie the transient attacks of ataxia characterizing EA1.

scholarly journals Immunocytochemical demonstration of alpha-tubulin modification during axonal maturation in the cerebellar cortex.

1984 ◽  
Vol 98 (1) ◽  
pp. 347-351 ◽  
Author(s):  
R Cumming ◽  
R D Burgoyne ◽  
N A Lytton
Keyword(s):  
Cerebellar Cortex ◽  
Time Course ◽  
Staining Pattern ◽  
Molecular Layer ◽  
Parallel Fiber ◽  
Differential Staining ◽  
Germinal Layer ◽  
Progressive Change ◽  
Parallel Fibers ◽  
Alpha Tubulin

Previous light microscopic immunocytochemical studies using two monoclonal antibodies that recognise alpha-tubulin (YOL/34 and YL1/2) but differ in their isotypic specificity have shown that the unmyelinated parallel fiber axons in the cerebellar cortex are labeled with only one of the antibodies (YOL/34). We now show that at 10 d postnatally the parallel fibers are labeled with both antibodies, and that during development YL1/2 (but not YOL/34) immunoreactivity disappears progressively from parallel fibers in the lower regions of the molecular layer upwards towards the external germinal layer. By approximately 28 d postnatally, the differential staining pattern of parallel fibers by the antibodies is established throughout the molecular layer. The time course, light microscopic, and ultrastructural staining distribution corresponds to a progressive change in alpha-tubulin immunoreactivity as the parallel fibers form synaptic contacts. This modification of alpha-tubulin (which was not observed in Purkinje cell dendrites or Bergmann glia) may be related to the formation of a basic isotype of alpha-tubulin within parallel fiber axons at maturation.

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