The Use of Microwave Heating to Inactivate Cholinesterase in the Rat Brain Prior to Analysis for Acetylcholine

Stavinoha, W. B.; Weintraub, Susan T.; Modak, A. T.

doi:10.1111/j.1471-4159.1973.tb12135.x

Cited by 202 publications

(35 citation statements)

References 10 publications

(1 reference statement)

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“…Identical results were obtained in multiple repetitions of this analysis. This pattern was not generated by postmortem enzymatic mechanisms since similar proximodistal changes were observed in mice killed by microwave irradiation in order to rapidly inactivate enzymatic activity (55,58).…”

Section: Distribution Of Nfps Along the Primary Optic Pathwaymentioning

confidence: 87%

Posttranslational modification of a neurofilament protein during axoplasmic transport: implications for regional specialization of CNS axons

Nixon¹,

Brown²,

Marotta³

1982

The Journal of Cell Biology

111

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The possibility that proteins are modified during axoplasmic transport in central nervous system axons was examined by analyzing neurofilament proteins (200,000, 140,000, and 70,000 mol wt) along the mouse primary optic pathway (optic nerve and optic tract). The major neurofilament proteins (NFPs) exhibited considerable microheterogeneity. At least three forms of the " 140,000" neurofilament protein differing in molecular weight by SDS PAGE (140,000-145,000 mol wt) were identified. The "140,000" proteins, and their counterparts in purified neurofilament preparations, displayed similar isoelectric points and the same peptide maps. The "140,000" NFPs exhibited regional heterogeneity when consecutive segments of the optic pathway were separately examined on polyacrylamide gels. Two major species (145,000 and 140,000 mol wt) were present along the entire length of the optic pathway. The third protein (143,000 mol wt) was absent proximally but became increasingly prominent in distal segments. After intravitreal injection of [(3)H]proline, newly synthesized radiolabeled proteins in the "140,000" mol wt region entered proximal mouse retinal ganglion cell (RGC) axons as two major species corresponding to the 145,000 and 14,000 mol wt NFPs observed on stained gels. When transported NFPs reached more distal axonal regions (30 d postinjection or longer), a 143,000 mol wt protein appeared that was similar in isoelectric point and peptide map to the 145,000 and 140,000 mol wt species. The results suggest that (a) the composition of CNS neurofilaments, particularly the "140,000" component, is more complex than previously recognized, that (b) retinal ganglion cell axons display regional differentiation with respect to these cytoskeletal proteins, and that (c) structural heterogeneity of "140,000" NFPs arises, at least in part, from posttranslational modification during axoplasmic transport. When excised but intact optic pathways were incubated in vitro at pH 7.4, a 143,000 NFP was rapidly formed by a calcium-dependent enzymatic process active at endogenous calcium levels. Changes in major proteins other than those in the 145,000-140,000 mol wt region were minimal. In optic pathways from mice injected intravitreally with L-[(3)H]proline, tritiated 143,000 mol wt NFP formed rapidly in vitro if radioactively labeled NFPs were present in distal RGC axonal regions (31 d postinjection). By contrast, no 143,000 mol wt NFP was generated if radioactively labeled NFPs were present proximally in RGC axons (6 d postinjection). The enzymatic process that generates 143,000 mol wt NFP in vitro, therefore, appears to have a nonuniform distribution along the RGC axons. The foregoing results and other observations, including the accompanying report (J. Cell Biol., 1982, 94:159-164), imply that CNS axons may be regionally specialized with respect to structure and function.

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Section: Distribution Of Nfps Along the Primary Optic Pathwaymentioning

confidence: 87%

Posttranslational modification of a neurofilament protein during axoplasmic transport: implications for regional specialization of CNS axons

Nixon¹,

Brown²,

Marotta³

1982

The Journal of Cell Biology

111

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“…Eight sham-operated and nine lesioned rats were sacrificed by focalized microwave irradiation (2.0 s; 6.3 kW; Sairem, Villeurbanne, France) in order to rapidly inactivate brain enzymes such as AChE (Stavinoha et al, 1973). The brains were prepared as described by Lehmann et al (2002b).…”

Section: Hplc Determination Of Ach Levelsmentioning

confidence: 99%

Long‐term effects of immunotoxic cholinergic lesions in the septum on acquisition of the cone‐field task and noncognitive measures in rats

et al. 2006

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In rats, nonspecific mechanical or neurotoxic lesions of the septum impair spatial memory in, e.g., Morris water- and radial-maze tasks. Unfortunately, the lack of specificity of such lesions limits inferences about the role of the cholinergic hippocampal projections in spatial cognition. We therefore tested the effects of septal lesions produced by 192 IgG-saporin in rats, which is highly selective for basal forebrain cholinergic neurons, on home cage activity, noncognitive tests (modified Irwin test, open field and forced swimming tests, and various sensorimotor tasks), and the cone-field spatial learning task. The immunotoxic lesion reduced acetylcholine (ACh) levels in the septum (-61%) and hippocampus (>-75%). Rats with lesions showed mild home-cage hyperactivity at 4 weeks postlesion, but no noncognitive deficits at 13 weeks postsurgery. In the cone-field task, rats with septal lesions made more working- and reference-memory errors than the controls, but acquisition curves were parallel in both groups. The speed of visiting cones was faster in the rats with lesions, indicative of disturbed attention or increased motivation. These data support the growing evidence that involvement of the septohippocampal cholinergic system in spatial learning and memory may have been overestimated in studies that used lesions with poor selectivity.

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“…We have used this technique to estimate the concentration of glucose in 20 regions of the mouse brain (39). After rapid inactivation of the enzymes responsible for glucose metabolism by microwave irradiation (2,3,28,29,34,47,48), the frozen brains were cut coronally in a cryostat. Each region on the section was removed.…”

Section: Combination Of Autoradiography and Biochemical Analysismentioning

confidence: 99%

Carbohydrate distribution in mammal; Whole-body autoradiographic approach.

Watanabe

Kihara

1986

Acta Histochem. Cytochem

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The Use of Microwave Heating to Inactivate Cholinesterase in the Rat Brain Prior to Analysis for Acetylcholine

Cited by 202 publications

References 10 publications

Posttranslational modification of a neurofilament protein during axoplasmic transport: implications for regional specialization of CNS axons

Posttranslational modification of a neurofilament protein during axoplasmic transport: implications for regional specialization of CNS axons

Long‐term effects of immunotoxic cholinergic lesions in the septum on acquisition of the cone‐field task and noncognitive measures in rats

Carbohydrate distribution in mammal; Whole-body autoradiographic approach.

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