Improved purification of rat brain choline acetyltransferase by using an immunoabsorbent

Rossier, Jean; Bauman, Annie; Benda, P

doi:10.1016/0014-5793(73)80839-7

Cited by 57 publications

(16 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…ChAT activity was measured as described by Fonnum (1968) modified by Rossier et al (1973). For the measurement of transmitter content, the oocytes were treated with trichloroacetic acid, homogenized and centrifuged.…”

Section: Activity Of Choline Acetyltransferase and Measurement Of Achmentioning

confidence: 99%

Release of acetylcholine by Xenopus oocytes injected with mRNAs from cholinergic neurons.

Eder-Colli

Dunant

et al. 1991

The EMBO Journal

View full text Add to dashboard Cite

Xenopus laevis oocytes were injected with poly(A)+ mRNAs extracted from the electric lobes of Torpedo marmorata. The electric lobes contain the perikarya of approximately 120,000 cholinergic neurons that innervate the electric organs and are homologous to motor neurons. The injected oocytes accumulated acetylcholine and were able to synthesize [14C]acetylcholine from 1‐[14C]acetate. With KCl depolarization and upon treatment with a Ca2+ ionophore, they released their endogenous as well as the radiolabelled neurotransmitter in a Ca(2+)‐dependent manner. No synthesis or release were obtained from control oocytes. With respect to their dependency upon Ca2+ concentration, the oocytes injected with Torpedo electric lobe mRNAs released acetylcholine in a manner which closely resembled that found in the native synapses. In contrast to the controls, primed oocytes were also able to release [14C]acetylcholine that was injected a few hours prior to the release trial. Immunoblot analysis demonstrated that the 15 kd proteolipid antigen of the purified mediatophore, a 200 kd presynaptic protein able to translocate acetylcholine, was expressed in the ACh‐releasing oocytes but not in the controls. The present observation may provide a useful approach for investigating the proteins involved in the release of acetylcholine and of other neurotransmitter substances.

show abstract

Section: Activity Of Choline Acetyltransferase and Measurement Of Achmentioning

confidence: 99%

Release of acetylcholine by Xenopus oocytes injected with mRNAs from cholinergic neurons.

Eder-Colli

Dunant

et al. 1991

The EMBO Journal

View full text Add to dashboard Cite

show abstract

“…The assay was carried out by the method of Rossier (1973) based on Fonnum's extraction technique (1969). The tissues were homogenized with a Teflon-glass homogenizer in 10 volumes of 0.2 M-sodium phosphate (pH 7.4) supplemented with 0.5% Triton X-100.…”

Section: Choline Acetyltransferase (Ec 2316)mentioning

confidence: 99%

Morphological and Biochemical Studies on the Cerebral Cortex from Reeler Mutant Mice: Development of Cortical Layers and Metabolic Mapping by the Deoxyglucose Method

Mikoshiba

Kohsaka

Takamatsu

et al. 1980

Journal of Neurochemistry

View full text Add to dashboard Cite

Cerebral cortex from reeler mutant mice was examined morphologically and biochemically. The sequential process of postnatal cell migration in the cerebral cortex of reeler (rl/rl) was examined morphologically. The dense cellular cortical plate lies below the molecular layer near the cerebral surface just after birth in normal mice while in reeler most of the cells are concentrated in the center of the cortex. In the cortex of adult reeler, the broad laminar structure of the neurons could be seen to form inverted positions in the cortical layers. The total wet weight, and the concentration of DNA and RNA in the pallium cerebri from reeler did not differ significantly from those in the control. As to the protein profiles of the pallium cerebri detected by SDS‐ polyacrylamide gel electrophoresis, no significant differences were observed. Activities of CNPase (2′,3′‐cyclic nucleotide 3′‐phosphohydrolase), which is a myelin enzyme of CNS, and choline acetyltransferase were at the same level in both the reeler and the control. Therefore, reeler mutation does not appear to affect the genetically determined cell numbers, number of cholinergic fibers, and myelination. By autoradiographic observation of the cerebral cortex after intraperitoneal injection of [14C]2‐deoxyglucose, it was revealed that 2‐deoxyglucose was incorporated intensively into the fourth layer (granular layer) of the cerebrum from the control. In reeler it was also incorporated into the granular layer but in a more widespread distribution. We conclude that terminals to the granular layer make metabolically active synapse, perhaps even in a manner inverted from normal.

show abstract

“…Antigen was prepared as previously described [3] : i) Rat brains (630 gm wet weight) were homogenized in a buffer solution adjusted to pH 5; ii) After centrifugation at 10 000 g, the supernatant was discarded. The pellet was rehomogenized in 200 mM NaCl.…”

Section: Antigen Purificationmentioning

confidence: 99%

“…The story of Ra # 27 was rather unique: the first few bleedings obtained during the semester following the start of immunization have already been described [3] and will be quoted from now on as bleedings 27 A. The latest bleedings ~ bleedings 27 B -obtained after the rabbit had been boosted again twice (see Materials and methods), still gave in agar gels multiple bands of precipitation with the ChAc preparation, similar to that shown previously in the analysis of bleedings 27 A (see fig.…”

Section: Obtention Of Bleeding 27 Band Its Analysismentioning

confidence: 99%

Antibodies to rat brain choline acetyltransferase: Species and organ specificity

Rossier¹,

Bauman²,

Benda³

1973

FEBS Letters

View full text Add to dashboard Cite

Improved purification of rat brain choline acetyltransferase by using an immunoabsorbent

Cited by 57 publications

References 8 publications

Release of acetylcholine by Xenopus oocytes injected with mRNAs from cholinergic neurons.

Release of acetylcholine by Xenopus oocytes injected with mRNAs from cholinergic neurons.

Morphological and Biochemical Studies on the Cerebral Cortex from Reeler Mutant Mice: Development of Cortical Layers and Metabolic Mapping by the Deoxyglucose Method

Antibodies to rat brain choline acetyltransferase: Species and organ specificity

Contact Info

Product

Resources

About