DOI: 10.1111/j.1471-4159.1993.tb03226.x. Measurement of Hippocampal Levels of Cellular Second Messengers Following In Situ Freezing

Measurement of Hippocampal Levels of Cellular Second Messengers Following In Situ Freezing

10.1111/j.1471-4159.1993.tb03226.x

Crossref journal-article

Wiley

Journal of Neurochemistry (311)

Abstract

Abstract: The in situ freezing technique has been widely used to fix labile metabolites and cellular second messengers in cerebral cortex. In this study, we isolated specific brain regions at 0°C from coronal sections of frozen heads following in situ brain freezing and measured regional concentrations of labile metabolites and cellular messengers. These levels in the cortex were compared with those in cortical punches obtained at freezing temperature (less than −40°C) from the same in situ frozen brains and those of cortex dissected from decapitated animals. In both isoflurane‐ and pentobarbital‐anesthetized animals, we observed that the levels of lactate, free fatty acids, inositol 1,4,5‐trisphosphate, and diacylglycerol, as well as the proportion of protein kinase C associated with the membrane fraction, were similar in cortical punches taken at freezing temperature and those dissected at 0°C. However, with animals decapitated at room temperature, cortical and hippocampal levels of lactate, free fatty acids, and inositol 1,4,5‐trisphosphate and the proportion of membrane protein kinase C were significantly higher than those of corresponding brain regions isolated at 0°C from in situ frozen brains (p < 0.05). These results indicate that dissection of cortex and hippocampus at 0°C following in situ freezing will eliminate decapitation‐induced production of artifacts and changes in the levels of cellular second messengers such as inositol 1,4,5‐trisphosphate, diacylglycerol, and protein kinase C. The present technique, used in conjunction with in situ freezing, will fix cellular second messengers and labile metabolites in several regions of brain and may facilitate accurate characterization of molecular and cellular mechanisms underlying CNS function.

Bibliography

Tzigaret, C., McIntosh, T. K., Okiyama, K., Jenkins, W. L., & Prasad, M. R. (1993). Measurement of Hippocampal Levels of Cellular Second Messengers Following In Situ Freezing. Journal of Neurochemistry, 60(3), 827â834. Portico.

Authors 5

Cezar Tzigaret (first)
Tracy K. McIntosh (additional)
Koichi Okiyama (additional)
W. L. Jenkins (additional)
M. Renuka Prasad (additional)

References 30 Referenced 23

10.1111/j.1471-4159.1987.tb04121.x
10.1016/0166-2236(87)90135-4
10.1042/bj2200345
10.1139/y59-099
10.1016/0006-291X(89)92204-3
10.1016/S0006-291X(88)80304-8
10.1146/annurev.pa.29.040189.000443
10.1111/j.1471-4159.1990.tb04566.x
10.1111/j.1471-4159.1990.tb05788.x
10.1016/S0021-9258(17)39482-6 / J. Biol. Chem. / Thyrotropin‐releasing hormone induces redistribution of protein kinase C in GH4C1 rat pituitary cells by Fearon C. W. (1985)
10.1038/jcbfm.1992.4
10.1111/j.1471-4159.1989.tb10915.x
{'key': 'e_1_2_2_14_1', 'first-page': '67', 'volume-title': 'Advances in Biochemical Psychopharmacology, Vol. 3: Role of Cyclic AMP in Cell Function', 'author': 'Goldberg N. D.', 'year': '1970'} / Advances in Biochemical Psychopharmacology, Vol. 3: Role of Cyclic AMP in Cell Function by Goldberg N. D. (1970)
10.1016/S0021-9258(18)68115-3 / J. Biol. Chem. / Biochemical characterization of rat brain protein kinase C isozymes by Huang K.‐P. (1988)
10.1111/j.1471-4159.1986.tb02839.x
10.1073/pnas.84.13.4418
10.1111/j.1471-4159.1977.tb03923.x
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{'key': 'e_1_2_2_20_1', 'first-page': '10', 'volume-title': 'Cerebral Metabolism and Neuronal Function', 'author': 'Lust W. D.', 'year': '1980'} / Cerebral Metabolism and Neuronal Function by Lust W. D. (1980)
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10.1111/j.1471-4159.1990.tb04890.x
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10.1016/0006-291X(91)91511-A
10.1139/y88-248
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10.1111/j.1471-4159.1978.tb07819.x
10.1016/0166-2236(88)90194-4
10.1073/pnas.84.23.8707
10.3171/jns.1985.63.4.0617

Dates

Type	When
Created	18 years, 10 months ago (Oct. 5, 2006, 11:43 p.m.)
Deposited	1 year, 10 months ago (Oct. 24, 2023, 11:45 p.m.)
Indexed	1 month, 4 weeks ago (July 7, 2025, 1:26 a.m.)
Issued	32 years, 6 months ago (March 1, 1993)
Published	32 years, 6 months ago (March 1, 1993)
Published Online	18 years, 10 months ago (Oct. 5, 2006)
Published Print	32 years, 6 months ago (March 1, 1993)

Funders 0

None

BibTeX

@article{Tzigaret_1993, title={Measurement of Hippocampal Levels of Cellular Second Messengers Following In Situ Freezing}, volume={60}, ISSN={1471-4159}, url={http://dx.doi.org/10.1111/j.1471-4159.1993.tb03226.x}, DOI={10.1111/j.1471-4159.1993.tb03226.x}, number={3}, journal={Journal of Neurochemistry}, publisher={Wiley}, author={Tzigaret, Cezar and McIntosh, Tracy K. and Okiyama, Koichi and Jenkins, W. L. and Prasad, M. Renuka}, year={1993}, month=mar, pages={827–834} }

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