Skip to main content
PMC home page
Journal of Anatomy logoLink to Journal of Anatomy
. 1999 Apr;194(Pt 3):423–431. doi: 10.1046/j.1469-7580.1999.19430423.x

The effect of the timing of ethanol exposure during early postnatal life on total number of Purkinje cells in rat cerebellum

TAKANORI MIKI 1 , SIMON HARRIS 2 , PETER WILCE 3 , YOSHIKI TAKEUCHI 1 , KULDIP S BEDI 2 ,✉
PMCID: PMC1467941  PMID: 10386779

Abstract

We have previously shown that exposing rats to a high dose of ethanol on postnatal d 5 can affect Purkinje cell numbers in the cerebellum whilst similar exposure on d 10 had no such effect. The question arose whether a longer period of ethanol exposure after d 10 could produce loss of Purkinje cells. We have examined this question by exposing young rats to a relatively high dose (∼420–430 mg/dl) of ethanol for 6 d periods between the ages of either 4 and 9 d or 10 and 15 d of age. Exposure was carried out by placing the rats in an ethanol vapour chamber for 3 h per day during the exposure period. Groups of ethanol-treated (ET), separation controls (SC) and mother-reared controls (MRC) were anaesthetised and killed when aged 30 d by perfusion with buffered 2.5% glutaraldehyde. Stereological methods were used to determine the numbers of Purkinje cells in the cerebellum of each rat. MRC, SC and rats treated with ethanol between 10–15 d of age each had, on average, about 254–258 thousand cerebellar Purkinje cells; the differences between these various groups were not statistically significant. However, the rats treated with ethanol vapour between 4–9 d of age had an average of only about 128000±20000 Purkinje cells per cerebellum. This value was significantly different from both the MRC and group-matched SC animals. It is concluded that the period between 4 and 9 d of age is an extremely vulnerable period during which the rat cerebellar Purkinje cells are particularly susceptible to the effects of a high dose of ethanol. However, a similar level and duration of ethanol exposure commencing after 10 d of age has no significant effect on Purkinje cell numbers.

Keywords: Cell death, stereology, fetal alcohol syndrome

Full Text

The Full Text of this article is available as a PDF (378.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barnes D. E., Walker D. W. Prenatal ethanol exposure permanently reduces the number of pyramidal neurons in rat hippocampus. Brain Res. 1981 Jun;227(3):333–340. doi: 10.1016/0165-3806(81)90071-7. [DOI] [PubMed] [Google Scholar]
  2. Bauer-Moffett C., Altman J. The effect of ethanol chronically administered to preweanling rats on cerebellar development: a morphological study. Brain Res. 1977 Jan 7;119(2):249–268. doi: 10.1016/0006-8993(77)90310-9. [DOI] [PubMed] [Google Scholar]
  3. Beattie J. O. Transplacental alcohol intoxication. Alcohol Alcohol. 1986;21(2):163–166. [PubMed] [Google Scholar]
  4. Bedi K. S., Campbell L. F., Mayhew T. M. A fractionator study of the effects of undernutrition during early life on rat Purkinje cell numbers (with a caveat on the use of nucleoli as counting units). J Anat. 1992 Oct;181(Pt 2):199–208. [PMC free article] [PubMed] [Google Scholar]
  5. Bedi K. S., Hall R., Davies C. A., Dobbing J. A stereological analysis of the cerebellar granule and Purkinje cells of 30-day-old and adult rats undernourished during early postnatal life. J Comp Neurol. 1980 Oct 15;193(4):863–870. doi: 10.1002/cne.901930404. [DOI] [PubMed] [Google Scholar]
  6. Bedi K. S., Thomas Y. M., Davies C. A., Dobbing J. Synapse-to-neuron ratios of the frontal and cerebellar cortex of 30-day-old and adult rats undernourished during early postnatal life. J Comp Neurol. 1980 Sep 1;193(1):49–56. doi: 10.1002/cne.901930104. [DOI] [PubMed] [Google Scholar]
  7. Bonthius D. J., Goodlett C. R., West J. R. Blood alcohol concentration and severity of microencephaly in neonatal rats depend on the pattern of alcohol administration. Alcohol. 1988 May-Jun;5(3):209–214. doi: 10.1016/0741-8329(88)90054-7. [DOI] [PubMed] [Google Scholar]
  8. Bonthius D. J., West J. R. Alcohol-induced neuronal loss in developing rats: increased brain damage with binge exposure. Alcohol Clin Exp Res. 1990 Feb;14(1):107–118. doi: 10.1111/j.1530-0277.1990.tb00455.x. [DOI] [PubMed] [Google Scholar]
  9. Bonthius D. J., West J. R. Permanent neuronal deficits in rats exposed to alcohol during the brain growth spurt. Teratology. 1991 Aug;44(2):147–163. doi: 10.1002/tera.1420440203. [DOI] [PubMed] [Google Scholar]
  10. Borges S., Lewis P. D. A study of alcohol effects on the brain during gestation and lactation. Teratology. 1982 Jun;25(3):283–289. doi: 10.1002/tera.1420250304. [DOI] [PubMed] [Google Scholar]
  11. Braendgaard H., Gundersen H. J. The impact of recent stereological advances on quantitative studies of the nervous system. J Neurosci Methods. 1986 Oct;18(1-2):39–78. doi: 10.1016/0165-0270(86)90112-3. [DOI] [PubMed] [Google Scholar]
  12. Clarren S. K., Alvord E. C., Jr, Sumi S. M., Streissguth A. P., Smith D. W. Brain malformations related to prenatal exposure to ethanol. J Pediatr. 1978 Jan;92(1):64–67. doi: 10.1016/s0022-3476(78)80072-9. [DOI] [PubMed] [Google Scholar]
  13. Clarren S. K., Smith D. W. The fetal alcohol syndrome. N Engl J Med. 1978 May 11;298(19):1063–1067. doi: 10.1056/NEJM197805112981906. [DOI] [PubMed] [Google Scholar]
  14. Dobbing J., Sands J. Comparative aspects of the brain growth spurt. Early Hum Dev. 1979 Mar;3(1):79–83. doi: 10.1016/0378-3782(79)90022-7. [DOI] [PubMed] [Google Scholar]
  15. Dobbing J., Sands J. Quantitative growth and development of human brain. Arch Dis Child. 1973 Oct;48(10):757–767. doi: 10.1136/adc.48.10.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Goodlett C. R., Marcussen B. L., West J. R. A single day of alcohol exposure during the brain growth spurt induces brain weight restriction and cerebellar Purkinje cell loss. Alcohol. 1990 Mar-Apr;7(2):107–114. doi: 10.1016/0741-8329(90)90070-s. [DOI] [PubMed] [Google Scholar]
  17. Gundersen H. J. Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones, in memory of William R. Thompson. J Microsc. 1986 Jul;143(Pt 1):3–45. [PubMed] [Google Scholar]
  18. Hamre K. M., West J. R. The effects of the timing of ethanol exposure during the brain growth spurt on the number of cerebellar Purkinje and granule cell nuclear profiles. Alcohol Clin Exp Res. 1993 Jun;17(3):610–622. doi: 10.1111/j.1530-0277.1993.tb00808.x. [DOI] [PubMed] [Google Scholar]
  19. Jones K. L., Smith D. W., Hanson J. W. The fetal alcohol syndrome: clinical delineation. Ann N Y Acad Sci. 1976;273:130–139. doi: 10.1111/j.1749-6632.1976.tb52873.x. [DOI] [PubMed] [Google Scholar]
  20. Jones K. L., Smith D. W. Recognition of the fetal alcohol syndrome in early infancy. Lancet. 1973 Nov 3;302(7836):999–1001. doi: 10.1016/s0140-6736(73)91092-1. [DOI] [PubMed] [Google Scholar]
  21. Jones K. L., Smith D. W., Ulleland C. N., Streissguth P. Pattern of malformation in offspring of chronic alcoholic mothers. Lancet. 1973 Jun 9;1(7815):1267–1271. doi: 10.1016/s0140-6736(73)91291-9. [DOI] [PubMed] [Google Scholar]
  22. Karanian J., Yergey J., Lister R., D'Souza N., Linnoila M., Salem N., Jr Characterization of an automated apparatus for precise control of inhalation chamber ethanol vapor and blood ethanol concentrations. Alcohol Clin Exp Res. 1986 Aug;10(4):443–447. doi: 10.1111/j.1530-0277.1986.tb05121.x. [DOI] [PubMed] [Google Scholar]
  23. Kelly S. J., Mahoney J. C., West J. R. Changes in brain microvasculature resulting from early postnatal alcohol exposure. Alcohol. 1990 Jan-Feb;7(1):43–47. doi: 10.1016/0741-8329(90)90059-l. [DOI] [PubMed] [Google Scholar]
  24. Kelly S. J., Pierce D. R., West J. R. Microencephaly and hyperactivity in adult rats can be induced by neonatal exposure to high blood alcohol concentrations. Exp Neurol. 1987 Jun;96(3):580–593. doi: 10.1016/0014-4886(87)90220-2. [DOI] [PubMed] [Google Scholar]
  25. Kyllerman M., Aronson M., Sabel K. G., Karlberg E., Sandin B., Olegård R. Children of alcoholic mothers. Growth and motor performance compared to matched controls. Acta Paediatr Scand. 1985 Jan;74(1):20–26. doi: 10.1111/j.1651-2227.1985.tb10915.x. [DOI] [PubMed] [Google Scholar]
  26. Little R. E., Anderson K. W., Ervin C. H., Worthington-Roberts B., Clarren S. K. Maternal alcohol use during breast-feeding and infant mental and motor development at one year. N Engl J Med. 1989 Aug 17;321(7):425–430. doi: 10.1056/NEJM198908173210703. [DOI] [PubMed] [Google Scholar]
  27. Marcus J. C. Neurological findings in the fetal alcohol syndrome. Neuropediatrics. 1987 Aug;18(3):158–160. doi: 10.1055/s-2008-1052471. [DOI] [PubMed] [Google Scholar]
  28. Marcussen B. L., Goodlett C. R., Mahoney J. C., West J. R. Developing rat Purkinje cells are more vulnerable to alcohol-induced depletion during differentiation than during neurogenesis. Alcohol. 1994 Mar-Apr;11(2):147–156. doi: 10.1016/0741-8329(94)90056-6. [DOI] [PubMed] [Google Scholar]
  29. Mayhew T. M. A review of recent advances in stereology for quantifying neural structure. J Neurocytol. 1992 May;21(5):313–328. doi: 10.1007/BF01191700. [DOI] [PubMed] [Google Scholar]
  30. Mayhew T. M. Accurate prediction of Purkinje cell number from cerebellar weight can be achieved with the fractionator. J Comp Neurol. 1991 Jun 8;308(2):162–168. doi: 10.1002/cne.903080203. [DOI] [PubMed] [Google Scholar]
  31. Meyer L. S., Kotch L. E., Riley E. P. Neonatal ethanol exposure: functional alterations associated with cerebellar growth retardation. Neurotoxicol Teratol. 1990 Jan-Feb;12(1):15–22. doi: 10.1016/0892-0362(90)90107-n. [DOI] [PubMed] [Google Scholar]
  32. Nairn J. G., Bedi K. S., Mayhew T. M., Campbell L. F. On the number of Purkinje cells in the human cerebellum: unbiased estimates obtained by using the "fractionator". J Comp Neurol. 1989 Dec 22;290(4):527–532. doi: 10.1002/cne.902900407. [DOI] [PubMed] [Google Scholar]
  33. Ogbuihi S., Cruz-Orive L. M. Estimating the total number of lymphatic valves in infant lungs with the fractionator. J Microsc. 1990 Apr;158(Pt 1):19–30. doi: 10.1111/j.1365-2818.1990.tb02973.x. [DOI] [PubMed] [Google Scholar]
  34. Pauli J., Wilce P., Bedi K. S. Acute exposure to alcohol during early postnatal life causes a deficit in the total number of cerebellar Purkinje cells in the rat. J Comp Neurol. 1995 Sep 25;360(3):506–512. doi: 10.1002/cne.903600311. [DOI] [PubMed] [Google Scholar]
  35. Phillips S. C., Cragg B. G. A change in susceptibility of rat cerebellar Purkinje cells to damage by alcohol during fetal, neonatal and adult life. Neuropathol Appl Neurobiol. 1982 Nov-Dec;8(6):441–454. doi: 10.1111/j.1365-2990.1982.tb00312.x. [DOI] [PubMed] [Google Scholar]
  36. Pierce D. R., Goodlett C. R., West J. R. Differential neuronal loss following early postnatal alcohol exposure. Teratology. 1989 Aug;40(2):113–126. doi: 10.1002/tera.1420400205. [DOI] [PubMed] [Google Scholar]
  37. Sampson P. D., Streissguth A. P., Bookstein F. L., Little R. E., Clarren S. K., Dehaene P., Hanson J. W., Graham J. M., Jr Incidence of fetal alcohol syndrome and prevalence of alcohol-related neurodevelopmental disorder. Teratology. 1997 Nov;56(5):317–326. doi: 10.1002/(SICI)1096-9926(199711)56:5<317::AID-TERA5>3.0.CO;2-U. [DOI] [PubMed] [Google Scholar]
  38. Streissguth A. P., Barr H. M., Martin D. C., Herman C. S. Effects of maternal alcohol, nicotine, and caffeine use during pregnancy on infant mental and motor development at eight months. Alcohol Clin Exp Res. 1980 Apr;4(2):152–164. doi: 10.1111/j.1530-0277.1980.tb05630.x. [DOI] [PubMed] [Google Scholar]
  39. Streissguth A. P., Landesman-Dwyer S., Martin J. C., Smith D. W. Teratogenic effects of alcohol in humans and laboratory animals. Science. 1980 Jul 18;209(4454):353–361. doi: 10.1126/science.6992275. [DOI] [PubMed] [Google Scholar]
  40. Thomas J. D., Goodlett C. R., West J. R. Alcohol-induced Purkinje cell loss depends on developmental timing of alcohol exposure and correlates with motor performance. Brain Res Dev Brain Res. 1998 Feb 10;105(2):159–166. doi: 10.1016/s0165-3806(97)00164-8. [DOI] [PubMed] [Google Scholar]
  41. Warren M. A., Bedi K. S. Synapse-to-neuron ratios in rat cerebellar cortex following lengthy periods of undernutrition. J Anat. 1990 Jun;170:173–182. [PMC free article] [PubMed] [Google Scholar]
  42. Warren M. A., Bedi K. S. The effects of a lengthy period of undernutrition from birth and subsequent nutritional rehabilitation on the granule-to-Purkinje cell ratio in the rat cerebellum. J Anat. 1988 Aug;159:147–153. [PMC free article] [PubMed] [Google Scholar]
  43. West J. R., Goodlett C. R., Bonthius D. J., Hamre K. M., Marcussen B. L. Cell population depletion associated with fetal alcohol brain damage: mechanisms of BAC-dependent cell loss. Alcohol Clin Exp Res. 1990 Dec;14(6):813–818. doi: 10.1111/j.1530-0277.1990.tb01820.x. [DOI] [PubMed] [Google Scholar]
  44. West J. R., Hamre K. M., Pierce D. R. Delay in brain growth induced by alcohol in artificially reared rat pups. Alcohol. 1984 May-Jun;1(3):213–222. doi: 10.1016/0741-8329(84)90101-0. [DOI] [PubMed] [Google Scholar]
  45. West J. R. Recent findings on the mechanisms by which alcohol damages the developing nervous system. Alcohol Alcohol Suppl. 1994;2:395–399. [PubMed] [Google Scholar]

Articles from Journal of Anatomy are provided here courtesy of Anatomical Society of Great Britain and Ireland

RESOURCES