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. 2008 Jul 24:1221:98-107.
doi: 10.1016/j.brainres.2008.05.015. Epub 2008 May 16.

Ethanol-related increases in degenerating bodies in the Purkinje neuron dendrites of aging rats

Cynthia A Dlugos  1
Affiliations

Ethanol-related increases in degenerating bodies in the Purkinje neuron dendrites of aging rats

Cynthia A Dlugos. Brain Res. .

Abstract

Chronic ethanol consumption in aging rats results in regression of Purkinje neuron (PN) dendritic arbors ([Pentney, 1995 Measurements of dendritic pathlengths provide evidence that ethanol-induced lengthening of terminal dendritic segments may result from dendritic regression. Alcohol Alcohol. 30, 87-96]), loss of synapses (Dlugos and Pentney, 1997), dilation of the smooth endoplasmic reticulum (SER), and the formation of degenerating bodies within PN dendrites ([Dlugos, C.A., 2006a. Ethanol-Related Smooth Endoplasmic Reticulum Dilation in Purkinje Dendrites of Aging Rats. Alcohol., Clin. Exp. Res. 30, 883-891,Dlugos, C.A., 2006b. Smooth endoplasmic reticulum dilation and degeneration in Purkinje neuron dendrites of aging ethanol-fed female rats. Cerebellum. 5, 155-162]). Dilation of the SER and the formation of degenerating bodies may be a predictor of dendritic regression. Ethanol-induced effects on mitochondria may be involved as mitochondria cooperate with the SER to maintain calcium homeostasis. The purpose of this study was to determine whether degenerating body number and mitochondrial density and structure are altered by chronic ethanol treatment in PN dendrites. Male, Fischer 344 rats, 12 months of age, were fed an ethanol or pair-fed liquid diet, or rat chow for a period of 10, 20, or 40 weeks (15 rats/treatment; 45 rats/treatment duration). Ethanol-fed rats received 35% of their calories as ethanol. At the end of treatment, all animals were euthanized, perfused, and tissue prepared for electron microscopy. The densities of degenerating bodies and mitochondria, mitochondrial areas, and the distance between the SER and the mitochondria were measured. Results showed that there was an ethanol-related increase in degenerating bodies compared to controls at 40 weeks. Ethanol-induced alterations to mitochondria were absent. Correlation of the present results with those of previous studies suggest that degenerating bodies may be formed from membrane reabsorption during dendritic regression or from degenerating SER whose function has been compromised by dilation.

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Figures

Fig 1
Fig 1
Normal Purkinje neuron morphology in F344 rats between 15 and 22 mo of age. A. Purkinje neuron with visible nucleus and nucleolus (arrow). Purkinje neuron soma (asterisk) becomes tapered at origin of primary dendrite (arrowhead). B. Low magnification of Purkinje neuron dendritic shaft (DSh). Several branchpoints (arrows) are visible. Mitochondria are shown as electron dense profiles within the shaft (arrowheads). C. Higher magnification of a large Purkinje neuron dendritic shaft (DSh) close to the primary dendrite containing many longitudinal profiles of microtubules. A mitochondrion (arrowhead) is found next to the smaller of two SER profiles (asterisks). D. A small dendritic branchlet (arrow) with two dendritic spines (asterisks). A mitochondrion (arrowhead) lies next to a small profile of SER. E. A slightly larger dendritic shaft (DSh) in which a large mitochondrion (arrow) lies close to an SER profile. F A large dendritic shaft at a branchpoint (arrow) with a dilated SER profile (asterisk) and degenerating bodies (arrowhead).
Fig 2
Fig 2
Degenerating bodies in dendritic shafts (DSh) of rats fed the pair-fed diet for 20 weeks (A), the ethanol diet for 20 weeks (B,C) and the ethanol diet for 40 weeks (D, E, F). In some cases the degenerating bodies (arrowheads) are filled with rounded vesicles (A, B, F) while in others they are lamellar in structure (C, D, E). Normal appearing mitochondria (arrowheads) are found in dendritic shafts A–E (DSh). In E a dilated SER profile ( asterisk) is found in close proximity to the degenerating structures. In F, there degenerating bodies (arrowheads) occupy a part of the dendritic shaft devoid of cytoskeleton.
Fig 3
Fig 3
Areal density (mm2) of degenerating bodies in posterior lobe of the cerebellum following 10. 20 and 40 weeks of treatment. ANOVA showed significant effects of treatment [F(2,126)=9.583, P<.001]. Post hoc analysis showed that at 40 weeks of treatment, EF rats had increased numbers of degenerating bodies compared to PF (P<.001)and CF (P=.015, ++) rats. At 20 weeks EF rats had increased degenerating bodies compared to the CF rats (P=.011, +). Comparison of EF to PF rats approached but did not reach significant levels (P=.085).
Fig 4
Fig 4
The significant interaction between treatment and duration of treatment [F(4,126) =3.834, P=.006] showed the pattern of the ethanol-related increases in degenerating bodies with increased treatment duration in Purkinje neuron dendrites. Although the numbers of degenerating structures are relatively the same among the treatment groups at 10 weeks, by 20 and 40 weeks, the density of degenerating structures in the EtOH-fed rats escalated at a higher rate than in any other treatment groups. The interaction occurred because the EtOH-fed group, with the smallest amount of degenerating structures at 10 weeks, far exceeded the other groups at 20 and 40 weeks.
Fig 5
Fig 5
Areal density of mitochondria (mm2) within Purkinje neuron dendrites of the posterior lobe of the cerebellum following 10, 20, and 40 weeks of chow-fed, pair-fed, or ethanol-fed dietary treatment. There was no effect of treatment (P=.650) or duration of treatment (P=.286) on mitochondrial density within PN dendrites.
Fig 6
Fig 6
Distance (um) between SER and mitochondria in 40 week Chow-fed, pair-fed, and ethanol-fed rats. SER and mitochondrial distances were measured at <1, 3. and 4 microns. There was not effect of treatment at <1 (P=.967), <3 (P=.998), and <4 (P=.838) micron distances.
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