Anat Cell Biol.  2013 Sep;46(3):191-197. 10.5115/acb.2013.46.3.191.

Stereological study of the effects of morphine consumption and abstinence on the number of the neurons and oligodendrocytes in medial prefrontal cortex of rats

• Rafati A
• Noorafshan A
• Torabi N

• KMID: 2263145
• DOI: http://doi.org/10.5115/acb.2013.46.3.191

Abstract

Quantitative studies to date on the effects of opioid consumption and abstinence on the nervous system using modern stereological methods have not received enough attention. In addition, they have yielded controversial results. The present study was conducted to investigate the effects of morphine, with or without abstinence, on the neurons and oligodendrocytes of the medial prefrontal cortex (MPFC) in rats using quantitative stereological methods. The male rats were divided into four groups: the first (saline [SAL]) and second (morphine [MOR]) groups were treated with saline and an escalating dose of morphine (5-20 mg/kg) for 30 days, respectively; the third (SAL+abstinence [ABS]) and fourth (MOR+ABS) groups were treated in the same manner as the previous groups plus they had a 30-day abstinence period. The results showed that the volume of the MPFC and its subdivisions decreased by approximately 15% in the MOR group compared with that in the SAL group (P<0.05). In addition, the volume decreased by approximately 24% in the MOR+ABS group compared with that in the SAL+ABS group (P<0.05). The number of neurons in the MOR and MOR+ABS groups decreased by approximately 44% and 35%, respectively, compared with that in their corresponding control groups. Moreover, the number of the oligodendrocytes in the MOR and MOR+ABS groups decreased by approximately 41% and 37%, respectively. No significant difference was noted in the number of cells in the MOR and MOR+ABS groups. In conclusion, morphine consumption leads to a permanent reduction in the number of neurons and oligodendrocytes, and no additional neuron and oligodendrocyte loss occurs after abstinence.

Keyword

Morphine; Cortex; Neurons; Oligodendroglia; Stereology; Rat

MeSH Terms

Animals
Humans
Male
Morphine
Nervous System
Neurons
Oligodendroglia
Prefrontal Cortex
Rats
Morphine

Figure

• 

  Fig. 1 The Cavalieri method. The volume of the medial prefrontal cortex (MPFC) and its subdivisions; anterior cingulated (Cg1), prelimbic (PrL), and infralimbic areas (IL) were estimated using Cavalieri's method. (A) The MPFC was sectioned executively. Nine to twelve sections were sampled (here 9). "t" is the distance between the sampled sections. (B) A grid of points was superimposed on the sampled sections. "a(p)" is the area per point and is calculated by multiplying "ΔX" by "ΔY". The arrow indicates the point that is the right upper corner of the cross. Cresyl-violet staining.

• 

  Fig. 2 Estimation of numerical density. (A) A schematic drawing of a microscopic slide with a section of the medial prefrontal cortex mounted on it. "h" is the height of the optical disector and "t" is the real thickness of the section. In each field, the first 5 µm of the section thickness was ignored (guard zone: gz), and cell counting was done in the next 10 µm of the section thickness "h". (B, C) Sampling of cells using optical disector. The unbiased counting frame was superimposed on the images. Just the cells whose nucleoli did not appear in the beginning of the disector height (B) and appeared at the following optical scan (C) were counted. The cells whose nucleoli were completely or partly inside the counting frame or touching the upper and right lines were counted as "ΣQ-". Cresyl-violet staining.

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