Anat Cell Biol.  2021 Mar;54(1):112-123. 10.5115/acb.20.267.

Nicotine impact on rat substantia nigra compacta

• Elgayar SAM
• Hussein OA
• Mubarak HA
• Ismaiel AM
• Gomaa AM

• KMID: 2514591
• DOI: http://doi.org/10.5115/acb.20.267

Abstract

Nicotine neuronal interactions exert an adverse potential in some brain regions and a significant link has been established between tobacco smoke/nicotine and vascular impairment. This work addresses nicotine impact on various components of the substantia nigra compacta (SNc) in rat. Twenty adult male Albino rats were divided equally into two groups: Group I, vehicle-control group (received saline [1 ml/kg body weight intra peritoneally] for 11 days). Group II; nicotine group (received 1.5 mg/kg body weight/day Sc) for 11 days. Nicotine levels were detected in the serum. Specimens were taken from the mid brain, processed and examined using biochemical, immunohistochemical, ultrastructural and morphometric techniques. In nicotine group, biochemical analysis revealed reduction in total antioxidant capacity (TAC), decrease in dopamine and malondialdehyde (MDA) levels. The mean number of light cells, and the mean surface area of nerve cells/field were significantly reduced, with an increase of dark cells were found in nicotine group compared to control. Immunoreactivity in nicotine group revealed an increase in neuronal α-synuclein, reduction in tyrosine hydroxylase enzyme, an increase in caspase 3 and ultrastructure changes suggestive of neuronal apopto. The blood capillaries were markedly affected. Nicotine induced endothelial and pericytic apoptotic changes, irregular lumena and indistinct endothelial junctional complex. Nicotine administered subcutaneously in a small dose may have a deleterious effect on SNc, mainly involving dopaminergic neurons and blood capillaries. This effect seems to be secondary to an oxidative stress that might be produced by reduced TAC and increased MDA levels.

Keyword

Nicotine; Substantia nigra; Immunohistochemistry

Figure

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  Fig. 1 Serum nicotine levels. Data are expressed as mean±standard deviation. *Statistically significant in comparison with the control group (P<0.05).

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  Fig. 2 (A) Serum TAC levels and (B) serum MDA levels. Data are expressed as mean±standard deviation. MDA, malondialdehyde; TAC, total antioxidant capacity. *Statistically significant in comparison with the control group (P<0.05).

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  Fig. 3 (A) Number of light and dark cells per field. (B) Surface area of cells/field µm2. SD, standard deviation.

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  Fig. 4 (A) SNc of control group showing a strong positive immune reaction to TH-antibody. (B) Higher magnification of the previous figure (□). (C) SNc of nicotine group showing a moderate positive immune reaction to TH-antibody. (D) Higher magnification of the previous figure. SNc, substantia nigra compacta; TH, tyrosine hydroxylase.

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  Fig. 5 (A) SNc of control group showing a negative immune reaction to caspase 3 antibody. (B) Higher magnification of the previous figure (□). (C) SNc of nicotine group showing numerous caspase 3 immune positive neurons. (D) Higher magnification of the previous figure (□). SNc, substantia nigra compacta; SNr, substantia nigra reticulata.

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  Fig. 6 (A) SNc of control group showing a negative immune reaction to α-synuclein antibody. (B) Higher magnification of the previous figure (□). (C) SNc of nicotine group showing a positive immune reaction to α-synuclein antibody. (D) Higher magnification of the previous figure (□). SNc, substantia nigra compacta; SNr, substantia nigra reticularis.

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  Fig. 7 (A) TEM of SNc of control group showing a neuron with euchromatic N. The cytoplasm contains numerous Nissl bodies and ordinary organelles. Inset: semithin section of SNc of control group sowing a flattened neuron with vesicular N and well-developed Nu. (B) TEM of SNc of nicotine group showing a neuron with euchromatic N and electron dense cytoplasm which contains numerous NBs. Inset: Semithin section of SNc of nicotine group showing a neuron with dark N and cytoplasm and prominent Nu. (C) TEM of SNc of nicotine group showing a neuron with euchromatic electron dense N, electron dense cytoplasm and irregular outlining (blebbing) (▲) surrounded by astrocytic end feet processes. Inset: semithin section for a neuron in a juxta-capillary position. (D) TEM of SNc of nicotine group showing a part of a neuron with euchromatic N. Note the presence of fragmented Golgi body (G) and non-membrane bound inclusion in the cytoplasm (▲). (E) TEM of SNc of nicotine group showing a part of a neuron with euchromatic N. Note the presence of secretory material within RER, shown at higher magnification in the right sided inset. Note non-membrane bound inclusion in the cytoplasm shown at a higher magnification in the left sided inset (curved arrow). (F) TEM of SNc of nicotine group showing an As and two adjacent N; one is dense and shrunken, the other possesses an irregular N and prominent Nu. (G) TEM of SNc of nicotine group showing longitudinally cut dendrites. Note the irregular outer surface (□) of the two adjacent neuronal processes shown at higher magnification in the insets. Spines of variable shapes (↑). (H) TEM of SNc of nicotine group showing a part of a longitudinally cut D1 in synaptic contact with another transversely cut D2. The synaptic terminal is enveloped by astocytic processes (*). As, astrocyte; BC, blood capillary; D, dendrite; N, nucleus; NBs, Nissl bodies; Nu, nucleolus; RER, rough endoplasmic reticulum; SNc, substantia nigra compacta.

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  Fig. 8 (A) TEM of SNc of control group showing an As with euchromatic ovoid N, surrounded by a scanty cytoplasm. Note the adjacent Og with moderated electron dense heterochromatic N and cytoplasm. (B) Shows another Og with high electron dense N and cytoplasm. (C) TEM of SNc of nicotine group showing an As with an abnormally large irregular N, prominent Nu surrounded by a thin rim of cytoplasm. (D) TEM of SNc of nicotine group showing an Og with an elongated N. The adjacent D reveals disorganized microtubules. (E) TEM of SNc of nicotine group showing Mg with a rod shape N, separated from the adjacent Ne by astrocytic processes. The cytoplasm contains numerous DB. As, astrocyte; D, dendrite; DB, dense bodies; Mg, microglia; N, nucleus; Og, oligodendrocyte; SNc, substantia nigra compacta.

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  Fig. 9 (A) TEM of SNc of nicotine group showing a blood capillary lined by an E with an abnormally large and irregular N. (B) TEM of SNc of nicotine group showing a blood capillary with an irregular Lu. The N of the lining E has marginated heterochromatin and the surrounding astrocytic processes has corrugated surface (▲). Inset: TEM of SNc of nicotine group showing a part of a blood capillary lining E with indistinct junctional complex. (C) TEM of SNc of nicotine group showing a blood capillary with Lu lined by a P with an apoptotic N (Pr). Inset: Semithin section showing apoptotic bodies (↑) between a BC and a dark Ne. (D) TEM of SNc of nicotine group showing a blood capillary with Lu, lined by an attenuated E with dense nucleus. The presence of the close by Mg with a rod-shaped nucleus and numerous apoptotic dense bodies can be detected. The capillary is surrounded by dilated astrocytic processes (*). Inset: Semithin section showing dilated astrocytic processes (*) around a blood capillary. Ap, apoptotic dense bodies; BC, blood capillary; E, endothelial cell; Lu, lumen; Mg, microglia; N, nucleus; Ne, neuron; P, pericytic process; Pr, pericyte; SNc, substantia nigra compacta; V, vacuole.

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