Anat Cell Biol.  2016 Mar;49(1):21-33. 10.5115/acb.2016.49.1.21.

Modulation of axonal sprouting along rostro-caudal axis of dorsal hippocampus and no neuronal survival in parahippocampal cortices by long-term post-lesion melatonin administration in lithium-pilocarpine model of temporal lobe epilepsy

• Kazemi M
• Shokri S
• Ganjkhani
• Ali R
• Iraj JA

• KMID: 2308929
• DOI: http://doi.org/10.5115/acb.2016.49.1.21

Abstract

Feature outcome of hippocampus and extra-hippocampal cortices was evaluated in melatonin treated lithium-pilocarpine epileptic rats during early and chronic phases of temporal lobe epilepsy (TLE). After status epilepticus (SE) induction, 5 and 20 mg/kg melatonin were administered for 14 days or 60 days. All animals were killed 60 days post SE induction and the histological features of the rosrto-caudal axis of the dorsal hippocampus, piriform and entorhinal cortices were evaluated utilizing Nissl, Timm, and synapsin I immunoflorescent staining. Melatonin (20 mg/kg) effect on CA1 and CA3 neurons showed a region-specific pattern along the rostro-caudal axis of the dorsal hippocampus. The number of counted granular cells by melatonin (20 mg/kg) treatment increased along the rostro-caudal axis of the dorsal hippocampus in comparison to the untreated epileptic group. The density of Timm granules in the inner molecular layer of the dentate gyrus decreased significantly in all melatonin treated groups in comparison to the untreated epileptic animals. The increased density of synapsin I immunoreactivity in the outer molecular layer of the dentate gyrus of untreated epileptic rats showed a profound decrease following melatonin treatment. There was no neuronal protection in the piriform and entorhinal cortices whatever the melatonin treatment. Long-term melatonin administration as a co-adjuvant probably could reduce the post-lesion histological consequences of TLE in a region-specific pattern along the rostro-caudal axis of the dorsal hippocampus.

Keyword

Status epilepticus; Melatonin; Temporal lobe epilepsy; Hippocampus

MeSH Terms

Animals
Axis, Cervical Vertebra*
Axons*
Dentate Gyrus
Entorhinal Cortex
Epilepsy, Temporal Lobe*
Hippocampus*
Melatonin*
Neurons*
Rats
Status Epilepticus
Synapsins
Temporal Lobe*
Melatonin
Synapsins

Figure

• 

  Fig. 1 (A) Location of circles placed sequentially along the junction between the stratum pyramidale and stratum oriens. If artifact or disrupted tissue was noted, the circles were placed on immediately adjacent tissue. (B) Location of circles placed over the dentate granule cell layer (top row of circles) and the band of the molecular layer. Scale bars=150 µm.

• 

  Fig. 2 Descriptive pattern of immune-densitometry process. (A) The synapsin stained taken image was converted to white and gray scale after omitting the background. DAPI stained image was merged with synapsin stained image to determine the boundary of granular layer. (B) A fixed number of circles with determined dimension was randomly placed in inner and outer molecular layers to determine the density of punctuate pattern of synapsin immunofluorescence. Scale bar=200 µm.

• 

  Fig. 3 (A–L) Descriptive representation of Timm staining in the caudal part of dorsal hippocampus and rostral pole of dorsal CA3 subfields. (A–F) Dentate gyrus (DG). (G–L) CA3 subfield. Control (A, G), epilepsy (B, H), 5 mg/kg for 60 days (C, I), 5 mg/kg for 14 days (D, J), 20 mg/kg for 60 days (E, K), and 20 mg/kg for 14 days (F, L). Scale bars=800 µm. (M) The density of sprouted axons in the both rostro-caudal pole of dorsal dentate gyrus in addition to CA3 subfields. The epilepsy group was compared to control one. The melatonin treated group was compared to the vehicle one. Mel, melatonin. *P≤0.05.

• 

  Fig. 4 (A–F) Representation of distribution of synapsin I immunoreactivity in the different layers of dorsal dentate gyrus. Control (A), epilepsy (B), 5 mg/kg melatonin for 60 days (C), 5 mg/kg melatonin for 14 days (D), 20 mg/kg melatonin for 60 days (E), and 20 mg/kg melatonin for 14 days (F). Scale bar=200 µm. (G) The quantitative densitometry of inner and outer molecular layers of dentate gyrus in the different groups. The epilepsy group was compared to control one. The melatonin treated groups were compared to sham one. Mel, melatonin. *P≤0.05.

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