Does Melatonin Protect The Cerebellum Against Structural Alterations In Diabetic Rats?
Mahmoud Mohamed Abdallah;
Abstract
The present work investigated the effects of Diabetes mellitus on the structure of cerebellum of both male and female rats and the possible protective role of oral intake of melatonin.
Forty eight adult albino rats, twenty four males and twenty four females, were used in this study. Animals were divided into four groups. Rats in group I Were used as control. Animals in group II were given alloxan intraperitoneal in a dose of 150 mg/ Kg bodyweight to induce diabetes mellitus. Those in group III were given oral melatonin only in a dose of 10 mg/kg bodyweight per day. Rats in group IV were given oral melatonin after induction of diabetes. Each group was further subdivided into male and female subgroup. After two weeks at the end of experiment, animals were sacrificed; the cerebella were excised. Midsagittal serial sections of half of the cerebellum were cut at 7 µm thickness and stained with Haematoxylin and Eosin, Cresyl violet stain and Glial Fibrillar Acid Protein (GFAP). The other half of the cerebella were fixed immediately in 2.5 % glutaraldehyde in phosphate buffer for 3 hours. Fixed tissue samples were washed with phosphate buffer and post fixed in 1 % osmium tetra oxide. Semi thin sections one um thick were cut using L.K.B. ultra microtome, picked up on a gelatinized glass slides and stained with toluidine blue. All sections were examined and photographed with light microscope.
Ultrathin sections were cut with a diamond knife with L.K.B. ultra microtome and picked on uncoated 300-mesh copper grids, stained with ultrastainer using Uranyl acetate and lead citrate and examined with transmission electron microscope.
Morphometric studies were also done. The GFAP percentage areas were measured in the GFAP immunohistochemistry stained sections using digital photomicrographs taken at magnification of 400. The number of apparently normal Purkinje cells was counted in cresyl violet sections using digital photomicrographs taken at magnification of 400.
In the present study, histological sections of the cerebella from alloxan-induced diabetic rats generally showed intact arrangement of the three cortical layers. However, considerable degenerative changes were observed in the three layers. Purkinje cells were mostly affected; Most of them were distorted in shape with irregular darkly stained nuclei with wide areas of pericellular halo surrounding them. They were few and some of them had pyknotic nuclei. They were distorted in shape and contained ill defined Nissl granules in cresyl violet sections. Examination of the GFAP stained sections of diabetic rat cerebella showed a strong positive reaction of GFAP immunostaining for astrocytes cell bodies and dendrites in all layers of cerebellum. In semithin sections
Forty eight adult albino rats, twenty four males and twenty four females, were used in this study. Animals were divided into four groups. Rats in group I Were used as control. Animals in group II were given alloxan intraperitoneal in a dose of 150 mg/ Kg bodyweight to induce diabetes mellitus. Those in group III were given oral melatonin only in a dose of 10 mg/kg bodyweight per day. Rats in group IV were given oral melatonin after induction of diabetes. Each group was further subdivided into male and female subgroup. After two weeks at the end of experiment, animals were sacrificed; the cerebella were excised. Midsagittal serial sections of half of the cerebellum were cut at 7 µm thickness and stained with Haematoxylin and Eosin, Cresyl violet stain and Glial Fibrillar Acid Protein (GFAP). The other half of the cerebella were fixed immediately in 2.5 % glutaraldehyde in phosphate buffer for 3 hours. Fixed tissue samples were washed with phosphate buffer and post fixed in 1 % osmium tetra oxide. Semi thin sections one um thick were cut using L.K.B. ultra microtome, picked up on a gelatinized glass slides and stained with toluidine blue. All sections were examined and photographed with light microscope.
Ultrathin sections were cut with a diamond knife with L.K.B. ultra microtome and picked on uncoated 300-mesh copper grids, stained with ultrastainer using Uranyl acetate and lead citrate and examined with transmission electron microscope.
Morphometric studies were also done. The GFAP percentage areas were measured in the GFAP immunohistochemistry stained sections using digital photomicrographs taken at magnification of 400. The number of apparently normal Purkinje cells was counted in cresyl violet sections using digital photomicrographs taken at magnification of 400.
In the present study, histological sections of the cerebella from alloxan-induced diabetic rats generally showed intact arrangement of the three cortical layers. However, considerable degenerative changes were observed in the three layers. Purkinje cells were mostly affected; Most of them were distorted in shape with irregular darkly stained nuclei with wide areas of pericellular halo surrounding them. They were few and some of them had pyknotic nuclei. They were distorted in shape and contained ill defined Nissl granules in cresyl violet sections. Examination of the GFAP stained sections of diabetic rat cerebella showed a strong positive reaction of GFAP immunostaining for astrocytes cell bodies and dendrites in all layers of cerebellum. In semithin sections
Other data
| Title | Does Melatonin Protect The Cerebellum Against Structural Alterations In Diabetic Rats? | Other Titles | هل يحمي الميلاتونين المخيخ من التغيرات التي تحدث للفئران المصابة بداء البول السكري؟ | Authors | Mahmoud Mohamed Abdallah | Issue Date | 2017 |
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