Staining of cerebellar cortex granular layer interneurons with natural dye of Madder.

Anneh Mohammad Gharravi

doi:10.1186/s40673-016-0050-6

Abstract

BackgroundThe objective of the present study was an investigation of root Rubia Tinctorum (Madder) as a natural dye to identification of granular layer interneurons of the rat cerebellum.MethodsSeven to ten micrometre sections were collected from the cerebellum and stained only with Madder for 2, 24 and 48 h. Other sections were stained with Madder then with hematoxyllin, cresyl violet, eosin, light green. Microscopic identification of cells was performed based on cell morphology, reaction and binding of with the dye. All data were expressed as mean ± SD in and significance was set at p ≤0.05.ResultsMadder with alum as mordant resulted a deep red staining of interneurons. Unipolar brush cells (UBCs) were observed with a cell body diameter intermediate between granule and Golgi cells in the superficial layer of the granular layer. Golgi cells were identified almost as large as Purkinje cells with irregular rounded or polygonal morphology. Lugaro cells were observed as spindle-shaped cells adjacent to Purkinje layer.ConclusionResults of the present study showed that mader could stain granular layer interneurons in cerebellum cortex of rat.

Highlights

The objective of the present study was an investigation of root Rubia Tinctorum (Madder) as a natural dye to identification of granular layer interneurons of the rat cerebellum
Our results indicate that madder with alum as mordant result a deep red staining of granular layer interneurons observed in acidic solution of madder
Unipolar brush cells (UBCs) were observed with a cell body diameter intermediate between granule and Golgi cells

Summary

Introduction

The objective of the present study was an investigation of root Rubia Tinctorum (Madder) as a natural dye to identification of granular layer interneurons of the rat cerebellum. There are three distinct interneurons in the granular layer; Golgi, Lugaro and UBCs [1, 2]. The cells are a unique type excitatory, glutamatergic interneurons that receive direct mossy fiber excitation, play a role in amplifying mossy fiber signals [4]. UBCs are intermediate in size between granule cells are found in abundance within the granule cell layer, in the uvula-nodulus [5]. Golgi cells are inhibitory (GABAergic) interneurons and sample mossy fiber activity, provide both feedforward and feedback inhibition of granule cells. The cells are the largest and most numerous interneurons within the granular layer [6, 7]. Granule cells receive glutamategic inputs from mossy fibers and UBCs, and glycinergic/GABA aergic inputs from Golgi cells. Lugaro cells have numerous axosomatic and axodendritic contacts

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