Visualization of Purkinje Neurons of Hypothyroid Brain under Golgi Cox Stain

Mariyah Hidayat, Inayatullah Khan, Tahira Hassan, Shire Chaudhry, Khalid P.Lone

Abstract


Background: Golgi staining was invented hundreds of years ago but it is still a reliable method to study the anatomy of neurons.
Objective: This experiment was conducted to study the structure of Purkinje neurons of cerebellum of neonate rats born from hypothyroid dams under golgi cox stain and compare it to the control group.
Material and Methods: For this purpose 10 female wistar rats were divided equally into 2 groups control (A) and hypothyroid (B) groups and allowed to conceive. For inducing hypothyroidism in dams, Propylthiouracyl (PTU) was administered in a dose of 15mg/kg/day orally mixed with chow daily a week before mating and throughout the period of gestation and weaning uptil 22nd day after delivery. On the 23rd day, 10 neonatal rats from each group were sacrificed and blood samples were immediately collected for evaluating serum levels of T3, T4 and TSH. The freshly extracted brains were placed in Golgi Cox solution in complete darkness for 18 to 21 days and the solution was
changed every alternate day. The brains were then removed from this stain, processed, infiltrated with parafin, cut into 80ìm thick sections by a microtome and mounted on gelatin coated slides, which were now incubated in 20% ammonium hydroxide for ten minutes.
They were then washed in distilled water for 2 minutes and passed through ascending ethanol series 70%, 95%, 100% (five minutes each) and two times xylene (10 minutes each). The sections were now coverslipped, visualized and photographed under a research microscope.
Results: Serum enzyme analysis revealed that group PP pups had significantly increased serum levels of TSH and control CC pups showed normal levels of this hormone. T3 and T4 levels were not significantly altered. The number of dendrites seen in purkinje neurons were less in group PP as compared to CC. Normal Purkinje cell count in both the groups were not significantly variable, but the apoptotic cells were significantly more in PP group.
Conclusion; Hypothyroidism decreases the number of dendrites ofpurkinje neurons in cerebellum, causing impaired neuronal
connectivity.

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