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Abstract

Objective: Pedunculopontine nucleus (PPN) is a brainstem nucleus with an obscure border and located in the caudal tegmentum. In recent years, the numbers of studies related to PPN have been increasing due to its role in sleep/waking, in behavior, learning, and attention as well as its role in generation and control of locomotion. Besides cholinergic neurons, there are GABAergic, glutamatergic, dopaminergic and noradrenergic neurons in the PPN. Recently, the PPN has been a target for treatment of axial semptoms in Parkinson disease (PD). Calbindin, a member of the calcium binding protein family, which is an important molecule during protein synthesis has been related to neurodegenerative diseases. In our study, we investigated the developmental differences of calbindin expression which is related to GABAergic and glutamatergic neurons.Materials and Methods: The borders of the PPN were determined on the basis of the distribution of cholinergic neurons. Calbindin is expressed by GABAergic and glutamatergic neurons. Wistar rats aged 7, 14 and 21 days were perfused through the heart with a 4% paraformaldehyde solution. Brains were removed and 40 µm plane of sections were cut via microtome. Free-floating slices were stained with anti choline asetyltransferase and calbindin. In our study, we used 4 rats for each group, totally 12 rats were utilized. The number, location and diameters of calbindin positive neurons in PPN were evaluated in 7, 14 and 21 day-old rats.Results: The mean numbers of neurons were 30 ±3 at 7 days, 11 ±1 at 14 days and 150±50 at 21 days rats. There were statistically significant differences between 7 and 14 days and 14 and 21 day-old rats (p=0.0075 ve p=0.0343 respectively). The diameters of the neurons were 13 ±1.4 µm at 7 days, 11±1.6 µm at 14 days; 13.5±1.1µm at 21days. Conclusion: This study indicates that critical developmental changes in PPN occur between these intervals. These findings support and contribute to the findings of previous studies

Keywords

• Tegmental pedunculopontine nucleus, Calbindin, GABAergic, Cholinergic

Nucleus pedunculopontinus tegmenti’de nörokimyasal özelliklerin gelişimsel olarak incelenmesi

Abstract

Amaç: Kaudal tegmentumda yerleşimli, sınırları belirsiz bir beyin sapı nükleusu olan nucleus pedunculopontinus (PPN), lokomotor hareketlerin oluşumunda ve kontrolünde rol oynar. Bunun yanı sıra, PPN, uyku/uyanıklık, davranış, öğrenme ve dikkat ile de ilişkilendirilmiştir. PPN içinde kolinerjik glutamaterjik, dopaminerjik, nöradrenerjik ve GABAerjik nöronlar tanımlanmıştır. PPN son dönemlerde Parkinson hastalığının aksial semptomlarının tedavisi için potansiyel bir hedef haline gelmiştir. Protein sentezi ve salınımı sırasında gerek duyulan önemli bir molekül olan kalsiyum bağlayıcı protein ailesi üyesi kalbindin, nörodejeneratif hastalıklarla ilişkilendirilmektedir. Amacımız, bu çalışma ile GABAerjik ve glutamaterjik nöronlarla yakından ilişkili olan kalbindinin PPN’de gelişimsel olarak sentezlenme farklılıklarını incelemektir. Gereç ve Yöntem: PPN sınırları, içerdiği kolinerjik nöronlara göre tanımlanmıştır. Kalbindin, glutamaterjik ve GABAerjik nöronlardan sentezlenir. Wistar sıçanlar doğumlarının 7.gün, 14. gün ve 21. günlerinde alınarak % 4’lük paraformaldehit ile transkardiyak perfüze edilip, beyinleri çıkarılmıştır. 40 µm’luk kesitler mikrotom kullanılarak alınmış ve kesitlere immunohistokimyasal anti kolinasetil transferaz boyama ile kalbindin boyama uygulanmıştır. Çalışmamızda, her bir hayvan grubu için 4’er tane olmak üzere 12 sıçan kullanılmıştır. 7 gün, 14 gün ve 21 günlük sıçanların PPN nüklesuna ait kalbindin pozitif nöronlarının sayı, lokasyon ve nükleus çapları araştırılmıştır. Bulgular: Ortalama nöron sayıları 7 günlük hayvan için 30±3, 14 günlük hayvan için 11±1 ve 21 günlük hayvan için 150±50 olarak sayılmıştır. Nöron sayıları açısından 7. ve 14. günler ile 14 ile 21. günler arasında anlamlı farklılıklar gözlenmiştir (p=0,0075 ve p=0,0343 ).Nöron çapları 7. günde 13±1,4 µm ;14.günde 11±1,6 µm ve 21. günde 13,5±1,1 µm olarak ölçülmüştür. Sonuç: Bu çalışma, PPN’deki gelişimsel farklılıkların ortaya çıktığı kritik dönemin yukarıda belirtilen zaman aralığında olduğunu göstermiştir. Elde ettiğimiz sonuçlar, daha önce yapılan çalışmaların sonuçlarını destekleyip, katkı sağlamaktadır.

Keywords

• Nucleus pedunculopontinus tegmenti, Kalbindin, GABAerjik, Kolinerjik

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