Kisspeptin’in Gelişimsel Döneme Bağlı Olarak Dişi Sıçanlarda Medial Preoptik Bölgede İn Vitro Noradrenalin Salıverilmesi Üzerine Etkisi

Abstract

Üçüncü ventrikülün rostral periventriküler bölgesinde lokalize olan kisspeptin nöronlarının, ovulasyon öncesi LH salıverilmesinden sorumlu olan GnRH nöronlarının major stimülatörü olduğu son yıllarda yapılan çalışmalar ile ortaya konmuştur. GnRH salıverilmesinin bir diğer ana modülatörü ise noradrenerjik sistemdir. Kisspeptinerjik ve noradrenerjik nöronların medial preoptik bölgedeki (MPB) yerleşimleri çok yakınlık göstermekle birlikte, bu iki sistem arasındaki ilişkinin yapılacak çalışmalar ile aydınlatılmasına ihtiyaç duyulmaktadır. Yapılan bu çalışma gelişim dönemi farklı dişi Sprague Dawley sıçanların MPB’sinden hazırlanan beyin dilimleri kullanılarak kisspeptinin noradrenalin (NA) salıverilmesi üzerine direk etkisinin olup olmadığını ortaya koymak amacı ile yapılmıştır. Oksijenlenmiş Krebs solüsyonu içeren inkübasyon kuyucuklarına yerleştirilen dilimler preinkübasyon dönemi ardından 60 dakika boyunca kisspeptin (40 ve 400 μM) ile inkübe edildi. İnkübasyon periyodu sonrasında inkübasyon ortamı salınan NA düzeylerinin belirlenmesi amacıyla kullanıldı. Salıverilmenin Ca2+ ile ilişkisini incelemek amacıyla Ca2+’suz Krebs solüsyonu ve hücre dışı Ca2+ şelasyonu için 400 μM BAPTA kullanıldı. Prepubertal, adölesan ve yetişkin dişi sıçanların MPB’den elde edilen dilimlerin 40 ve 400 μM kisspeptin ile inkübasyonu prepubertal dönemdeki dilimlerden NA salıverilmesini etkilemezken, adölesan ve yetişkin sıçanlarda ise salıverilmenin anlamlı olarak arttığı gözlendi. Ca2+’un ortamdan uzaklaştırılması kisspeptin kaynaklı NA salıverilmesinde anlamlı bir düşüşe (p‹0.05) neden olması veziküler salım mekanizmasının ekstrasellüler Ca+2 iyonlarına bağımlı olduğunu göstermiştir. Kisspeptinin NA salıverilmesini direkt olarak uyarabildiğini gösteren bu bulgular, söz konusu peptidin NA salıverilmesi üzerinden GnRH salıverilmesini indirekt olarak modüle edebileceğini düşündürmektedir.

Keywords

• GnRH
• Kisspeptin
• medial preoptik alan
• noradrenalin

Kisspeptin Stimulates In Vitro Noradrenaline Release in Medial Preoptic Area of the Female Rats Depending on Developmental Stage

Abstract

Kisspeptin neurons localized in the rostral periventricular area of the third ventricle are considered to provide a major stimulatory input to the GnRH neuronal network which is responsible for triggering the preovulatory LH surge. Noradrenaline (NA) is one of the main modulators of GnRH release. Although NA fibers are found in close opposition to kisspeptin neurons, interaction between kisspeptin and NA neurons needs to be clarify. Thus, present study was undertaken to determine whether kisspeptin can directly regulate NA release from brain slices prepared from medial preoptic area (MPOA) the rats. Brain slices prepared from female prepubertal, adolescents or adult Sprague Dawley rats were placed incubation wells containing oxygenated Krebs. After preincubation period, slices were incubated with two concentrations of kisspeptin (40 and 400 μM) for 60 min. At the end of kisspeptin incubation, incubation medium was used for determination of NA released from the slices. When used Ca+2-free medium, 400 μM of BAPTA was also added into the medium to chelate the extracellular Ca+2. Incubation of the brain slices prepared from MPOA of the adolescent and adult female rats with 40 and 400 μM of kisspeptin caused significant increase in NA release, unlike prepubertal rat. Since removal of the Ca+2 from the medium caused significant decline in kisspeptin-induced NA release (p‹0.05), a vesicular release mechanism seems to be likely involved. The results presented here probably indicate that in addition to direct effect of kisspeptin, it may also modulate GnRH release indirectly by increasing NA release from MPOA of hypothalamus.

Keywords

• Kisspeptin
• noradrenaline
• GnRH
• medial preoptic area

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