Lloyd R V, Gilles P A, Karavolas H J
Steroids. 1979 Jan;33(1):97-113. doi: 10.1016/s0039-128x(79)80009-4.
Rat medial basal hypothalami (MBH) and sections of cerebral cortex (CC) were dissociated with trypsin to prepare single cells and subcellular fractions. They were then separated into four fractions on a discontinuous sucrose gradient. The small neurons in Fraction D were highly purified. Fraction A had synaptosomes, myelin and other cell particulates. Fraction B had glial cells, neurons and a few synaptosomes. Fraction C had large neurons and red blood cells. All four fractions contained LHRH, but most (62.5%) of this hormone was present in Fraction A. Dissociated cell suspensions were incubated with [3H]-steroids, with and without a 100-fold excess of unlabeled steroids, then separated on sucrose gradients. In most fractions the total uptake and specific uptake of [3H]-progesterone, [3H]-5 alpha-pregnane-3,20-dione (5 alpha-dihydroprogesterone) and [3H]-17 beta-estradiol were greater for the dissociated cells from the MBH than the CC. The dissociated cells and cell particulates in all four fractions from the MBH and CC metabolized progesterone, 5 alpha-dihydroprogesterone and 17 beta-estradiol. These results indicate that hypothalamic neurons contain small amounts of LHRH and retain the ability to take up and metabolize progesterone, 5 alpha-dihydroprogesterone and 17 beta-estradiol.
将大鼠内侧基底部下丘脑(MBH)和大脑皮质(CC)切片用胰蛋白酶解离以制备单细胞和亚细胞组分。然后将它们在不连续蔗糖梯度上分离成四个组分。组分D中的小神经元高度纯化。组分A含有突触体、髓磷脂和其他细胞颗粒。组分B含有神经胶质细胞、神经元和一些突触体。组分C含有大神经元和红细胞。所有四个组分均含有促性腺激素释放激素(LHRH),但该激素的大部分(62.5%)存在于组分A中。将解离的细胞悬液与[3H] - 类固醇一起孵育,有无100倍过量的未标记类固醇,然后在蔗糖梯度上分离。在大多数组分中,来自MBH的解离细胞对[3H] - 孕酮、[3H] - 5α - 孕烷 - 3,20 - 二酮(5α - 双氢孕酮)和[3H] - 17β - 雌二醇的总摄取和特异性摄取均高于CC。来自MBH和CC的所有四个组分中的解离细胞和细胞颗粒均代谢孕酮、5α - 双氢孕酮和17β - 雌二醇。这些结果表明,下丘脑神经元含有少量LHRH,并保留摄取和代谢孕酮、5α - 双氢孕酮和17β - 雌二醇的能力。