Khalil Mona H, Silverman Ann-Judith, Silver Rae
Department of Biochemistry and Molecular Biophysics, Columbia University College of Physicians and Surgeons, New York, New York 10032, USA.
J Neurobiol. 2003 Aug;56(2):113-24. doi: 10.1002/neu.10220.
Mast cells occur in the brain and their number changes with reproductive status. While it has been suggested that brain mast cells contain the mammalian hypothalamic form of gonadotropin-releasing hormone (GnRH-I), it is not known whether mast cells synthesize GnRH-I de novo. In the present study, mast cells in the rat thalamus were immunoreactive to antisera generated against GnRH-I and the GnRH-I associated peptide (GAP); mast cell identity was confirmed by the presence of heparin, a molecule specific to mast cells, or serotonin. To test whether mast cells synthesize GnRH-I mRNA, in situ hybridization was performed using a GnRH-I cRNA probe, and the signal was identified as being within mast cells by the binding of avidin to heparin. GnRH-I mRNA was also found, using RT-PCR, in mast cells isolated from the peritoneal cavity. Given the function of GnRH-I in the regulation of reproduction, changes in the population of brain GnRH-I mast cells were investigated. While housing males with sexually receptive females for 2 h or 5 days resulted in a significant increase in the number of brain mast cells, the proportion of mast cells positive for GnRH-I was similar to that in males housed with a familiar male. These findings represent the first report showing that mast cells synthesize GnRH-I and that the mast cell increase seen in a reproductive context is the result of a parallel increase in GnRH-I positive and non-GnRH-I positive mast cells.
肥大细胞存在于大脑中,其数量会随着生殖状态的变化而改变。虽然有人提出大脑中的肥大细胞含有哺乳动物下丘脑形式的促性腺激素释放激素(GnRH-I),但尚不清楚肥大细胞是否能从头合成GnRH-I。在本研究中,大鼠丘脑中的肥大细胞对针对GnRH-I和GnRH-I相关肽(GAP)产生的抗血清具有免疫反应性;通过存在肝素(一种肥大细胞特有的分子)或血清素证实了肥大细胞的身份。为了测试肥大细胞是否合成GnRH-I mRNA,使用GnRH-I cRNA探针进行原位杂交,并通过抗生物素蛋白与肝素的结合将信号鉴定为在肥大细胞内。使用RT-PCR还在从腹腔分离的肥大细胞中发现了GnRH-I mRNA。鉴于GnRH-I在生殖调节中的作用,研究了大脑中GnRH-I肥大细胞群体的变化。虽然将雄性与处于性接受期的雌性同居2小时或5天会导致大脑肥大细胞数量显著增加,但GnRH-I阳性肥大细胞的比例与与熟悉的雄性同居的雄性相似。这些发现首次表明肥大细胞能合成GnRH-I,并且在生殖环境中观察到的肥大细胞增加是GnRH-I阳性和非GnRH-I阳性肥大细胞平行增加的结果。