Groome J R, Pryma D, Donahue R M
Biology Department, Harvey Mudd College, 1250 North Dartmouth Avenue, Claremont, CA 91711 USA.
Invert Neurosci. 2001 Oct;4(2):95-103. doi: 10.1007/s101580100011. Epub 2001 Aug 15.
The presence and distribution of immunoreactivity to the cyclic AMP response element binding protein (CREB) were determined in the central nervous system (CNS) and in peripheral tissues of the medicinal leech Hirudo. Western blots revealed several CREB-immunoreactive (CREB-IR) bands including one whose molecular weight (43-44 kDa) was similar to mammalian CREB. The 43-44 kDa CREB-like protein was detected in nuclear extracts of the ventral nerve cord and was not observed following preincubation of the primary antiserum with the epitope sequence. CREB-like immunoreactivity was detected in extracts from each of six regions of the leech CNS, and in extracts from leech body wall musculature, crop, intestine, jaw musculature, pharynx, and salivary tissues. Whole mounts of leech ganglia revealed specific CREB-IR in a restricted population of neurons distributed throughout the leech CNS. Apparent homologues to a pair of CREB-IR dorsolateral neurons were observed in most ganglia along the ventral nerve cord. Several CREB-IR neurons exhibited segmental specificity. A number of neurons stained with an antiserum to the cyclic AMP response element modulator (CREM). These neurons showed no overlap in location with CREB-IR neurons, and this staining was not eliminated with a preabsorption control. Possible roles for a CREB-like protein in the leech are discussed.
在药用水蛭(Hirudo)的中枢神经系统(CNS)和外周组织中,测定了环磷酸腺苷反应元件结合蛋白(CREB)免疫反应性的存在和分布。蛋白质免疫印迹法显示了几条CREB免疫反应性(CREB-IR)条带,其中一条分子量(43 - 44 kDa)与哺乳动物CREB相似。在腹神经索的核提取物中检测到43 - 44 kDa的CREB样蛋白,而在用表位序列预孵育一抗后未观察到该蛋白。在水蛭中枢神经系统的六个区域中的每个区域的提取物中,以及在水蛭体壁肌肉组织、嗉囊、肠道、颌部肌肉组织、咽和唾液组织的提取物中,均检测到CREB样免疫反应性。水蛭神经节的整装标本显示,在分布于整个水蛭中枢神经系统的有限神经元群体中存在特异性的CREB-IR。在腹神经索沿线的大多数神经节中,观察到一对CREB-IR背外侧神经元的明显同源物。几个CREB-IR神经元表现出节段特异性。一些神经元用环磷酸腺苷反应元件调节剂(CREM)的抗血清染色。这些神经元在位置上与CREB-IR神经元没有重叠,并且这种染色不会因预吸收对照而消除。本文讨论了水蛭中一种CREB样蛋白的可能作用。