Bombardi Cristiano, Grandis Annamaria, Chiocchetti Roberto, Lucchi Maria Luisa
Department of Veterinary Morphophysiology and Animal Productions, Faculty of Veterinary Medicine, University of Bologna, Via Tolara di Sopra, 50, 40064, Ozzano dell'Emilia (BO), Italy.
Anat Embryol (Berl). 2006 Nov;211(6):707-20. doi: 10.1007/s00429-006-0133-x. Epub 2006 Oct 18.
This study describes calbindin-D28k (CB), neuronal nitric oxide synthase (nNOS), and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) expression in the lateral nucleus of the sheep amygdaloid complex. Double immunofluorescence protocol was used in order to determine whether there is colocalization of CB and nNOS. The CB-immunoreactive (IR) neuronal population was composed especially of non-pyramidal neurons, but a few pyramidal cells were also present. The non-pyramidal neurons showed a multipolar and, occasionally, a fusiform morphology. The comparison between single-labeled CB-IR non-pyramidal neurons and cells belonging to CB-IR neuronal population showed they were identical for morphology, mean size, and distribution. The single-labeled CB-IR non-pyramidal neurons were only the 17.8% of the total non-pyramidal neurons counted. The nNOS-IR neuronal population was represented by non-pyramidal multipolar and fusiform neurons. Single-labeled nNOS-IR non-pyramidal neurons had the same morphology, mean area, and distribution as cells belonging to nNOS-IR neuronal population. Single-labeled nNOS-IR non-pyramidal neurons were more numerous than single-labeled CB-IR, and represented the 73.7% of total non-pyramidal neurons counted. NADPH-d-positive cells had the same morphology and distribution as the nNOS-IR neurons. Double immunolabeling (CB/nNOS) was found mostly in non-pyramidal multipolar neurons and only in a few non-pyramidal fusiform cells. These neurons had a mean perikaryal area significantly higher and significantly smaller than that of single-labeled nNOS and single-labeled CB-IR non-pyramidal neurons, respectively. CB and nNOS coexist only in a minority of non-pyramidal neurons (8.5%). The 32.4% of all CB-IR non-pyramidal neurons were nNOS-positive; only 10.4% of nNOS-IR non-pyramidal neurons were CB-positive. These results indicate that CB and nNOS are expressed by selective neurons and that the majority of nNOS-IR non-pyramidal neurons are lacking in CB.
本研究描述了绵羊杏仁核复合体外侧核中钙结合蛋白-D28k（CB）、神经元型一氧化氮合酶（nNOS）和烟酰胺腺嘌呤二核苷酸磷酸黄递酶（NADPH-d）的表达情况。采用双重免疫荧光法来确定CB和nNOS是否存在共定位。CB免疫反应阳性（IR）神经元群体尤其由非锥体神经元组成，但也存在少数锥体细胞。非锥体神经元呈现多极形态，偶尔也呈梭形。对单标记CB-IR非锥体神经元与属于CB-IR神经元群体的细胞进行比较，结果显示它们在形态、平均大小和分布上相同。单标记CB-IR非锥体神经元仅占所计数的非锥体神经元总数的17.8%。nNOS-IR神经元群体由非锥体多极和梭形神经元组成。单标记nNOS-IR非锥体神经元与属于nNOS-IR神经元群体的细胞具有相同的形态、平均面积和分布。单标记nNOS-IR非锥体神经元比单标记CB-IR非锥体神经元数量更多，占所计数的非锥体神经元总数的73.7%。NADPH-d阳性细胞与nNOS-IR神经元具有相同的形态和分布。双重免疫标记（CB/nNOS）主要见于非锥体多极神经元，仅在少数非锥体梭形细胞中发现。这些神经元的平均核周面积分别显著大于和小于单标记nNOS和单标记CB-IR非锥体神经元的平均核周面积。CB和nNOS仅在少数非锥体神经元（8.5%）中共存。所有CB-IR非锥体神经元中有32.4%为nNOS阳性；nNOS-IR非锥体神经元中只有10.4%为CB阳性。这些结果表明，CB和nNOS由选择性神经元表达，并且大多数nNOS-IR非锥体神经元缺乏CB。
