Laterality of spinocerebellar neurons in the chicken spinal cord

The aim in this study is to elucidate the laterality of chicken spinocerebellar (SC) neurons that originate from the caudal cervical to caudal lumbosacral spinal cord. SC neurons in the spinal segment (SS) 17-20 consisted of a mixture of crossed and uncrossed axons. SC neurons in the more cranial and caudal SS than SS 17-20 (transitional zone) were generally uncrossed and crossed, respectively. In the transitional zone, SC neurons in spinal border cells and ventral border cells of the ventral horn changed dramatically from an uncrossed to a crossed type between SS 17 and SS 18. Chicken SC neurons are markedly different in laterality from mammalian SC neurons.     

神经肽Y免疫阳性神经元在江汉鸡延髓内的分布

为了探明江汉鸡延髓内神经肽Y（neuropeptide Y，NPY）免疫反应神经元的分布状况，采用石蜡切片和免疫组织化学链霉亲合素-生物素-过氧化物酶复合法（SABC法），对10羽江汉鸡的延髓进行了研究，并与北京鸭、乌鸡、肉鸡及大鼠的相关研究结果进行了比较。在光镜下观察分析了阳性神经元的分布状况，并用图像分析系统进行半定量分析。免疫组织化学染色结果显示，NPY阳性神经元主要存在于延髓下橄榄核中，孤束核及中缝核也有少量表达。表明，江汉鸡延髓NPY阳性神经元的分布与肉鸡、乌鸡、大鼠的大体相似；下橄榄核可能是NPY在延髓-小脑传递通路中的一个重要枢纽。     

Distribution of parvalbumin in specific fibre types of chicken skeletal muscles

1. The distribution of parvalbumin (PA), which functions as a relaxing factor in the skeletal muscles, was examined in slow anterior latissimus dorsi (ALD), fast posterior latissimus dorsi (PLD), mixed sartorius (SA), pectoralis superficialis (PS) and pectoralis profundus (PP) muscles from chickens.

2. The biochemical characteristics of these muscles were confirmed by the assay of total lactate dehydrogenase (LDH) activity as well as the LDH isozymes for anaerobic metabolism, and by the photometrical analysis of myoglobin for aerobic metabolism.

3. PA in individual muscles was determined by a sandwich ELISA and was demonstrated by 2‐dimensional polyacrylamide‐gel electrophoresis.

4. Because of poor myoglobin and higher LDH activity or M‐type isozyme pattern, the PLD was confirmed as containing primarily fast‐twitch glycolytic and oxidative‐glycolytic (FG/FOG) fibres, while the SA was shown to be composed mosdy of FOG fibres because of the highest myoglobin content and the intermediate LDH activity or H and M‐type isozyme pattern. PA content was high and variable in both PLD and SA.

5. PA was undetectable in the ALD which appears to contain exclusively slow‐tonic (ST) fibres, being verified by its myoglobin‐rich nature and lowest LDH activity or predominant H‐type isozyme characteristics. PA was absent from the PS and PP, which probably contain predominantly FG fibres because of negligible amounts of myoglobin and the highest LDH activities or M‐type isozyme pattern.     

真核表达质粒pTracer-CMV2FIL2在鸡体组织中表达及分布规律

将3周龄的三黄鸡肌肉接种200 μg真核表达质粒pTraeer-CMV2FIL2,巢式PCR法检测血清、心、脾脏、肺脏、法氏囊、脑、骨髓、胸腺和肌肉接种部位中pTracer-CMV2FIL2出现和存留的时间;荧光定量PCR法检测其在血液中的动态变化;RT-巢式PCR法检测其在上述部位的表达情况.结果显示,接种后2～3 h血清中可检测到pTracer-CMV2FIL2,6 h时在血液中的含量最高,7 d时已经检测不到;4 h后在上述器官中可陆续检测到pTracer-CMV2FIL2的出现,13 d后陆续消失;1 d后可陆续检测到质粒所转录的mRNA,13 d后陆续消失;接种后2 h可在接种部位肌肉的细胞中检测到pTracer-cMV2FIL2的出现,10 h可检测到mRNA,150 d时仍然可以在肌肉接种部位的细胞中检测到质粒和mRNA存在.     

Distribution and developmental change of lymphoid tissues in the chicken proventriculus

In the chicken proventricular mucosa, aggregations of lymphocytes were localized in three different sites of the lamina propria, namely, underneath the surface epithelium, near the duct orifice of the deep proventricular gland, and in the gland tissue itself. In the lymphoid masses underneath the surface epithelium and in those near the duct orifice, CD4+ T lymphocytes and TCR2+ T lymphocytes occupied their central part, and B lymphocytes were localized in the periphery. CD8+ T lymphocytes and TCR1+ lymphocytes were evenly distributed in the masses. Infiltration of lymphocytes into these sites was first observed on the 20th embryonic day. At 1 week after hatching, CD3+ lymphocytes began to occupy the central area of the masses and His-C1+ B lymphocytes tended to be located in the periphery. Ultrastructurally, M cells were found neither in the epithelium of the mucosa nor in that of the excretory duct close to the lymphoid masses. In the deep proventricular gland, the lymphoid masses had a germinal center consisting of B lymphocytes, surrounded by the T lymphocyte-rich periphery. These masses were first recognized at the 3rd post-hatching week, presumably being formed against possible antigens invading into the lumen of the proventricular gland. On the other hand, the lymphoid masses beneath the surface epithelium and those near the duct orifice existing before the hatching period were considered to be prepared to establish the local mucosal immune barriers against the expectant antigenic invasion.     

Distribution of immunoreactive vasotocin and mesotocin in the chicken gastrointestinal tract

Immunoreactive arginine vasotocin (AVT) and mesotocin (MT) were measured in heart, breast muscle, adrenals, testes, and different parts of the gastrointestinal tract in adult male chickens. Neither of the peptides were detected in liver, testis, heart and breast muscle. The amounts of AVT and MT in the adrenals were 167 +/- 25 and 669 +/- 198 pg/gland, respectively. Considerable amounts of immunoreactive peptides were found in the gastrointestinal tract with the highest concentration in the proventriculus (4.18 +/- 0.31 ng AVT and 16.58 +/- 0.86 ng MT per organ). Dose-response curves of duodenal and proventriculus extracts were parallel with synthetic AVT and MT standards.     

支配鸡肝的交感传出神经元分布规律的研究

为了解支配鸡肝的交感传出神经元的分布规律，将CT—HRP溶液注入鸡肝门周围区，动物存活3～4d后，经左心室灌流固定，取内脏神经节、肾上腺神经节及双侧胸、腰和荐段交感干神经节，制成50μm厚的连续冰冻切片，TMB法呈色反应，明视野显微镜下观察统计。结果发现，支配鸡肝的交感传出神经元胞体位于内脏神经节（占41．9％）、肾上腺神经节（占41．7％）和T2～T7交感干神经节（占16．4％）。在交感干神经节中，标记细胞的峰值位于T5、T6交感干神经节。     

Distribution of neurons supplying the porcine mammary gland

The distribution of neurons projecting to the mammary gland was investigated by using the retrograde tracing method in juvenile pigs (n = 12). Fluorescent retrograde tracer Fast Blue (FB) was injected into the nipple (n = 3) or parenchyma (n = 3) of the second, right thoracic mamma or into the nipple (n = 3) and parenchyma (n = 3) of the last, right abdominal mamma. FB-positive (FB+) mammary gland-projecting neurons were found in some right dorsal root ganglia (DRG) and sympathetic chain ganglia (SChG) only. After injection of the tracer into the second, right thoracic mamma, FB+ neurons were observed in Th9-Th12 DRG but most of them were located in Th11 and Th12 ganglia. As concerns SChG, FB+ neurons were found in Th1-Th4, Th7-Th14 and L1-L4 ganglia. The vast majority of them were located in Th10 and Th11 SChG, which appeared to be the main sources of efferent innervation of this mamma. Neurons projecting to the last right abdominal mamma were found in L1-L3 DRG and L1-L4 SChG but most of them were located in L1-L2 ganglia and L1-L2 ganglia, respectively. This study for the first time has disclosed the localization of neurons supplying the mammary gland in larger domestic animal species, the pig, by using the retrograde tracing method.     

Location of sympathetic postganglionic and sensory neurons innervating the testis in the male chicken

Sympathetic postganglionic and sensory neurons were labelled by injections of horseradish peroxidase into the testis of the male chicken. The total number of labelled neurons in the paravertebral, prevertebral, dorsal root and nodose ganglia was 943 on average for five chickens. Sympathetic postganglionic neurons were located in the paravertebral ganglia T3-LS3 (10% of the total number of labelled neurons), especially in T6 and T7, and in the prevertebral ganglia adjacent to the adrenal glands and aorta (19%). They were found almost ipsilaterally. No labelled neurons were observed in the dorsal motor nucleus of the vagus. Sensory neurons were found bilaterally in the dorsal root ganglia T2-LS3 (71%), especially in T5 to T7. Over a quarter of labelled sensory neurons were located in the contralateral dorsal root ganglia. In the nodose ganglia, only a few labelled sensory neurons were observed (much less than 1%). These results indicate that, unlike the ovary, the testis of the chicken tends to be innervated by ipsilaterally located sympathetic postganglionic and sensory neurons, with the sensory neurons being more numerous than the sympathetic postganglionic neurons.     

SPF鸡各组织中流感病毒唾液酸受体的分布

为了解SPF鸡体内各组织中流感病毒唾液酸受体分布状况,本研究采用凝集素免疫荧光组织染色的方法,系统检测了流感病毒唾液酸受体在SPF鸡呼吸系统、消化系统以及其它系统各组织中的分布,并进行半定量分析。结果表明,大部分组织中均同时存在唾液酸α2,3-半乳糖受体与唾液酸α2,6-半乳糖受体,只有少部分组织仅存在其中一种受体,这种受体分布的广泛性与流感病毒组织噬性相一致;但鼻甲与喉头中唾液酸受体明显低于许多其他组织,这也表明不能简单的通过唾液酸受体的有无或者密度高低来判定病毒在各组织中的实际分布与复制增殖情况,或许还有其他机制参与了病毒入侵与增殖的过程。     

Distribution of nitric oxide synthase (NOS) immunoreactivity in the chicken stomach

NOS-immunoactive nerve structures were studied in the domestic hen (Gallus domesticus) stomach using single-labelling immunofluorescence. The examined tissues were collected from 10-12 months old laying hens (n=11), Isabrown and Zielononózka races. The distribution and number of the nerve fibres was examined considering differences between the proventriculus and gizzard, and also between particular layers of the stomach wall. Large or moderate numbers of nerve fibres showing NOS-immunoreactivity (NOS-IR) were found in all major layers of both parts of the stomach. In the muscular layer of the gizzard, NOS-positive nerve cell bodies were also observed. Small or moderate numbers of nerve terminals were distributed around blood vessels. NOS-IR nerve fibres associated with blood vessels were slightly more numerous in the gizzard than in the proventriculus. The widespread occurrence of NOS in both parts of the hen stomach can prove its important role in the control of digestive processes. It seems that there is a reasonable necessity of further, more detailed studies on morphological characteristics of the nitrergic nerve structures the bird alimentary tract, their functional significance and plasticity in different physiological and pathological conditions.     

Neuroma formation following partial beak amputation (beak trimming) in the chicken

Branches of the trigeminal nerve, which innervates the beak in the chicken, were damaged after partial beak amputation (beak trimming) and degeneration in the nerve extended from 2 to 3 mm proximal to the cut stumps. Within 10 days the damaged nerves showed regeneration and axon sprouts had been formed. This neural regeneration was rapid so that by 20 to 30 days bundles of regenerating fibres were present and neuromas formed adjacent to the scar tissue of the stump of the beak. These neuromas became more extensive over the 70 day observation period.     

辣根过氧化物酶标记法研究鸡直肠传入神经元的分布

采用辣根过氧化物酶（HRP）逆行追踪法研究了鸡直肠感觉传入神经元的分布，证明鸡直肠脊神经感觉传入神经元集中分布于双侧T5～LS3和LS7～LSl2脊神经节，峰值位于T7和LS10脊神经节；LS7-LS12区段脊神经节内标记细胞明显多于T5～LS3区段脊神经节内的标记细胞；标记细胞为椭圆形和圆形。     

A quantitative study of the purkinje cells in the cerebellum and the inferior olivary neurons in the chicken

A single olivocerebellar fiber branches off several climbing fibers. One Purkinje cell receives input from only one climbing fiber. A single inferior olivary neuron, therefore, synapses with several Purkinje cells, so that there are more Purkinje cells than the inferior olivary neurons. We aimed to elucidate the numerical ratio of the inferior olivary neurons to Purkinje cells in the chicken. The total numbers were 353,834 +/- 5,274 in the Purkinje cells per the cerebellum and 21,553 +/- 904 in the inferior olivary neurons of both sides. The numerical ratio of inferior olivary neurons to Purkinje cells was 1:16. The ratio of those neurons in mammals is about 1:4-17, so that the ratio in the chicken is within the range of mammals.     

Mitochondrial inclusion bodies (intracytoplasmic acidophilic droplets) in neurons of chicken spinal cords increase with age

We studied the pathologic features of neurons that contain intracytoplasmic acidophilic droplets (IADs) in chicken spinal cords. The IADs were lustrous spheroid bodies scattered in the cytoplasm of neurons, variable in size, and protein-rich bodies stained eosinophilic with hematoxylin-eosin, acidophilic with Azan, blue indigo with phosphotungstic acid hematoxylin, and yellow-green with Elastica van Gieson stain histopathologically. Ultrastructurally, almost all IADs were observed as homogeneous highly electron-dense spheroid bodies enclosed by double-limited membranes. Small IADs were observed in mitochondria. Anatomically, IAD-CNs were observed only in the ventral horn of the spinal cord between the fourth sacral and third lumbal vertebrae, and they were particularly frequent in the third sacral vertebrae. Their appearance and accumulative amount were likely to increase with age, while the clinical and pathologic significances of IAD-CNs remain unclear.     

Distribution of chicken cathepsins B and L,cystatin and ovalbumin in extra-embryonic fluids during embryogenesis

1. Concentrations of chicken cathepsin B, cathepsin L, cystatin and ovalbumin were determined in the allantoic fluid, amniotic fluid and extracts of chorioallantoic membranes during days 6 to 12 of embryogenesis.

2. Similar trends for cystatin and ovalbumin were observed in the allantoic fluid with maximum concentrations of cystatin on day 7 (12?±?4?µg/ml) and ovalbumin on day 8 (～19?±?2·5?µg/ml) of embryonic development. The highest concentrations of cathepsin B was found on day 7 and of cathepsin L on day 10, but were significantly lower than those of cystatin and ovalbumin.

3. In the allantoic fluid, especially on day 7, considerable proportions of cystatin and ovalbumin were phosphorylated and contained phosphorylated serine.

4. Concentrations of cathepsin B and L, cystatin and ovalbumin in the amniotic fluid were variable but were comparable to those in allantoic fluid.