牙鲆生长抑素分泌细胞的免疫细胞化学研究

采用链霉亲合素-生物素-过氧化物酶复合物(SABC)免疫细胞化学染色技术,利用兔抗哺乳动物生长抑素血清对牙鲆(Paralichthys olivaceus)消化道(包括食道、胃、幽门垂、肠道全长及肛门)、肝胰脏、肾脏、脾脏以及生殖腺等器官的生长抑素(SS)分泌细胞进行定位和鉴别。结果表明,生长抑素分泌细胞仅存在于胃和胰岛中,其他检测部位均未见分布。在胃中,生长抑素分泌细胞主要散在分布于胃黏膜上皮、胃小凹底部上皮和胃腺的颈部周围。在胃的不同部位,生长抑素分泌细胞的分布密度存有差异,以胃体部数量最多,贲门部次之,幽门部最少。参照消化道内分泌细胞与胃肠腔或腺腔有无直接联系,以及基部有无胞质突起的分类方法,将SS分泌细胞分为4种不同的类型,这4种类型的SS分泌细胞在胃中均有出现,且不同部位出现的几率不同。其中,贲门部以第Ⅲ、Ⅰ型居多;胃体部以第Ⅳ、Ⅰ型细胞为主;幽门部仅有第Ⅰ、Ⅱ型细胞。提示,牙鲆的胃是一个复杂的内分泌器官。在胰脏中,生长抑素分泌细胞零散地分布于胰岛细胞之间,数量很少。     

A rapid multiple immunofluorescence method for the simultaneous detection of CD20 and CD3 in canine and feline cytological samples

CD20 and CD3 are considered reliable markers for B and T cells, respectively. This study aimed to develop a rapid multiple immunofluorescence (RMIF) method for the detection of CD20 and CD3 on a single cytology slide. Air-dried smears were prepared using samples collected from dogs (n=26) and cats (n=6). Immunosignal detection using the newly developed method required 60 min. Clear immunosignals for CD20 and CD3 were detected in 24 of 26 samples in dogs and in all 6 cats. As the RMIF (CD20/CD3) method can detect markers of both B and T cells simultaneously on a single cytology smear, it would be an efficient tool for the immunophenotyping of canine and feline lymphoma samples.     

Identification of FXPRLamide Family Neuropeptides from the Japanese Oak Silkworm, Antheraea yamamai Using Immunocytochemistry Methods

In the present study, zooblooting, ELISA, and whole-mount immunocytochemistry methods were used to identify the FXPRLamide family neuropeptides from the Japanese oak silkworm, Antheraea yamamai. The results showed that the genomic DNA from A. yamamai showed positive bands after being hybridized with the fragment of DH-PBAN cDNA from Samia cynthia ricini, which was labeled with [α-32p]-dCTP. The SG showed highest FXPRLamide peptides titer in neural organs. Using an antiserum against Helicoverpa armigera PBAN, PBAN-like immunoreactivity was detected in the SG and TG of A. yamamai by whole-mount immunocytochemistry, and there were three cluster cells in the SG which shows positive PBAN-like immunoreactivity. The titers of FXPRLamide peptides immunoreactivity in the hemolymph were kept at a steady level. During pupation, the titer was increased promptly, but then decreased to a low level after the early pupal stage. The above-mentioned results demonstrate the existence of FXPRLamide family peptides in A. yamamai, but its function needs to be further investigated in the future.     

Distinct expression of GnRH genes in the red seabream brain

This paper reports the molecular cloning of a cDNA encoding the precursor of seabream gonadotropin-releasing hormone (prepro-sbGnRH) and the localization of salmon GnRH (sGnRH) and seabream GnRH (sbGnRH) expressing neurons in the brain of the red seabream (Pagrus major). The cloned prepro-sbGnRH cDNA has a 285 bps open reading frame encoding a 23 amino acid signal peptide, a 10 amino acid sbGnRH, the cleavage site (Gly-Lys-Arg), and a 59 amino acid GnRH-associated peptide. The expression of sGnRH and sbGnRH peptides, and prepro-sGnRH and prepro-sbGnRH mRNA were studied using immunocytochemistry and non-radioactive in situ hybridization, respectively. We found cell bodies that reacted positively with both the sGnRH cRNA probe and anti-sGnRH serum, but not with the sbGnRH cRNA probe or anti-sbGnRH serum in the ganglion of the terminal nerve. Cell bodies that reacted positively with the sbGnRH cRNA probe, anti-sbGnRH serum, and anti-sGnRH serum, but negatively with the sGnRH cRNA probe were found in the preoptic area (POA). Immunocytochemistry showed that a distinct bundle of axons arises in the POA which projected to the pituitary gland. These results suggest that sbGnRH is the most relevant hypophysiotropic form of GnRH.     

Subcutaneous embryonal rhabdomyosarcoma in a dog: cytologic, immunocytochemical, histologic, and ultrastructural features

Abstract: A subcutaneous mass on the left antebrachium of an 11‐year‐old intact female English Pointer dog was evaluated presurgically by cytologic examination and immunocytochemical staining. The sample consisted of discrete, variably sized, markedly pleomorphic neoplastic cells that expressed vimentin with diffuse cytoplasmic staining, desmin with focal paranuclear staining, and myoglobin with diffuse cytoplasmic staining, consistent with a diagnosis of rhabdomyosarcoma. Lymphocytic and histiocytic markers were negative. Aspirates of the enlarged ipsilateral prescapular lymph node were positive for metastatic disease. Surgical excision of the tumor and lymph node were followed by histologic and electron microscopic examination. Histomorphologic appearance of neoplastic cells from the mass and the lymph node paralleled cytologic findings; the histologic diagnosis was round cell variant of embryonal rhabdomyosarcoma. By ultrastructural evaluation, cells contained numerous mitochondria and masses of cytoplasmic tangled myofilaments, features typical of rhabdomyoblasts. The dog received doxorubicin (30 mg/m²) every 3 weeks for 5 treatments. Local recurrence developed 6 months after resection but was not treated. Despite a guarded prognosis and untreated local recurrence, the dog was still alive 18 months after surgery. Cytologic evaluation and immunocytochemical staining were pivotal for the presurgical diagnosis of rhabdomyosarcoma.     

6 种胃肠激素样内分泌细胞在细鳞鲑消化道的定位

采用免疫细胞化学ABC方法,选择5-羟色胺(5-hydroxytryptamine,5-HT)、生长抑素(somatostatin,SS)、胰多肽(pancreatic polypeptide,PP)、胃泌素(gastrin,GAS)和P物质(substance P,SP)6种特异性哺乳类胃肠激素抗血清,对不同年龄段(1龄、2龄和3龄)细鳞鲑(Brachymystax lenok)的消化道内分泌细胞进行了免疫细胞化学定位研究.结果表明,仅在食管、胃贲门、胃体和胃幽门检出有5-HT、SS和PP阳性细胞的分布(除1龄鱼食道),且这3种内分泌细胞均大量定位于胃部;在前肠、中肠、后肠和直肠中均未检测到这3种内分泌细胞的阳性反应.在3个年龄段的细鳞鲑胃肠各部位均未检测到GAS、GLU和SP阳性细胞.细鳞鲑的5-HT、SS和PP与其他有胃鱼类的内分泌细胞一样,可分为2种类型,即开放型和闭合型,这类细胞主要通过腔分泌和旁分泌两种方式释放激素.5-HT、SS和PP这3种细胞在1龄幼鱼消化道内就已经发育成熟,其对胃肠道活动的调节作用已经与成鱼没有差别,细胞的分布密度随着细鳞鲑的年龄增长不断增加.本研究揭示了不同生长阶段细鳞鲑消化道中这6种胃肠激素内分泌细胞的发育特征,并阐明了这些胃肠激素细胞在细鳞鲑幼鱼消化道的分布、形态以及生理作用.     

Organotypic slice culture of the hypothalamic paraventricular nucleus of rat

Organotypic slice cultures have been developed as an alternative to acute brain slices because the neuronal viability and synaptic connectivity in these cultures can be preserved well for a prolonged period of time. This study evaluated a stationary organotypic slice culture developed for the hypothalamic paraventricular nucleus (PVN) of rat. The results showed that the slice cultures maintain the typical shape of the nucleus, the immunocytochemical signals for oxytocin, vasopressin, and corticotropin-releasing hormone, and the electrophysiological properties of PVN neurons for up to 3 weeks in vitro. The PVN neurons in the culture expressed the green fluorescent protein gene that had been delivered by the adenoviral vectors. The results indicate that the cultured slices preserve the properties of the PVN neurons, and can be used in longterm studies on these neurons in vitro.