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Immunohistochemical Study of Serum Proteins in Normal and Cadmium-Treated Mouse Testis by in vivo Cryotechnique

Journal of Reproduction and Contraception 2005年第3期16卷 137-146页

作者： nobuo terada Nobuhiko OHNO Yasuhisa FUJII Shinichi OHNO Department of Anatomy Interdisciplinary Graduate School of Medicine and Engineering University of Yamanashi Yamanashi 409-3898 Japan

Objective To investigate time-dependent changes of serum proteins permeability in the normal and cadmium（Cd）-treated mouse testis, reflecting tight junctional （TJ） barriers of Sertoli cells Methods The serum proteins, albumin and immunoglobulin-G（IgG）, in the seminiferous tubules were firstly immobilized by the “in vivo cryotechnique”, in which the dynamic blood circulation was always kept. The cryofixed testicular tissues were then processed for the freeze-substitution method, and embedded in the paraffin wax. Serial sections of 5μm thickness were immunostained by anti-mouse albumin or IgG antibody with peroxidase immunostaining, and also stained with hematoxylin-eosine （HE）for morphological observation. Results In normal seminiferous tubules, albumin immunoreaction products were localized around peritubular myoid cells and among Leydig cells, as well as in blood vessels. They were also localized as arch-like patterns around some spermatogonia in basal compartments. The number of the immunopositive arch structures was different according to developmental stages of the seminiferous cycle, judging from the arrangement of germ cells by HE-staining. The patterns of localization of IgG immunostaining in normal mouse testis were similar to that of albumin. In 24 h after Cd-treatment, some enlarged spaces and vesicular formation in the seminiferous epithelium were observed on the paraffin sections by HE-staining. The albumin or IgGimmunolocalization was seen not only in the basal compartments, but also in the adluminal compartments between *** and germ cells. Conclusion The structural changes of inter-Sertoli TJ barriers in vivo, such as different immunostaining patterns of serum proteins between the normal and Cd-treated mouse seminiferous tubules, could be clearly detected by the “in vivo cryotechnique” with albumin or IgG immunohistochemistry.

关键词： immunohistochemistry in vivo cryotechnique serum proteins testis cadmium

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体内冷冻的正常及镉处理小鼠睾丸内血清蛋白质的免疫组织化学研究

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生殖与避孕 2006年第3期26卷 135-141页

作者：廖晓岗 nobuo terada 李子龙 Nobuhiko OHNO Yasuhisa FUJII Shinichi OHNO 重庆医科大学电子显微镜室重庆400016 山梨大学医学工学综合研究部解剖学系中国医科大学第一附属医院沈阳110001

目的:了解正常和镉(Cd)处理小鼠睾丸内血清蛋白质渗透性的时相性变化,揭示支持细胞间紧密连接的屏障功能。方法:保持正常动力学的血循环下,运用活体内冷冻技术对曲细精管内白蛋白和IgG进行快速冷冻固定。冷冻固定的睾丸行冷冻置换后作... 详细信息

目的:了解正常和镉(Cd)处理小鼠睾丸内血清蛋白质渗透性的时相性变化,揭示支持细胞间紧密连接的屏障功能。方法:保持正常动力学的血循环下,运用活体内冷冻技术对曲细精管内白蛋白和IgG进行快速冷冻固定。冷冻固定的睾丸行冷冻置换后作石蜡包埋。5μm厚连续切片分别用于抗鼠血清白蛋白和IgG的免疫组化染色和H.E.染色。结果:正常曲细精管,白蛋白免疫反应产物主要定位于管周肌样细胞周围,以及间质细胞之间和毛细血管内,部分免疫反应产物呈拱型出现在曲细精管上皮基底室内,沿精原细胞表面分布。在依据生精细胞排列而区分的曲细精管上皮周期的不同发育时相,拱型免疫反应产物的数量不同。正常小鼠睾丸内IgG免疫反应的定位型式与白蛋白的免疫定位相似。Cd处理后24h,部分曲细精管上皮内空泡形成和管腔增大,白蛋白和IgG免疫反应产物不仅出现在基底室,而且也见于近腔室的支持细胞和生精细胞之间。结论:运用活体内快速冷冻技术结合血清白蛋白和IgG的免疫组化方法,能清楚地揭示正常和Cd处理小鼠曲细精管内血清蛋白质的不同免疫染色型式,反映体内支持细胞间紧密连接结构的变化。

关键词：免疫组化活体内冷冻技术血清蛋白质睾丸镉(Cd)

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活体冷冻固定及冷冻置换的小鼠睾丸组织内血清白蛋白和IgG的免疫组织化学定位

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中国组织化学与细胞化学杂志 2006年第3期15卷 251-255页

作者：廖晓岗 nobuo terada 李子龙 Nobuhiko Ohno Yasuhisa Fujii Shinichi Ohno 重庆医科大学电子显微镜室重庆400016 山梨大学医学工学综合研究部解剖学讲座中国医科大学第一附属医院沈阳110001

目的了解小鼠睾丸组织内血清白蛋白和IgG渗透性的时相性变化,揭示支持细胞间紧密连接的屏障功能。方法运用活体内冷冻固定技术对小鼠睾丸进行快速冷冻固定和冷冻置换,并作石蜡包埋。5μm厚连续切片用于抗鼠血清白蛋白和IgG的免疫组化染... 详细信息

目的了解小鼠睾丸组织内血清白蛋白和IgG渗透性的时相性变化,揭示支持细胞间紧密连接的屏障功能。方法运用活体内冷冻固定技术对小鼠睾丸进行快速冷冻固定和冷冻置换,并作石蜡包埋。5μm厚连续切片用于抗鼠血清白蛋白和IgG的免疫组化染色和H.E染色。结果H.E染色切片显示距冷冻组织表面300-400μm深度范围内的精曲小管结构保存完好,无明显冰晶形成所致的人工损伤。血清白蛋白和IgG免疫反应产物主要定位于精曲小管管周肌样细胞周围,以及间质细胞之间和毛细血管内,部分免疫反应产物呈拱型出现在精曲小管上皮内,并严格限定在基底室,沿精原细胞表面分布。拱型免疫反应产物的数量与精曲小管上皮周期的发育时相密切相关。结论活体内快速冷冻固定和冷冻置换结合血清白蛋白和IgG的免疫组化方法,可很好地保存精曲小管组织结构并清楚地揭示支持细胞间紧密连接结构的时相变化。

关键词：免疫组化活体内冷冻技术冷冻置换白蛋白 IgG 睾丸

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Intramanchette transport during primate spermiogenesis： expression of dynein, myosin Va, motor recruiter myosin Va, VIIa-Rab27a/b interacting protein, and Rab27b in the manchette during human and monkey spermiogenesis

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Asian Journal of Andrology 2008年第4期10卷 561-568页

作者： Shinichi Hayasaka Yukihiro terada Kichiya Suzuki Haruo Murakawa Ikuo Tachibana Tadashi Sankai Takashi Murakami nobuo Yaegashi Kunihiro Okamura Department of Obstetrics and Gynecology Tohoku University School of Medicine Sendai Miyagi 980-8574 Japan Suzuki Memorial Hospital lwanuma Miyagi 989-2481 Japan Tsukuba Primate Research Center National Institute of Biomedical Innovation Tukuba Ibaraki 305-0843 Japan

Aim： To show whether molecular motor dynein on a microtubule track, molecular motor myosin Va, motor recruiter myosin Va, VIIa-Rab27a/b interacting protein （MyRIP）, and vesicle receptor Rab27b on an F-actin track were present during human and monkey spermiogenesis involving intramanchette transport （IMT）. Methods： Spermiogenic cells were obtained from three men with obstructive azoospermia and normal adult cynomolgus monkey （Macacafascieularis）. Immunocytochemical detection and reverse transcription-polymerase chain reaction （RT-PCR） analysis of the proteins were carried out. Samples were analyzed by light microscope. Results： Using RT-PCR, we found that dynein, myosin Va, MyRIP and Rab27b were expressed in monkey testis. These proteins were localized to the manchette, as shown by immunofluorescence, particularly during human and monkey spermiogenesis. Conclusion： We speculate that during primate spermiogenesis, those proteins that compose microtubule-based and actin-based vesicle transport systems are actually present in the manchette and might possibly be involved in intramanchette transport.

关键词： intramanchette transport manchette spermiogenesis

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Treatment Outcomes in Head and Neck Cancer Patients 80 Years Old and over

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International Journal of Otolaryngology and Head & Neck Surgery 2015年第6期4卷 401-408页

作者： Tomonori terada Nobuhiro Uwa Kosuke Sagawa Takeshi Mohri nobuo Saeki Kota Kida Kenzo Tsuzuki Masafumi Sakagami Department of Otolaryngology—Head and Neck Surgery Hyogo College of Medicine Nishinomiya City Japan

Background: With the recent aging of society, the need for medical treatment of elderly patients with head and neck cancer seems to have been increasing. Method: The present study analyzed all 103 patients with head and neck cancer ≥80 years, and we compared results with those of the previous generation (Group P;range: 75 - 79 years) comprising 104 patients treated in the same period. Results: We provided treatment just as wanted and could not choose it often. The reasons were oncological factors such as unresectable tumor or distant metastasis, refusal of treatment, and physical factors such as poor PS or number of comorbidities. Conclusion: Treatment choices should be based on the wishes and motivations of the patient and the medical assessment of physical function. When a patient ≥80 years old is treated, the high incidence of complications and severity of the disease should be considered.

关键词： 80 Years Old and over Elderly Patients Head and Neck Cancer Treatment

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