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Immunohistochemical study of osteopontin in boar testis 总被引：3，自引：0，他引：3
The expression of osteopontin (OPN) in boar testis was studied. Western blot analysis detected 66- and 32-kDa OPN immunopositive bands in the testes of adult boars. In postnatal piglets, the 66-kDa OPN band was detected in the testes, but not the 32-kDa band. In the newborn testis, OPN immunostaining was seen in gonocytes and in some supporting cells in the seminiferous tubules, as well as in interstitial Leydig cells. In the adult boar testis, OPN immunoreactivity was detected in seminiferous tubules with varying intensities. Intense OPN immunostaining was seen in the residual bodies and acrosomes in the spermatids while, occasionally, OPN immunostaining was seen in spermatogonia and various stage of spermatocytes but in few Sertoli cells in the seminiferous tubules. In addition, Leydig cells in adult boars were weakly immunostained with OPN. These findings suggest that OPN is detected in the majority of germ cells and is involved in spermatogenesis in boar testis. 相似文献
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Futoshi YAZAMA Haruki SATO Tomoko SONODA 《The Journal of reproduction and development》2015,61(5):399-406
An experimental ischemia (EI)-induced mouse model was used to
analyze pathological and biochemical alterations in testes. Initial
morphological changes were observed in Sertoli cells of EI testes at
the light microscopic level. Examination of the ultrastructure using
transmission electron microscopy confirmed that Sertoli cells were
partially detached from the basement membrane of the seminiferous
epithelium and that the cell membranes of adjacent Sertoli cells were
not joined. The functional integrity of the blood-testis barrier (BTB)
was assessed using the lanthanum tracer technique. Lanthanum had
penetrated into the spaces between adjacent Sertoli cells in the
adluminal compartment up to the lumen of the seminiferous epithelium
in EI testes. Proteome analysis showed that the expression of heat
shock protein (HSP) 70 was significantly upregulated in EI testes.
Western blot analysis confirmed that the expression of HSP70 increased
in a time-dependent manner after the EI procedure. HSP70
immunostaining was observed in spermatocytes and in round and
elongated spermatids in EI testes. Our results suggest that a change
in the junctions between adjacent Sertoli cells on the basal
compartment is involved in the BTB disruption in EI testes. Therefore,
male infertility caused by the BTB disruption could be associated with
heat stress induced by ischemia. 相似文献
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Detection of infectious laryngotracheitis virus infected cells with cloned DNA probes. 总被引：1，自引：1，他引：0 
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下载免费PDF全文 E Nagy 《Canadian journal of veterinary research》1992,56(1):34-40
A genomic library of infectious laryngotracheitis virus (ILTV) DNA BamH1 fragments was prepared and two cloned fragments were evaluated for their potential as probes for the detection of ILTV infected cells. The virus was purified by a modified sucrose density gradient procedure for the isolation of pure ILTV DNA. A genomic library was constructed using BamH1-digested ILTV DNA and pGEM7 as a vector. A 1.1 kb cloned BamH1 fragment of ILTV DNA was tested in a slot or dot blot assay for the detection of ILTV infected cells. The limit of detection for this probe was at least 0.12 ng of pure ILTV DNA. The probe was able to identify both chicken embryo liver (CELi) cells and choriallantoic membranes infected with ILTV. Chicken embryo liver cells infected with several field isolates and a vaccine strain of ILTV were positive by dot blot analysis using this probe. Some qualitative differences in the degree of hybridization to cells infected by different ILTV isolates were observed. Uninfected cells and cells infected with fowlpox virus, turkey herpesvirus, Marek's disease virus or Newcastle disease virus were negative by the same assay. Compared with the 1.1 kb fragment, a larger 6 kb cloned BamH1 fragment of ILTV DNA showed a stronger hybridization signal to DNA from ILTV infected cells. 相似文献
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To confirm ability forming the basement membrane of the regenerated laminar epidermis
(rLE) in chronic laminitis, expression of type VII and type XVII collagen mRNAs in the rLE
was studied applying sequences of two type of murine collagens. On northern blot analysis,
complement DNA (cDNA) probes adjusted from the murine type VII and type XVII collagen
could hybridize with the equine mRNAs, and each signal was detected as single-bands at
approximately 9.5 kb and 5.6 kb, respectively. Contrasting with the expression level of
equine glyceraldehyde-3-phosphate dehydrogenease mRNA, the band of type VII collagen mRNA
in laminitis was stronger than normal, but the type XVII collagen mRNA in laminitis was
less than normal. By in situ hybridization, positive signals in response to the murine
type VII and type XVII collagen mRNA probes could be detected in the equine laminitic rLE
region. From these results, it is concluded that the keratinocytes constructing the rLE in
chronic stage of laminitis can express type VII and type XVII collagen mRNAs and these
expression patterns were different from the normal. 相似文献
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Hicks BA Etter SJ Carnahan KG Joyce MM Assiri AA Carling SJ Kodali K Johnson GA Hansen TR Mirando MA Woods GL Vanderwall DK Ott TL 《Journal of animal science》2003,81(6):1552-1561
Pregnancy and interferon-tau (IFN tau) upregulate uterine Mx gene expression in ewes; however, the only known role for Mx is in the immune response to viral infection. We hypothesize that Mx functions as a conceptus-induced component of the anti-luteolytic mechanism and/or regulator of endometrial secretion or uterine remodeling during early pregnancy. This study was conducted to determine the effects of early pregnancy on uterine Mx expression in domestic farm species with varied mechanisms of pregnancy recognition. Endometrium from cows, gilts, and mares was collected during the first 20 d of the estrous cycle or pregnancy, and total messenger RNA (mRNA) and protein were analyzed for steady-state levels of Mx mRNA and protein. Northern blot analysis of Mx mRNA detected an approximately 2.5 Kb of mRNA in endometrium from each species. In pregnant cows, steady-state levels of Mx mRNA increased 10-fold (P < 0.05) above levels observed in cyclic cows by d 15 to 18. In cyclic gilts, slot blot analysis indicated that endometrial Mx mRNA levels did not change between d 5 and 18 of the cycle. However, in pregnant gilts, Mx levels tended (P = 0.06) to be elevated two-fold on d 16 only, and in situ hybridization indicated that this increase occurred in the stroma. In mares, Mx mRNA was low, but detectable, and did not change between ovulation (d 0) and d 20, regardless of reproductive status. Western blot analysis revealed multiple immunoreactive Mx protein bands in each species. One band was specific to pregnancy in cows. As in ewes, in situ hybridization analysis indicated that Mx mRNA was strongly expressed in the luminal epithelium, stroma, and myometrium by d 18 in cows. However, on d 14 in gilts, Mx was expressed primarily in the stroma, and on d 14 in mares, low levels of Mx expression were confined largely to the luminal epithelium. The uteruses of cows, gilts, and mares express Mx, and expression is upregulated during pregnancy in cows and gilts--animals whose conceptuses secrete interferons during early pregnancy, but that possess different mechanisms for pregnancy recognition. 相似文献
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Antimicrobial resistance patterns and plasmid profiles of Streptococcus suis isolates. 总被引：5，自引：0，他引：5
M Cantin J Harel R Higgins M Gottschalk 《Journal of veterinary diagnostic investigation》1992,4(2):170-174
Streptococcus suis isolates recovered from diseased animals in Quebec and western Canada and from human cases in Europe were tested for their susceptibility to different antimicrobial agents and screened for their plasmid content. Most isolates from Quebec were clindamycin, erythromycin, and tetracycline resistant; animal isolates from western Canada were notably less resistant to clindamycin and erythromycin, whereas human isolates were considerably more susceptible to most antimicrobials tested. More than 60% of isolates had plasmids that ranged from 1.5 to 35 kilobases (kb). Of the 7 plasmid profiles found, 2 were particularly frequent in isolates from Quebec and western Canada, suggesting the presence of epidemic strains in the swine population. A particular plasmid band of about 5 kb was present in most Canadian isolates. When this band was used as a probe in colony and Southern blot hybridization, most isolates harboring the 5-kb plasmid hybridized, even though their plasmid profiles were different. Human isolates from Europe differed in their plasmid content from Canadian isolates of animal origin. Although a high degree of antimicrobial resistance was associated with the presence of plasmids in most isolates, it was not possible to establish a causative relationship. 相似文献
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N Tsunekawa T Nishida H Fujimoto 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》1999,61(4):381-388
The anatomical location of testes in mammals ranges from a location close to that observed in the embryo to a lower position usually involving a pendant scrotum. In scrotal mammals, the abdominal position of the cryptorchid testis, which elevates its temperature, is detrimental to spermatogenesis and causes infertility. Spermatocytes are sensitive but late spermatids are relatively resistant to thermal stress suggesting that the latter might be protected in some way. In general, most organisms express Hsp70 proteins, which play a crucial role in the protection of cells against thermal stress. We have found previously that the Hsc70t protein, a member of the Hsp70 family of proteins, is constitutively expressed in the late spermatids of mice. Here, we have utilized immunohistochemistry with anti-mouse Hsc70t antiserum to examine the expression of the spermatid-specific Hsp70 antigen in the testes of several mammalian species with different degrees of testes migration. Our data indicate that the antigen is conserved in the mammals including marsupials. We also examined whether antigens of Hsp70-related proteins were expressed in non-mammalian vertebrates including not only homoiothermal but also poikilothermal animals. The spermatid-specific Hsp70 antigens were not detectable in the testes of the animals examined. From results of immunohistochemistry with BRM22 monoclonal antibody which reacts broadly with Hsp70 family proteins, however, we revealed constitutive expression of antigens of Hsp70-related proteins in spermatogenic cells of the vertebrates. These results suggest that the expression of spermatid-specific Hsp70 protein may be involved in the developmental pathway during spermiogenesis in mammals rather than in thermotolerance. 相似文献
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N Miyoshi S Tateyama K Ogawa D Nosaka T Ohashi B Sunyasootcharee 《American journal of veterinary research》1991,52(6):940-944
To provide information about oncogenes for molecular biological studies of tumors in domestic animals, the proto-oncogenes homologous to the c-myc, c-erbB-2, c-ros-1, c-yes-1, v-myc, v-Ki-ras, and v-Ha-ras oncogenes of genomic DNA in cattle, horses, pigs, dogs, cats, and chickens were investigated by Southern blot hybridization. High molecular weight genomic DNA in each of the animals contained proto-oncogenes that had a certain homology with the oncogenes used, but the extent of nucleotide homology of the proto-oncogenes differed in number and molecular weight: ie, 1 or 2 bands at 1.6 to 22.0 kilobase (kb) in the c-myc probe, 1 or 2 bands at 1.1 to 16.0 kb in the c-ros-1 probe, 1 to 3 bands at 0.7 to 23.0 kb in the c-erbB-2 probe, 1 to 4 bands at 0.6 to 18.0 kb in the c-yes-1 probe, 1 to 3 bands at 1.6 to 30.0 kb in the v-myc probe, 1 to 7 bands at 1.0 to 36.0 kb in the v-Ki-ras probe, and 1 to 4 bands at 1.0 to 27.0 kb in the v-Ha-ras probe. Furthermore, signal strength of each band, as determined by autoradiography, was not always the same for each probe in the various animals. Our findings indicate that these proto-oncogenes are well conserved with species specificities in each animal. 相似文献
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Ochiai H Morishita T Onda K Sugiyama H Maruo T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2012,74(7):917-922
A full-length cDNA sequence of canine L-type amino acid transporter 1 (Lat1) was determined from a canine brain. The sequence was 1828 bp long and was predicted to encode 485 amino acid polypeptides. The deduced amino acid sequence of canine Lat1 showed 93.2% and 91.1% similarities to those of humans and rats, respectively. Northern blot analysis detected Lat1 expression in the cerebellum at 4 kb, and Western blot analysis showed a single band at 40 kDa. RT-PCR analysis revealed a distinct expression of Lat1 in the pancreas and testis in addition to the cerebrum and cerebellum. Notably, Lat1 expression was observed in the tissues of thyroid cancer, melanoma and hemangiopericytoma. Although the cancer samples examined were not enough, Lat1 may serve as a useful biomarker of cancer cells in veterinary clinic. 相似文献
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T. Komatsu Y. Yamamoto Y. Atoji T. Tsubota Y. Suzuki 《Anatomia, histologia, embryologia》1998,27(3):209-213
The seasonal changes of the cytoskeletal protein expressions were immunohistochemically investigated in the testes of Japanese black bear, Ursus thibetanus japonicus. A strong immunoreaction for α-smooth muscle actin is restricted to the vascular smooth muscle cells and the peritubular cells which surround the seminiferous tubules by several layers throughout the year. Weak immunoreactions for B4 antigen and desmin were observed in the vascular smooth muscle cells and in a part of peritubular cells throughout the year. A strong immunoreaction for vimentin was also detected in the fibroblasts and Leydig cells, in addition to the vascular smooth muscle and epithelial cells and the peritubular cells throughout the year. A strong α-tubulin immunoreaction was detected in the elongating spermatids during the acrosome phase of spermiogenesis in May and June. The cytoplasm of several Sertoli cells was faintly immunoreacted for vimentin in the basal and lateral region, while an intense α-tubulin reaction was seen in the entire cytoplasm in May, April and June. In November, January and March, the immunoreactions for vimentin and α-tubulin strongly accumulate in a perinuclear region of Sertoli cells when developmental spermatids are not seen in the seminiferous tubules. These accumulations in the immunoreactions for vimentin and α-tubulin seem to be caused by the reduction in size of Sertoli cells cytoplasm with season. However, the seasonal changes of distributions in the cytoskeletal proteins are obscure in the bear testes. These results suggest that the contents of cytoskeletal proteins may not change in relation to the morphological differences with season in the testes of the seasonal breeders. 相似文献
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Rin YANAI Yudai YAMASHITA Kohei UMEZU Yuuki HIRADATE Kenshiro HARA Kentaro TANEMURA 《The Journal of reproduction and development》2021,67(1):59
The structure of microtubules is essential for the fertilizing ability of spermatozoa. Acetylation of α-tubulin plays an important role in flagellar elongation and spermatozoa motility. Previous reports have suggested that alpha-tubulin N-acetyltransferase 1 (ATAT1) is the main acetyltransferase involved in the acetylation of α-tubulin. Although ATAT1 is reported to express in the testis, no information is available regarding its expression in elongated spermatids, epididymis, and mature spermatozoa. Hence, it remains unclear whether ATAT1 is involved in spermatozoa maturation and capacitation. Therefore, we evaluated the expression of ATAT1 in the mouse male reproductive system using immunostaining and western blotting. Our results showed that ATAT1 was expressed in spermatids during spermiogenesis in mouse testes, but its expression varied according to the seminiferous tubule stage. We observed ATAT1 in the cytoplasm of round spermatids, the flagella of elongated spermatids, and in the cytoplasm of step 16 spermatids, just before its release into the lumen. In addition, ATAT1 was expressed in epithelial cells of the epididymis. In spermatozoa of the cauda epididymis, ATAT1 expression was primarily observed in the midpiece of the spermatozoa. The localization of ATAT1 protein in the male germline was observed during spermiogenesis as well as during spermatozoa maturation. Our results suggest that ATAT1 may be involved in the formation of flagella and in the acetylation process, which has attracted attention in recent years regarding male infertility. 相似文献
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A synthetic deoxyribonucleotide probe for virulent Listeria monocytogenes, designated ADO7, was evaluated for its ability to identify restriction fragments of L. monocytogenes with nucleic acid sequences homologous with the beta-hemolysin gene by Southern blot hybridization of clinical and food isolates. The synthetic probe hybridized with three restriction fragments (approximately 1.1, 0.86, and 0.76 kb) of the serotype 1/2A isolates. Southern blot hybridization of the serogroup 4B isolates indicated that the nucleic acid sequences homologous with the beta-hemolysin gene probe were limited to a single restriction fragment of approximately 1 kb. 相似文献
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K. Han H. W. Seo Y. Oh I. Kang C. Park J. H. Han S.-H. Kim C. Chae 《Veterinary research communications》2013,37(2):155-162
The objective of this study was to determine the pathogenesis of experimental infection with a type 1 porcine reproductive and respiratory syndrome virus (PRRSV) by defining the sites of viral replication and apoptosis in male gonads from infected boars for a period of 21 days after intranasal inoculation. Microscopically, hypospermatogenesis and abundant germ cell depletion and death were observed in the testes. Such germ cell death occurs by apoptosis, as determined by a characteristic histological patterns and evidence of massive DNA fragment detected in situ terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) reaction. PRRSV was detected in the testicular tissue of infected boars only. Viral nucleic acid was localized in spermatogonia, spermatocytes and spermatids but not in the vesicular and bulbourethral gland. In serial sections, PRRSV-positive cells did not co-localized with apoptotic cells. TUNEL-positive apoptotic cells were more numerous than PRRSV-positive cells in testicular sections. The present study demonstrated that type 1 PRRSV infects the spermatogonia and their progeny, and induces apoptosis in these germ cells. 相似文献
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Masamichi Kurohmaru Toshiyasu Matsui Hitomi Igarashi Shosaku Hattori Yoshihiro Hayashi 《Anatomia, histologia, embryologia》2021,50(2):417-421
The postnatal testicular development and actin distribution in the seminiferous epithelium were examined by light microscopy, using the testes of the Habu (Trimeresurus flavoviridis; snake) from 0-year-old to 3-year-old. At 0-year-old (about 1 month after birth), the testis was quite small in size, and the seminiferous epithelium was composed of only Sertoli cells and large spermatogonia. Actin immunoreactivity was observed in the peritubular myoid cells, but could not be detected in the seminiferous epithelium. At 1-year-old (about 10 months after birth), the testicular size increased to a great degree. In the seminiferous epithelium, spermatocytes newly appeared. Actin could still not be detected in the seminiferous epithelium. At 2-year-old (about 1 year and 10 months after birth), the testes continued to develop in size. In the seminiferous epithelium, elongate spermatids and round spermatids were frequently seen, in addition to Sertoli cells, spermatogonia and spermatocytes. Thus, active spermatogenesis was clearly recognized at this age. Moreover, the actin distribution in the seminiferous epithelium was observed at the site between Sertoli cells and spermatids, as well as that at adult stage. The immunoreactivity of actin in the peritubular myoid cells gradually increased from 0-year-old to 2-year-old. Conclusively, it seems likely that spermatogenesis in the Habu initiates at 2-year-old, accompanying with the appearance of actin in the seminiferous epithelium. 相似文献