The Experts below are selected from a list of 312 Experts worldwide ranked by ideXlab platform
Li Lai - One of the best experts on this subject based on the ideXlab platform.
-
Pharmacokinetic Behaviors of Intravenously Administered siRNA in Glandular Tissues
Theranostics, 2016Co-Authors: Yuanyu Huang, Qiang Cheng, Shuquan Zheng, Lingwei Meng, Deyao Zhao, Xiaoxia Wang, Li LaiAbstract:The pharmacokinetics of small interfering RNAs (siRNAs) is a pivotal issue for siRNA-based drug development. In this study, we comprehensively investigated the behavior of siRNAs in vivo in various tissues and demonstrated that intravenously-injected naked siRNA accumulated remarkably in the submandibular Gland, Bulbourethral Gland, and pancreas, with a respective half-life of ~22.7, ~45.6, and ~30.3 h. This was further confirmed by gel separation of tissue homogenates and/or supernatants. In vivo imaging and cryosectioning suggested that delivery carriers significantly influence the distribution and elimination profiles of siRNA. Gene-silencing assays revealed that neither naked nor liposome-formulated siRNA resulted in gene knockdown in the submandibular and Bulbourethral Glands after systemic administration, suggesting that these Glands function as drug reservoirs that enable slow siRNA release into the circulation. But robust gene-silencing was achieved by local injection of liposome-encapsulated siRNA into the submandibular Gland. Our results enhance understanding of the pharmacokinetic properties of siRNAs and we believe that they will facilitate the development of siRNA therapy, especially for the submandibular Gland.
-
Pharmacokinetic Behaviors of Intravenously Administered siRNA in Glandular Tissues
THERANOSTICS, 2016Co-Authors: Huang Yuanyu, Cheng Qiang, Ji Jia-li, Zheng Shuquan, Du Lili, Meng Lingwei, Wu Yidi, Zhao Deyao, Wang Xiaoxia, Li LaiAbstract:The pharmacokinetics of small interfering RNAs (siRNAs) is a pivotal issue for siRNA-based drug development. In this study, we comprehensively investigated the behavior of siRNAs in vivo in various tissues and demonstrated that intravenously-injected naked siRNA accumulated remarkably in the submandibular Gland, Bulbourethral Gland, and pancreas, with a respective half-life of similar to 22.7, similar to 45.6, and similar to 30.3 h. This was further confirmed by gel separation of tissue homogenates and/or supernatants. In vivo imaging and cryosectioning suggested that delivery carriers significantly influence the distribution and elimination profiles of siRNA. Gene-silencing assays revealed that neither naked nor liposome-formulated siRNA resulted in gene knockdown in the submandibular and Bulbourethral Glands after systemic administration, suggesting that these Glands function as drug reservoirs that enable slow siRNA release into the circulation. But robust gene-silencing was achieved by local injection of liposome-encapsulated siRNA into the submandibular Gland. Our results enhance understanding of the pharmacokinetic properties of siRNAs and we believe that they will facilitate the development of siRNA therapy, especially for the submandibular Gland.National Natural Science Foundation of China [81402863]; Postdoctoral Science Foundation of China [2014M550008, 2015T80016]; National High-tech R&D Program of China [2014AA021103, 2012AA022501]; National Drug Program of China [2015ZX09102-023-002, 2014ZX09304313-001]SCI(E)PubMedARTICLEyyhuang@pku.edu.cn; liangz@pku.edu.cn101528-1541
E. M. Garcia - One of the best experts on this subject based on the ideXlab platform.
-
Localization and expression of spermadhesin PSP-I/PSP-II subunits in the reproductive organs of the boar
International journal of andrology, 2008Co-Authors: E. M. Garcia, Inmaculada Parrilla, Emilio A. Martinez, Jordi Roca, Juan M. Vazquez, Maria D. Ortega, Juan J. Calvete, Libia Sanz, Heriberto Rodriguez-martinezAbstract:The epithelial localization and expression of the spermadhesin PSP-I and PSP-II subunits were determined in the testis, ductus epididymes (caput, corpus and cauda), seminal vesicles and Bulbourethral Glands of mature boars, using immunohistochemical, western blotting and RT-PCR methods. Immunohistochemistry showed positive labelling for PSP-I and PSP-II antibodies in the epithelium of seminal vesicles in all males tested. Positive immunolabelling, but with variable intensity, was also present in the epididymal epithelium (caput, corpus and cauda), although varying largely among segments and boars. Immunoreactivity was nearly or completely absent in the seminiferous epithelium and the Bulbourethral Gland, although SDS-PAGE and western blotting revealed the presence of PSP-I and PSP-II immunoreactive bands in all the tissue extracts, including the testis and the Bulbourethral Gland. mRNA amplification by RT-PCR using primers specific for PSP-I and PSP-II showed a trend similar to that observed for western blotting, i.e. intensity variation between tissues (even between segments of the same epididymis) and among boars. Our results indicate that the seminal vesicles are the main source of PSP-I and PSP-II spermadhesins, although epididymal segments, testis and the Bulbourethral Gland also participate in the expression of both proteins.
-
323 EXPRESSION OF PSP-I AND PSP-II IN THE REPRODUCTIVE TRACT OF THE BOAR BY IMMUNOHISTOCHEMISTRY, WESTERN BLOTTING, AND RT-PCR
Reproduction Fertility and Development, 2007Co-Authors: E. M. Garcia, Inmaculada Parrilla, Emilio A. Martinez, Jordi Roca, Juan M. Vazquez, Juan J. Calvete, Libia Sanz, H. Rordríguez-martínezAbstract:In boar seminal plasma, PSP-I and PSP-II form a noncovalent heterodimer, which is known to have a beneficial effect on highly extended boar spermatozoa. These proteins are mainly produced by the seminal vesicles of the male reproductive tract and are mixed with the spermatozoa during ejaculation. This study assessed the epithelial localization and expression of spermadhesin PSP-I and PSP-II subunits using immunohistochemical, western blotting, and RT-PCR methods in porcine testis, epididymis (caput, corpus, and caudal), seminal vesicles, and Bulbourethral Glands. Tissues were collected from 10 mature boars (Swedish Yorkshire) of proven fertility, frozen or fixed, and embedded in paraffin. Sections of 5 µm were mounted on poly-l-lysine-coated glass slides for immunohistochemistry, and 50 mg from the frozen counterparts were homogenized to isolate proteins (Western blotting study) or RNA (RT-PCR study). Polyclonal antibodies (antiPSP-I and antiPSP-II) against proteins (PSP-I and PSP-II) were used. The immunohistochemistry showed positive labelling for both antibodies in the epithelium of seminal vesicles in all males. Positive immunolabelling, but of variable intensity, was present in the epididymal epithelium (caput, corpus, and caudal), with variation among segments and boars. No labelling was found in the seminiferous epithelium or Bulbourethral Glands of any boar. After SDS-PAGE and Western blotting, immunoreactive bands were obtained in the extracts of all tissues for both PSP proteins. Among tissues, the highest intensity corresponded to the seminal vesicle lane for both proteins. Epididymis (caput, corpus, and caudal), Bulbourethral Gland, and testis showed more feeble bands, yet present. The intensity of the bands varied among boars. Amplification products from mRNA were obtained in all tissues explored by RT-PCR, using specific primers for PSP-I and PSP-II. The same trend was obtained regarding intensity bands, corresponding with the intensity obtained by Western blotting. The results indicate that PSP-I and PSP-II are mainly expressed in seminal vesicles and epididymal segments and, in lower amount, in the testis and Bulbourethral Gland. This work was supported by Formas, Stockholm, and Fundacion Seneca, Murcia, Spain.
Emilio A. Martinez - One of the best experts on this subject based on the ideXlab platform.
-
Western blot detection of GPX-5 in boar genital tract.
2016Co-Authors: Isabel Barranco, Asta Tvarijonaviciute, Cristina Perez-patiño, Alejandro Vicente-carrillo, Inmaculada Parrilla, Jose J. Ceron, Emilio A. Martinez, Heriberto Rodriguez-martinez, Jordi RocaAbstract:L1: protein ladder; L2: blood vessel (positive control); L3: testis. L4: epididymis. L5: cauda epididymis. L6: seminal vesicle. L7: prostate. L8: Bulbourethral Gland.
-
Immunolocalization of glutathione peroxidase-5 (GPX5) in boar genital organs.
2016Co-Authors: Isabel Barranco, Asta Tvarijonaviciute, Cristina Perez-patiño, Alejandro Vicente-carrillo, Inmaculada Parrilla, Jose J. Ceron, Emilio A. Martinez, Heriberto Rodriguez-martinez, Jordi RocaAbstract:Fig 4a depicts a representative positive control section depicting immunostained blood vessels with a major restricted endothelial GPX-5 localization (arrows). A representative negative control prostate section (primary Ab omitted) is depicted in Fig 4b. Fig 4c shows a representative section of boar testis, showing immunostaining in the interstitium (blood vessels and Leydig cells, asterisk) and the seminiferous tubules, with a marked immunostaining in the elongated spermatids (arrows). The immunostaining was more marked in the lining epithelium of the ductus epididymides (Fig 4d, cauda segment) where both the principal epithelial cells (arrows point to the apical cell region, base of the stereocilia) and the luminal spermatozoa (spz) appeared stained, while the surrounding smooth muscle appears only faibly stained (m), lu: lumen. Fig 4e-g depict accessory sexual Glands secretory epithelia of the prostate (4e), the seminal vesicles (4f) and the Bulbourethral Gland (4g). While prostate depicted both nuclear and cytoplasmic staining (4e), the seminal vesicles depicted mainly cytoplasmic staining (arrows) and a clear staining in the luminal secretion (lu), lp: lamina propria. The Bulbourethral Gland (4g) was immunostained in the blood vessel dominated interstitial with some staining in the latero-basal epithelial membrane (arrows). Not-counterstained sections, viewed with phase-contrast optics. Bars: 50μm.
-
Localization and expression of spermadhesin PSP-I/PSP-II subunits in the reproductive organs of the boar
International journal of andrology, 2008Co-Authors: E. M. Garcia, Inmaculada Parrilla, Emilio A. Martinez, Jordi Roca, Juan M. Vazquez, Maria D. Ortega, Juan J. Calvete, Libia Sanz, Heriberto Rodriguez-martinezAbstract:The epithelial localization and expression of the spermadhesin PSP-I and PSP-II subunits were determined in the testis, ductus epididymes (caput, corpus and cauda), seminal vesicles and Bulbourethral Glands of mature boars, using immunohistochemical, western blotting and RT-PCR methods. Immunohistochemistry showed positive labelling for PSP-I and PSP-II antibodies in the epithelium of seminal vesicles in all males tested. Positive immunolabelling, but with variable intensity, was also present in the epididymal epithelium (caput, corpus and cauda), although varying largely among segments and boars. Immunoreactivity was nearly or completely absent in the seminiferous epithelium and the Bulbourethral Gland, although SDS-PAGE and western blotting revealed the presence of PSP-I and PSP-II immunoreactive bands in all the tissue extracts, including the testis and the Bulbourethral Gland. mRNA amplification by RT-PCR using primers specific for PSP-I and PSP-II showed a trend similar to that observed for western blotting, i.e. intensity variation between tissues (even between segments of the same epididymis) and among boars. Our results indicate that the seminal vesicles are the main source of PSP-I and PSP-II spermadhesins, although epididymal segments, testis and the Bulbourethral Gland also participate in the expression of both proteins.
-
323 EXPRESSION OF PSP-I AND PSP-II IN THE REPRODUCTIVE TRACT OF THE BOAR BY IMMUNOHISTOCHEMISTRY, WESTERN BLOTTING, AND RT-PCR
Reproduction Fertility and Development, 2007Co-Authors: E. M. Garcia, Inmaculada Parrilla, Emilio A. Martinez, Jordi Roca, Juan M. Vazquez, Juan J. Calvete, Libia Sanz, H. Rordríguez-martínezAbstract:In boar seminal plasma, PSP-I and PSP-II form a noncovalent heterodimer, which is known to have a beneficial effect on highly extended boar spermatozoa. These proteins are mainly produced by the seminal vesicles of the male reproductive tract and are mixed with the spermatozoa during ejaculation. This study assessed the epithelial localization and expression of spermadhesin PSP-I and PSP-II subunits using immunohistochemical, western blotting, and RT-PCR methods in porcine testis, epididymis (caput, corpus, and caudal), seminal vesicles, and Bulbourethral Glands. Tissues were collected from 10 mature boars (Swedish Yorkshire) of proven fertility, frozen or fixed, and embedded in paraffin. Sections of 5 µm were mounted on poly-l-lysine-coated glass slides for immunohistochemistry, and 50 mg from the frozen counterparts were homogenized to isolate proteins (Western blotting study) or RNA (RT-PCR study). Polyclonal antibodies (antiPSP-I and antiPSP-II) against proteins (PSP-I and PSP-II) were used. The immunohistochemistry showed positive labelling for both antibodies in the epithelium of seminal vesicles in all males. Positive immunolabelling, but of variable intensity, was present in the epididymal epithelium (caput, corpus, and caudal), with variation among segments and boars. No labelling was found in the seminiferous epithelium or Bulbourethral Glands of any boar. After SDS-PAGE and Western blotting, immunoreactive bands were obtained in the extracts of all tissues for both PSP proteins. Among tissues, the highest intensity corresponded to the seminal vesicle lane for both proteins. Epididymis (caput, corpus, and caudal), Bulbourethral Gland, and testis showed more feeble bands, yet present. The intensity of the bands varied among boars. Amplification products from mRNA were obtained in all tissues explored by RT-PCR, using specific primers for PSP-I and PSP-II. The same trend was obtained regarding intensity bands, corresponding with the intensity obtained by Western blotting. The results indicate that PSP-I and PSP-II are mainly expressed in seminal vesicles and epididymal segments and, in lower amount, in the testis and Bulbourethral Gland. This work was supported by Formas, Stockholm, and Fundacion Seneca, Murcia, Spain.
Inmaculada Parrilla - One of the best experts on this subject based on the ideXlab platform.
-
Western blot detection of GPX-5 in boar genital tract.
2016Co-Authors: Isabel Barranco, Asta Tvarijonaviciute, Cristina Perez-patiño, Alejandro Vicente-carrillo, Inmaculada Parrilla, Jose J. Ceron, Emilio A. Martinez, Heriberto Rodriguez-martinez, Jordi RocaAbstract:L1: protein ladder; L2: blood vessel (positive control); L3: testis. L4: epididymis. L5: cauda epididymis. L6: seminal vesicle. L7: prostate. L8: Bulbourethral Gland.
-
Immunolocalization of glutathione peroxidase-5 (GPX5) in boar genital organs.
2016Co-Authors: Isabel Barranco, Asta Tvarijonaviciute, Cristina Perez-patiño, Alejandro Vicente-carrillo, Inmaculada Parrilla, Jose J. Ceron, Emilio A. Martinez, Heriberto Rodriguez-martinez, Jordi RocaAbstract:Fig 4a depicts a representative positive control section depicting immunostained blood vessels with a major restricted endothelial GPX-5 localization (arrows). A representative negative control prostate section (primary Ab omitted) is depicted in Fig 4b. Fig 4c shows a representative section of boar testis, showing immunostaining in the interstitium (blood vessels and Leydig cells, asterisk) and the seminiferous tubules, with a marked immunostaining in the elongated spermatids (arrows). The immunostaining was more marked in the lining epithelium of the ductus epididymides (Fig 4d, cauda segment) where both the principal epithelial cells (arrows point to the apical cell region, base of the stereocilia) and the luminal spermatozoa (spz) appeared stained, while the surrounding smooth muscle appears only faibly stained (m), lu: lumen. Fig 4e-g depict accessory sexual Glands secretory epithelia of the prostate (4e), the seminal vesicles (4f) and the Bulbourethral Gland (4g). While prostate depicted both nuclear and cytoplasmic staining (4e), the seminal vesicles depicted mainly cytoplasmic staining (arrows) and a clear staining in the luminal secretion (lu), lp: lamina propria. The Bulbourethral Gland (4g) was immunostained in the blood vessel dominated interstitial with some staining in the latero-basal epithelial membrane (arrows). Not-counterstained sections, viewed with phase-contrast optics. Bars: 50μm.
-
Localization and expression of spermadhesin PSP-I/PSP-II subunits in the reproductive organs of the boar
International journal of andrology, 2008Co-Authors: E. M. Garcia, Inmaculada Parrilla, Emilio A. Martinez, Jordi Roca, Juan M. Vazquez, Maria D. Ortega, Juan J. Calvete, Libia Sanz, Heriberto Rodriguez-martinezAbstract:The epithelial localization and expression of the spermadhesin PSP-I and PSP-II subunits were determined in the testis, ductus epididymes (caput, corpus and cauda), seminal vesicles and Bulbourethral Glands of mature boars, using immunohistochemical, western blotting and RT-PCR methods. Immunohistochemistry showed positive labelling for PSP-I and PSP-II antibodies in the epithelium of seminal vesicles in all males tested. Positive immunolabelling, but with variable intensity, was also present in the epididymal epithelium (caput, corpus and cauda), although varying largely among segments and boars. Immunoreactivity was nearly or completely absent in the seminiferous epithelium and the Bulbourethral Gland, although SDS-PAGE and western blotting revealed the presence of PSP-I and PSP-II immunoreactive bands in all the tissue extracts, including the testis and the Bulbourethral Gland. mRNA amplification by RT-PCR using primers specific for PSP-I and PSP-II showed a trend similar to that observed for western blotting, i.e. intensity variation between tissues (even between segments of the same epididymis) and among boars. Our results indicate that the seminal vesicles are the main source of PSP-I and PSP-II spermadhesins, although epididymal segments, testis and the Bulbourethral Gland also participate in the expression of both proteins.
-
323 EXPRESSION OF PSP-I AND PSP-II IN THE REPRODUCTIVE TRACT OF THE BOAR BY IMMUNOHISTOCHEMISTRY, WESTERN BLOTTING, AND RT-PCR
Reproduction Fertility and Development, 2007Co-Authors: E. M. Garcia, Inmaculada Parrilla, Emilio A. Martinez, Jordi Roca, Juan M. Vazquez, Juan J. Calvete, Libia Sanz, H. Rordríguez-martínezAbstract:In boar seminal plasma, PSP-I and PSP-II form a noncovalent heterodimer, which is known to have a beneficial effect on highly extended boar spermatozoa. These proteins are mainly produced by the seminal vesicles of the male reproductive tract and are mixed with the spermatozoa during ejaculation. This study assessed the epithelial localization and expression of spermadhesin PSP-I and PSP-II subunits using immunohistochemical, western blotting, and RT-PCR methods in porcine testis, epididymis (caput, corpus, and caudal), seminal vesicles, and Bulbourethral Glands. Tissues were collected from 10 mature boars (Swedish Yorkshire) of proven fertility, frozen or fixed, and embedded in paraffin. Sections of 5 µm were mounted on poly-l-lysine-coated glass slides for immunohistochemistry, and 50 mg from the frozen counterparts were homogenized to isolate proteins (Western blotting study) or RNA (RT-PCR study). Polyclonal antibodies (antiPSP-I and antiPSP-II) against proteins (PSP-I and PSP-II) were used. The immunohistochemistry showed positive labelling for both antibodies in the epithelium of seminal vesicles in all males. Positive immunolabelling, but of variable intensity, was present in the epididymal epithelium (caput, corpus, and caudal), with variation among segments and boars. No labelling was found in the seminiferous epithelium or Bulbourethral Glands of any boar. After SDS-PAGE and Western blotting, immunoreactive bands were obtained in the extracts of all tissues for both PSP proteins. Among tissues, the highest intensity corresponded to the seminal vesicle lane for both proteins. Epididymis (caput, corpus, and caudal), Bulbourethral Gland, and testis showed more feeble bands, yet present. The intensity of the bands varied among boars. Amplification products from mRNA were obtained in all tissues explored by RT-PCR, using specific primers for PSP-I and PSP-II. The same trend was obtained regarding intensity bands, corresponding with the intensity obtained by Western blotting. The results indicate that PSP-I and PSP-II are mainly expressed in seminal vesicles and epididymal segments and, in lower amount, in the testis and Bulbourethral Gland. This work was supported by Formas, Stockholm, and Fundacion Seneca, Murcia, Spain.
Jordi Roca - One of the best experts on this subject based on the ideXlab platform.
-
Western blot detection of GPX-5 in boar genital tract.
2016Co-Authors: Isabel Barranco, Asta Tvarijonaviciute, Cristina Perez-patiño, Alejandro Vicente-carrillo, Inmaculada Parrilla, Jose J. Ceron, Emilio A. Martinez, Heriberto Rodriguez-martinez, Jordi RocaAbstract:L1: protein ladder; L2: blood vessel (positive control); L3: testis. L4: epididymis. L5: cauda epididymis. L6: seminal vesicle. L7: prostate. L8: Bulbourethral Gland.
-
Immunolocalization of glutathione peroxidase-5 (GPX5) in boar genital organs.
2016Co-Authors: Isabel Barranco, Asta Tvarijonaviciute, Cristina Perez-patiño, Alejandro Vicente-carrillo, Inmaculada Parrilla, Jose J. Ceron, Emilio A. Martinez, Heriberto Rodriguez-martinez, Jordi RocaAbstract:Fig 4a depicts a representative positive control section depicting immunostained blood vessels with a major restricted endothelial GPX-5 localization (arrows). A representative negative control prostate section (primary Ab omitted) is depicted in Fig 4b. Fig 4c shows a representative section of boar testis, showing immunostaining in the interstitium (blood vessels and Leydig cells, asterisk) and the seminiferous tubules, with a marked immunostaining in the elongated spermatids (arrows). The immunostaining was more marked in the lining epithelium of the ductus epididymides (Fig 4d, cauda segment) where both the principal epithelial cells (arrows point to the apical cell region, base of the stereocilia) and the luminal spermatozoa (spz) appeared stained, while the surrounding smooth muscle appears only faibly stained (m), lu: lumen. Fig 4e-g depict accessory sexual Glands secretory epithelia of the prostate (4e), the seminal vesicles (4f) and the Bulbourethral Gland (4g). While prostate depicted both nuclear and cytoplasmic staining (4e), the seminal vesicles depicted mainly cytoplasmic staining (arrows) and a clear staining in the luminal secretion (lu), lp: lamina propria. The Bulbourethral Gland (4g) was immunostained in the blood vessel dominated interstitial with some staining in the latero-basal epithelial membrane (arrows). Not-counterstained sections, viewed with phase-contrast optics. Bars: 50μm.
-
Localization and expression of spermadhesin PSP-I/PSP-II subunits in the reproductive organs of the boar
International journal of andrology, 2008Co-Authors: E. M. Garcia, Inmaculada Parrilla, Emilio A. Martinez, Jordi Roca, Juan M. Vazquez, Maria D. Ortega, Juan J. Calvete, Libia Sanz, Heriberto Rodriguez-martinezAbstract:The epithelial localization and expression of the spermadhesin PSP-I and PSP-II subunits were determined in the testis, ductus epididymes (caput, corpus and cauda), seminal vesicles and Bulbourethral Glands of mature boars, using immunohistochemical, western blotting and RT-PCR methods. Immunohistochemistry showed positive labelling for PSP-I and PSP-II antibodies in the epithelium of seminal vesicles in all males tested. Positive immunolabelling, but with variable intensity, was also present in the epididymal epithelium (caput, corpus and cauda), although varying largely among segments and boars. Immunoreactivity was nearly or completely absent in the seminiferous epithelium and the Bulbourethral Gland, although SDS-PAGE and western blotting revealed the presence of PSP-I and PSP-II immunoreactive bands in all the tissue extracts, including the testis and the Bulbourethral Gland. mRNA amplification by RT-PCR using primers specific for PSP-I and PSP-II showed a trend similar to that observed for western blotting, i.e. intensity variation between tissues (even between segments of the same epididymis) and among boars. Our results indicate that the seminal vesicles are the main source of PSP-I and PSP-II spermadhesins, although epididymal segments, testis and the Bulbourethral Gland also participate in the expression of both proteins.
-
323 EXPRESSION OF PSP-I AND PSP-II IN THE REPRODUCTIVE TRACT OF THE BOAR BY IMMUNOHISTOCHEMISTRY, WESTERN BLOTTING, AND RT-PCR
Reproduction Fertility and Development, 2007Co-Authors: E. M. Garcia, Inmaculada Parrilla, Emilio A. Martinez, Jordi Roca, Juan M. Vazquez, Juan J. Calvete, Libia Sanz, H. Rordríguez-martínezAbstract:In boar seminal plasma, PSP-I and PSP-II form a noncovalent heterodimer, which is known to have a beneficial effect on highly extended boar spermatozoa. These proteins are mainly produced by the seminal vesicles of the male reproductive tract and are mixed with the spermatozoa during ejaculation. This study assessed the epithelial localization and expression of spermadhesin PSP-I and PSP-II subunits using immunohistochemical, western blotting, and RT-PCR methods in porcine testis, epididymis (caput, corpus, and caudal), seminal vesicles, and Bulbourethral Glands. Tissues were collected from 10 mature boars (Swedish Yorkshire) of proven fertility, frozen or fixed, and embedded in paraffin. Sections of 5 µm were mounted on poly-l-lysine-coated glass slides for immunohistochemistry, and 50 mg from the frozen counterparts were homogenized to isolate proteins (Western blotting study) or RNA (RT-PCR study). Polyclonal antibodies (antiPSP-I and antiPSP-II) against proteins (PSP-I and PSP-II) were used. The immunohistochemistry showed positive labelling for both antibodies in the epithelium of seminal vesicles in all males. Positive immunolabelling, but of variable intensity, was present in the epididymal epithelium (caput, corpus, and caudal), with variation among segments and boars. No labelling was found in the seminiferous epithelium or Bulbourethral Glands of any boar. After SDS-PAGE and Western blotting, immunoreactive bands were obtained in the extracts of all tissues for both PSP proteins. Among tissues, the highest intensity corresponded to the seminal vesicle lane for both proteins. Epididymis (caput, corpus, and caudal), Bulbourethral Gland, and testis showed more feeble bands, yet present. The intensity of the bands varied among boars. Amplification products from mRNA were obtained in all tissues explored by RT-PCR, using specific primers for PSP-I and PSP-II. The same trend was obtained regarding intensity bands, corresponding with the intensity obtained by Western blotting. The results indicate that PSP-I and PSP-II are mainly expressed in seminal vesicles and epididymal segments and, in lower amount, in the testis and Bulbourethral Gland. This work was supported by Formas, Stockholm, and Fundacion Seneca, Murcia, Spain.
