Production of transgenic rats by ooplasmic injection of spermatogenic cells exposed to exogenous DNA: A preliminary study

2004 ◽  
Vol 69 (2) ◽  
pp. 153-158 ◽  
Author(s):  
Megumi Kato ◽  
Ayako Ishikawa ◽  
Ryosuke Kaneko ◽  
Takeshi Yagi ◽  
Shinichi Hochi ◽  
...  
Keyword(s):  
Spermatogenic Cells ◽  
Transgenic Rats ◽  
Exogenous Dna ◽  
Preliminary Study

scholarly journals 317 FACTORS AFFECTING PRODUCTION EFFICIENCY OF TRANSGENIC RATS BY ICSI-MEDIATED DNA TRANSFER

2005 ◽  
Vol 17 (2) ◽  
pp. 309
Author(s):  
M. Kato ◽  
S. Hochi ◽  
M. Hirabayashi
Keyword(s):  
Production Efficiency ◽  
Dna Transfer ◽  
Egfp Expression ◽  
Transgenic Rat ◽  
Sprague Dawley ◽  
Transgenic Rats ◽  
Rat Strains ◽  
Exogenous Dna ◽  
Factors Affecting ◽  
Rat Offspring

Successful DNA transfer via intracytoplasmic sperm injection (ICSI) was first reported in mice (Perry et al. 1999, Science 284, 1180), and was recently extended to rats (Kato et al. 2004, Mol. Reprod. Dev. 69, 153). In the present study, factors affecting the production efficiency of transgenic rats by the ICSI-mediated DNA transfer were investigated. Cauda epididymal spermatozoa from Sprague-Dawley rats were sonicated (SO) and/or frozen-thawed (FT) for tail-cutting and membrane-disrupting. The sperm heads were exposed for 1 min to different concentrations (0.02–2.5 ng/μL) of 3.0 kb EGFP DNA solution, and then microinjected into denuded F1 (Donryu × LEW) rat oocytes. The optimal concentration of EGFP DNA was 0.1 ng/μL, as determined by the in vitro developmental competence into morulae/blastocysts and the EGFP expression of the ICSI oocytes. The presumptive 1- or 2-cell stage zygotes were transferred into oviducts of pseudopregnant Wistar females, and the presence of EGFP DNA in the offspring was examined by fluorescence under the 480 nm UV light. The production efficiency of transgenic rat offspring was 2.8% (2/71 zygotes transferred), 1.6% (1/63), and 3.3% (2/61) in the oocytes into which SO-, FT-, and SO+ FT-treated sperm heads were injected, respectively. The founder transgenic rats carrying EGFP DNA transmitted the transgenes to their progeny according to the Mendelian fashion (43.8–54.8%), suggesting the stable incorporation of the transgenes into rat genomes. Four rat strains (F344, LEW, Donryu, and Sprague-Dawley) were compared for their suitability as sperm/oocyte donors in the production of transgenic rats by ICSI with SO + FT-treated and 0.1 ng/μL EGFP DNA-exposed sperm heads. The production efficiency of the transgenic rats in the Sprague-Dawley strain (8.2%, 8/98) was significantly higher than that in LEW strain (0.9%, 1/114), while those in F344 (4.3%, 4/92) and Donryu (4.4%, 5/114) strains were intermediate. Attempts were made to introduce three other DNA constructs (5.0 kb plasmid and 208 kb BAC, both with Fyn gene, and 186 kb BAC with Svet1/IRES-Cre gene) into rat genomes by ICSI with SO + FT-treated and 0.1 ng/μL DNA-exposed sperm heads. PCR analysis showed that the Fyn, Fyn/BAC, and Svet1/IRES-Cre DNA constructs were successfully introduced into Sprague-Dawley rat offspring via ICSI, with production efficiencies of 2.8% (3/109), 0.9% (1/109), and 2.4% (3/125), respectively. These results indicate that transgenic rats can be produced by ICSI-mediated DNA transfer using the various types of exogenous DNA and rat strains with different genetic backgrounds.

The Ultrastructure of Metastatic Human Mammary Carcinoma After Prolonged Estrogen Therapy

1967 ◽  
Vol 25 ◽  
pp. 180-181
Author(s):  
John H.L. Watson ◽  
John L. Swedo ◽  
R.W. Talley
Keyword(s):  
Mammary Carcinoma ◽  
Osmium Tetroxide ◽  
Estrogen Therapy ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Human Mammary Carcinoma ◽  
Surgical Biopsy ◽  
Lead Hydroxide ◽  
Preliminary Study ◽  
Hormonal Therapies

A preliminary study of human mammary carcinoma on the ultrastructural level is reported for a metastatic, subcutaneous nodule, obtained as a surgical biopsy. The patient's tumor had responded favorably to a series of hormonal therapies, including androgens, estrogens, progestins, and corticoids for recurring nodules over eight years. The pertinent nodule was removed from the region of the gluteal maximus, two weeks following stilbestrol therapy. It was about 1.5 cms in diameter, and was located within the dermis. Pieces from it were fixed immediately in cold fixatives: phosphate buffered osmium tetroxide, glutaraldehyde, and paraformaldehyde. Embedment in each case was in Vestopal W. Contrasting was done with combinations of uranyl acetate and lead hydroxide.

Annulate lamellae in the Sertoli cells of guinea pig testis after unilateral torsion of the spermatic cord

1984 ◽  
Vol 42 ◽  
pp. 196-197
Author(s):  
J. Chakraborty ◽  
A. P. Sinha Hikim ◽  
J. S. Jhunjhunwala
Keyword(s):  
Sertoli Cells ◽  
Guinea Pigs ◽  
Spermatic Cord ◽  
Present Report ◽  
Cell Types ◽  
Annulate Lamellae ◽  
Spermatogenic Cells ◽  
Electron Microscopic ◽  
Structural Details ◽  
First Time

Although the presence of annulate lamellae was noted in many cell types, including the rat spermatogenic cells, this structure was never reported in the Sertoli cells of any rodent species. The present report is based on a part of our project on the effect of torsion of the spermatic cord to the contralateral testis. This paper describes for the first time, the fine structural details of the annulate lamellae in the Sertoli cells of damaged testis from guinea pigs.One side of the spermatic cord of each of six Hartly strain adult guinea pigs was surgically twisted (540°) under pentobarbital anesthesia (1). Four months after induction of torsion, animals were sacrificed, testes were excised and processed for the light and electron microscopic investigations. In the damaged testis, the majority of seminiferous tubule contained a layer of Sertoli cells with occasional spermatogonia (Fig. 1). Nuclei of these Sertoli cells were highly pleomorphic and contained small chromatinic clumps adjacent to the inner aspect of the nuclear envelope (Fig. 2).

Fine structure of early degenerating myofibers in the tibialis anterior of young ‘MDX’ mouse

1988 ◽  
Vol 46 ◽  
pp. 322-323
Author(s):  
H.D. Geissinger ◽  
C.K. McDonald-Taylor
Keyword(s):  
Fine Structure ◽  
Muscle Regeneration ◽  
Tibialis Anterior ◽  
Genetic Defect ◽  
Mdx Mouse ◽  
Mdx Mice ◽  
Histopathological Changes ◽  
Electron Microscopic ◽  
Dystrophic Mouse ◽  
Preliminary Study

A new strain of mice, which had arisen by mutation from a dystrophic mouse colony was designated ‘mdx’, because the genetic defect, which manifests itself in brief periods of muscle destruction followed by episodes of muscle regeneration appears to be X-linked. Further studies of histopathological changes in muscle from ‘mdx’ mice at the light microscopic or electron microscopic levels have been published, but only one preliminary study has been on the tibialis anterior (TA) of ‘mdx’ mice less than four weeks old. Lesions in the ‘mdx’ mice vary between different muscles, and centronucleation of fibers in all muscles studied so far appears to be especially prominent in older mice. Lesions in young ‘mdx’ mice have not been studied extensively, and the results appear to be at variance with one another. The degenerative and regenerative aspects of the lesions in the TA of 23 to 26-day-old ‘mdx’ mice appear to vary quantitatively.

Ultrastructural observations of the aorta of calves implanted with a mechanical left ventricular assist device

1990 ◽  
Vol 48 (3) ◽  
pp. 834-835
Author(s):  
J P Cassella ◽  
V Salih ◽  
T R Graham
Keyword(s):  
Left Ventricular Assist Device ◽  
Ventricular Assist Device ◽  
Left Ventricular ◽  
Aortic Tissue ◽  
Assist Device ◽  
Ventricular Assist ◽  
Transmission Electron ◽  
Left Ventricular Assist ◽  
Preliminary Study

Left ventricular assist systems are being developed for eventual long term or permanent implantation as an alternative to heart transplantation in patients unsuitable for or denied the transplant option. Evaluation of the effects of these devices upon normal physiology is required. A preliminary study was conducted to evaluate the morphology of aortic tissue from calves implanted with a pneumatic Left Ventricular Assist device-LVAD. Two 3 month old heifer calves (calf 1 and calf 2) were electively explanted after 128 days and 47 days respectively. Descending thoracic aortic tissue from both animals was removed immediately post mortem and placed into karnovsky’s fixative. The tissue was subsequently processed for transmission electron microscopy (TEM). Some aortic tissue was fixed in neutral buffered formalin and processed for routine light microscopy.

A preliminary study on the potential of front face fluorescence spectroscopy for the discrimination of Moroccan virgin olive oils and the prediction of their quality

2021 ◽  
Author(s):  
Hicham Zaroual ◽  
El Mestafa El Hadrami ◽  
Romdhane Karoui
Keyword(s):  
Fluorescence Spectroscopy ◽  
Olive Oil ◽  
Virgin Olive Oil ◽  
Front Face ◽  
Virgin Olive Oils ◽  
Olive Oils ◽  
Preliminary Study

This study examines the feasibility of using front face fluorescence spectroscopy (FFFS) to authenticate 41 virgin olive oil (VOO) samples collected from 5 regions in Morocco during 2 consecutive crop seasons.

Dysarthria Profiles in Adults With Hereditary Ataxia

2019 ◽  
Vol 28 (2S) ◽  
pp. 915-924 ◽  
Author(s):  
Kristie A. Spencer ◽  
Mallory Dawson
Keyword(s):  
Rating Scales ◽  
Feedforward Control ◽  
Hereditary Ataxia ◽  
Severity Rating ◽  
Visual Analog Scales ◽  
Speech Characteristics ◽  
Ataxic Dysarthria ◽  
Preliminary Study

Purpose This preliminary study examined whether speech profiles exist for adults with hereditary ataxia based on 2 competing frameworks: a pattern of instability/inflexibility or a pattern of differential subsystem involvement. Method Four dysarthria experts rated the speech samples of 8 adults with dysarthria from hereditary ataxia using visual analog scales and presence/severity rating scales of speech characteristics. Speaking tasks included diadochokinetics, sustained phonation, and a monologue. Results Speech profiles aligned with the instability/inflexibility framework, with the pattern of instability being the most common. Speech profiles did not emerge for the majority of speakers using the differential subsystem framework. Conclusions The findings extend previous research on pure ataxic dysarthria and suggest a possible framework for understanding the speech heterogeneity associated with the ataxias. The predominance of the instability profile is consistent with the notion of impaired feedforward control in speakers with cerebellar disruption.

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