Archive for the ‘Spermatogenic’ Category

In Vivo Gene Transfer to Mouse Spermatogenic Cells: DISCUSSION(4)

DISCUSSION(4)

A previous study has revealed that undifferentiated progenitor type-A spermatogonia originating from a spermatogenic stem cell proliferate and extend horizontally onto the basement membrane. Serial distribution along the tubules of sper-matogenic stem cells has recently been reported in the testes of transgenic mice carrying a lacZ reporter driven by a specific promoter of a mouse retrotransposon gene that is specifically expressed in the spermatogenic stem cell. Judging from these data, we believe that our EP transfection method is sufficiently efficient for introducing a foreign gene even into spermatogenic stem cells, and also that the transgene can be transmitted to the progenitor spermat-ogenic cells as development proceeds. ventolin 100 mcg

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In Vivo Gene Transfer to Mouse Spermatogenic Cells: DISCUSSION(3)

In the mouse, one spermatogenic cycle from spermatogonia to mature spermatozoa is estimated to take about 35 days. Therefore, in about 1 mo all the differentiating germ cells in the premeiotic as well as postmeiotic stages finish maturing to spermatozoa and are released from the seminiferous epithelium to the epididymis. Nevertheless, our finding that p-gal-positive cells were still detected in a clump of spermatogenic germ cells even after 1 mo of CMV-lacZ transfection (Fig. 3B) suggests that beyond the blood-testis barrier the foreign gene was transferred into the spermatogenic stem cells and the proliferating spermatogonia residing on the basal membrane. buy birth control online

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In Vivo Gene Transfer to Mouse Spermatogenic Cells: DISCUSSION(2)

DISCUSSION(2)

EP is the easiest and most economical method for gene transfer. Another advantage is that it can be used for any type of tissue or cell. Recently, due to its efficient infection into numerous cell types, including nonproliferating tissue cells, adenovirus-mediated in vivo transfer has been regarded as the most attractive tool for treating a disease using gene therapy. Blanchard and Boe-kelheide have carried out adenovirus-mediated gene transfer to adult rat testes. They showed that expression of the SV40-lacZ transgene used in their study was detected only in Sertoli cells and Leydig cells but not in germ cells, indicating that adenovirus-mediated gene transfer to the testes has a strict cell-type preference and cannot deliver a transgene into all types of spermatogenic germ cells. flovent inhaler

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In Vivo Gene Transfer to Mouse Spermatogenic Cells: DISCUSSION(1)

In this study, we have shown that the combination of intratubular DNA injection followed by in vivo EP results in the efficient delivery of foreign genes into various types of spermatogenic cells. Mammalian spermatogenesis is an excellent model system with which to study specific gene expression during differentiation of a defined cell lineage as well as to study the molecular mechanism responsible for switching from mitotic to meiotic cell division. However, because of unsolved difficulties in culturing and manipulating spermatogenic cells in vitro, we have no choice but to make transgenic mice for the functional analyses of such spermatogenic genes. buy cipro

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In Vivo Gene Transfer to Mouse Spermatogenic Cells: RESULTS(4)

RESULTS(4)

A majority of the positive cells observed as dot-like localizations along the seminiferous tubules turned out to be Sertoli cells, which are nonproliferating somatic cells (Fig. 3A). Another pattern of staining was observed as a relatively long linear mass along the tubules, where p-gal-expressing cells were found as a clump of spermatogenic germ cells present from the basal membrane to the inner lumen of the tubule (Fig. 3B), indicating that the CMV-lacZ transgene was incorporated into undifferentiated germ cells at the time of EP transfection. buy ventolin inhalers

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In Vivo Gene Transfer to Mouse Spermatogenic Cells: RESULTS(3)

Considering that the population of round spermatid cells that specifically transcribe the Prm-1 gene is about 30% of all testicular cells, the relatively lower activity of the Prm-1 enhancer appeared to be mainly due to the difference in the ratio of cells expressing each transgene. Moreover, when immature mouse testes (18-20 days after birth), which rarely have haploid cells, were used for the same experiments, the Prm-1 enhancer vector showed a small enhancing activity of about 0.9-fold compared with that of the SV40-enhancer vector. These results indicate that the transcriptional effect of the spermatogenic stage-specific Prm-1 enhancer is transiently detectable by this EP transfection method. buy yasmin online

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In Vivo Gene Transfer to Mouse Spermatogenic Cells: RESULTS(2)

RESULTS(2)

Detection of Enhancer Activity of the Spermatogenic Gene

The enhancer activity of a spermatid-specific gene using a luciferase reporting system was studied to examine the possible application of an intratubular injection and in vivo EP as a new transient expression assay for spermatogenic specific genes. The Prm-1 gene encodes a sperm-specific chromosomal protein that replaces histone proteins during spermiogenesis. The well-defined enhancer element Prm-1 was monitored as a typical indicator of sper-matogenic stage-specific expression. buy flovent inhaler

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